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WO2025049419A1 - Pyrazolyl compounds as emopamil binding protein inhibitors - Google Patents

Pyrazolyl compounds as emopamil binding protein inhibitors Download PDF

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Publication number
WO2025049419A1
WO2025049419A1 PCT/US2024/043947 US2024043947W WO2025049419A1 WO 2025049419 A1 WO2025049419 A1 WO 2025049419A1 US 2024043947 W US2024043947 W US 2024043947W WO 2025049419 A1 WO2025049419 A1 WO 2025049419A1
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compound
pharmaceutically acceptable
acceptable salt
mmol
phenyl
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John Ziqi Jiang
Sungtaek Lim
Gregory H. Merriman
Paul Justin Mueller
Sukanthini Thurairatnam
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Genzyme Corp
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Genzyme Corp
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/08—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing alicyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings

Definitions

  • the present disclosure relates to inhibitors of Emopamil Binding Protein (EBP) useful for treating demyelinating diseases such as multiple sclerosis.
  • EBP Emopamil Binding Protein
  • this disclosure describes compounds and compositions for inhibiting EBP, methods of treating demyelinating diseases, and methods of synthesizing these compounds.
  • Myelin is an insulating layer, or sheath, that wraps around nerve cell axons. This lipid- rich insulating material protects the axons and makes possible the saltatory conduction, which speeds axonal electric impulse (Williamson et al., Front Cell Neurosci., 2018, 12: 424). Myelin is formed in the central nervous system (CNS) by glial cells called oligodendrocytes and in the peripheral nervous system (PNS) by glial cells called Schwann cells.
  • CNS central nervous system
  • PNS peripheral nervous system
  • Demyelinating diseases cause damage to the myelin sheath and such diseases include multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), and Guillain- Barre syndrome.
  • MS multiple sclerosis
  • NOSD neuromyelitis optica spectrum disorder
  • CIDP chronic inflammatory demyelinating polyneuropathy
  • Guillain- Barre syndrome a complex neurological disease characterized by deterioration of CNS myelin (Ghasemi et al., Cell J., 2017, 19(1): 1-10).
  • the myelin sheaths surrounding neuronal axons are damaged or destroyed, leading to dysfunction of propagated signals, axonal injury, neuronal loss, accumulation of lesion load, and overt neurological disabilities in patients.
  • the CNS has the capacity to regenerate myelin sheaths after injury through the proliferation, migration, and differentiation of a population of adult progenitor cells referred to as oligodendrocyte precursor cells (OPCs) into new myelinating oligodendrocytes to generate new myelin sheaths, or to a lesser extent, by generation of new myelin sheaths by existing oligodendrocytes (Kuhn et al., Cells, 2019, 8(11): 1424).
  • OPCs oligodendrocyte precursor cells
  • Emopamil Binding Protein also referred to as A8A7 isomerase, 3-beta- hydroxysteroid-Delta(8),Delta(7)-isomerase, human sterol isomerase (HSI), CDPX2, CH02, CPX, or CPXD) is the enzyme in the cholesterol biosynthesis pathway which catalyzes the conversion of zymosterol and zymostenol to dehyolathosterol and lathosterol (Silve et al., The Journal of Biological Chemistry, 1996, 271(37): 22434-22440).
  • targeting EBP is one strategy for increasing OPC differentiation.
  • compounds and therapeutic methods capable of targeting EBP to induce differentiation of OPCs to enhance generation of new oligodendrocytes and increase myelination and/or remyelination in demyelinated lesions are examples of the enzyme in the cholesterol biosynthesis pathway.
  • provided herein are compounds which inhibit EBP for use in treating demyelinating diseases such as multiple sclerosis.
  • EBP Emopamil Binding Protein
  • Embodiment 1 A compound of Formula (I): or a pharmaceutically acceptable salt thereof, wherein:
  • X 1 is C and X 2 is N, or X 1 is N and X 2 is C;
  • R 1 is Ci-C 6 alkyl, C 3 -C 6 cycloalkyl, -CN, Ci-C 6 haloalkyl, -(Ci-C 6 alkylene)-O-(Ci-C 6 alkyl), -(Ci-Ce alkylene)-O-(Ci-C6 haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl);
  • L 1 is a bond, O, or -CH2-;
  • Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing 1 or 2 nitrogen atoms, or Ce-Cio aryl; each R 2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, -O(Ci-Ce alkyl), or
  • L 2 is a bond or O
  • Ring A is , 9- to 11-membered spiro heterocyclylene, or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms;
  • Y 1 is N or CH
  • Y 2 is N or CH; x is 0, 1, or 2; y is 0 or 1; m is 0-5; each R 3 is independently Ci-Ce alkyl, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl;
  • L 3 is a bond, -CH(R a )-, -CH(R a )CH(R a )-, -OCH(R a )CH(R a )-, -CH(R a )CH(R a )N(R a )-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms; each R a is independently H or Ci-Ce alkyl;
  • Z is N or CH; r is 0, 1, or 2; s is 0 or 1; each R 4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group; and n is 0-5.
  • Embodiment 2 The compound of embodiment 1, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 3 The compound of embodiment 1, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 4 The compound of any one of embodiments 1-3, or a pharmaceutically acceptable salt thereof, wherein:
  • R 1 is C1-C6 alkyl, C3-C5 cycloalkyl, -CN, C1-C3 haloalkyl, -(C1-C3 alkylene)-O-(Ci-C 3 alkyl), -(C1-C3 alkylene)-O-(Ci-C3 haloalkyl), or -(C1-C3 alkylene)(C3-Ce cycloalkyl).
  • Embodiment 5 The compound of embodiment 4, or a pharmaceutically acceptable salt thereof, wherein:
  • R 1 is -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH 3 ) 2 , -C(CH 3 )3, -CH2CH2CH2CH3, -CH 2 CH(CH 3 )2, -CH(CH 3 )CH 2 CH3, -CH 2 CH 2 CH(CH3)2, -CH2OCH3, -CH2CH2OCH3, -CH(CH 3 )OCH 3 , -CH(CH 3 )CH 2 OCH3, -CH2CHF2, -CF3, -CHF2,
  • Embodiment 6 The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein: L 1 is a bond.
  • Embodiment 7 The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein: L 1 is O.
  • Embodiment 8 The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 9 The compound of any one of embodiments 1-8, or a pharmaceutically acceptable salt thereof, wherein:
  • Ring B is C4-C6 cycloalkyl, pyridinyl, pyrazinyl, pyrimidinyl, or phenyl.
  • Embodiment 10 The compound of any one of embodiments 1-9, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 11 The compound of any one of embodiments 1-10, or a pharmaceutically acceptable salt thereof, wherein: each R 2 is independently C1-C4 alkyl, C1-C3 haloalkyl, halo, -O(Ci-C3 alkyl), or
  • Embodiment 12 The compound of embodiment 11, or a pharmaceutically acceptable salt thereof, wherein: each R 2 is independently -CF3, -CF2CH3, -CH2CHF2, -CHF2, F, Cl, Br, -OCF3, -OCHF2, -OCH3, or -C(CH 3 ) 3 , or two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from F or -CF3.
  • Embodiment 13 The compound of any one of embodiments 1-12, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 14 The compound of any one of embodiments 1-13, or a pharmaceutically acceptable salt thereof, wherein:
  • L 2 is a bond
  • Embodiment 15 The compound of any one of embodiments 1-13, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 16 The compound of any one of embodiments 1-15, or a pharmaceutically acceptable salt thereof, wherein:
  • Ring Y 1 is N or CH
  • Y 2 is N or CH; x is 0, 1, or 2; and y is 0 or 1.
  • Embodiment 17 The compound of any one of embodiments 1-16, or a pharmaceutically acceptable salt thereof, wherein:
  • Ring A is 9- to 11 -membered spiro heterocyclylene or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms.
  • Embodiment 18 The compound of any one of embodiments 1-17, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 19 The compound of any one of embodiments 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 0.
  • Embodiment 20 The compound of any one of embodiments 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 1-3.
  • Embodiment 21 The compound of any one of embodiments 1-18 and 20, or a pharmaceutically acceptable salt thereof, wherein: each R 3 is independently C1-C3 alkyl, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl.
  • Embodiment 22 The compound of embodiment 21, or a pharmaceutically acceptable salt thereof, wherein: each R 3 is independently -CEE, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro cyclopropyl.
  • Embodiment 23 The compound of any one of embodiments 1-22, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 24 The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein: L 3 is a bond.
  • Embodiment 25 The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein:
  • L 3 is -CH(R a )-, -CH(R a )CH(R a )-, -OCH(R a )CH(R a )-, or -CH(R a )CH(R a )N(R a )-; and each R a is independently H or C1-C3 alkyl.
  • Embodiment 26 The compound of embodiment 25, or a pharmaceutically acceptable salt thereof, wherein:
  • L 3 is -CH2, -CH2CH2-, -CH(CH 3 )CH 2 -, -CH 2 CH(CH 3 )-, -OCH2CH2-, or -CH2CH2N(CH 2 CH 3 )-.
  • Embodiment 27 The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein:
  • L 3 is 5- to 6-membered heterocyclylene or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms.
  • Embodiment 28 The compound of embodiment 27, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 29 The compound of any one of embodiments 1-28, or a pharmaceutically acceptable salt thereof, wherein:
  • W is O, CH2, or N(H).
  • Embodiment 30 The compound of any one of embodiments 1-28, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 31 The compound of any one of embodiments 1-30, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 32 The compound of any one of embodiments 1-30, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 33 The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 1.
  • Embodiment 34 The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 0.
  • Embodiment 35 The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 1; and s is 0.
  • Embodiment 36 The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 2; and s is 1.
  • Embodiment 37 The compound of any one of embodiments 1-36, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 38 The compound of any one of embodiments 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 0.
  • Embodiment 39 The compound of any one of embodiments 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 1-3.
  • Embodiment 40 The compound of any one of embodiments 1-37 and 39, or a pharmaceutically acceptable salt thereof, wherein: each R 4 is independently halo, -OH, C1-C3 alkyl, or C1-C3 haloalkyl, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
  • Embodiment 41 The compound of embodiment 40, or a pharmaceutically acceptable salt thereof, wherein: each R 4 is independently F, -OH, -CH3, -CH(CH3)2, or -CF3, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
  • Embodiment 42 The compound of any one of embodiments 1-41, or a pharmaceutically acceptable salt thereof, wherein:
  • Embodiment 43 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-a), (I-b), or (I-c):
  • Embodiment 44 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-d) or (I-e):
  • Embodiment 45 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), or
  • Embodiment 46 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Ila) or (lib) :
  • Embodiment 47 The compound of embodiment 46, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (lie):
  • Embodiment 48 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Illa) or (Illb) :
  • Embodiment 49 The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IVa) or (IVb): nitrogen atoms.
  • Embodiment 50 A compound selected from the compounds of Table 1 or a pharmaceutically acceptable salt thereof.
  • Embodiment 52 A compound selected from the compounds of Table 3 or a pharmaceutically acceptable salt thereof.
  • Embodiment 53 A pharmaceutical composition comprising the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
  • Embodiment 54 A method of inhibiting Emopamil Binding Protein (EBP) comprising contacting EBP with an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
  • Embodiment 55 A method of remyelinating a neuronal axon comprising contacting the neuronal axon with an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
  • Embodiment 56 A method of treating a demyelinating disease in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
  • Embodiment 57 The method of embodiment 56, wherein the demyelinating disease is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain- Barre syndrome.
  • MS multiple sclerosis
  • NOSD neuromyelitis optica spectrum disorder
  • CIDP chronic inflammatory demyelinating polyneuropathy
  • Guillain- Barre syndrome is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain- Barre syndrome.
  • Embodiment 58 A method of treating multiple sclerosis (MS) in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
  • MS multiple sclerosis
  • ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 pL” means “about 5 pL” and also “5 pL .” Generally, the term “about” includes an amount that would be expected to be within experimental error, such as for example, within 15%, 10%, or 5%.
  • alkyl refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (i.e., C1-C20 alkyl), 1 to 10 carbon atoms (i.e., C1-C10 alkyl), 1 to 6 carbon atoms (i.e., Ci-Ce alkyl) or 1 to 3 carbon atoms (i.e., C1-C3 alkyl).
  • alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl and 3 -methylpentyl.
  • butyl includes n-butyl (i.e., -(CEhjsCEE), isobutyl (i.e., -CH2CH(CH3)2), sec-butyl (i.e.,
  • alkylene refers to a divalent unbranched or branched saturated hydrocarbon chain. As used herein, alkylene has 1 to 20 carbon atoms (i.e., C1-C20 alkylene), 1 to 10 carbon atoms (i.e., C1-C10 alkylene), 1 to 6 carbon atoms (i.e., Ci-Ce alkylene) or 1 to 3 carbon atoms (i.e., Ci- C3 alkylene). Examples of alkyl groups include methylene, ethylene, propylene, butylene, pentylene, and hexylene.
  • C4 alkylene includes -(CEhjsCEh- , -CH 2 C(CH 3 )2-, and
  • C3 alkylene includes -(CH 2 ) 2 CH2- and -C(CH 3 )2-.
  • aryl group is an aromatic carbocyclic group of from 6 to 14 carbon atoms (Ce-Cu aryl) having a single ring (e.g., phenyl or Ce aryl) or multiple condensed rings (e.g., naphthyl or anthryl).
  • aryl groups contain 6-14 carbons (C6-C14 aryl), and in others from 6 to 12 (C6-C12 aryl) or even 6 to 10 carbon atoms (Ce-Cio aryl) in the ring portions of the groups.
  • Particular aryls include phenyl, biphenyl, naphthyl and the like.
  • Cycloalkyl refers to a saturated or partially unsaturated cyclic alkyl group having a single ring or multiple rings including fused, bridged and spiro ring systems.
  • the term “cycloalkyl” includes cycloalkenyl groups (i.e., the cyclic group having at least one double bond).
  • cycloalkyl has from 3 to 20 ring carbon atoms (i.e., C3-C20 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C3-C10 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C3-C6 cycloalkyl).
  • Cycloalkyl also includes “spiro cycloalkyl” when there are two positions for substitution on the same carbon atom.
  • Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
  • Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl and the like.
  • the term cycloalkyl is intended to encompass any non-aromatic ring which may be fused to an aryl ring, regardless of the attachment to the remainder of the molecule.
  • Haloalkyl refers to an unbranched or branched alkyl group as defined above, wherein one or more hydrogen atoms are replaced by a halogen.
  • Ci-Ce haloalkyl refers to a Ci-Ce alkyl which is substituted by one or more halogen atoms.
  • a Ci haloalkyl refers to a methyl group that may be substituted by 1-3 halo groups
  • a C2 haloalkyl refers to an ethyl group that may be substituted by 1-5 halo groups
  • a C3 haloalkyl refers to a propyl group that may be substituted by 1-7 halo groups, etc.
  • haloalkyl examples include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like.
  • a haloalkyl may contain one or more halo atoms that are the same (i.e., all fluoro) or a mixture of halo atoms (i.e, chloro and fluoro).
  • Heteroaryl refers to an aromatic group (e.g., a 5-14 membered ring system) having a single ring, multiple rings, or multiple fused rings, with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur.
  • heteroaryl includes 1 to 10 ring carbon atoms and 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur within the ring.
  • heteroaryl groups include pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl and thiophenyl (i.e., thienyl).
  • Heterocyclyl refers to a saturated or unsaturated cyclic alkyl group, with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur.
  • the term “heterocyclyl” includes heterocycloalkenyl groups (i.e., the heterocyclyl group having at least one double bond), bridged-heterocyclyl groups, fused-heterocyclyl groups and spiro- heterocyclyl groups.
  • any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of the attachment (i.e., can be bound through a carbon atom or a heteroatom).
  • heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of the attachment to the remainder of the molecule.
  • heterocyclyl has 1 to 10 ring carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms, and 1 to 5 ring heteroatoms, 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 to 2 heteroatoms independently selected from nitrogen, sulfur and oxygen.
  • heterocyclyl groups include dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl and 1,1-
  • Cyano refers to the group -CN.
  • Halogen or “halo” includes fluoro, chloro, bromo, and iodo.
  • Haldroxy refers to the group -OH.
  • a divalent group such as a divalent “phenyl” group, a divalent “heteroaryl” group, a divalent “heterocyclyl” group etc., may also be referred to as a “phenylene” group, a “heteroarylene” group, or a “heterocyclylene” group, respectively.
  • the term “optionally substituted” refers to any one or more hydrogen atoms on the designated atom or group which may or may not be replaced by a moiety other than hydrogen.
  • the substituted group may be substituted with one or more substituents, such as e.g., 1, 2, 3, 4, or 5 substituents.
  • the substituents are selected from the functional groups provided herein.
  • any compound or formula described herein is intended to represent unlabeled forms as well as isotopically labeled forms of the compounds.
  • Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number.
  • isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine and iodine, such as 2 H, 3 H, n C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 36 C1, 123 I and 125 I, respectively.
  • isotopically labeled compounds of the present disclosure for example those into which radioactive isotopes, such as 2 H, 3 H, 13 C, and 14 C are incorporated, are included in this disclosure.
  • isotopically labelled compounds may be useful in metabolic studies, reaction kinetic studies, detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays or in radioactive treatment of patients.
  • PET positron emission tomography
  • SPECT single-photon emission computed tomography
  • the disclosure also includes “deuterated analogs” of compounds described herein in which from 1 to n hydrogens attached to a carbon atom is/are replaced by deuterium, in which n is the number of hydrogens in the molecule.
  • the deuterium atoms may be on the same portion of the molecule (for example, on a single alkyl group or on a single ring) or on different portions of the molecule (for example, on separate alkyl groups or separate rings).
  • Such compounds may exhibit increased resistance to metabolism and thus may be useful for increasing the half-life of any compound when administered to a mammal, particularly a human.
  • “Pharmaceutically acceptable” refers to compounds, salts, compositions, dosage forms, and other materials which are useful in preparing a pharmaceutical composition that is suitable for veterinary or human pharmaceutical use.
  • the term “pharmaceutically acceptable salt” of a given compound refers to salts that retain the biological effectiveness and properties of the given compound and which are not biologically or otherwise undesirable.
  • “Pharmaceutically acceptable salts” include, for example, salts with inorganic acids and salts with an organic acid.
  • the free base can be obtained by basifying a solution of the acid salt.
  • an addition salt, particularly a pharmaceutically acceptable addition salt may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds.
  • Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.
  • Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p- toluene-sulfonic acid, salicylic acid and the like.
  • pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts.
  • Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines.
  • suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri(isopropyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine and the like.
  • reference to a particular salt, such as hydrochloride or formate may refer to a single salt, such as monohydrochloride or monoformate, or may refer to a multiple salt, such a dihydrochloride or diformate.
  • the compounds disclosed herein, or their pharmaceutically acceptable salts may include an asymmetric center and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (5)- or, as (D)- or (L)- for amino acids.
  • the disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms.
  • Optically active (+) and (-), (R)- and (5)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization.
  • a “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable.
  • the present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are nonsuperimposable mirror images of one another.
  • “Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.
  • “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” or “excipient” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • Effective amount or dose of a compound or a composition refers to that amount of the compound or the composition that results in an intended result as desired based on the disclosure herein. Effective amounts can be determined by standard pharmaceutical procedures in cell cultures or experimental animals including, without limitation, by determining the LD50 (the dose lethal to 50% of the population) and the EDso (the dose therapeutically effective in 50% of the population).
  • “Therapeutically effective amount” or dose of a compound or a composition refers to that amount of the compound or the composition that results in reduction or inhibition of symptoms or a prolongation of survival in a subject (i.e., a human patient). The results may require multiple doses of the compound or the composition.
  • Treating” or “treatment” of a disease in a subject refers to 1) preventing the disease from occurring in a patient that is predisposed or does not yet display symptoms of the disease; 2) inhibiting the disease or arresting its development; or 3) ameliorating or causing regression of the disease.
  • treatment or “treating” is an approach for obtaining beneficial or desired results including clinical results.
  • beneficial or desired results include, but are not limited to, one or more of the following: decreasing one or more symptoms resulting from the disease or disorder, diminishing the extent of the disease or disorder, stabilizing the disease or disorder (e.g., preventing or delaying the worsening of the disease or disorder), delaying the occurrence or recurrence of the disease or disorder, delay or slowing the progression of the disease or disorder, ameliorating the disease or disorder state, providing a remission (whether partial or total) of the disease or disorder, decreasing the dose of one or more other medications required to treat the disease or disorder, enhancing the effect of another medication used to treat the disease or disorder, delaying the progression of the disease or disorder, increasing the quality of life, and/or prolonging survival of a subject.
  • treatment is a reduction of pathological consequence of the disease or disorder. The methods of the invention contemplate any one or more of these aspects of treatment.
  • the terms “subject(s)” and “patient(s)” mean any mammal.
  • the mammal is a human.
  • the mammal is a non-human, such as a primate, dog, cat, rabbit, or rodent. None of the terms require or are limited to situations characterized by the supervision (e.g., constant or intermittent) of a health care worker (e.g., a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker).
  • a health care worker e.g., a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker.
  • composition or “medicament” refer to a composition suitable for pharmaceutical use in a subject, e.g., as an EBP inhibitor.
  • X 1 is C and X 2 is N, or X 1 is N and X 2 is C;
  • R 1 is Ci-C 6 alkyl, C 3 -C 6 cycloalkyl, -CN, Ci-C 6 haloalkyl, -(Ci-C 6 alkylene)-O-(Ci-C 6 alkyl), -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl);
  • L 1 is a bond, O, or -CH2-;
  • Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing 1 or 2 nitrogen atoms, or Ce-Cio aryl; each R 2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, -O(Ci-Ce alkyl), or -O(Ci-Ce haloalkyl), or two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl; L 2 is a bond or O;
  • Ring A is , 9- to 11-membered spiro heterocyclylene, or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms;
  • Y 1 is N or CH
  • Y 2 is N or CH; x is 0, 1, or 2; y is 0 or 1; m is 0-5; each R 3 is independently Ci-Ce alkyl, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl;
  • L 3 is a bond, -CH(R a )-, -CH(R a )CH(R a )-, -OCH(R a )CH(R a )-, -CH(R a )CH(R a )N(R a )-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms; each R a is independently H or Ci-Ce alkyl;
  • Z is N or CH; r is 0, 1, or 2; s is 0 or 1; each R 4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group; and n is 0-5.
  • R 1 is Ci-Ce alkyl, C3-C6 cycloalkyl, -CN, Ci-Ce haloalkyl, -(Ci- Ce alkylene)-O-(Ci-Ce alkyl), -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl).
  • R 1 is Ci-Ce alkyl, C3-C5 cycloalkyl, -CN, C1-C3 haloalkyl, - (Ci-C 3 alkylene)-O-(Ci-C 3 alkyl), -(C1-C3 alkylene)-O-(Ci-C 3 haloalkyl), or -(Ci-C 3 alkylene)(C 3 -C6 cycloalkyl).
  • R 1 is Ci-Ce alkyl. In some embodiments, R 1 is C1-C5 alkyl. In some embodiments, R 1 is C1-C4 alkyl. In some embodiments, R 1 is Ci-C 3 alkyl. In some embodiments, R 1 is C1-C2 alkyl. In some embodiments, R 1 is -CH 3 , -CFbCHs, -CFbCFbCHs, -CH(CH 3 ) 2 , -C(CH 3 ) 3 , -CH 2 CH2CH 2 CH 3 , -CH 2 CH(CH 3 ) 2 , -CH(CH 3 )CH 2 CH 3 , or -CH2CH 2 CH(CH 3 )2.
  • R 1 is C 3 -Ce cycloalkyl. In some embodiments, R 1 is C 3 -Cs cycloalkyl. In some embodiments, R 1 is C 3 -C4 cycloalkyl. In some embodiments, R 1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, R 1 is cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, R 1 is cyclopropyl. In some embodiments, R 1 is cyclobutyl.
  • R 1 is -CN.
  • R 1 is Ci-Ce haloalkyl. In some embodiments, R 1 is Ci-Ce haloalkyl containing 1-13 halogen atoms. In some embodiments, R 1 is Ci-C 3 haloalkyl. In some embodiments, R 1 is Ci-C 3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 1 is -CF 3 , -CHF2, -CH2F, -CC13, -CHCh, -CH2CI, -CF2CI, -CFCh, -CH 2 CF 3 , -CH2CHF2, or - CFFCCh.
  • R 1 is -CH2CHF2, -CF 3 , or -CHF2. In some embodiments, R 1 is -CF 3 . In some embodiments, R 1 is -CH2CHF2. In some embodiments, R 1 is -CHF2.
  • R 1 is -(Ci-Ce alkylene)-O-(Ci-Ce alkyl). In some embodiments, R 1 is -(Ci-C 3 alkylene)-O-(Ci-C 3 alkyl). It is understood that -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6. For example, -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6.
  • -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6.
  • R 1 is -(Ci-C 3 alkylene)-O-(CH 3 ), -(Ci-C 3 alkylene)-O- (CFbCHs), or -(Ci-C 3 alkylene)-O-(CH2CH2CH 3 ).
  • R 1 is-CFbOCHs, - CH 2 CH 2 OCH 3 ,
  • R 1 is -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl). In some embodiments, R 1 is -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), wherein Ci-Ce haloalkyl contains 1- 13 halogen atoms. In some embodiments, R 1 is -(Ci-C 3 alkylene)-O-(Ci-C 3 haloalkyl). In some embodiments, R 1 is -(Ci-C 3 alkylene)-O-(Ci-C 3 haloalkyl), wherein Ci-C 3 haloalkyl contains 1- 7 halogen atoms.
  • -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6.
  • -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6.
  • -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6.
  • R 1 is -(C1-C3 alkylene)-O-(CF3), -(C1-C3 alkylene)-O-(CHF2), or -(C1-C3 alkylene)-O-(CH2F). In some embodiments, R 1 is -(CH2)-O-(CI-C3 haloalkyl), -(CH2CH2XO- (C1-C3 haloalkyl), or
  • R 1 is -(Ci-Ce alkylene)(C3-Ce cycloalkyl). In some embodiments, R 1 is -(C1-C3 alkylene)(C3-Ce cycloalkyl). It is understood that -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6. For example, -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6.
  • -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6.
  • R 1 is -(C1-C3 alkylene)(cyclopropyl), -(C1-C3 alkylene)(cyclobutyl), or -(C1-C3 alkylene)(cyclopentyl).
  • R 1 is - (CH2XC3-C6 cycloalkyl), -(CH2CH2XC3-C6 cycloalkyl), or-(CH2CH2CH2)(C3-Ce cycloalkyl).
  • R 1 is -(CFbXcyclopropyl), -(CFbXcyclobutyl), or -(CFbXcyclopentyl). In some embodiments, R 1 is -(CFbXcyclopropyl).
  • R 1 is -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH 3 ) 2 , - C(CH 3 ) 3 ,
  • L 1 is a bond, O, or -CH2-. In some embodiments, L 1 is a bond. In some embodiments, L 1 is O. In some embodiments, L 1 is -CH2-.
  • Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing
  • Ring B is C4-C6 cycloalkyl, pyridinyl, pyrazinyl, pyrimidinyl, or phenyl.
  • Ring B is C3-C6 cycloalkyl. In some embodiments, Ring B is C4-C6 cycloalkyl. In some embodiments, Ring B is cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, Ring B is cyclopropyl. In some embodiments, Ring B is cyclobutyl. In some embodiments, Ring B is cyclopentyl. In some embodiments, Ring B is cyclohexyl.
  • Ring B is 6-membered heteroaryl containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is 6-membered heteroaryl containing 1 nitrogen atom. In some embodiments, Ring B is 6-membered heteroaryl containing 2 nitrogen atoms. In some embodiments, Ring B is pyridinyl, pyrazinyl, or pyrimidinyl. In some embodiments, Ring B is
  • Ring B is Ce-Cio aryl. In some embodiments, Ring B is Ce aryl. In some embodiments, Ring B is phenyl. In some embodiments, Ring B is Cio aryl. In some embodiments, Ring B is naphthalenyl.
  • each R 2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, - O(Ci-Ce alkyl), or -O(Ci-Ce haloalkyl), or two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl.
  • each R 2 is independently C1-C4 alkyl, C1-C3 haloalkyl, halo, - O(Ci-C 3 alkyl), or
  • R 2 is Ci-Ce alkyl. In some embodiments, R 2 is C1-C4 alkyl. In some embodiments, R 2 is methyl, ethyl, propyl, or butyl. In some embodiments, R 2 is isopropyl. In some embodiments, R 2 is -C(CH3)3.
  • R 2 is Ci-Ce haloalkyl. In some embodiments, R 2 is Ci-Ce haloalkyl containing 1-13 halogen atoms. In some embodiments, R 2 is C1-C3 haloalkyl. In some embodiments, R 2 is C1-C3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R 2 is -CF 3 , -CHF 2 , -CH2F, -CCh, -CHCh, -CH 2 C1, -CF 2 C1, -CFCh, -CF 2 CH 3 , -CH 2 CF 3 , -CH 2 CHF 2 , or
  • R 2 is -CF3, -CF 2 CH3, -CH 2 CHF 2 , or -CHF 2 .
  • R 2 is halo. In some embodiments, R 2 is F, Cl, Br, or I. In some embodiments, R 2 is F, Cl, or Br. In some embodiments, R 2 is F. In some embodiments, R 2 is Cl. In some embodiments, R 5 is Br.
  • R 2 is -O(Ci-Ce alkyl). In some embodiments, R 2 is -O(Ci-C3 alkyl). In some embodiments, R 2 is -O(CH3), -O(CH 2 CH3), or -O(CH 2 CH 2 CH3). In some embodiments, R 2 is -O(CH3). In some embodiments, R 2 is -O(CH 2 CH3),
  • R 2 is -O(Ci-Ce haloalkyl). In some embodiments, R 2 is -O(Ci- Ce haloalkyl), wherein Ci-Ce haloalkyl contains 1-13 halogen atoms. In some embodiments, R 2 is
  • R 2 is -O(Ci-C3 haloalkyl).
  • R 2 is -O(Ci-C3 haloalkyl), wherein C1-C3 haloalkyl contains 1-7 halogen atoms.
  • R 2 is -OCF3, -OCHF 2 , -OCH 2 F, -OCCI3, -OCHCh, -OCH 2 C1, -OCF 2 C1, -OCFCh, -OCH 2 CF 3 , -OCH 2 CHF 2 , or -OCH 2 CC1 3 .
  • R 2 is -OCF3 or -OCHF 2 .
  • R 2 is -OCF3.
  • R 2 is -OCHF 2 .
  • two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl.
  • two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl.
  • two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-3 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-2 groups selected from F and -CF3.
  • two R 2 groups on adjacent carbon atoms are taken together to form a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, which is optionally substituted by 1-4 groups selected from halo and C1-C3 haloalkyl.
  • two R 2 groups on adjacent carbon atoms are taken together to form a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, which is optionally substituted by 1-4 groups selected from F and -CF3.
  • each R 2 is independently -CF3, -CF2CH3, -CH2CHF2, -CHF2, F, Cl, Br, -OCF3, -OCHF2, -OCH3, or -C(CH3)3, or two R 2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from F or -CF3.
  • L 2 is a bond or O. In some embodiments, L 2 is a bond. In some embodiments, L 2 is O. -Y 1 Y 2 - ⁇ -
  • Ring A is , wherein Y 1 is N or CH; Y 2 is N or
  • Y 1 is N and Y 2 is N. In some embodiments, Y 1 is N and Y 2 is CH. In some embodiments, Y 1 is CH and Y 2 is N. In some embodiments, Y 1 is CH and Y 2 is CH. In some embodiments, x is 0. In some embodiments, x is
  • Ring A is 9- to 11 -membered spiro heterocyclylene, or 8- to 10- membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 1- 2 nitrogen atoms. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 1 nitrogen atom. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 2 nitrogen atoms.
  • Ring A is 8- to 10- membered bicyclic fused heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, Ring A is 8- to 10-membered bicyclic fused heterocyclylene containing 1 nitrogen atom. In some embodiments, Ring A is 8- to 10-membered bicyclic fused heterocyclylene containing 2 nitrogen atoms.
  • — (R 3 )m is drawn across two rings, either ring or both rings can be substituted with m R 3 groups.
  • m is 0-5. In some embodiments, m is 0. In some embodiments, m is 1-3. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5.
  • each R 3 is independently Ci-Ce alkyl, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl.
  • each R 3 is independently C1-C3 alkyl, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl.
  • R 3 is Ci-Ce alkyl. In some embodiments, R 3 is C1-C3 alkyl. In some embodiments, R 3 is methyl, ethyl, w-propyl, or isopropyl. In some embodiments, R 3 is methyl, ethyl, or isopropyl. In some embodiments, R 3 is methyl. In some embodiments, R 3 is ethyl. In some embodiments, R 3 is isopropyl.
  • two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group. In some embodiments, two R 3 groups are taken together to form a bridging - CH2-group. In some embodiments, two R 3 groups are taken together to form a bridging - CH2CH2- group.
  • two R 3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl. In some embodiments, two R 3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl. In some embodiments, two R 3 groups on the same carbon atom are taken together to form a spiro cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, two R 3 groups on the same carbon atom are taken together to form a spiro cyclopropyl. In some embodiments, two R 3 groups on the same carbon atom are taken together to form a spiro cyclobutyl.
  • each R 3 is independently -CH3, or two R 3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 3 groups on the same carbon atom are taken together to form a spiro cyclopropyl.
  • L 3 is a bond, -CH(R a )-, -CH(R a )CH(R a )-, -OCH(R a )CH(R a )-, -CH(R a )CH(R a )N(R a )-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms, and each R a is independently H or Ci-Ce alkyl.
  • L 3 is a bond
  • L 3 is -CH(R a )-, -CH(R a )CH(R a )-, -OCH(R a )CH(R a )-, or -CH(R a )CH(R a )N(R a )-.
  • each R a is independently H or C1-C3 alkyl.
  • each R a is independently H, methyl, ethyl, or propyl.
  • each R a is independently H or methyl.
  • L 3 is -CH2, - CH2CH2-, -CH(CH 3 )CH 2 -,
  • L 3 is 5- to 6-membered heterocyclylene or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms.
  • L 3 is 5- to 6-membered heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, L 3 is 5- to 6-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L 3 is 5- to 6-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L 3 is 5-membered heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, L 3 is 5-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L 3 is 5-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L 3 is 6-membered heterocyclylene containing 1-2 nitrogen atoms.
  • L 3 is 6-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L 3 is 6-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L 3 is pyrrolidinylene, piperidinylene, or piperazinylene.
  • L 3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms. In some embodiments, L 3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1 nitrogen atom. In some embodiments, L 3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 2 nitrogen atoms. In some embodiments, L 3 is -O-(azetidinylene) or -O-(diazetidinylene).
  • L 3 is:
  • Z is N or CH. In some embodiments, Z is N. In some embodiments, Z is CH.
  • r is 0, 1, or 2. In some embodiments, r is 0. In some embodiments, r is 1. In some embodiments, r is 2.
  • s is 0 or 1. In some embodiments, s is 0. In some embodiments, s is 1.
  • r and s are each 1. In some embodiments, r and s are each 0. In some embodiments, r is 1 and s is 0. In some embodiments, r is 2 and s is 1.
  • each R 4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
  • each R 4 is independently halo, -OH, C1-C3 alkyl, or C1-C3 haloalkyl, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5 -membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
  • each R 4 is independently F, -OH, -CH3, -CH(CH3)2, or -CF3, or two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
  • R 4 is halo. In some embodiments, R 4 is F, Cl, Br, or I. In some embodiments, R 4 is F, Cl, or Br. In some embodiments, R 4 is F or Cl. In some embodiments, R 4 is F. In some embodiments, R 4 is Cl.
  • R 4 is -OH.
  • two R 4 groups are taken together to form a bridging -CH2- or -CH2CH2- group. In some embodiments, two R 4 groups are taken together to form a bridging - CH2- group. In some embodiments, two R 4 groups are taken together to form a bridging - CH2CH2- group.
  • two R 4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom.
  • two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group. In some embodiments, two R 4 groups on the same carbon atom are taken together to form a spiro 4- membered heterocyclyl containing 1 oxygen atom. In some embodiments, two R 4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 SO2 group.
  • n is 0-5. In some embodiments, n is 0. In some embodiments, n is 1-3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.
  • the compound of Formula (I) is a compound of Formula (I-a),
  • the compound of Formula (I) is a compound of Formula (I-f),
  • the compound of Formula (I) is a compound of Formula (lie): wherein R 1 , R 2 , R 3 , R 4 , m, and n are as described for Formula (I).
  • the compound of Formula (I) is a compound of Formula (Illa) or (Illb): wherein for Formula (I); and wherein is 9- to 11 -membered spiro heterocyclylene containing 1-2 nitrogen atoms.
  • a compound selected from the compounds in Table 1 or a pharmaceutically acceptable salt thereof is provided.
  • certain compounds described in the present disclosure, including in Table 1 are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 1, are herein described.
  • certain compounds described in the present disclosure are presented as specific salts, it is understood that any pharmaceutically acceptable salt of any of the compounds of the present disclosure are herein described. It is further understood that although certain compounds described in the present disclosure are presented as specific salts, the free form of the compounds of the present disclosure are also herein described.
  • &1 denotes that the absolute stereochemistry was not determined; “abs” denotes that the absolute stereochemistry was determined or a pharmaceutically acceptable salt thereof.
  • a compound selected from the compounds in Table 2 or a pharmaceutically acceptable salt thereof is provided herein.
  • certain compounds described in the present disclosure, including in Table 2 are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 2, are herein described.
  • a compound selected from the compounds in Table 3 or a pharmaceutically acceptable salt thereof is also provided herein.
  • certain compounds described in the present disclosure, including in Table 3 are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 3, are herein described.
  • Piperazinyl-pyrazole compounds of Formula B may be prepared according to the general reactions shown in Scheme 2. Scheme 2. base
  • Compounds of Formula B-l can be treated with an acid, such as TFA, to afford compounds of Formula B-2, which can then be alkylated to give compounds of Formula B.
  • compounds of Formula B-2 can be alkylated with 2-chloroacetaldehyde followed by reductive coupling with HN(R a )(R b ) using a coupling agent such as NaBH(OAc)3.
  • compounds of Formula B-2 can be alkylated with an amine-based alkylchloride to provide compounds of Formula B-2.
  • Piperazinyl amino-pyrazole compounds of Formula C may be prepared according to the general reaction shown in Scheme 3.
  • Compounds of Formula D-l can be arylated by coupling with compounds of Formula D-2 using, for example, Cu(OAc)2, to give compounds of Formula D-3, which can then be alkylated using D-4 under Pd-catalyzed reaction conditions, followed by Pt-catalyzed hydrogenation, to give compounds of Formula D-5.
  • Pd-catalyzed coupling of compounds of Formula D-5 with amines of Formula D-6 affords compounds of Formula D.
  • Compounds of Formula E-l can be treated with compound of Formula E-2 in the presence of an alkoxide to afford compounds of Formula E-3, which can then be treated with hydrazine hydrate to yield compounds of Formula E-4. Subsequent treatment with PtCh and an acid (such as HC1) affords compounds of Formula E-5, which can then be reacted with BOC2O to provide compounds of Formula E.
  • Compounds of Formula E can be arylated with compounds of Formula F-l using, for example, Cu(OAc)2, to give compounds of Formula F-2. Subsequent deprotection with an acid, such as TFA, affords compounds of Formula F-3, which can then be alkylated to afford compounds of Formula F.
  • compounds of Formula F-3 can be reductively coupled with an aldehyde using a coupling agent such as NaBH(OAc)3 to provide compounds of Formula F.
  • compounds of Formula F-3 can be treated with an alkylchloride to provide compounds of Formula F.
  • compositions of the compounds of Formula (I) or a pharmaceutically acceptable salt thereof include pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.
  • Pharmaceutical compositions according to the disclosure may take a form suitable for oral, buccal, sublingual, parenteral (subcutaneous, intramuscular, intravenous, or intrathecal), nasal, topical, vaginal, rectal, intracerebral, intradermal, intravitreal, intraosseous infusion, intraperitoneal, or inhalation administration.
  • Pharmaceutical compositions of the present disclosure comprise a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.
  • a compound described herein can be used in the preparation of a pharmaceutical composition by combining the compound as an active ingredient with a pharmaceutically acceptable excipient.
  • materials which can serve as pharmaceutically acceptable excipients include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; surfactants, such as polysorbate 80 (i.e., Tween 80); powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as
  • wetting agents, emulsifiers and lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.
  • antioxidants examples include: water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.
  • water soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like
  • oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), lecithin, prop
  • the pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy.
  • the amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated and the particular mode of administration.
  • the amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, this amount will range from about 1% to about 99% of active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.
  • a pharmaceutical composition of the present disclosure comprises an excipient selected from the group consisting of cyclodextrins, liposomes, micelle forming agents, e.g., bile acids and polymeric carriers, e.g., polyesters and polyanhydrides; and a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • the pharmaceutical composition renders orally bioavailable a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • compositions of the disclosure suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules or as a solution or a suspension in an aqueous or non-aqueous liquid or as an oil-in-water or water-in-oil liquid emulsion or as an elixir or syrup or as pastilles (using an inert base, such as gelatin and glycerin or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as an active ingredient.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof may also be administered as a bolus, electuary, or paste.
  • the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol, glycerol
  • compositions may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-shelled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
  • a tablet may be made by compression or molding, optionally with one or more accessory ingredients.
  • Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent.
  • Molded tablets may be made in a suitable machine in which a mixture of the powdered compound is moistened with an inert liquid diluent.
  • the tablets and other solid dosage forms of the pharmaceutical compositions of the present disclosure may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be formulated for rapid release, e.g., freeze-dried.
  • compositions may be sterilized by, for example, filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other sterile injectable medium immediately before use.
  • These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner.
  • embedding compositions that can be used include polymeric substances and waxes.
  • the active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.
  • Liquid dosage forms for oral administration of the compounds of Formula (I), or a pharmaceutically acceptable salt thereof include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.
  • the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan and mixtures thereof.
  • inert diluents commonly used in the art, such as, for example, water or other
  • the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
  • Suspensions in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth and mixtures thereof.
  • suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth and mixtures thereof.
  • compositions of the disclosure for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the disclosure with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
  • suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
  • Dosage forms for the topical or transdermal administration of a compound of this disclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.
  • the active compound i.e., a compound of Formula (I) or a pharmaceutically acceptable salt thereof
  • the ointments, pastes, creams, and gels may contain, in addition to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures thereof.
  • excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures thereof.
  • Powders and sprays can contain, in addition to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances.
  • Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
  • compositions of this disclosure suitable for parenteral administration comprise one or more compounds of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
  • sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents
  • aqueous and nonaqueous carriers examples include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils, such as olive oil and injectable organic esters, such as ethyl oleate.
  • polyols such as glycerol, propylene glycol, polyethylene glycol and the like
  • vegetable oils such as olive oil
  • injectable organic esters such as ethyl oleate.
  • Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.
  • the pharmaceutical compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms upon the subject compounds may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenyl sorbic acid and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
  • the absorption of the drug in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
  • Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.
  • biodegradable polymers such as polylactide-polyglycolide.
  • Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.
  • Compounds of Formula (I), or a pharmaceutically acceptable salt thereof, and pharmaceutical compositions comprising compounds of Formula (I), or a pharmaceutically acceptable salt thereof, may be used in methods of administration and treatment as provided herein.
  • the compounds and pharmaceutical compositions may also be used in in vitro methods, such as in vitro methods of administering a compound or pharmaceutical composition to cells for screening purposes and/or for conducting quality control assays.
  • EBP Emopamil Binding Protein
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof inhibits the activity of EBP by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
  • a compound of formula (I) inhibits the activity of EBP by about 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5- 95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5- 35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof binds EBP with an ICso of less than about 10 pM, such as about 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM, or 0.5 pM.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof binds EBP with an ICso from about 0.01 pM to about 5 pM, from about 0.01 pM to about 4 pM, from about 0.01 pM to about 3 pM, from about 0.01 pM to about 2 pM, from about 0.01 pM to about 1 pM, from about 0.01 pM to about 0.05 pM, from about 0.1 pM to about 5 pM, from about 0.1 pM to about 4 pM, from about 0.1 pM to about 3 pM, from about 0.1 pM to about 2 pM, from about 0.1 pM to about 1 pM, from about 0.5 pM to about 5 pM, from about 0.5 pM to about 4 pM, from about 0.5 pM to about 3 pM, from about 0.5 pM to about 2 pM, from about 0.5 pM to about 1 pM, from about
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof binds EBP with an ICso from about 0.01 pM to about 1 pM, from about 0.01 pM to about 0.9 pM, from about 0.01 pM to about 0.8 pM, from about 0.01 pM to about 0.7 pM, from about 0.01 pM to about 0.6 pM, from about 0.01 pM to about 0.5 pM, from about 0.01 pM to about 0.4 pM, from about 0.01 pM to about 0.3 pM, from about 0.01 pM to about 0.2 pM, from about 0.01 pM to about 0.1 pM, from about 0.1 pM to about 1 pM, from about 0.1 pM to about 0.9 pM, from about 0.1 pM to about 0.8 pM, from about 0.1 pM to about 0.7 pM, from about 0.1 pM to about 0.6 pM
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof inhibits EBP and promotes low hERG toxicity.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof binds hERG with an ICso of less than about 30 pM, such as about 25 pM, 20 pM, 15 pM, 10 pM, 5 pM, 2 pM, or 1 pM.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof binds hERG with an ICso from about 0.05 pM to about 30 pM, such as from about 0.05 pM to about 25 pM, from about 0.05 pM to about 20 pM, from about 0.05 pM to about 15 pM, from about 0.05 pM to about 10 pM, from about 0.05 pM to about 5 pM, from about 0.05 pM to about 2 pM, from about 0.05 pM to about 1 pM, from about 1 pM to about 30 pM, such as from about 1 pM to about 25 pM, from about 1 pM to about 20 pM, from about 1 pM to about 15 pM, from about 1 pM to about 10 pM, from about 1 pM to about 5 pM, from about 1 pM to about 2 pM, from about 5 pM to about 30 pM,
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof stimulates remyelination of neuronal axons.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof stimulates remyelination of neuronal axons by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
  • a compound of formula (I) stimulates remyelination of neuronal axons by about 1-100%, 5-100%, 10-100%, 15-100%, 20- 100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65- 100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5- 80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5- 20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.
  • the neuronal axons are artificial, such as those used in a laboratory setting (i.e., cell culture).
  • the neuronal axons are endogenous (i.e., naturally occuring inside a subject, such as a subject having multiple sclerosis (MS)).
  • remyelination of neuronal axons is achieved by oligodendrocyte precursor cells (OPCs), which repair and limit the damage associated with MS.
  • OPCs oligodendrocyte precursor cells
  • administering a compound of Formula (I), or a salt thereof, to a subject having multiple sclerosis (MS) stimulates remyelination to enhance the repair of damaged myelin and to preserve neuronal function.
  • a method for treating a demyelinating disease comprising administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • a method for preventing a demyelinating in a subject in need thereof comprising administering to the subject an effective amount of a compound of Formula (I).
  • the demyelinating disease is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain-Barre syndrome.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof diminishes the extent of the demyelinating disease in the subject.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof stabilizes the demyelinating disease (for example, prevents or delays the worsening of the disease).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the occurrence or recurrence of the demyelinating disease.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof slows the progression of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prevents relapse of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof decreases the dose of one or more other medications required to treat the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof enhances the effect of another medication used to treat the demyelinating disease.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the progression of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof increases the quality of life of the subject having the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prolongs survival of a subject having the demyelinating disease.
  • provided herein is a method of slowing progression of a demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • a method of stabilizing a demyelinating disease in a subject the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • the method prevents the progression of the demyelinating disease.
  • the method delays the progression of the demyelinating disease.
  • a method of delaying the occurrence or recurrence of a demyelinating disease in a subject comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • provided herein is a method of decreasing the dose of one or more other medications required to treat the demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • a method of enhancing the effect of another medication used to treat the demyelinating disease in a subject the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • Also provided here is a method of delaying the progression of the demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, the method increases the quality of life of the subject having the demyelinating disease. In some embodiments, the method prolongs survival of the subject having the demyelinating disease.
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use in treating a demyelinating disease. In other aspects, provided herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the manufacture of medicament for treating a demyelinating disease.
  • a method for treating multiple sclerosis (MS) with EBP inhibition in a subject in need thereof comprising administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
  • a method for preventing multiple sclerosis (MS) in a subject in need thereof comprising administering to the subject an effective amount of a compound of Formula (I).
  • the type of multiple sclerosis is relapsing remitting MS, in which an individual having MS will have episodes of new or worsening symptoms, known as relapses.
  • the type of multiple sclerosis (MS) is primary progressive MS, in which an individual having MS will experience worsening symptoms and/or accumulating symptoms and with no periods of remission.
  • the type of multiple sclerosis (MS) is secondary progressive MS, in which an individual having MS experiences nerve damage or loss, leading to a general worsening of the disease.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof diminishes the extent of the disease multiple sclerosis (MS) (for example, loss of myelin coating of brain and spinal cord nerves, development of plaque around brain and spinal cord nerves, neuroinflammation) in the subject.
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof stabilizes the disease multiple sclerosis (MS) (for example, prevents or delays the worsening of MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the occurrence or recurrence of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof slows the progression of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prevents relapse of the disease multiple sclerosis (MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof decreases the dose of one or more other medications required to treat the disease multiple sclerosis (MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof enhances the effect of another medication used to treat the disease multiple sclerosis (MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the progression of the disease multiple sclerosis (MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof increases the quality of life of the subject having the disease multiple sclerosis (MS).
  • administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prolongs survival of a subject having the disease multiple sclerosis (MS).
  • a method of slowing progression of multiple sclerosis (MS) in a subject comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • a method of stabilizing multiple sclerosis (MS) in a subject comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • the method prevents the progression of multiple sclerosis (MS).
  • the method delays the progression of multiple sclerosis (MS).
  • a method of delaying the occurrence or recurrence of multiple sclerosis (MS) in a subject comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • provided herein is a method of decreasing the dose of one or more other medications required to treat the disease multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • a method of enhancing the effect of another medication used to treat multiple sclerosis (MS) in a subject the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • Also provided here is a method of delaying the progression of the disease multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
  • the method increases the quality of life of the subject having multiple sclerosis (MS).
  • the method prolongs survival of the subject having multiple sclerosis (MS).
  • a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use in treating multiple sclerosis (MS).
  • parenteral administration and “administered parenterally” as used herein mean modes of administration other than enteral and topical administration, usually by injection and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion.
  • systemic administration means the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient’s system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
  • These compounds may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intraci stemally and topically, as by powders, ointments or drops, including buccally and sublingually.
  • the compounds of the present disclosure, or the pharmaceutical compositions of the present disclosure are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art.
  • Actual dosage levels of the active ingredients in the pharmaceutical compositions of this disclosure may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration, without being toxic to the patient.
  • the selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present disclosure employed or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated and like factors well known in the medical arts. A daily, weekly or monthly dosage (or other time interval) can be used.
  • a physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required.
  • the physician or veterinarian could start doses of the compounds of the disclosure employed in the pharmaceutical composition at levels lower than that required to achieve the desired therapeutic effect and then gradually increasing the dosage until the desired effect is achieved.
  • a suitable daily dose of a compound of the disclosure will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect (e.g., inhibit necrosis). Such an effective dose will generally depend upon the factors described above. Generally, doses of the compounds of this disclosure for a patient, when used for the indicated effects, will range from about 0.0001 to about 100 mg per kg of body weight per day. Preferably the daily dosage will range from 0.001 to 50 mg of compound per kg of body weight and even more preferably from 0.01 to 10 mg of compound per kg of body weight.
  • the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.
  • the compounds of the present disclosure are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (such as 0.5% to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.
  • the compounds of the present application or the pharmaceutical compositions thereof may be administered once, twice, three, or four times daily, using any suitable mode described above. Also, administration or treatment with the compounds may be continued for a number of days; for example, commonly treatment would continue for at least 7 days, 14 days, or 28 days, for one cycle of treatment. Treatment cycles are well known and are frequently alternated with resting periods of about 1 to 28 days, commonly about 7 days or about 14 days, between cycles. The treatment cycles, in certain embodiments, may also be continuous.
  • the total daily dosage for a human subject may be between about 1 mg and 1,000 mg, between about 1,000-2,000 mg/day, between about 10-500 mg/day, between about 50-300 mg/day, between about 75-200 mg/day or between about 100-150 mg/day.
  • the daily dosage may also be described as a total amount of a compound described herein administered per dose or per day.
  • Daily dosage of a compound may be between about 1 mg and 4,000 mg, between about 2,000 to 4,000 mg/day, between about 1 to 2,000 mg/day, between about 1 to 1,000 mg/day, between about 10 to 500 mg/day, between about 20 to 500 mg/day, between about 50 to 300 mg/day, between about 75 to 200 mg/day or between about 15 to 150 mg/day.
  • the method comprises administering to the subject an initial daily dose of about 1 to 800 mg of a compound described herein and increasing the dose by increments until clinical efficacy is achieved. Increments of about 5, 10, 25, 50 or 100 mg can be used to increase the dose. The dosage can be increased daily, every other day, twice per week or once per week.
  • a compound or pharmaceutical preparation is administered orally.
  • the compound or pharmaceutical preparation is administered intravenously.
  • Alternative routes of administration include sublingual, intramuscular and transdermal administrations.
  • the preparations of the present disclosure may be given orally, parenterally, topically, or rectally. They are, of course, given in forms suitable for each administration route. For example, they are administered in tablets or capsule form; by injection, inhalation, eye lotion, ointment, suppository, infusion, inhalation, etc.; topical by lotion or ointment; and rectal by suppositories.
  • the administration is oral.
  • kits and articles of manufacture for use with one or more compounds, compositions, or methods described herein.
  • Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein.
  • Suitable containers include, for example, bottles, vials, syringes, and test tubes.
  • the containers are formed from a variety of materials such as glass or plastic.
  • a kit typically includes labels listing contents and/or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.
  • a label is on or associated with the container.
  • a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert.
  • a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.
  • the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein.
  • the pack for example, contains metal or plastic foil, such as a blister pack.
  • the pack or dispenser device is accompanied by instructions for administration.
  • the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for drugs, or the approved product insert.
  • compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.
  • Step 1 Synthesis of tert-butyl 4-(3-oxobutanoyl) piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.32% (214 nm), Mass: found peak 293.3 (M+23) at 1.039 min.
  • Step 2 Synthesis of tert-butyl 4-(3-oxobutanethioyl) piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
  • Step 3 Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine-1- carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.86% (214 nm), Mass: found peak 267.3 (M+1) at 1.022 min.
  • Step 4 Synthesis of tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine-1- carboxylate.
  • the filtrate was purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine- 1 -carboxylate (0.54 g, yield 70%) as a yellow solid.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 343.4 (M+l) at 1.395 min.
  • Step 5 Synthesis of l-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.07% (214 nm) Mass: found peak 243.3 (M+l) at 0.928 min.
  • Step 6 Synthesis of l-(5-methyl-l-phenyl-pyrazol-3-yl)-4-(tetrahydropyran-4- ylmethyl) piperazine (Compound 1).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 341.3 (M+l) at 1.813 min.
  • the filtrate was purified by flash chromatography (Biotage, 40 g silica gel column @50 mL/min, eluting with 5-40% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4- [5 -methyl- 1 -(2 -naphthyl) pyrazol-3-yl] piperazine-l-carboxylate (160 mg, yield 54.3%) as a yellow oil.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 393.3 (M+l) at 1.503 min.
  • Step 2 Synthesis of l-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine.
  • Step 3 Synthesis of 4-[2-[4-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazin-1- yl] ethyl] morpholine (Compound 2).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 406.3 (M+l) at 1.751 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 355.3 (M+l) at 1.794 min.
  • Step 1 Synthesis of l-(4-Pyridinyl)-l,3-butanedione.
  • LCMS method Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3min, 95%B for 0.7min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 164.1(M+1) at 0.791 min.
  • Step 2 Synthesis of 4-(3-methyl-lH-pyrazol-5-yl) pyridine.
  • LCMS method Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3min, 95%B for 0.7min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 90% (214 nm), Mass: found peak 160.1 (M+l) at 0.303 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B); Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Column: X-BRIDGE C18, 4.6x50 mm, 3.5 pm; Temperature: 50 °C; LC purity: 90% (214 nm), Mass: found peak 166.1(M+1) at 0.592 min.
  • Step 4 Synthesis of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl) piperidine-1- carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile(B); Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Column: X-BRIDGE C18, 4.6x50 mm, 3.5 pm; Temperature: 50 °C; LC purity: 77% (214 nm), Mass: found peak 210.2 (M-55) + at 1.690 min.
  • Step 5 Synthesis of tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl)piperidine-l- carboxylate.
  • reaction mixture was concentrated in vacuo to dryness and purified by prep-HPLC (water/ammonium hydrogen carbonate/acetonitrile) to afford the desired product tert-butyl 4-(5-methyl-l-phenyl- pyrazol-3-yl)piperidine-l -carboxylate (420 mg, yield 48%) as a yellow oil.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2 min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 95.53% (214 nm), Mass: found peak 342.2 (M-55) + at 1.374 min.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
  • Step 7 Synthesis of 4-(5-methyl-l-phenyl-pyrazol-3-yl)-l-(tetrahydropyan-4- ylmethyl) piperidine (Compound 4).
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3min, 95% B for 1.7min; Flow rate: 2mL/min;
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3min, 95% B for 1.7min; Flow rate: 2mL/min;
  • Example S6 Synthesis of 4-[2-[4-[l-[4-fluoro-3-(trifluoromethoxy) phenyl] -5-methyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 6).
  • Step 2 Synthesis of [4-fluoro-3-(trifluoromethoxy) phenyl] boronic acid.
  • the filtrate was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[l-[4-fluoro-3 -(trifluoromethoxy) phenyl]-5-methyl-pyrazol-3- yl] piperazine- 1 -carboxylate (160 mg, yield 91.1%) as a yellow solid.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 95.04% (214 nm) Mass: found peak 445.3 (M+l) at 1.527 min.
  • Step 4 Synthesis of l-[l-[4-fluoro-3-(trifluoromethoxy) phenyl]-5-methyl-pyrazol- 3-yl] piperazine.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95.22% (214 nm) Mass: found peak 345.1 (M+l) at 1.989 min.
  • Step 5 Synthesis of 4-[2-[4-[l-[4-fluoro-3-(trifluoromethoxy) phenyl]-5-methyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 6).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 458.1 (M+l) at 2.029 min.
  • Example S7 Synthesis of 4-[2-[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] pyrrolidin-3-yl] oxyethyl] morpholine (Compound 7). [00337] Compound 7 was prepared as outlined below.
  • Step 1 Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] pyr rolidin-3-ol.
  • reaction mixture was directly purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-80% ethyl acetate in petroleum ether) to afford the desired product l-[5-methyl-l- [4-(trifluoromethoxy) phenyl]pyrazol-3-yl]pyrrolidin-3-ol (97 mg, yield 17.9%) as a yellow oil.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 93.81% (214 nm) Mass: found peak 328.2 (M+l) at 1.164 min.
  • Step 2 Synthesis of 4-[2-[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] pyrrolidin-3-yl] oxyethyl] morpholine (Compound 7). [00342] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]pyrrolidin-3- ol (97 mg, 0.296 mmol) in dry DMF (9 mL) was added sodium hydride (60%, 237 mg, 5.93 mmol). The reaction was stirred at 60°C for 2h.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
  • Step 1 Synthesis of methyl (2S,4R)-l-[2-(tert-butoxycarbonylamino)acetyl]-4- hydroxy-pyrrolidine-2-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.42% (214 nm) Mass: found peak 325.1 (M+23)+ at 1.567 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 171 (M+l) at 0.349 min.
  • Step 3 Synthesis of (7R,8aS)-l,2,3,4,6,7,8,8a-octahydropyrrolo[l,2-a]pyrazin-7-ol.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B with 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; Mass: found peak 143 (M+l) at 0.367 min.
  • Step 4 Synthesis of (7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-ol.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90.87% (214 nm) Mass: found peak 383.1 (M+l) at 1.885 min.
  • Step 5 Synthesis of [(7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-yl] 4-methylbenzenesulfonate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94.66% (214 nm) Mass: found peak 537.1 (M+l) at 2.262 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
  • Step 1 Synthesis of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo [3.1.1] heptane-6-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 76 % (214 nm); Mass: found peak 439.3 (M + H) at 2.16 min.
  • Step 2 Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo-[3.1.1]-heptane.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 78 % (214 nm); Mass: found peak 339.2 (M + H) at 1.55 min.
  • Step 3 Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-3, 6-diazabicyclo [3.1.1] heptan-6-yl] ethyl] morpholine (Compound 9).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 452.2 (M + H) at 2.00 min.
  • Step 1 Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] -2, 5-diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100 % (214 nm); Mass: found peak 439.3 (M + H) at 2.15 min.
  • Step 2 Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2, 5-diazabicyclo [2.2.1] heptane.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
  • Step 3 Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-2, 5-diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine (Compound 10).
  • LCMS method Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 452.1 (M + H) at 1.92 min.
  • Step 1 Synthesis of 3,5-dibromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl) pyrazole.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 93 % (214 nm); Mass: found peak 382.9 (M + H) at 1.48 min.
  • Step 2 Synthesis of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropenyl- pyrazole.
  • the filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)- 5-isopropenyl-pyrazole (91 mg, yield: 29.6%) as a colorless oil.
  • LCMS method Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 92 % (214 nm); Mass: found peak 343.0 (M + H) at 2.25 min.
  • Step 3 Synthesis of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl- pyrazole.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 85 % (214 nm); Mass: found peak 345.0(M + H) at 2.27 min.
  • Step 4 Synthesis of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5- isopropyl-pyrazol-3-yl] piperazine-l-carboxylate.
  • LCMS method Column: HALO C18 2.7pm 4.6*30mm. Mobile phase: Water (0.01%TFA) (A) / ACN (0.01% TFA) (B). Elution program: Gradient from 5 to 95% of B in l.Omin at 2.2 mL/min. Temperature: 40°C. LC purity: 85% (214 nm), Mass: found peak 451.3 (M+l) at 1.55 min.
  • Step 5 Synthesis of l-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl-pyrazol-
  • Step 6 Synthesis of 4-[2-[4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 11).
  • LCMS method Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 464.1 (M + H) at 2.05 min.
  • Step 1 Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol- 3-yl] piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 411.1 (M + H) at 2.27 min.
  • Step 2 Synthesis of l-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine.
  • Step 3 Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 12).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 424.2 (M + H) at 2.01 min.
  • Step 1 Synthesis of tert-butyl 4-(3-oxobutanoyl)piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 215.1 (M-56)+ at 1.30 min.
  • Step 2 Synthesis of tert-butyl 4-(3-oxobutanethioyl)piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.32% (254 nm) Mass: found peak 287.0 (M+l) at 1.51 min.
  • Step 3 Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.32% (254 nm) Mass: found peak 287.0 (M+l) at 1.51 min. LC purity: 98.33% (254 nm) Mass: found peak 267.1 (M+l) at 1.33 min.
  • Step 4 Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
  • Step 5 Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
  • Step 1 Synthesis of tert-butyl 4-(2-morpholinoethoxy) piperidine-l-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95% (214 nm), Mass: found peak 315.2 (M+l) at 1.861 min.
  • Step 2 Synthesis of 4- [2-(4-piperidyloxy)ethyl] morpholine.
  • the filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]cyclohexoxy] ethyl]morpholine (11.7 mg, yield 19%) as a colorless oil.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98% (214 nm), Mass: found peak 455.2 (M+l) at 2.108 min.
  • Step 1 Synthesis of tert-butyl 3-(4-pyridyloxy)azetidine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 70% (214 nm), Mass: found peak 251.2 (M+l) at 0.871 min.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 50% (214 nm), Mass: found peak 257.4 (M+l) at 0.87 min.
  • Step 4 Synthesis of 4-(azetidin-3-yloxy)-l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperidine.
  • [00458] To a solution of tert-butyl 3-[[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-4-piperidyl] oxy] azetidine- 1 -carboxylate (160 mg, 0.3 mmol) in dichloromethane (10 mL) was added TFA (3 mL).
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 86% (214 nm), Mass: found peak 397.2 (M+l) at 1.075 min.
  • Step 5 Synthesis of l-[5-methyl-l-[4 (trifluoromethoxy) phenyl]pyrazol-3-yl]-4-(l- tetrahydropyran-4-ylazetidin-3-yl)oxy-piperidine (Compound 14).
  • the filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product l-[5-methyl-l-[4 (trifluoromethoxy) phenyl]pyrazol-3-yl]-4-(l-tetrahydropyran-4-ylazetidin-3-yl)oxy-piperidine (96.5 mg, yield 62.7 %) as a yellow oil.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm), Mass: found peak 481.2 (M+l) at 2.096 min.
  • Example S15 Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2- tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (Compound 15).
  • Step 1 Synthesis of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2,7-diazaspiro [3.5] nonane-2-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 66% (214 nm), Mass: found peak 367.3 (M+l) at 1.077 min.
  • Step 3 Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2- tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (Compound 15).
  • the filtrate was purified by prep- HPLC (ammonium hydrogen carbonate) to afford the desired product 7-[5-methyl-l-[4- (trifluoromethoxy) phenyl] pyrazol-3-yl]-2-tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (33.5 mg, yield 34.1%) as a yellow solid.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93% (214 nm), Mass: found peak 451.2 (M+l) at 2.144 min.
  • Step 1 Synthesis of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl] pyrazole. CHCI 3 , RT, o/n
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 95% (214 nm); Mass: found peak 387.0 (M+l) at 1.493 min.
  • Step 2 Synthesis of 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole. dioxane/H2O(10:1 )
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 67% (214 nm); Mass: found peak 347.1 (M+l) at 1.511 min.
  • Step 3 Synthesis of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazole.
  • LCMS method Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 66% (214 nm) Mass: found peak 349.1 (M+l) at 2.089 min.
  • Step 4 Synthesis of tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate. Boc THF, 60°c, o/n
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 455.3 (M+l) at 1.562 min.
  • Step 5 Synthesis of tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
  • Step 6 Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 16).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm), Mass: found peak 468.2 (M+l) at 2.115 min.
  • Step 1 Synthesis of tert-butyl thiomorpholine-4-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min 95%B for 1.7min at 2.0 mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50 °C; LC purity: 97% (214 nm) Mass: found peak 226.3 (M+23) at 1.266min.
  • Step 2 Synthesis of tert-butyl l-oxo-l,4-thiazinane-4-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3min 95%B for 1.7min at 2.0mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 369.2(M+1) at 1.250 min.
  • Step 4 Synthesis of benzyl N-(l-oxo-l,4-thiazinan-l-ylidene) carbamate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 97.91% (214 nm), Mass: found peak 269.2 (M+l) + , at 0.867 min.
  • Step 5 Synthesis of benzyl N-[4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l-yl]ethyl]-l-oxo-l,4-thiazinan-l-ylidene]carbamate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.68% (214 nm) Mass: found peak 621.3 (M+l) at 1.148 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 487.1 (M+l) at 1.861 min.
  • Step 1 Synthesis of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
  • LCMS method Column: X Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B in 1.5 min at 1.5 mL/minTemperature: 50 °C; LC purity: 98.8% (214 nm) Mass: found peak 510.2 (M+l) at 2.441 min.
  • LCMS method Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: from 5% increase to 95%B within 1.4 min, 95%B for 1.6 min. Flow Rate: 1.8 mL/min; Column: X Bridge C18, 4.6*50mm, 3.5pm; Column Temperature: 45 °C; LC purity: 83.4% (214 nm) Mass: found peak 409.9 (M+l) at 1.845 min.
  • Step 3 Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy- 3-tetrahydropyran-4-yl-3-azaspiro[5.5] undecane (Compound 18).
  • LCMS method Column: Sun fire, 50*4.6mm, 3.5pm; Mobile phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Elution program: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min Flow Rate: 2 mL/min; Temperature: 50°C; LC purity: 91.0% (214 nm) Mass: found peak 494.0 (M+l) at 1.650 min.
  • LCMS method Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Gradient: 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 88.0% (214 nm) Mass: found peak 141.3 (M+l) at 0.963 min.
  • Step 3 Synthesis of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol.
  • LCMS method Column: HALO C18 2.7pm 4.6*30mm; Mobile phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Elution program: Gradient from 5% to 95% of B in 1.0 min at 2.2 mL/min; Temperature: 40°C; LC purity: 98.4% (214 nm) Mass: found peak 259.2 (M+l) at 1.204 min.
  • Step 4 Synthesis of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] oxy-3-azaspiro [5.5] undecane-3-carboxylate.
  • LCMS method Column: X Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B ini.5 min at 1.5 mL/min. Temperature: 50 °C; LC purity: 98.7% (214 nm) Mass: found peak 426.1 (M-56+l)+ at 2.364 min.
  • LCMS method Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 10% to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 87.3% (214 nm) Mass: found peak 382.1 (M+l) at 1.907 min.
  • LCMS method Column: Sun fire, 50*4.6mm, 3.5pm; Mobile phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Elution program: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min Flow Rate : 2 mL/min; Temperature: 50°C; LC purity: 97.0% (214 nm) Mass: found peak 466.0 (M+l) at 1.610 min.
  • Step 1 Synthesis of tert-butyl 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]oxy-7-azaspiro[3.5]nonane-7-carboxylate.
  • LCMS method Column: X-Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10% to 95% of B in 1.5 min at 1.8 mL/min; Temperature: 50°C; LC purity: 92.3% (214 nm) Mass: found peak 426.1 (M-56+l)+ at 2.374 min.
  • Step 2 Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]oxy- 7-azaspiro [3.5] nonane.
  • LCMS method Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 5% to 95% of B in 1.5 min at 1.5 mL/min; Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 52.0% (214 nm) Mass: found peak 382.1 (M+l) at 2.086 min.
  • LCMS method Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10% to 95% of B in 1.5 min at 1.8 mL/min; Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 466.1 (M+l) at 2.146 min.
  • Step 1 Synthesis of tert-butyl 4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5- methyl-pyrazol-3-yl]piperazine-l-carboxylate.
  • LCMS method Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Gradient: from 5% to 95% of B ini.5 min at 1.5 mL/min, Column: X Bridge C18 (4.6x 50 mm, 3.5pm), Column Temperature: 50 °C, LC purity: 100% (254 nm) Mass: found peak 427.1 (M+l) at 2.179 min.
  • Step 2 Synthesis of l-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl-pyrazol- 3-yl] piperazine.
  • LCMS method Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B), Gradient: from 5% to 95% of B in 1.0 min at 2.2 mL/min, Column: HALO C18 2.7pm 4.6*30mm, Column Temperature: 40 °C, LC purity: 74.6% (214 nm) Mass: found peak 327.5 (M+l) at 0.954 min.
  • Step 3 Synthesis of 4-[2-[4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 21).
  • LCMS method Mobile Phase: WATER (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Gradient: from 5% to 95% of B ini.5 min at 1.5 mL/min, Column: X Bridge C18 (4.6x 50 mm, 3.5pm), Column Temperature: 50 °C, LC purity: 100% (214 nm) Mass: found peak 440.2 (M+l) at 1.900 min.
  • Example S22 Synthesis of 5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethoxy)phenyl] pyrazole-3-carbonitrile (Compound 22) and 5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethoxy)phenyl]pyrazole-3-carboxamide (Compound a3).
  • Step 1 Synthesis of trimethyl-[2-(pyrazol-l-ylmethoxy)ethyl]silane. THF 0 C-RT, 16 h
  • Step 3 Synthesis of 5-iodo-2-(2-trimethylsilylethoxymethyl)pyrazole-3- carbonitrile.
  • M (2 eq) SEM h _ ⁇ NC ⁇ _N CgH 18 CI 2 MgN.Li (1 .5 eq) THF, -16 °c, 2 h
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 80.28% (214 nm) Mass: found peak 382.2 (M-55) + at 2.297 min.
  • Step 7 Synthesis of 5-piperazin-l-yl-2-[4-(trifluoromethoxy)phenyl] pyrazole-3- carbonitrile.
  • Step 8 Synthesis of 5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethoxy)phenyl] pyrazole-3-carbonitrile (Compound 22) and 5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethoxy)phenyl]pyrazole-3-carboxamide (Compound a3).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C.
  • Step 1 Synthesis of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
  • Step 2 Synthesis of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl] piperazine.
  • Step 3 Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 23).
  • Step 1 Synthesis of tert-butyl 4-(3-oxobutanoyl) piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97.84% (214 nm) Mass: found peak 215.3 (M-55) + at 1.48 min.
  • Step 2 Synthesis of tert-butyl 4-(3-oxobutanethioyl) piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
  • Step 3 Synthesis of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97.73% (214 nm) Mass: found peak 422.9 (M + 1)+ at 2.036 min.
  • Step 4 Synthesis of l-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl] piperazine.
  • LCMS method Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 323.2 (M + 1)+ at 1.644 min.
  • Step 5 Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.56 % (214 nm); Mass: found peak 436.2 (M + H) at 1.758 min.
  • Step 1 Synthesis of tert-butyl 4-[l-(4-fluoro-3-formyl-phenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: CH 3 CN (0.01%TFA);
  • Step 2 Synthesis of tert-butyl 4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: CH3CN; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X- Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96 % (214 nm); Mass: found peak 411.2 (M + H) at 2.15 min.
  • Step 3 Synthesis of l-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-3- yl] piperazine.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95 % (214 nm); Mass: found peak 323.2 (M + H) at 1.49 min.
  • Step 4 Synthesis of 4-[2-[4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 24).
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.5 min, 95%B for 1.5 min, back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column: Sunfire C18 ,4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 423.9 (M + H) at 1.71 min.
  • Example S25 Synthesis of 4-[(7R,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido [l,2-a]pyrazin-7-yl]morpholine (Compound 25). [00636] Compound 25 was prepared as outlined below.
  • Step 1 Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; Mass: found peak 160 (M + H) at 0.399 mm.
  • Step 2 Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 100% (214 nm); Mass: found peak 160 (M -100) + at 1.061 min.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak Mass: found peak 158.3 (M- 100)+ at 1.111 min.
  • Step 4 Synthesis of 1-tert-butyl 2-methyl (2S)-5-morpholinopiperidine-l,2- dicarboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 329.2(M+1) at 1.1850 min.
  • Step 5 Synthesis of methyl (2S)-5-morpholinopiperidine-2-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: 229.1 [M-100]+ at 1.358 min.
  • Step 6 Synthesis of methyl l-[2-(l, 3-dioxoisoindolin-2-yl) ethyl]-4-morpholino- piperidine-2-carboxylate.
  • LCMS method Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 94.33% (214 nm); Mass: found peak 402.2 (M + H) at 0.855 min.
  • Step 7 Synthesis of (9aS)-7-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2- a]pyrazin-l-one.
  • reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 25-P1 4-[(7R,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido [l,2-a]pyrazin-7-yl]morpholine (36.5 mg, yield 10%) as a yellow solid and product 25-P2 4-[(7S,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido
  • 25-P1 LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.62% (214 nm) Mass: found peak 465.9 (M+l) at 1.747 min.
  • 25-P2 LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 465.9 (M+l) at 1.907 min.
  • Step 1 Synthesis of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-4,7-diazaspiro [2.5] octane-4-carboxylate.
  • reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0- 10% EA in PE) to afford the desired product tert-butyl 7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7-diazaspiro[2.5]octane-4-carboxylate (160 mg, yield 34.7%) as a yellow solid.
  • LCMS method Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.78% (214 nm) Mass: found peak 414.9 (M+l) at 2.055 min.
  • reaction was cooled to room temperature, directly purified by prep-HPLC (ammonium hydrogen carbonate/ water/ acetonitrile) to afford the desired product 4-[2-[7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7-diazaspiro[2.5]octan-4-yl]ethyl]-l,4-thiazinane 1,1- dioxide (16.5 mg, yield 12.1%) as a brown solid.
  • prep-HPLC ammonium hydrogen carbonate/ water/ acetonitrile

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Abstract

Provided herein are compounds and pharmaceutical compositions thereof for inhibiting Emopamil Binding Protein (EBP) and their use in the treatment of demyelinating diseases such as multiple sclerosis.

Description

PYRAZOLYL COMPOUNDS AS EMOPAMIL BINDING PROTEIN INHIBITORS
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Application No. 63/579,111, filed on August 28, 2023, the disclosure of which is hereby incorporated by reference in its entirety.
FIELD OF INVENTION
[0002] The present disclosure relates to inhibitors of Emopamil Binding Protein (EBP) useful for treating demyelinating diseases such as multiple sclerosis. Specifically, this disclosure describes compounds and compositions for inhibiting EBP, methods of treating demyelinating diseases, and methods of synthesizing these compounds.
BACKGROUND OF THE DISCLOSURE
[0003] Myelin is an insulating layer, or sheath, that wraps around nerve cell axons. This lipid- rich insulating material protects the axons and makes possible the saltatory conduction, which speeds axonal electric impulse (Williamson et al., Front Cell Neurosci., 2018, 12: 424). Myelin is formed in the central nervous system (CNS) by glial cells called oligodendrocytes and in the peripheral nervous system (PNS) by glial cells called Schwann cells.
[0004] Demyelinating diseases cause damage to the myelin sheath and such diseases include multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), and Guillain- Barre syndrome. Multiple sclerosis (MS) is a complex neurological disease characterized by deterioration of CNS myelin (Ghasemi et al., Cell J., 2017, 19(1): 1-10). In the pathogenesis of MS, the myelin sheaths surrounding neuronal axons are damaged or destroyed, leading to dysfunction of propagated signals, axonal injury, neuronal loss, accumulation of lesion load, and overt neurological disabilities in patients.
[0005] The CNS has the capacity to regenerate myelin sheaths after injury through the proliferation, migration, and differentiation of a population of adult progenitor cells referred to as oligodendrocyte precursor cells (OPCs) into new myelinating oligodendrocytes to generate new myelin sheaths, or to a lesser extent, by generation of new myelin sheaths by existing oligodendrocytes (Kuhn et al., Cells, 2019, 8(11): 1424). While such endogenous remyelination by OPCs and oligodendrocytes can repair and limit the damage in early stages of MS, this process becomes inefficient during later stages of disease progression (Chari, Int Rev Neurobiol., 2007, 79: 589-620). Therefore, stimulating remyelination by enhancing the repair of damaged myelin to preserve neuronal function is a focus for developing new therapeutics for the treatment of MS.
[0006] Recent work established inhibition of specific cholesterol biosynthesis pathway enzymes as a functional mechanism by which many small molecules can enhance oligodendrocyte formation (Hubler et al., Nature, 2018, 560(7718): 372-376). These molecules enhanced oligodendrocyte formation by inhibiting the enzymes and causing accumulation of the 8,9-unsaturated sterol substrates of these enzymes, such as zymosterol and zymostenol.
[0007] Emopamil Binding Protein (EBP, also referred to as A8A7 isomerase, 3-beta- hydroxysteroid-Delta(8),Delta(7)-isomerase, human sterol isomerase (HSI), CDPX2, CH02, CPX, or CPXD) is the enzyme in the cholesterol biosynthesis pathway which catalyzes the conversion of zymosterol and zymostenol to dehyolathosterol and lathosterol (Silve et al., The Journal of Biological Chemistry, 1996, 271(37): 22434-22440). As such, targeting EBP is one strategy for increasing OPC differentiation. However, there exists a need for compounds and therapeutic methods capable of targeting EBP to induce differentiation of OPCs to enhance generation of new oligodendrocytes and increase myelination and/or remyelination in demyelinated lesions.
[0008] Accordingly, in one aspect, provided herein are compounds which inhibit EBP for use in treating demyelinating diseases such as multiple sclerosis.
SUMMARY OF THE DISCLOSURE
[0009] Described herein, in certain embodiments, are compounds and compositions thereof for inhibiting Emopamil Binding Protein (EBP) for treating demyelinating diseases such as multiple sclerosis.
[0010] The following embodiments are encompassed.
[0011] Embodiment 1. A compound of Formula (I):
Figure imgf000003_0001
or a pharmaceutically acceptable salt thereof, wherein:
X1 is C and X2 is N, or X1 is N and X2 is C;
= is a single bond or a double bond, provided that a double bond and a single bond; R1 is Ci-C6 alkyl, C3-C6 cycloalkyl, -CN, Ci-C6 haloalkyl, -(Ci-C6 alkylene)-O-(Ci-C6 alkyl), -(Ci-Ce alkylene)-O-(Ci-C6 haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl);
L1 is a bond, O, or -CH2-;
Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing 1 or 2 nitrogen atoms, or Ce-Cio aryl; each R2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, -O(Ci-Ce alkyl), or
-O(Ci-C6 haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl;
L2 is a bond or O;
Ring A is
Figure imgf000004_0001
, 9- to 11-membered spiro heterocyclylene, or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms;
Y1 is N or CH;
Y2 is N or CH; x is 0, 1, or 2; y is 0 or 1; m is 0-5; each R3 is independently Ci-Ce alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl;
L3 is a bond, -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, -CH(Ra)CH(Ra)N(Ra)-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms; each Ra is independently H or Ci-Ce alkyl;
W is O, CH2, SO2, S(O)=NH, SO, or N(H);
Z is N or CH; r is 0, 1, or 2; s is 0 or 1; each R4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group; and n is 0-5.
[0012] Embodiment 2. The compound of embodiment 1, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000005_0001
[0013] Embodiment 3. The compound of embodiment 1, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000005_0002
[0014] Embodiment 4. The compound of any one of embodiments 1-3, or a pharmaceutically acceptable salt thereof, wherein:
R1 is C1-C6 alkyl, C3-C5 cycloalkyl, -CN, C1-C3 haloalkyl, -(C1-C3 alkylene)-O-(Ci-C3 alkyl), -(C1-C3 alkylene)-O-(Ci-C3 haloalkyl), or -(C1-C3 alkylene)(C3-Ce cycloalkyl).
[0015] Embodiment 5. The compound of embodiment 4, or a pharmaceutically acceptable salt thereof, wherein:
R1 is -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -CH2CH2CH(CH3)2, -CH2OCH3, -CH2CH2OCH3, -CH(CH3)OCH3, -CH(CH3)CH2OCH3, -CH2CHF2, -CF3, -CHF2,
Figure imgf000005_0003
[0016] Embodiment 6. The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein: L1 is a bond.
[0017] Embodiment 7. The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein: L1 is O.
[0018] Embodiment 8. The compound of any one of embodiments 1-5, or a pharmaceutically acceptable salt thereof, wherein:
L1 is -CH2-. [0019] Embodiment 9. The compound of any one of embodiments 1-8, or a pharmaceutically acceptable salt thereof, wherein:
Ring B is C4-C6 cycloalkyl, pyridinyl, pyrazinyl, pyrimidinyl, or phenyl.
[0020] Embodiment 10. The compound of any one of embodiments 1-9, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000006_0001
[0021] Embodiment 11. The compound of any one of embodiments 1-10, or a pharmaceutically acceptable salt thereof, wherein: each R2 is independently C1-C4 alkyl, C1-C3 haloalkyl, halo, -O(Ci-C3 alkyl), or
-O(Ci-C3 haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl.
[0022] Embodiment 12. The compound of embodiment 11, or a pharmaceutically acceptable salt thereof, wherein: each R2 is independently -CF3, -CF2CH3, -CH2CHF2, -CHF2, F, Cl, Br, -OCF3, -OCHF2, -OCH3, or -C(CH3)3, or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from F or -CF3.
[0023] Embodiment 13. The compound of any one of embodiments 1-12, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000007_0001
[0024] Embodiment 14. The compound of any one of embodiments 1-13, or a pharmaceutically acceptable salt thereof, wherein:
L2 is a bond.
[0025] Embodiment 15. The compound of any one of embodiments 1-13, or a pharmaceutically acceptable salt thereof, wherein:
L2 is O.
[0026] Embodiment 16. The compound of any one of embodiments 1-15, or a pharmaceutically acceptable salt thereof, wherein:
Ring
Figure imgf000007_0002
Y1 is N or CH;
Y2 is N or CH; x is 0, 1, or 2; and y is 0 or 1.
[0027] Embodiment 17. The compound of any one of embodiments 1-16, or a pharmaceutically acceptable salt thereof, wherein:
Ring A is 9- to 11 -membered spiro heterocyclylene or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms.
[0028] Embodiment 18. The compound of any one of embodiments 1-17, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000008_0001
Figure imgf000009_0001
[0029] Embodiment 19. The compound of any one of embodiments 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 0.
[0030] Embodiment 20. The compound of any one of embodiments 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 1-3.
[0031] Embodiment 21. The compound of any one of embodiments 1-18 and 20, or a pharmaceutically acceptable salt thereof, wherein: each R3 is independently C1-C3 alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl.
[0032] Embodiment 22. The compound of embodiment 21, or a pharmaceutically acceptable salt thereof, wherein: each R3 is independently -CEE, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro cyclopropyl.
[0033] Embodiment 23. The compound of any one of embodiments 1-22, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000009_0002
Figure imgf000010_0001
[0034] Embodiment 24. The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein: L3 is a bond.
[0035] Embodiment 25. The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein:
L3 is -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, or -CH(Ra)CH(Ra)N(Ra)-; and each Ra is independently H or C1-C3 alkyl.
[0036] Embodiment 26. The compound of embodiment 25, or a pharmaceutically acceptable salt thereof, wherein:
L3 is -CH2, -CH2CH2-, -CH(CH3)CH2-, -CH2CH(CH3)-, -OCH2CH2-, or -CH2CH2N(CH2CH3)-.
[0037] Embodiment 27. The compound of any one of embodiments 1-23, or a pharmaceutically acceptable salt thereof, wherein:
L3 is 5- to 6-membered heterocyclylene or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms.
[0038] Embodiment 28. The compound of embodiment 27, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000010_0002
[0039] Embodiment 29. The compound of any one of embodiments 1-28, or a pharmaceutically acceptable salt thereof, wherein:
W is O, CH2, or N(H).
[0040] Embodiment 30. The compound of any one of embodiments 1-28, or a pharmaceutically acceptable salt thereof, wherein:
W is SO2, S(O)=NH, or SO. [0041] Embodiment 31. The compound of any one of embodiments 1-30, or a pharmaceutically acceptable salt thereof, wherein:
Z is N.
[0042] Embodiment 32. The compound of any one of embodiments 1-30, or a pharmaceutically acceptable salt thereof, wherein:
Z is CH.
[0043] Embodiment 33. The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 1.
[0044] Embodiment 34. The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 0.
[0045] Embodiment 35. The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 1; and s is 0.
[0046] Embodiment 36. The compound of any one of embodiments 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 2; and s is 1.
[0047] Embodiment 37. The compound of any one of embodiments 1-36, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000011_0001
Figure imgf000012_0001
[0048] Embodiment 38. The compound of any one of embodiments 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 0.
[0049] Embodiment 39. The compound of any one of embodiments 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 1-3.
[0050] Embodiment 40. The compound of any one of embodiments 1-37 and 39, or a pharmaceutically acceptable salt thereof, wherein: each R4 is independently halo, -OH, C1-C3 alkyl, or C1-C3 haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
[0051] Embodiment 41. The compound of embodiment 40, or a pharmaceutically acceptable salt thereof, wherein: each R4 is independently F, -OH, -CH3, -CH(CH3)2, or -CF3, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
[0052] Embodiment 42. The compound of any one of embodiments 1-41, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000013_0001
[0053] Embodiment 43. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-a), (I-b), or (I-c):
Figure imgf000013_0002
[0054] Embodiment 44. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-d) or (I-e):
Figure imgf000014_0001
[0055] Embodiment 45. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), or
Figure imgf000014_0002
(I-j)
Figure imgf000015_0001
wherein
Figure imgf000015_0002
is 5- to 6-membered heterocyclylene and
Figure imgf000015_0003
is 4-membered heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms.
[0056] Embodiment 46. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Ila) or (lib) :
Figure imgf000015_0004
(Ila) (lib)
[0057] Embodiment 47. The compound of embodiment 46, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (lie):
Figure imgf000015_0005
[0058] Embodiment 48. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Illa) or (Illb) :
Figure imgf000016_0001
wherein
Figure imgf000016_0002
is 9- to 11 -membered spiro heterocyclylene containing 1-2 nitrogen atoms.
[0059] Embodiment 49. The compound of any one of embodiments 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IVa) or (IVb):
Figure imgf000016_0003
nitrogen atoms.
[0060] Embodiment 50. A compound selected from the compounds of Table 1 or a pharmaceutically acceptable salt thereof.
[0061] Embodiment 51. A compound selected from the compounds of Table 2 or a pharmaceutically acceptable salt thereof.
[0062] Embodiment 52. A compound selected from the compounds of Table 3 or a pharmaceutically acceptable salt thereof.
[0063] Embodiment 53. A pharmaceutical composition comprising the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
[0064] Embodiment 54. A method of inhibiting Emopamil Binding Protein (EBP) comprising contacting EBP with an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53. [0065] Embodiment 55. A method of remyelinating a neuronal axon comprising contacting the neuronal axon with an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53. [0066] Embodiment 56. A method of treating a demyelinating disease in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
[0067] Embodiment 57. The method of embodiment 56, wherein the demyelinating disease is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain- Barre syndrome.
[0068] Embodiment 58. A method of treating multiple sclerosis (MS) in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of embodiments 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of embodiment 53.
DETAILED DESCRIPTION OF THE DISCLOSURE
Definitions
[0069] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. To the extent any material incorporated herein by reference is inconsistent with the express content of this disclosure, the express content controls. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and/or” unless the context requires otherwise. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting.
[0070] Reference in the specification to “some embodiments”, “an embodiment”, “one embodiment” or “other embodiments” means that a particular feature, structure, or characteristic described in connection with the embodiments is included in at least some embodiments, but not necessarily all embodiments, of the inventions.
[0071] As used herein, ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 pL” means “about 5 pL” and also “5 pL .” Generally, the term “about” includes an amount that would be expected to be within experimental error, such as for example, within 15%, 10%, or 5%.
[0072] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.
[0073] “Alkyl” refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (i.e., C1-C20 alkyl), 1 to 10 carbon atoms (i.e., C1-C10 alkyl), 1 to 6 carbon atoms (i.e., Ci-Ce alkyl) or 1 to 3 carbon atoms (i.e., C1-C3 alkyl). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl and 3 -methylpentyl. When an alkyl residue having a specific number of carbons is named by chemical name or identified by molecular formula, all positional isomers having that number of carbons may be encompassed; thus, for example, “butyl” includes n-butyl (i.e., -(CEhjsCEE), isobutyl (i.e., -CH2CH(CH3)2), sec-butyl (i.e.,
-CH(CH3)CH2CH3), and tert-butyl (i.e., -C(CH )3); and “propyl” includes n-propyl
(i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).
[0074] “Alkylene” refers to a divalent unbranched or branched saturated hydrocarbon chain. As used herein, alkylene has 1 to 20 carbon atoms (i.e., C1-C20 alkylene), 1 to 10 carbon atoms (i.e., C1-C10 alkylene), 1 to 6 carbon atoms (i.e., Ci-Ce alkylene) or 1 to 3 carbon atoms (i.e., Ci- C3 alkylene). Examples of alkyl groups include methylene, ethylene, propylene, butylene, pentylene, and hexylene. When an alkylene residue having a specific number of carbons is named by chemical name or identified by molecular formula, all positional isomers having that number of carbons may be encompassed; thus, for example, “C4 alkylene” includes -(CEhjsCEh- , -CH2C(CH3)2-, and
-CH(CH3)CH2CH2-); and “C3 alkylene” includes -(CH2)2CH2- and -C(CH3)2-.
[0075] An “aryl” group is an aromatic carbocyclic group of from 6 to 14 carbon atoms (Ce-Cu aryl) having a single ring (e.g., phenyl or Ce aryl) or multiple condensed rings (e.g., naphthyl or anthryl). In some embodiments, aryl groups contain 6-14 carbons (C6-C14 aryl), and in others from 6 to 12 (C6-C12 aryl) or even 6 to 10 carbon atoms (Ce-Cio aryl) in the ring portions of the groups. Particular aryls include phenyl, biphenyl, naphthyl and the like.
[0076] “Cycloalkyl” refers to a saturated or partially unsaturated cyclic alkyl group having a single ring or multiple rings including fused, bridged and spiro ring systems. The term “cycloalkyl” includes cycloalkenyl groups (i.e., the cyclic group having at least one double bond). As used herein, cycloalkyl has from 3 to 20 ring carbon atoms (i.e., C3-C20 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C3-C10 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C3-C6 cycloalkyl). Cycloalkyl also includes “spiro cycloalkyl” when there are two positions for substitution on the same carbon atom. Monocyclic radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic radicals include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl and the like. Further, the term cycloalkyl is intended to encompass any non-aromatic ring which may be fused to an aryl ring, regardless of the attachment to the remainder of the molecule.
[0077] “Haloalkyl” refers to an unbranched or branched alkyl group as defined above, wherein one or more hydrogen atoms are replaced by a halogen. For example, “Ci-Ce haloalkyl” refers to a Ci-Ce alkyl which is substituted by one or more halogen atoms. A Ci haloalkyl refers to a methyl group that may be substituted by 1-3 halo groups, a C2 haloalkyl refers to an ethyl group that may be substituted by 1-5 halo groups, a C3 haloalkyl refers to a propyl group that may be substituted by 1-7 halo groups, etc. Examples of haloalkyl include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, and the like. A haloalkyl may contain one or more halo atoms that are the same (i.e., all fluoro) or a mixture of halo atoms (i.e, chloro and fluoro).
[0078] “Heteroaryl” refers to an aromatic group (e.g., a 5-14 membered ring system) having a single ring, multiple rings, or multiple fused rings, with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. As used herein, heteroaryl includes 1 to 10 ring carbon atoms and 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur within the ring. Examples of heteroaryl groups include pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl and thiophenyl (i.e., thienyl).
[0079] “Heterocyclyl” refers to a saturated or unsaturated cyclic alkyl group, with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. The term “heterocyclyl” includes heterocycloalkenyl groups (i.e., the heterocyclyl group having at least one double bond), bridged-heterocyclyl groups, fused-heterocyclyl groups and spiro- heterocyclyl groups. A heterocyclyl may be a single ring or multiple rings wherein the multiple rings may be fused, bridged or spiro, and may comprise one or more oxo (C=O) or N-oxide (N- O-) moieties. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of the attachment (i.e., can be bound through a carbon atom or a heteroatom). Further, the term heterocyclyl is intended to encompass any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of the attachment to the remainder of the molecule. As used herein, heterocyclyl has 1 to 10 ring carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms, and 1 to 5 ring heteroatoms, 1 to 4 heteroatoms, 1 to 3 heteroatoms, or 1 to 2 heteroatoms independently selected from nitrogen, sulfur and oxygen. Examples of heterocyclyl groups include dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl and 1,1- dioxo-thiomorpholinyl.
[0080] “Cyano” refers to the group -CN.
[0081] “Halogen” or “halo” includes fluoro, chloro, bromo, and iodo.
[0082] “Hydroxy” refers to the group -OH.
[0083] “ Oxo” refers to the atom (=0) or (O).
[0084] Certain commonly used alternative chemical names may be used. For example, a divalent group such as a divalent “phenyl” group, a divalent “heteroaryl” group, a divalent “heterocyclyl” group etc., may also be referred to as a “phenylene” group, a “heteroarylene” group, or a “heterocyclylene” group, respectively.
[0085] The terms “optional” or “optionally” means that the subsequently described event or circumstance may or may not occur and that the description includes instances where said event or circumstance occurs and instances in which it does not.
[0086] Also, the term “optionally substituted” refers to any one or more hydrogen atoms on the designated atom or group which may or may not be replaced by a moiety other than hydrogen. The substituted group may be substituted with one or more substituents, such as e.g., 1, 2, 3, 4, or 5 substituents. In some embodiments, the substituents are selected from the functional groups provided herein.
[0087] Any compound or formula described herein is intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. Isotopically labeled compounds have structures depicted by the formulas given herein except that one or more atoms are replaced by an atom having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine and iodine, such as 2H, 3H, nC, 13C, 14C, 13N, 15N, 15O, 17O, 18O, 31P, 32P, 35S, 18F, 36C1, 123I and 125I, respectively. Various isotopically labeled compounds of the present disclosure, for example those into which radioactive isotopes, such as 2H, 3H, 13C, and 14C are incorporated, are included in this disclosure. Such isotopically labelled compounds may be useful in metabolic studies, reaction kinetic studies, detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) including drug or substrate tissue distribution assays or in radioactive treatment of patients.
[0088] The disclosure also includes “deuterated analogs” of compounds described herein in which from 1 to n hydrogens attached to a carbon atom is/are replaced by deuterium, in which n is the number of hydrogens in the molecule. When multiple deuterium atoms are present in a compound, the deuterium atoms may be on the same portion of the molecule (for example, on a single alkyl group or on a single ring) or on different portions of the molecule (for example, on separate alkyl groups or separate rings). Such compounds may exhibit increased resistance to metabolism and thus may be useful for increasing the half-life of any compound when administered to a mammal, particularly a human. See, for example, Foster, “Deuterium Isotope Effects in Studies of Drug Metabolism,” Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogens have been replaced by deuterium.
[0089] “Pharmaceutically acceptable” refers to compounds, salts, compositions, dosage forms, and other materials which are useful in preparing a pharmaceutical composition that is suitable for veterinary or human pharmaceutical use.
[0090] The term “pharmaceutically acceptable salt” of a given compound refers to salts that retain the biological effectiveness and properties of the given compound and which are not biologically or otherwise undesirable. “Pharmaceutically acceptable salts” include, for example, salts with inorganic acids and salts with an organic acid. In addition, if the compounds described herein are obtained as an acid addition salt, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, may be produced by dissolving the free base in a suitable organic solvent and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from base compounds. Those skilled in the art will recognize various synthetic methodologies that may be used to prepare nontoxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts may be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.
Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p- toluene-sulfonic acid, salicylic acid and the like. Likewise, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, such as alkyl amines. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri(isopropyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine and the like. It is understood that reference to a particular salt, such as hydrochloride or formate, may refer to a single salt, such as monohydrochloride or monoformate, or may refer to a multiple salt, such a dihydrochloride or diformate.
[0091] The compounds disclosed herein, or their pharmaceutically acceptable salts, may include an asymmetric center and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined, in terms of absolute stereochemistry, as (R)- or (5)- or, as (D)- or (L)- for amino acids. The disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (5)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation/isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC).
[0092] “ Tautomer” refers to alternate forms of a compound that differ in the position of a proton, such as enol-keto and imine-enamine tautomers, or the tautomeric forms of heteroaryl groups containing a ring atom attached to both a ring -NH-moiety and a ring=N moiety such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. All tautomeric forms of the compounds described herein are intended to be included.
[0093] A “stereoisomer” refers to a compound made up of the same atoms bonded by the same bonds but having different three-dimensional structures, which are not interchangeable. The present disclosure contemplates various stereoisomers and mixtures thereof and includes “enantiomers”, which refers to two stereoisomers whose molecules are nonsuperimposable mirror images of one another.
[0094] “Diastereoisomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.
[0095] As used herein, “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” or “excipient” includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
[0096] “Effective amount” or dose of a compound or a composition refers to that amount of the compound or the composition that results in an intended result as desired based on the disclosure herein. Effective amounts can be determined by standard pharmaceutical procedures in cell cultures or experimental animals including, without limitation, by determining the LD50 (the dose lethal to 50% of the population) and the EDso (the dose therapeutically effective in 50% of the population).
[0097] “Therapeutically effective amount” or dose of a compound or a composition refers to that amount of the compound or the composition that results in reduction or inhibition of symptoms or a prolongation of survival in a subject (i.e., a human patient). The results may require multiple doses of the compound or the composition.
[0098] “Treating” or “treatment” of a disease in a subject refers to 1) preventing the disease from occurring in a patient that is predisposed or does not yet display symptoms of the disease; 2) inhibiting the disease or arresting its development; or 3) ameliorating or causing regression of the disease. As used herein, “treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. For the purposes of this disclosures, beneficial or desired results include, but are not limited to, one or more of the following: decreasing one or more symptoms resulting from the disease or disorder, diminishing the extent of the disease or disorder, stabilizing the disease or disorder (e.g., preventing or delaying the worsening of the disease or disorder), delaying the occurrence or recurrence of the disease or disorder, delay or slowing the progression of the disease or disorder, ameliorating the disease or disorder state, providing a remission (whether partial or total) of the disease or disorder, decreasing the dose of one or more other medications required to treat the disease or disorder, enhancing the effect of another medication used to treat the disease or disorder, delaying the progression of the disease or disorder, increasing the quality of life, and/or prolonging survival of a subject. Also encompassed by “treatment” is a reduction of pathological consequence of the disease or disorder. The methods of the invention contemplate any one or more of these aspects of treatment.
[0099] As used herein, the terms “subject(s)” and “patient(s)” mean any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human, such as a primate, dog, cat, rabbit, or rodent. None of the terms require or are limited to situations characterized by the supervision (e.g., constant or intermittent) of a health care worker (e.g., a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker).
[00100] As used herein, the terms “pharmaceutical composition” or “medicament” refer to a composition suitable for pharmaceutical use in a subject, e.g., as an EBP inhibitor.
[00101] Although various features of the invention may be described in the context of a single embodiment, the features may also be provided separately or in any suitable combination. Conversely, although the invention may be described herein in the context of separate embodiments for clarity, the invention may also be implemented in a single embodiment.
Compounds
[00102] In one aspect, provided herein is a compound of Formula (I):
Figure imgf000024_0001
or a pharmaceutically acceptable salt thereof, wherein:
X1 is C and X2 is N, or X1 is N and X2 is C;
= is a single bond or a double bond, provided that a double bond and a single bond;
R1 is Ci-C6 alkyl, C3-C6 cycloalkyl, -CN, Ci-C6 haloalkyl, -(Ci-C6 alkylene)-O-(Ci-C6 alkyl), -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl);
L1 is a bond, O, or -CH2-;
Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing 1 or 2 nitrogen atoms, or Ce-Cio aryl; each R2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, -O(Ci-Ce alkyl), or -O(Ci-Ce haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl; L2 is a bond or O;
Ring A is
Figure imgf000025_0001
, 9- to 11-membered spiro heterocyclylene, or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms;
Y1 is N or CH;
Y2 is N or CH; x is 0, 1, or 2; y is 0 or 1; m is 0-5; each R3 is independently Ci-Ce alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl;
L3 is a bond, -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, -CH(Ra)CH(Ra)N(Ra)-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms; each Ra is independently H or Ci-Ce alkyl;
W is O, CH2, SO2, S(O)=NH, SO, or N(H);
Z is N or CH; r is 0, 1, or 2; s is 0 or 1; each R4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group; and n is 0-5.
[00103] In some embodiments, X1 is C and X2 is N, wherein a double bond and a single bond. In some embodiments, X1 is N and X2 is C, wherein a double bond and a single bond.
[00104] In some embodiments,
Figure imgf000026_0001
some embodiments,
Figure imgf000026_0002
[00105] In some embodiments,
Figure imgf000026_0003
some embodiments,
Figure imgf000026_0004
[00106] In some embodiments, R1 is Ci-Ce alkyl, C3-C6 cycloalkyl, -CN, Ci-Ce haloalkyl, -(Ci- Ce alkylene)-O-(Ci-Ce alkyl), -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl). In some embodiments, R1 is Ci-Ce alkyl, C3-C5 cycloalkyl, -CN, C1-C3 haloalkyl, - (Ci-C3 alkylene)-O-(Ci-C3 alkyl), -(C1-C3 alkylene)-O-(Ci-C3 haloalkyl), or -(Ci-C3 alkylene)(C3-C6 cycloalkyl).
[00107] In some embodiments, R1 is Ci-Ce alkyl. In some embodiments, R1 is C1-C5 alkyl. In some embodiments, R1 is C1-C4 alkyl. In some embodiments, R1 is Ci-C3 alkyl. In some embodiments, R1 is C1-C2 alkyl. In some embodiments, R1 is -CH3, -CFbCHs, -CFbCFbCHs, -CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, or -CH2CH2CH(CH3)2.
[00108] In some embodiments, R1 is C3-Ce cycloalkyl. In some embodiments, R1 is C3-Cs cycloalkyl. In some embodiments, R1 is C3-C4 cycloalkyl. In some embodiments, R1 is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, R1 is cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, R1 is cyclopropyl. In some embodiments, R1 is cyclobutyl.
[00109] In some embodiments, R1 is -CN.
[00110] In some embodiments, R1 is Ci-Ce haloalkyl. In some embodiments, R1 is Ci-Ce haloalkyl containing 1-13 halogen atoms. In some embodiments, R1 is Ci-C3 haloalkyl. In some embodiments, R1 is Ci-C3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R1 is -CF3, -CHF2, -CH2F, -CC13, -CHCh, -CH2CI, -CF2CI, -CFCh, -CH2CF3, -CH2CHF2, or - CFFCCh. In some embodiments, R1 is -CH2CHF2, -CF3, or -CHF2. In some embodiments, R1 is -CF3. In some embodiments, R1 is -CH2CHF2. In some embodiments, R1 is -CHF2.
[00111] In some embodiments, R1 is -(Ci-Ce alkylene)-O-(Ci-Ce alkyl). In some embodiments, R1 is -(Ci-C3 alkylene)-O-(Ci-C3 alkyl). It is understood that -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6. For example, -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6. In some embodiments, -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6. In some embodiments, R1 is -(Ci-C3 alkylene)-O-(CH3), -(Ci-C3 alkylene)-O- (CFbCHs), or -(Ci-C3 alkylene)-O-(CH2CH2CH3). In some embodiments, R1 is-CFbOCHs, - CH2CH2OCH3,
-CH(CH3)OCH3, or -CH(CH3)CH2OCH3.
[00112] In some embodiments, R1 is -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl). In some embodiments, R1 is -(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), wherein Ci-Ce haloalkyl contains 1- 13 halogen atoms. In some embodiments, R1 is -(Ci-C3 alkylene)-O-(Ci-C3 haloalkyl). In some embodiments, R1 is -(Ci-C3 alkylene)-O-(Ci-C3 haloalkyl), wherein Ci-C3 haloalkyl contains 1- 7 halogen atoms. It is understood that -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6. For example, -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6. In some embodiments, -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6. In some embodiments, R1 is -(C1-C3 alkylene)-O-(CF3), -(C1-C3 alkylene)-O-(CHF2), or -(C1-C3 alkylene)-O-(CH2F). In some embodiments, R1 is -(CH2)-O-(CI-C3 haloalkyl), -(CH2CH2XO- (C1-C3 haloalkyl), or
-(CH2CH2CH2)-O-(CI-C3 haloalkyl).
[00113] In some embodiments, R1 is -(Ci-Ce alkylene)(C3-Ce cycloalkyl). In some embodiments, R1 is -(C1-C3 alkylene)(C3-Ce cycloalkyl). It is understood that -(Ci-Ce alkylene)- includes optional substitution by Ci-Ce alkyl groups such that the total number of carbon atoms is no more than 6. For example, -(Ci-Ce alkylene)- includes -(C1-C5 alkylene)- substituted by methyl, such that the total number of carbon atoms is 2-6. In some embodiments, -(Ci-Ce alkylene)- includes -(C1-C2 alkylene)- substituted by C1-C4 alkyl, such that the total number of carbon atoms is 2-6. In some embodiments, R1 is -(C1-C3 alkylene)(cyclopropyl), -(C1-C3 alkylene)(cyclobutyl), or -(C1-C3 alkylene)(cyclopentyl). In some embodiments, R1 is - (CH2XC3-C6 cycloalkyl), -(CH2CH2XC3-C6 cycloalkyl), or-(CH2CH2CH2)(C3-Ce cycloalkyl). In some embodiments, R1 is -(CFbXcyclopropyl), -(CFbXcyclobutyl), or -(CFbXcyclopentyl). In some embodiments, R1 is -(CFbXcyclopropyl).
[00114] In some embodiments, R1 is -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, - C(CH3)3,
-CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -CH2CH2CH(CH3)2, -CH2OCH3, -CH2CH2OCH3, -CH(CH3)OCH3, -CH(CH3)CH2OCH3, -CH2CHF2, -CF3, -CHF2,
Figure imgf000028_0001
[00115] In some embodiments, L1 is a bond, O, or -CH2-. In some embodiments, L1 is a bond. In some embodiments, L1 is O. In some embodiments, L1 is -CH2-.
[00116] In some embodiments, Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing
1 or 2 nitrogen atoms, or Ce-Cio aryl. In some embodiments, Ring B is C4-C6 cycloalkyl, pyridinyl, pyrazinyl, pyrimidinyl, or phenyl.
[00117] In some embodiments, Ring B is C3-C6 cycloalkyl. In some embodiments, Ring B is C4-C6 cycloalkyl. In some embodiments, Ring B is cyclobutyl, cyclopentyl, or cyclohexyl. In some embodiments, Ring B is cyclopropyl. In some embodiments, Ring B is cyclobutyl. In some embodiments, Ring B is cyclopentyl. In some embodiments, Ring B is cyclohexyl.
[00118] In some embodiments, Ring B is 6-membered heteroaryl containing 1 or 2 nitrogen atoms. In some embodiments, Ring B is 6-membered heteroaryl containing 1 nitrogen atom. In some embodiments, Ring B is 6-membered heteroaryl containing 2 nitrogen atoms. In some embodiments, Ring B is pyridinyl, pyrazinyl, or pyrimidinyl. In some embodiments, Ring B is
Figure imgf000029_0001
[00119] In some embodiments, Ring B is Ce-Cio aryl. In some embodiments, Ring B is Ce aryl. In some embodiments, Ring B is phenyl. In some embodiments, Ring B is Cio aryl. In some embodiments, Ring B is naphthalenyl.
Figure imgf000029_0002
[00121] In some embodiments, each R2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, - O(Ci-Ce alkyl), or -O(Ci-Ce haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl. In some embodiments, each R2 is independently C1-C4 alkyl, C1-C3 haloalkyl, halo, - O(Ci-C3 alkyl), or
-O(Ci-C3 haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl.
[00122] In some embodiments, R2 is Ci-Ce alkyl. In some embodiments, R2 is C1-C4 alkyl. In some embodiments, R2 is methyl, ethyl, propyl, or butyl. In some embodiments, R2 is isopropyl. In some embodiments, R2 is -C(CH3)3.
[00123] In some embodiments, R2 is Ci-Ce haloalkyl. In some embodiments, R2 is Ci-Ce haloalkyl containing 1-13 halogen atoms. In some embodiments, R2 is C1-C3 haloalkyl. In some embodiments, R2 is C1-C3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R2 is -CF3, -CHF2, -CH2F, -CCh, -CHCh, -CH2C1, -CF2C1, -CFCh, -CF2CH3, -CH2CF3, -CH2CHF2, or
-CH2CCh. In some embodiments, R2 is -CF3, -CF2CH3, -CH2CHF2, or -CHF2.
[00124] In some embodiments, R2 is halo. In some embodiments, R2 is F, Cl, Br, or I. In some embodiments, R2 is F, Cl, or Br. In some embodiments, R2 is F. In some embodiments, R2 is Cl. In some embodiments, R5 is Br.
[00125] In some embodiments, R2 is -O(Ci-Ce alkyl). In some embodiments, R2 is -O(Ci-C3 alkyl). In some embodiments, R2 is -O(CH3), -O(CH2CH3), or -O(CH2CH2CH3). In some embodiments, R2 is -O(CH3). In some embodiments, R2 is -O(CH2CH3),
[00126] In some embodiments, R2 is -O(Ci-Ce haloalkyl). In some embodiments, R2 is -O(Ci- Ce haloalkyl), wherein Ci-Ce haloalkyl contains 1-13 halogen atoms. In some embodiments, R2 is
-O(Ci-C3 haloalkyl). In some embodiments, R2 is -O(Ci-C3 haloalkyl), wherein C1-C3 haloalkyl contains 1-7 halogen atoms. In some embodiments, R2 is -OCF3, -OCHF2, -OCH2F, -OCCI3, -OCHCh, -OCH2C1, -OCF2C1, -OCFCh, -OCH2CF3, -OCH2CHF2, or -OCH2CC13. In some embodiments, R2 is -OCF3 or -OCHF2. In some embodiments, R2 is -OCF3. In some embodiments, R2 is -OCHF2.
[00127] In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-3 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl optionally substituted by 1-2 groups selected from F and -CF3. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, which is optionally substituted by 1-4 groups selected from halo and C1-C3 haloalkyl. In some embodiments, two R2 groups on adjacent carbon atoms are taken together to form a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, which is optionally substituted by 1-4 groups selected from F and -CF3. [00128] In some embodiments, each R2 is independently -CF3, -CF2CH3, -CH2CHF2, -CHF2, F, Cl, Br, -OCF3, -OCHF2, -OCH3, or -C(CH3)3, or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from F or -CF3.
Figure imgf000031_0001
[00130] In some embodiments, L2 is a bond or O. In some embodiments, L2 is a bond. In some embodiments, L2 is O. -Y1 Y2-^-
[00131] In some embodiments, Ring A is , wherein Y1 is N or CH; Y2 is N or
CH; x is 0, 1, or 2; and y is 0 or 1. In some embodiments, Y1 is N and Y2 is N. In some embodiments, Y1 is N and Y2 is CH. In some embodiments, Y1 is CH and Y2 is N. In some embodiments, Y1 is CH and Y2 is CH. In some embodiments, x is 0. In some embodiments, x is
1. In some embodiments, x is 2. In some embodiments, y is 0. In some embodiments, y is 1. [00132] In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene, or 8- to 10- membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 1- 2 nitrogen atoms. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 1 nitrogen atom. In some embodiments, Ring A is 9- to 11 -membered spiro heterocyclylene containing 2 nitrogen atoms. In some embodiments, Ring A is 8- to 10- membered bicyclic fused heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, Ring A is 8- to 10-membered bicyclic fused heterocyclylene containing 1 nitrogen atom. In some embodiments, Ring A is 8- to 10-membered bicyclic fused heterocyclylene containing 2 nitrogen atoms.
Figure imgf000032_0001
Figure imgf000033_0001
— (R3)m is drawn across two rings, either ring or both rings can be substituted with m R3 groups. [00134] In some embodiments, m is 0-5. In some embodiments, m is 0. In some embodiments, m is 1-3. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments, m is 5.
[00135] In some embodiments, each R3 is independently Ci-Ce alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl. In some embodiments, each R3 is independently C1-C3 alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl.
[00136] In some embodiments, R3 is Ci-Ce alkyl. In some embodiments, R3 is C1-C3 alkyl. In some embodiments, R3 is methyl, ethyl, w-propyl, or isopropyl. In some embodiments, R3 is methyl, ethyl, or isopropyl. In some embodiments, R3 is methyl. In some embodiments, R3 is ethyl. In some embodiments, R3 is isopropyl.
[00137] In some embodiments, two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group. In some embodiments, two R3 groups are taken together to form a bridging - CH2-group. In some embodiments, two R3 groups are taken together to form a bridging - CH2CH2- group.
[00138] In some embodiments, two R3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl. In some embodiments, two R3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl. In some embodiments, two R3 groups on the same carbon atom are taken together to form a spiro cyclopropyl, cyclobutyl, or cyclopentyl. In some embodiments, two R3 groups on the same carbon atom are taken together to form a spiro cyclopropyl. In some embodiments, two R3 groups on the same carbon atom are taken together to form a spiro cyclobutyl.
[00139] In some embodiments, each R3 is independently -CH3, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro cyclopropyl.
Figure imgf000034_0001
[00141] In some embodiments, L3 is a bond, -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, -CH(Ra)CH(Ra)N(Ra)-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms, and each Ra is independently H or Ci-Ce alkyl.
[00142] In some embodiments, L3 is a bond.
[00143] In some embodiments, L3 is -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, or -CH(Ra)CH(Ra)N(Ra)-. In some embodiments, each Ra is independently H or C1-C3 alkyl. In some embodiments, each Ra is independently H, methyl, ethyl, or propyl. In some embodiments, each Ra is independently H or methyl. In some embodiments, L3 is -CH2, - CH2CH2-, -CH(CH3)CH2-,
-CH2CH(CH3)-, -OCH2CH2-, or -CH2CH2N(CH2CH3)-.
[00144] In some embodiments, L3 is 5- to 6-membered heterocyclylene or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms.
[00145] In some embodiments, L3 is 5- to 6-membered heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, L3 is 5- to 6-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L3 is 5- to 6-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L3 is 5-membered heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, L3 is 5-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L3 is 5-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L3 is 6-membered heterocyclylene containing 1-2 nitrogen atoms. In some embodiments, L3 is 6-membered heterocyclylene containing 1 nitrogen atom. In some embodiments, L3 is 6-membered heterocyclylene containing 2 nitrogen atoms. In some embodiments, L3 is pyrrolidinylene, piperidinylene, or piperazinylene.
[00146] In some embodiments, L3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms. In some embodiments, L3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1 nitrogen atom. In some embodiments, L3 is -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 2 nitrogen atoms. In some embodiments, L3 is -O-(azetidinylene) or -O-(diazetidinylene).
[00147] In some embodiments, L3 is:
Figure imgf000035_0001
[00148] In some embodiments, W is O, CH2, SO2, S(O)=NH, SO, or N(H). In some embodiments, W is O, CH2, or N(H). In some embodiments, W is is SO2, S(O)=NH, or SO. In some embodiments, W is O. In some embodiments, W is CH2. In some embodiments, W is
SO2. In some embodiments, W is S(O)=NH. In some embodiments, W is SO. In some embodiments, W is N(H).
[00149] In some embodiments, Z is N or CH. In some embodiments, Z is N. In some embodiments, Z is CH.
[00150] In some embodiments, r is 0, 1, or 2. In some embodiments, r is 0. In some embodiments, r is 1. In some embodiments, r is 2.
[00151] In some embodiments, s is 0 or 1. In some embodiments, s is 0. In some embodiments, s is 1.
[00152] In some embodiments, r and s are each 1. In some embodiments, r and s are each 0. In some embodiments, r is 1 and s is 0. In some embodiments, r is 2 and s is 1.
Figure imgf000035_0002
Figure imgf000036_0001
understood that when — (R4)n is drawn across two rings, either ring or both rings can be substituted with n R4 groups.
[00154] In some embodiments, each R4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group. In some embodiments, each R4 is independently halo, -OH, C1-C3 alkyl, or C1-C3 haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5 -membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group. In some embodiments, each R4 is independently F, -OH, -CH3, -CH(CH3)2, or -CF3, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
[00155] In some embodiments, R4 is halo. In some embodiments, R4 is F, Cl, Br, or I. In some embodiments, R4 is F, Cl, or Br. In some embodiments, R4 is F or Cl. In some embodiments, R4 is F. In some embodiments, R4 is Cl.
[00156] In some embodiments, R4 is -OH.
[00157] In some embodiments, R4 is Ci-Ce alkyl. In some embodiments, R4 is C1-C3 alkyl. In some embodiments, R4 is methyl, ethyl, w-propyl, or isopropyl. In some embodiments, R46 is methyl, ethyl, or isopropyl. In some embodiments, R4 is methyl. In some embodiments, R4 is ethyl. In some embodiments, R4 is isopropyl. [00158] In some embodiments, R4 is Ci-Ce haloalkyl. In some embodiments, R4is Ci-Ce haloalkyl containing 1-13 halogen atoms. In some embodiments, R4 is C1-C3 haloalkyl. In some embodiments, R4 is C1-C3 haloalkyl containing 1-7 halogen atoms. In some embodiments, R4 is -CF3, -CHF2, -CH2F, -CCI3, -CHCh, -CH2CI, -CF2CI, -CFCh, -CH2CF3, -CH2CHF2, or - CH2CCI3. In some embodiments, R4 is -CF3.
[00159] In some embodiments, two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group. In some embodiments, two R4 groups are taken together to form a bridging - CH2- group. In some embodiments, two R4 groups are taken together to form a bridging - CH2CH2- group.
[00160] In some embodiments, two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom.
[00161] In some embodiments, two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group. In some embodiments, two R4 groups on the same carbon atom are taken together to form a spiro 4- membered heterocyclyl containing 1 oxygen atom. In some embodiments, two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 SO2 group.
[00162] In some embodiments, n is 0-5. In some embodiments, n is 0. In some embodiments, n is 1-3. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5.
Figure imgf000037_0001
Figure imgf000038_0001
[00164] In some embodiments, the compound of Formula (I) is a compound of Formula (I-a),
(I-b), or (I-c):
Figure imgf000038_0002
wherein Ring A, Ring B, X1, X2, L2, L3, R1, R2, R3, R4, Z, W, m, n, r, s, and = are as described for Formula (I).
[00165] In some embodiments, the compound of Formula (I) is a compound of Formula (I-d) or
Figure imgf000038_0003
wherein Ring A, Ring B, X1, X2, L1, L3, R1, R2, R3, R4, Z, W, m, n, r, s, and = are as described for Formula (I).
[00166] In some embodiments, the compound of Formula (I) is a compound of Formula (I-f),
(I-g), (I-h), (I-i), (I-j), (I-k), or (1-1):
Figure imgf000038_0004
Figure imgf000039_0001
wherein Ring A, Ring B, X1, X2, L1, L2, R1, R2, R3, R4, Ra, Z, W, m, n, r, s, and = are as described for Formula (I); wherein
Figure imgf000039_0002
is 5- to 6-membered heterocyclylene; and wherein
Figure imgf000039_0003
is 4-membered heterocyclylene containing 1-2 nitrogen atoms.
[00167] In some embodiments, the compound of Formula (I) is a compound of Formula (Ila) or (lib):
Figure imgf000040_0001
wherein X1, X2, Y1, Y2, R1, R2, R3, R4, Z, W, m, n, r, s, and = are as described for Formula
(I).
[00168] In some embodiments, the compound of Formula (I) is a compound of Formula (lie):
Figure imgf000040_0002
wherein R1, R2, R3, R4, m, and n are as described for Formula (I).
[00169] In some embodiments, the compound of Formula (I) is a compound of Formula (Illa) or (Illb):
Figure imgf000040_0003
wherein
Figure imgf000040_0004
for Formula (I); and wherein
Figure imgf000040_0005
is 9- to 11 -membered spiro heterocyclylene containing 1-2 nitrogen atoms.
[00170] In some embodiments, the compound of Formula (I) is a compound of Formula (IVa) or (IVb):
Figure imgf000041_0001
wherein X1, X2, R1, R2, R3, R4, Z, W, m, n, r, s, and = are as described for Formula (I); and wherein ^ZZCZZ”^ js 8- to 10-membered bicyclic fused heterocyclylene containing 1-2 nitrogen atoms.
[00171] In the descriptions herein, it is understood that every description, variation, embodiment, or aspect of a moiety may be combined with every description, variation, embodiment, or aspect of other moieties the same as if each and every combination of descriptions is specifically and individually listed. For example, every description, variation, embodiment, or aspect provided herein with respect to Ring A of Formula (I) may be combined with every description, variation, embodiment, or aspect of X1, X2, Y1, Y2, R1, R2, R3, R4, Ra, L1, L2, L3, W, Z, Ring B, x, y, m, r, s, and n, the same as if each and every combination were specifically and individually listed. It is also understood that all descriptions, variations, embodiments, or aspects of Formula (I), where applicable, apply equally to other formulae detailed herein, and are equally described, the same as if each and every description, variation, embodiment, or aspect were separately and individually listed for all formulae. For example, all descriptions, variations, embodiments, or aspects of Formula (I), where applicable, apply equally to any of the formulae as detailed herein, such as Formulae (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (Lg), (Lh), (Li), (I-j), (Lk), (1-1), (Ila), (lib), (lie), (Illa), (Illb), (IVa), and (IVb), are equally described, the same as if each and every description, variation, embodiment, or aspect were separately and individually listed for all formulae.
[00172] In some embodiments, provided is a compound selected from the compounds in Table 1 or a pharmaceutically acceptable salt thereof. Although certain compounds described in the present disclosure, including in Table 1, are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 1, are herein described. Similarly, although certain compounds described in the present disclosure are presented as specific salts, it is understood that any pharmaceutically acceptable salt of any of the compounds of the present disclosure are herein described. It is further understood that although certain compounds described in the present disclosure are presented as specific salts, the free form of the compounds of the present disclosure are also herein described.
Table 1.
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
Figure imgf000050_0001
Figure imgf000051_0001
Figure imgf000052_0001
Figure imgf000053_0001
Figure imgf000054_0001
Figure imgf000055_0001
“&1” denotes that the absolute stereochemistry was not determined; “abs” denotes that the absolute stereochemistry was determined or a pharmaceutically acceptable salt thereof.
[00173] Also provided herein is a compound selected from the compounds in Table 2 or a pharmaceutically acceptable salt thereof. Although certain compounds described in the present disclosure, including in Table 2, are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 2, are herein described.
Table 2.
Figure imgf000056_0001
Figure imgf000057_0001
or a pharmaceutically acceptable salt thereof.
[00174] Also provided herein is a compound selected from the compounds in Table 3 or a pharmaceutically acceptable salt thereof. Although certain compounds described in the present disclosure, including in Table 3, are presented as specific stereoisomers and/or in a nonstereochemical form, it is understood that any or all stereochemical forms, including any enantiomeric or diastereomeric forms, and any tautomers or other forms of any of the compounds of the present disclosure, including in Table 3, are herein described.
Table 3.
Figure imgf000058_0001
Figure imgf000059_0001
[00175] It is understood that in the present description, combinations of substituents and/or variables of the depicted formulae are permissible only if such contributions result in stable compounds.
[00176] Furthermore, all compounds disclosed herein, such as compounds of Formula (I), that exist in free base or acid form can be converted to their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid by methods known to one skilled in the art. Salts of the compounds can be converted to their free base or acid form by standard techniques. Methods of Synthesis
[00177] In a further aspect, provided herein are methods of preparing compounds of Formula (I) or pharmaceutically acceptable salts thereof. Also provided herein are intermediate compounds useful for preparing compounds of Formula (I) or pharmaceutically acceptable salts thereof.
[00178] Intermediate compounds, such as intermediates of Formula A, useful for preparing compounds of Formula (I) can be prepared as shown in Scheme 1.
Scheme 1.
Figure imgf000060_0001
wherein R1 and R2 are as described for Formula (I), and -N(R’)(R”) represents Ring A. [00179] Starting compounds A-l can be reacted with amines of Formula A-2 to form compounds of Formula A-3, which can then be treated with Lawesson’s reagent to afford compounds of Formula A-4. Treatment of compounds of Formula A-4 with hydrazine hydrate provides compounds of Formula A-5, which can then be coupled with compounds of Formula A-6 using, for example copper(II) acetate, to afford intermediates of Formula A.
[00180] Piperazinyl-pyrazole compounds of Formula B may be prepared according to the general reactions shown in Scheme 2. Scheme 2.
Figure imgf000061_0005
base
Figure imgf000061_0001
B-1
Figure imgf000061_0002
wherein R1 and R2 are as described for Formula (I), and -N(Ra)(Rb) represents the moiety
Figure imgf000061_0003
[00181] Compounds of Formula B-l can be treated with an acid, such as TFA, to afford compounds of Formula B-2, which can then be alkylated to give compounds of Formula B. For example, compounds of Formula B-2 can be alkylated with 2-chloroacetaldehyde followed by reductive coupling with HN(Ra)(Rb) using a coupling agent such as NaBH(OAc)3.
Alternatively, compounds of Formula B-2 can be alkylated with an amine-based alkylchloride to provide compounds of Formula B-2.
[00182] Piperazinyl amino-pyrazole compounds of Formula C may be prepared according to the general reaction shown in Scheme 3.
Scheme 3.
Figure imgf000061_0004
wherein R1 and R2 are as described for Formula (I), and -N(R’)(R”) represents Ring A optionally coupled to L3 and the Z-containing moiety of Formula (I).
[00183] Compounds of Formula C-1 can be coupled with amines of Formula C-2 under palladium-catalyzed coupling conditions to afford compounds of Formula C. [00184] Piperazinyl amino isopropyl-pyrazole compounds of Formula D may be prepared according to the general reactions shown in Scheme 4.
Scheme 4.
Figure imgf000062_0001
wherein R2 is as described for Formula (I), and -N(R’)(R”) represents Ring A optionally coupled to L3 and the Z-containing moiety of Formula (I).
[00185] Compounds of Formula D-l can be arylated by coupling with compounds of Formula D-2 using, for example, Cu(OAc)2, to give compounds of Formula D-3, which can then be alkylated using D-4 under Pd-catalyzed reaction conditions, followed by Pt-catalyzed hydrogenation, to give compounds of Formula D-5. Next, Pd-catalyzed coupling of compounds of Formula D-5 with amines of Formula D-6 affords compounds of Formula D.
[00186] The synthesis of additional intermediates useful for the preparation of compounds of Formula (I) is outlined in Scheme 5.
Scheme 5.
Figure imgf000062_0002
wherein R1 is as described for Formula (I).
[00187] Compounds of Formula E-l can be treated with compound of Formula E-2 in the presence of an alkoxide to afford compounds of Formula E-3, which can then be treated with hydrazine hydrate to yield compounds of Formula E-4. Subsequent treatment with PtCh and an acid (such as HC1) affords compounds of Formula E-5, which can then be reacted with BOC2O to provide compounds of Formula E.
[00188] Intermediates of Formula E can be used to prepare pyrazole-piperidinyl compounds of Formula F as shown in Scheme 6.
Scheme 6.
Figure imgf000063_0001
wherein R1 and R2 are as described for Formula (I), and Rc represents the moiety
Figure imgf000063_0002
Formula (I).
[00189] Compounds of Formula E can be arylated with compounds of Formula F-l using, for example, Cu(OAc)2, to give compounds of Formula F-2. Subsequent deprotection with an acid, such as TFA, affords compounds of Formula F-3, which can then be alkylated to afford compounds of Formula F. For example, compounds of Formula F-3 can be reductively coupled with an aldehyde using a coupling agent such as NaBH(OAc)3 to provide compounds of Formula F. Alternatively, compounds of Formula F-3 can be treated with an alkylchloride to provide compounds of Formula F.
[00190] It is understood that the synthetic processes disclosed herein may be modified to arrive at various compounds of the present disclosure by selection of appropriate reagents and starting materials.
[00191] All compounds of Formula (I) or any variation thereof as described herein which exist in free base or acid form can be converted to their pharmaceutically acceptable salts by treatment with the appropriate inorganic or organic base or acid by methods known to one skilled in the art. Salts of the compounds of the disclosure can be converted to their free base or acid form by standard techniques.
Pharmaceutical Compositions
[00192] In another aspect, provided herein are pharmaceutical compositions of the compounds of Formula (I) or a pharmaceutically acceptable salt thereof. Thus, the present disclosure includes pharmaceutical compositions comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Pharmaceutical compositions according to the disclosure may take a form suitable for oral, buccal, sublingual, parenteral (subcutaneous, intramuscular, intravenous, or intrathecal), nasal, topical, vaginal, rectal, intracerebral, intradermal, intravitreal, intraosseous infusion, intraperitoneal, or inhalation administration. Pharmaceutical compositions of the present disclosure comprise a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent, or excipient.
[00193] A compound described herein can be used in the preparation of a pharmaceutical composition by combining the compound as an active ingredient with a pharmaceutically acceptable excipient. Some examples of materials which can serve as pharmaceutically acceptable excipients include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; surfactants, such as polysorbate 80 (i.e., Tween 80); powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffered solutions; polyesters, polycarbonates and/or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations. Pharmaceutical formulations may be prepared by known pharmaceutical methods. Suitable formulations can be found in, for example, Remington: The Science and Practice of Pharmacy, Lippincott Williams & Wilkins, 21st ed. (2005), which is incorporated herein by reference.
[00194] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.
[00195] Examples of pharmaceutically-acceptable antioxidants include: water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid and the like.
[00196] The pharmaceutical compositions may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated and the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, this amount will range from about 1% to about 99% of active ingredient, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.
[00197] In certain embodiments, a pharmaceutical composition of the present disclosure comprises an excipient selected from the group consisting of cyclodextrins, liposomes, micelle forming agents, e.g., bile acids and polymeric carriers, e.g., polyesters and polyanhydrides; and a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In certain embodiments, the pharmaceutical composition renders orally bioavailable a compound of Formula (I) or a pharmaceutically acceptable salt thereof.
[00198] Pharmaceutical compositions of the disclosure suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules or as a solution or a suspension in an aqueous or non-aqueous liquid or as an oil-in-water or water-in-oil liquid emulsion or as an elixir or syrup or as pastilles (using an inert base, such as gelatin and glycerin or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as an active ingredient. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, may also be administered as a bolus, electuary, or paste.
[00199] In solid dosage forms of the disclosure for oral administration (capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate and/or any of the following: fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol and/or silicic acid; binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; humectants, such as glycerol; disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates and sodium carbonate; solution retarding agents, such as paraffin; absorption accelerators, such as quaternary ammonium compounds; wetting agents, such as, for example, cetyl alcohol, glycerol monostearate and non-ionic surfactants; absorbents, such as kaolin and bentonite clay; lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof; and coloring agents. In the case of capsules, tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-shelled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.
[00200] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made in a suitable machine in which a mixture of the powdered compound is moistened with an inert liquid diluent.
[00201] The tablets and other solid dosage forms of the pharmaceutical compositions of the present disclosure, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be formulated for rapid release, e.g., freeze-dried. They may be sterilized by, for example, filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.
[00202] Liquid dosage forms for oral administration of the compounds of Formula (I), or a pharmaceutically acceptable salt thereof, include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan and mixtures thereof.
[00203] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.
[00204] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth and mixtures thereof. [00205] Pharmaceutical compositions of the disclosure for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the disclosure with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.
[00206] Dosage forms for the topical or transdermal administration of a compound of this disclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound (i.e., a compound of Formula (I) or a pharmaceutically acceptable salt thereof) may be mixed under sterile conditions with a pharmaceutically- acceptable carrier and with any preservatives, buffers or propellants which may be required. [00207] The ointments, pastes, creams, and gels may contain, in addition to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or mixtures thereof.
[00208] Powders and sprays can contain, in addition to a compound of Formula (I), or a pharmaceutically acceptable salt thereof, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.
[00209] Pharmaceutical compositions of this disclosure suitable for parenteral administration comprise one or more compounds of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically-acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.
[00210] Examples of suitable aqueous and nonaqueous carriers, which may be employed in the pharmaceutical compositions of the disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like) and suitable mixtures thereof, vegetable oils, such as olive oil and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.
[00211] The pharmaceutical compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms upon the subject compounds may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenyl sorbic acid and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminum monostearate and gelatin.
[00212] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally-administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.
[00213] Injectable depot forms are made by forming microencapsule matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions, which are compatible with body tissue.
Methods of Treatment
[00214] Compounds of Formula (I), or a pharmaceutically acceptable salt thereof, and pharmaceutical compositions comprising compounds of Formula (I), or a pharmaceutically acceptable salt thereof, may be used in methods of administration and treatment as provided herein. The compounds and pharmaceutical compositions may also be used in in vitro methods, such as in vitro methods of administering a compound or pharmaceutical composition to cells for screening purposes and/or for conducting quality control assays.
[00215] In some embodiments, provided herein is a method of inhibiting Emopamil Binding Protein (EBP) comprising contacting either an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or an effective amount of a pharmaceutical composition provided herein, with EBP.
[00216] In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, inhibits the activity of EBP by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, a compound of formula (I) inhibits the activity of EBP by about 1-100%, 5-100%, 10-100%, 15-100%, 20-100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65-100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5- 95%, 5-90%, 5-85%, 5-80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5- 35%, 5-30%, 5-25%, 5-20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%.
[00217] In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, binds EBP with an ICso of less than about 10 pM, such as about 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2 pM, 1 pM, or 0.5 pM. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, binds EBP with an ICso from about 0.01 pM to about 5 pM, from about 0.01 pM to about 4 pM, from about 0.01 pM to about 3 pM, from about 0.01 pM to about 2 pM, from about 0.01 pM to about 1 pM, from about 0.01 pM to about 0.05 pM, from about 0.1 pM to about 5 pM, from about 0.1 pM to about 4 pM, from about 0.1 pM to about 3 pM, from about 0.1 pM to about 2 pM, from about 0.1 pM to about 1 pM, from about 0.5 pM to about 5 pM, from about 0.5 pM to about 4 pM, from about 0.5 pM to about 3 pM, from about 0.5 pM to about 2 pM, from about 0.5 pM to about 1 pM, from about 1 pM to about 5 pM, from about 1 pM to about 4 pM, from about 1 pM to about 3 pM, from about 1 pM to about 2 pM, from about 2 pM to about 5 pM, from about 2 pM to about 4 pM, from about 2 pM to about 3 pM, from about 3 pM to about 5 pM, from about 3 pM to about 4 pM, or from about 4 pM to about 5 pM. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, binds EBP with an ICso from about 0.01 pM to about 1 pM, from about 0.01 pM to about 0.9 pM, from about 0.01 pM to about 0.8 pM, from about 0.01 pM to about 0.7 pM, from about 0.01 pM to about 0.6 pM, from about 0.01 pM to about 0.5 pM, from about 0.01 pM to about 0.4 pM, from about 0.01 pM to about 0.3 pM, from about 0.01 pM to about 0.2 pM, from about 0.01 pM to about 0.1 pM, from about 0.1 pM to about 1 pM, from about 0.1 pM to about 0.9 pM, from about 0.1 pM to about 0.8 pM, from about 0.1 pM to about 0.7 pM, from about 0.1 pM to about 0.6 pM, from about 0.1 pM to about 0.5 pM, from about 0.1 pM to about 0.4 pM, from about 0.1 pM to about 0.3 pM, from about 0.1 pM to about 0.2 pM, from about 0.2 pM to about 1 pM, from about 0.2 pM to about 0.9 pM, from about 0.2 pM to about 0.8 pM, from about 0.2 pM to about 0.7 pM, from about 0.2 pM to about 0.6 pM, from about 0.2 pM to about 0.5 pM, from about 0.2 pM to about 0.4 pM, from about 0.2 pM to about 0.3 pM, from about 0.3 pM to about 1 pM, from about 0.3 pM to about 0.9 pM, from about 0.3 pM to about 0.8 pM, from about 0.3 pM to about 0.7 pM, from about 0.3 pM to about 0.6 pM, from about 0.3 pM to about 0.5 pM, from about 0.3 pM to about 0.4 pM, from about 0.4 pM to about 1 pM, from about 0.4 pM to about 0.9 pM, from about 0.4 pM to about 0.8 pM, from about 0.4 pM to about 0.7 pM, from about 0.4 pM to about 0.6 pM, from about 0.4 pM to about 0.5 pM, from about 0.5 pM to about 1 pM, from about 0.5 pM to about 0.9 pM, from about 0.5 pM to about 0.8 pM, from about 0.5 pM to about 0.7 pM, from about 0.5 pM to about 0.6 pM, from about 0.6 pM to about 1 pM, from about 0.6 pM to about 0.9 pM, from about 0.6 pM to about 0.8 pM, from about 0.6 pM to about 0.7 pM, from about 0.7 pM to about 1 pM, from about 0.7 pM to about 0.9 pM, from about 0.7 pM to about 0.8 pM, from about 0.8 pM to about 1 pM, from about 0.8 pM to about 0.9 pM, or from about 0.9 pM to about 1 pM.
[00218] In some instances, increased inhibition of EBP can be associated with increased human Ether-a-go-go-Related Gene (hERG) toxicity. Accordingly, in some embodiments of the present disclosure, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, inhibits EBP and promotes low hERG toxicity. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, binds hERG with an ICso of less than about 30 pM, such as about 25 pM, 20 pM, 15 pM, 10 pM, 5 pM, 2 pM, or 1 pM. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, binds hERG with an ICso from about 0.05 pM to about 30 pM, such as from about 0.05 pM to about 25 pM, from about 0.05 pM to about 20 pM, from about 0.05 pM to about 15 pM, from about 0.05 pM to about 10 pM, from about 0.05 pM to about 5 pM, from about 0.05 pM to about 2 pM, from about 0.05 pM to about 1 pM, from about 1 pM to about 30 pM, such as from about 1 pM to about 25 pM, from about 1 pM to about 20 pM, from about 1 pM to about 15 pM, from about 1 pM to about 10 pM, from about 1 pM to about 5 pM, from about 1 pM to about 2 pM, from about 5 pM to about 30 pM, from about 5 pM to about 25 pM, from about 5 pM to about 20 pM, from about 5 pM to about 15 pM, from about 5 pM to about 10 pM, from about 10 pM to about 30 pM, from about 10 pM to about 25 pM, from about 10 pM to about 20 pM, from about 10 pM to about 15 pM, from about 15 pM to about 30 pM, from about 15 pM to about 25 pM, from about 15 pM to about 20 pM, from about 20 pM to about 30 pM, or from about 20 pM to about 25 pM.
[00219] In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, stimulates remyelination of neuronal axons. In some embodiments, a compound of Formula (I), or a pharmaceutically acceptable salt thereof, stimulates remyelination of neuronal axons by about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, a compound of formula (I) stimulates remyelination of neuronal axons by about 1-100%, 5-100%, 10-100%, 15-100%, 20- 100%, 25-100%, 30-100%, 35-100%, 40-100%, 45-100%, 50-100%, 55-100%, 60-100%, 65- 100%, 70-100%, 75-100%, 80-100%, 85-100%, 90-100%, 95-100%, 5-95%, 5-90%, 5-85%, 5- 80%, 5-75%, 5-70%, 5-65%, 5-60%, 5-55%, 5-50%, 5-45%, 5-40%, 5-35%, 5-30%, 5-25%, 5- 20%, 5-15%, 5-10%, 10-90%, 20-80%, 30-70%, or 40-60%. [00220] In some embodiments, the neuronal axons are artificial, such as those used in a laboratory setting (i.e., cell culture). In some embodiments, the neuronal axons are endogenous (i.e., naturally occuring inside a subject, such as a subject having multiple sclerosis (MS)). In some embodiments, remyelination of neuronal axons is achieved by oligodendrocyte precursor cells (OPCs), which repair and limit the damage associated with MS. In some embodiments, administering a compound of Formula (I), or a salt thereof, to a subject having multiple sclerosis (MS) stimulates remyelination to enhance the repair of damaged myelin and to preserve neuronal function.
[00221] In one aspect, provided herein is a method for treating a demyelinating disease, comprising administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, provided herein is a method for preventing a demyelinating in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula (I). In some embodiments, the demyelinating disease is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain-Barre syndrome.
[00222] In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof diminishes the extent of the demyelinating disease in the subject. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof stabilizes the demyelinating disease (for example, prevents or delays the worsening of the disease). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the occurrence or recurrence of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof slows the progression of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prevents relapse of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof decreases the dose of one or more other medications required to treat the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof enhances the effect of another medication used to treat the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the progression of the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof increases the quality of life of the subject having the demyelinating disease. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prolongs survival of a subject having the demyelinating disease.
[00223] In some aspects, provided herein is a method of slowing progression of a demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, provided herein is a method of stabilizing a demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, the method prevents the progression of the demyelinating disease. In some embodiments, the method delays the progression of the demyelinating disease.
[00224] In another aspect, provided herein is a method of delaying the occurrence or recurrence of a demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
[00225] In further aspects, provided herein is a method of decreasing the dose of one or more other medications required to treat the demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, provided herein is a method of enhancing the effect of another medication used to treat the demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
[00226] Also provided here is a method of delaying the progression of the demyelinating disease in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, the method increases the quality of life of the subject having the demyelinating disease. In some embodiments, the method prolongs survival of the subject having the demyelinating disease. [00227] In some aspects, provided herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in treating a demyelinating disease. In other aspects, provided herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the manufacture of medicament for treating a demyelinating disease.
[00228] In another aspect, provided herein is a method for treating multiple sclerosis (MS) with EBP inhibition in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, provided herein is a method for preventing multiple sclerosis (MS) in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula (I).
[00229] In some embodiments, the type of multiple sclerosis (MS) is relapsing remitting MS, in which an individual having MS will have episodes of new or worsening symptoms, known as relapses. In some embodiments, the type of multiple sclerosis (MS) is primary progressive MS, in which an individual having MS will experience worsening symptoms and/or accumulating symptoms and with no periods of remission. In some embodiments, the type of multiple sclerosis (MS) is secondary progressive MS, in which an individual having MS experiences nerve damage or loss, leading to a general worsening of the disease.
[00230] In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof diminishes the extent of the disease multiple sclerosis (MS) (for example, loss of myelin coating of brain and spinal cord nerves, development of plaque around brain and spinal cord nerves, neuroinflammation) in the subject. In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof stabilizes the disease multiple sclerosis (MS) (for example, prevents or delays the worsening of MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the occurrence or recurrence of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof slows the progression of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prevents relapse of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof decreases the dose of one or more other medications required to treat the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof enhances the effect of another medication used to treat the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof delays the progression of the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof increases the quality of life of the subject having the disease multiple sclerosis (MS). In some embodiments, administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof prolongs survival of a subject having the disease multiple sclerosis (MS). [00231] In some aspects, provided herein is a method of slowing progression of multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, provided herein is a method of stabilizing multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, the method prevents the progression of multiple sclerosis (MS). In some embodiments, the method delays the progression of multiple sclerosis (MS).
[00232] In another aspect, provided herein is a method of delaying the occurrence or recurrence of multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
[00233] In further aspects, provided herein is a method of decreasing the dose of one or more other medications required to treat the disease multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, provided herein is a method of enhancing the effect of another medication used to treat multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject.
[00234] Also provided here is a method of delaying the progression of the disease multiple sclerosis (MS) in a subject, the method comprising administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, to the subject. In some embodiments, the method increases the quality of life of the subject having multiple sclerosis (MS). In some embodiments, the method prolongs survival of the subject having multiple sclerosis (MS).
[00235] In some aspects, provided herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in treating multiple sclerosis (MS). In other aspects, provided herein is a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the manufacture of medicament for treating multiple sclerosis (MS).
Dosing and Method of Administration
[00236] The phrases “parenteral administration” and “administered parenterally” as used herein mean modes of administration other than enteral and topical administration, usually by injection and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular, subarachnoid, intraspinal and intrastemal injection and infusion. [00237] The phrases “systemic administration,” “administered systemically,” “peripheral administration” and “administered peripherally” as used herein mean the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient’s system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.
[00238] These compounds may be administered to humans and other animals for therapy by any suitable route of administration, including orally, nasally, as by, for example, a spray, rectally, intravaginally, parenterally, intraci stemally and topically, as by powders, ointments or drops, including buccally and sublingually.
[00239] Regardless of the route of administration selected, the compounds of the present disclosure, or the pharmaceutical compositions of the present disclosure, are formulated into pharmaceutically acceptable dosage forms by conventional methods known to those of skill in the art.
[00240] Actual dosage levels of the active ingredients in the pharmaceutical compositions of this disclosure may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration, without being toxic to the patient.
[00241] The selected dosage level will depend upon a variety of factors including the activity of the particular compound of the present disclosure employed or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated and like factors well known in the medical arts. A daily, weekly or monthly dosage (or other time interval) can be used.
[00242] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the disclosure employed in the pharmaceutical composition at levels lower than that required to achieve the desired therapeutic effect and then gradually increasing the dosage until the desired effect is achieved.
[00243] In general, a suitable daily dose of a compound of the disclosure will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect (e.g., inhibit necrosis). Such an effective dose will generally depend upon the factors described above. Generally, doses of the compounds of this disclosure for a patient, when used for the indicated effects, will range from about 0.0001 to about 100 mg per kg of body weight per day. Preferably the daily dosage will range from 0.001 to 50 mg of compound per kg of body weight and even more preferably from 0.01 to 10 mg of compound per kg of body weight.
[00244] If desired, the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.
[00245] When the compounds of the present disclosure are administered as pharmaceuticals, to humans and animals, they can be given per se or as a pharmaceutical composition containing, for example, 0.1% to 99.5% (such as 0.5% to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.
[00246] The compounds of the present application or the pharmaceutical compositions thereof may be administered once, twice, three, or four times daily, using any suitable mode described above. Also, administration or treatment with the compounds may be continued for a number of days; for example, commonly treatment would continue for at least 7 days, 14 days, or 28 days, for one cycle of treatment. Treatment cycles are well known and are frequently alternated with resting periods of about 1 to 28 days, commonly about 7 days or about 14 days, between cycles. The treatment cycles, in certain embodiments, may also be continuous.
[00247] When administered orally, the total daily dosage for a human subject may be between about 1 mg and 1,000 mg, between about 1,000-2,000 mg/day, between about 10-500 mg/day, between about 50-300 mg/day, between about 75-200 mg/day or between about 100-150 mg/day.
[00248] The daily dosage may also be described as a total amount of a compound described herein administered per dose or per day. Daily dosage of a compound may be between about 1 mg and 4,000 mg, between about 2,000 to 4,000 mg/day, between about 1 to 2,000 mg/day, between about 1 to 1,000 mg/day, between about 10 to 500 mg/day, between about 20 to 500 mg/day, between about 50 to 300 mg/day, between about 75 to 200 mg/day or between about 15 to 150 mg/day.
[00249] In certain embodiments, the method comprises administering to the subject an initial daily dose of about 1 to 800 mg of a compound described herein and increasing the dose by increments until clinical efficacy is achieved. Increments of about 5, 10, 25, 50 or 100 mg can be used to increase the dose. The dosage can be increased daily, every other day, twice per week or once per week.
[00250] In certain embodiments, a compound or pharmaceutical preparation is administered orally. In certain embodiments, the compound or pharmaceutical preparation is administered intravenously. Alternative routes of administration include sublingual, intramuscular and transdermal administrations. [00251] The preparations of the present disclosure may be given orally, parenterally, topically, or rectally. They are, of course, given in forms suitable for each administration route. For example, they are administered in tablets or capsule form; by injection, inhalation, eye lotion, ointment, suppository, infusion, inhalation, etc.; topical by lotion or ointment; and rectal by suppositories. In certain embodiments, the administration is oral.
Kits/Article of Manufacture
[00252] Disclosed herein, in certain embodiments, are kits and articles of manufacture for use with one or more compounds, compositions, or methods described herein. Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In one embodiment, the containers are formed from a variety of materials such as glass or plastic.
[00253] A kit typically includes labels listing contents and/or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included.
[00254] In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself, a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In one embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein.
[00255] In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition. EXAMPLES
[00256] The examples and preparations provided below further illustrate and exemplify the compounds of the present disclosure and methods for testing such compounds. It is to be understood that the scope of the present disclosure is not limited in any way by the scope of the following examples.
[00257] The chemical reactions in the Examples described can be readily adapted to prepare a number of other compounds disclosed herein, and alternative methods for preparing the compounds of this disclosure are deemed to be within the scope of this disclosure. For example, the synthesis of non-exemplified compounds according to the present disclosure can be performed by modifications apparent to those skilled in the art, for example by appropriately protecting interfering groups, by utilizing other suitable reagents known in the art other than those described, or by making routine modification of reaction conditions, reagents, and starting materials. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the present disclosure.
[00258] The following abbreviations may be relevant for the application.
Abbreviations
ACN acetonitrile aq aqueous
CDI 1 , 1 '-carbonyl diimidazole
CV column volume(s)
DAST diethylaminosulfur trifluoride
DCE di chloroethane
DCM dichloromethane
DIAD diisopropyl azodi carb oxy late
DIEA, DIPEA N,N-Diisopropylethylamine
DMF dimethylformamide
DMP Dess-Martin periodinane
DMS dimethyl sulfide
DMSO dimethylsulfoxide
EA, EtOAc ethylacetate
Et2O di ethyl ether
EtOH ethanol h, hr hour(s)
HBTU 3-[bis(dimethylamino)methyliumyl]-3J/-benzotriazol-l-oxide hexafluorophosphate HPLC high-performance liquid chromatography
LCMS liquid chromatography-mass spectrometry
LiHMDS, LHMDS lithium Hexamethyldisilazide
MeOD-d4 deuterated methanol (CD3OD)
MeOH methanol min. minute(s)
MW micro wave
NIS N-iodosuccinimide
NMP N -methyl -2-py rroli done
Pd2(dba)3 tris(dibenzylideneacetone)dipalladium(0)
PdC12(dppf) [1, l'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride
Pd PEPPSI-IPENT dichlorofl, 3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3- chloropyridyl)palladium(II)
PE petroleum ether
RT, rt room temperature sat. saturated
SGC Silica Gel Chromatography tBu tert-butyl tBuXPhos Pd G3 [(2-di-tert-butylphosphino-2 ',4 6 ' -tri i sopropyl- 1 , 1 '-biphenyl)-2-(2 '- amino- 1 , 1 '-biphenyl)] palladium(II) methanesulfonate
TEA triethylamine
TFA trifluoroacetic acid
THF tetrahydrofuran xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene
Synthetic Examples
Example SI. Synthesis of l-(5-Methyl-l-Phenyl-Pyrazol-3-yl)-4-(Tetrahydropyran-4- ylmethyl) Piperazine (Compound 1).
[00259] Compound 1 was prepared as outlined below.
Figure imgf000080_0006
Figure imgf000080_0001
Figure imgf000080_0007
Figure imgf000080_0002
[00260] Step 1. Synthesis of tert-butyl 4-(3-oxobutanoyl) piperazine-l-carboxylate.
Figure imgf000080_0003
[00261] A mixture of tert-butyl piperazine-l-carboxylate (6.0 g, 32.2 mmol) and tert-butyl acetoacetate (5.61 g, 35.4 mmol) in toluene (100 mL) was heated at 100°C for 16 h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether) to afford the desired product tert-butyl 4-(3 -oxobutanoyl) piperazine-l- carboxylate (8.4 g, yield 93%) as a yellow oil.
[00262] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.32% (214 nm), Mass: found peak 293.3 (M+23) at 1.039 min.
[00263] ‘H NMR (500 MHz, CDC13) 5 3.63-3.59 (m, 2H), 3.58 (s, 2H), 3.46-3.42 (m, 4H), 3.41-3.36 (m, 2H), 2.28 (s, 3H), 1.47 (s, 9H) ppm.
[00264] Step 2. Synthesis of tert-butyl 4-(3-oxobutanethioyl) piperazine-l-carboxylate.
Lawesson's reagent
(0.5 eq) toluene, 75°c, 16h
Figure imgf000080_0004
Figure imgf000080_0005
[00265] To a solution of tert-butyl 4-(3 -oxobutanoyl) piperazine-l-carboxylate (4.0 g, 14.8 mmol) in toluene (100 mL) was added Lawesson's reagent (2.99 g, 7.4 mmol) and the mixture was heated at 75°C for 16 h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether) to afford the desired crude product tert-butyl 4-(3 -oxobutanethioyl) piperazine- 1 -carboxylate (2.33 g) as a brown oil.
[00266] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 2mL/min;
Column: SunFire, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 78.09% (214 nm), Mass: found peak 287.2 (M+1) at 1.744 min; LC purity: 20.64% (214 nm) Mass: found peak 287.1 (M+1) at 1.975 min.
[00267] Step 3. Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine-1- carboxylate.
Figure imgf000081_0001
[00268] To a solution of tert-butyl 4-(3 -oxobutanethioyl) piperazine- 1 -carboxylate (2.33 g, 8.14 mmol) in toluene (60 mL) was added hydrazine monohydrate (1.21 mL, 24.4 mmol) and the mixture was stirred at 70°C for 16 h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-6% MeOH in DCM) to afford the desired product tertbutyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine- 1 -carboxylate (1.59 g, yield 38.6% over 2 steps) as a yellow solid. LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.86% (214 nm), Mass: found peak 267.3 (M+1) at 1.022 min.
[00269] 'H NMR (400 MHz, CDCh) 5 5.52 (s, 1H), 3.54 (t, J = 4.8 Hz, 4H), 3.14 (t, J = 4.8 Hz, 4H), 2.25 (s, 3H), 1.48 (s, 9H) ppm.
[00270] Step 4. Synthesis of tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine-1- carboxylate.
Boc
Figure imgf000081_0002
[00271] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine- 1 -carboxylate (0.6 g, 2.25mmol) in dichloromethane (30 mL) was added phenylboronic acid (561 mg, 4.51 mmol), anhydrous copper acetate (614 mg, 3.38 mmol), pyridine (0.363 mL, 4.51 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 64 h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine- 1 -carboxylate (0.54 g, yield 70%) as a yellow solid.
[00272] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 343.4 (M+l) at 1.395 min.
[00273] Step 5. Synthesis of l-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine.
Figure imgf000082_0001
[00274] To a solution of tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine-1- carboxylate (0.62 g, 1.81 mmol) in dichloromethane (15 mL) was added TFA (3 mL, 40.4 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-(5-methyl-l-phenyl-pyrazol- 3-yl) piperazine (420 mg, yield 92%) as a yellow solid.
[00275] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.07% (214 nm) Mass: found peak 243.3 (M+l) at 0.928 min.
[00276] Step 6. Synthesis of l-(5-methyl-l-phenyl-pyrazol-3-yl)-4-(tetrahydropyran-4- ylmethyl) piperazine (Compound 1).
Figure imgf000083_0001
[00277] To a solution of l-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine (0.15 g, 0.619 mmol) in 1,2-dichloroethane (8 mL) was added tetrahydropyran-4-carbaldehyde (84.8 mg, 0.743 mmol), follow by Sodium triacetoxyborohydride (0.262 g, 1.24 mmol), 4A molecular sieve (0.5 g) and acetic acid (2 drops). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered. The filtrate was treated with MeOH (2 mL), concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-(5-methyl-l-phenyl-pyrazol-3-yl)- 4-(tetrahydropyran-4-ylmethyl) piperazine (126.5 mg, yield 60%) as a white solid.
[00278] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 341.3 (M+l) at 1.813 min.
[00279] 'H NMR (400 MHz, DMSO-d6) 5 7.52-7.42 (m, 4H), 7.33-7.27 (m, 1H), 5.84 (s, 1H), 3.82 (dd, J = 11.2, 2.8 Hz, 2H), 3.28 (td, J = 11.6, 1.6 Hz, 2H), 3.11 (t, J = 4.8 Hz, 4H), 2.43 (t, J = 4.8 Hz, 4H), 2.28 (s, 3H), 2.16 (d, J = 7.6 Hz, 2H), 1.82-1.71 (m, 1H), 1.61 (d, J = 11.2 Hz, 2H), 1.12 (qd, J = 12.0, 4.0 Hz, 2H) ppm.
Example S2. Synthesis of 4-[2-[4-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazin-1- yl] ethyl] morpholine (Compound 2)
[00280] Compound 2 was prepared as outlined below.
Figure imgf000083_0002
[00281] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine-l-carboxylate.
Boc
Figure imgf000084_0001
[00282] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine-l-carboxylate (0.2 g, 0.751 mmol) in dichloromethane (10 mL) was added 2-naphthylboronic acid (264 mg, 1.5 mmol), anhydrous copper acetate (205 mg, 1.13 mmol), pyridine (0.121 mL, 1.5 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40 g silica gel column @50 mL/min, eluting with 5-40% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4- [5 -methyl- 1 -(2 -naphthyl) pyrazol-3-yl] piperazine-l-carboxylate (160 mg, yield 54.3%) as a yellow oil.
[00283] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 393.3 (M+l) at 1.503 min.
[00284] Step 2. Synthesis of l-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine.
Boc
Figure imgf000084_0002
[00285] To a solution of tert-butyl 4-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine-1- carboxylate (0.24 g, 0.611 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 26.9 mmol). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine (157 mg, yield 84.9%) as a yellow solid. [00286] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 96.72% (214 nm), Mass: found peak 293.2 (M+l) at 1.073 min.
[00287] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazin-1- yl] ethyl] morpholine (Compound 2).
Figure imgf000085_0001
[00288] To a solution of l-[5-methyl-l-(2-naphthyl) pyrazol-3-yl] piperazine (130 mg, 0.445 mmol), potassium carbonate (184 mg, 1.33 mmol) and KI (10 mg, 0.06 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (99 mg, 0.534 mmol). The reaction was stirred at 90 °C for 3 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- methyl-l-(2-naphthyl) pyrazol-3-yl] piperazin- 1 -yl]ethyl] morpholine (63 mg, yield 34.9%) as a white solid.
[00289] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 406.3 (M+l) at 1.751 min.
[00290] 'H NMR (400 MHz, DMSO-d6) 5 8.03-7.92 (m, 4H), 7.70 (dd, J = 8.8, 2.0 Hz, 1H), 7.58-7.49 (m, 2H), 5.90 (s, 1H), 3.56 (t, J = 4.8 Hz, 4H), 3.14 (t, J = 4.4 Hz, 4H), 2.56-2.51 (m, 3H), 2.48-2.32 (m, 12H) ppm.
Example S3. Synthesis of l-(5-methyl-l-phenyl-pyrazol-3-yl)-4-(2-tetrahydropyran-4- ylethyl) piperazine (Compound 3).
[00291] Compound 3 was prepared as outlined below.
Figure imgf000086_0001
[00292] To a solution of l-(5-methyl-l-phenyl-pyrazol-3-yl) piperazine (0.14 g, 0.578 mmol) in 1,2-dichloroethane (8 mL) was added 2-tetrahydropyran-4-ylacetaldehyde (88.9 mg, 0.693 mmol), follow by sodium triacetoxyborohydride (0.245 g, 1.16 mmol), 4A molecular sieve (0.5 g) and acetic acid (2 drops). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered. The filtrate was treated with MeOH (2 mL), concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-(5-methyl-l-phenyl-pyrazol-3-yl)- 4-(2-tetrahydropyran-4-ylethyl) piperazine (95 mg, yield 46.4%) as a white solid.
[00293] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min O.Olmin; Flow Rate: 1.8mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 355.3 (M+l) at 1.794 min.
[00294] 'H NMR (400 MHz, DMSO-d6) 5 7.50-7.42 (m, 4H), 7.34-7.26 (m, 1H), 5.84 (s, 1H), 3.81 (dd, J = 10.4, 3.2 Hz, 2H), 3.26 (td, J = 11.6, 1.6 Hz, 2H), 3.11 (t, J = 4.4 Hz, 4H), 2.44 (d, J = 4.8 Hz, 4H), 2.32 (t, J = 7.2 Hz, 2H), 2.28 (s, 3H), 1.60-1.45 (m, 3H), 1.39 (dd, J = 14.8, 7.2 Hz, 2H), 1.22-1.08 (m, 2H) ppm.
Example S4. Synthesis of 4-(5-methyl-l-phenyl-pyrazol-3-yl)-l-(tetrahydropyan-4- ylmethyl) piperidine (Compound 4).
[00295] Compound 4 was prepared as outlined below.
Figure imgf000087_0001
[00296] Step 1. Synthesis of l-(4-Pyridinyl)-l,3-butanedione.
Figure imgf000087_0002
[00297] To a 500 mL flask with sodium methoxide (715 mg, 13.2 mmol) and anhydrous ether (15 mL) was sequentially added ethyl pyridine-4-carboxylate (2.0 g, 13.2 mmol), and a solution of acetone (3.84 g, 66.2 mmol) in ether (15 mL). The suspension was stirred at reflux for 6 h, cooled, and filtered. The isolated solid was washed with ether and dissolved in water (40 mL). Glacial acetic acid (5.2 mL) was added, and the mixture was extracted with chloroform (40 mL x 2). The organic extracts were dried over anhydrous Sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (PE: EA= 1 : 1) to afford the desired product l-(4-pyridinyl)- 1,3 -butanedi one (1.55 g, yield 72%) as a brown solid.
[00298] LCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3min, 95%B for 0.7min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 164.1(M+1) at 0.791 min.
[00299] Step 2. Synthesis of 4-(3-methyl-lH-pyrazol-5-yl) pyridine.
Figure imgf000087_0003
[00300] l-(4-Pyridinyl)-l,3-butanedione (1.55 g, 9.5 mmol) was suspended in ethanol (10 mL), hydrazinium hydroxide (476 mg, 9.5 mmol) was added and the mixture was stirred at 85°C for 48 h. The reaction mixture was diluted with saturated Sodium bicarbonate solution (pH 7-8) and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-(3-methyl-lH-pyrazol-5-yl) pyridine (1.2 g, 79.4%) as beige solid.
[00301] LCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3min, 95%B for 0.7min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 90% (214 nm), Mass: found peak 160.1 (M+l) at 0.303 min.
[00302] Step 3. Synthesis of 4-(3-methyl-lH-pyrazol-5-yl) piperidine hydrochloride.
Figure imgf000088_0001
[00303] 4-(3-methyl-lH-pyrazol-5-yl) pyridine (1.1 g, 6.91 mmol) was dissolved in ethanol (30 mL), hydrochloric acid solution (1 M, 13.8 mL, 1.38 mmol) was added and the mixture was hydrogenated with platinum oxide hydrate (80% Pt, 150 mg) at atmospheric pressure and room temperature for 16 h. The reaction mixture was filtered and concentrated in vacuo to afford the desired product 4-(3-methyl-lH-pyrazol-5-yl) piperidine hydrochloride (1.0 g, yield 71.7%) as a yellow solid.
[00304] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B); Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Column: X-BRIDGE C18, 4.6x50 mm, 3.5 pm; Temperature: 50 °C; LC purity: 90% (214 nm), Mass: found peak 166.1(M+1) at 0.592 min.
[00305] Step 4. Synthesis of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl) piperidine-1- carboxylate.
BOC2Q(1 .0 eq)
N-N / — \ NaHCO3(3 o eq)
II ) — \ H T - - - — Boc
\ / dioxane, H2O RT 2h
/ HCI
Figure imgf000088_0002
[00306] 4-(3-methyl-lH-pyrazol-5-yl)piperidine hydrochloride (1.0 g, 6.6 mmol) was dissolved in water (10 mL). Sodium bicarbonate (1.68 g, 20 mmol) and di-tertbutyldicarbonate (1.45 g, 6.6 mmol) in dioxane (15 mL) were added and the mixture was stirred for 2 h at room temperature. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (PE: EA= 3: 1) to afford the desired product tert-butyl 4- (3-methyl-lH-pyrazol-5-yl) piperidine- 1 -carboxylate (900 mg, yield 43.2%) as a colorless oil.
[00307] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile(B); Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Column: X-BRIDGE C18, 4.6x50 mm, 3.5 pm; Temperature: 50 °C; LC purity: 77% (214 nm), Mass: found peak 210.2 (M-55)+ at 1.690 min.
[00308] Step 5. Synthesis of tert-butyl 4-(5-methyl-l-phenyl-pyrazol-3-yl)piperidine-l- carboxylate.
Figure imgf000089_0001
[00309] Tert-butyl 4-(3 -methyl- lH-pyrazol-5-yl)piperidine-l -carboxylate (980 mg, 3.7 mmol), phenylboronic acid (980 mg, 7.4 mmol), anhydrous copper acetate (1.34 g, 7.4 mmol) and molecular sieves 4 A was suspended in anhydrous dichloromethane (30 mL). Pyridine (584 mg, 7.4 mmol) was added and the mixture was stirred at ambient temperature for 48h. The reaction mixture was concentrated in vacuo to dryness and purified by prep-HPLC (water/ammonium hydrogen carbonate/acetonitrile) to afford the desired product tert-butyl 4-(5-methyl-l-phenyl- pyrazol-3-yl)piperidine-l -carboxylate (420 mg, yield 48%) as a yellow oil.
[00310] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2 min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 40 °C; LC purity: 95.53% (214 nm), Mass: found peak 342.2 (M-55)+ at 1.374 min.
[00311] Step 6. Synthesis of 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperidine.
Figure imgf000089_0002
[00312] Hydrogen chloride solution (4.0 M in dioxane, 5.0 mL) was added to tert-butyl 4-(5- methyl-1 phenyl-pyrazol-3-yl) piperidine- 1 -carboxylate (600 mg, 1.76 mmol) in 1,4-dioxane (10 mL) and stirred for 2 h at ambient temperature. The reaction was concentrated in vacuo. The saturated Sodium bicarbonate solution was added to adjust to pH 7-8 and extracted with dichloromethane (50 mL x 2). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-(5-methyl-l-phenyl-pyrazol- 3-yl) piperidine (380 mg, yield 89.6%) as yellow oil.
[00313] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2 min; Flow Rate: 2.2mL/min;
Column: Chromolith Fast gradient RP-18e, 50mm*3mm; Column Temperature: 50 °C; LC purity: 98.27% (214 nm), Mass: found peak 242.3 (M+H)+ at 0.863 min.
[00314] Step 7. Synthesis of 4-(5-methyl-l-phenyl-pyrazol-3-yl)-l-(tetrahydropyan-4- ylmethyl) piperidine (Compound 4).
Figure imgf000090_0001
[00315] A solution of 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperidine (100 mg, 0.41 mmol), tetrahydropyran-4-carbaldehyde (47.3 mg, 0.41 mmol) and sodium triacetoxyborohydride (176 mg, 8.3 mmol) in 1,2-di chloroethane (5 mL) was stirred for overnight at room temperature. The reaction mixture was filtered, and concentrated in vacuo. The residue was purified by prep- HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-(5- methyl-l-phenyl-pyrazol-3-yl)-l-(tetrahydropyan-4-ylmethyl) piperidine (71.6 mg, yield 50.9%) as a yellow oil.
[00316] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3min, 95% B for 1.7min; Flow rate: 2mL/min;
Column: Sunfire, 50*4.6mm, 3.5pm; Column temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 340.2 (M+l) at 1.359 min.
[00317] 'H NMR (400 MHz, DMSO-d6) d 7.49 (d, J = 4.4 Hz, 4H), 7.42-7.31 (m, 1H), 6.13 (s, 1H), 3.82 (dd, J = 11.2, 2.8 Hz, 2H), 3.32-3.24 (m, 2H), 2.87 (d, J = 11.2 Hz, 2H), 2.53 (s, 1H), 2.30 (s, 3H), 2.14 (d, J = 6.8 Hz, 2H), 1.96 (t, J = 11.2 Hz, 2H), 1.85(d, J = 11.6 Hz, 2H) 1.78- 1.72 (m, 1H), 1.59-1.58 (m, 4H), 1.19-1.03 (m, 2H) ppm.
Example S5. Synthesis of 4-(5-methyl-l-phenyl-pyrazol-3-yl)-l-(2-tetrahydropyran-4- ylethyl) piperidine (Compound 5).
[00318] Compound 5 was prepared as outlined below.
Figure imgf000091_0001
[00319] A solution of 4-(5-methyl-l-phenyl-pyrazol-3-yl) piperidine (90 mg, 0.37 mmol), 2- (tetrahydro-2H-pyran-4-yl) acetaldehyde (47.3 mg, 0.41 mmol) and sodium triacetoxyborohydride (158 mg, 0.74 mmol) in 1,2-dichloroethane (5 mL) was stirred for overnight at room temperature. The reaction mixture was filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-(5-methyl-l-phenyl-pyrazol-3-yl)-l-(2-tetrahydropyran-4-ylethyl) piperidine (42.7 mg, yield 32.4 %) as a yellow solid.
[00320] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3min, 95% B for 1.7min; Flow rate: 2mL/min;
Column: Sunfire, 50*4.6mm, 3.5pm; Column temperature: 50 °C; LC purity: 100% (214 nm), found peak 354.3 (M+H)+ at 1.388 min.
[00321] 'H NMR (400 MHz, DMSO-d6) d 7.49 (d, J = 4.4 Hz, 4H), 7.42-7.31 (m, 1H), 6.13 (s, 1H), 3.82 (dd, J = 10.8, 3.2 Hz, 2H), 3.32-3.24 (m, 2H), 2.9 (d, J = 11.6 Hz, 2H), 2.53 (s, 1H), 2.32-2.80 (m, 5H), 1.96 (t, J = 11.6 Hz, 2H), 1.85 (d, J = 11.2 Hz, 2H), 1.60-1.40 (m, 5H), 1.35 (m, 2H), 1.20-1.13 (m ,2H) ppm.
Example S6. Synthesis of 4-[2-[4-[l-[4-fluoro-3-(trifluoromethoxy) phenyl] -5-methyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 6).
[00322] Compound 6 was prepared as outlined below.
Figure imgf000091_0002
[00323] Step 1. Sythesis of 2-[4-fluoro-3-(trifluoromethoxy) phenyl]-4,4,5,5-tetramethyl- 1,3,2-dioxaborolane.
Figure imgf000092_0002
[00324] Under argon atmosphere, a mixture of 4-bromo-l-fluoro-2-(trifluorom ethoxy) benzene (1.5 g, 5.79 mmol), l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (1.47 g, 5.79 mmol), PdC12(dppf) (315 mg, 0.386 mmol) and potassium acetate (1.14 g, 11.6 mmol) in 1,4-dioxane (30 mL) was stirred at 80 °C for 16h. The reaction mixture was cooled to room temperature, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 0-5% ethyl acetate in petroleum ether) to afford the desired product 2- [4-fluoro-3 -(trifluoromethoxy) phenyl]-4,4,5,5-tetramethyl-l,3,2- dioxaborolane (910 mg, yield 63.1%) as a yellow oil.
[00325] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 81.98% (214 nm) Mass: found peak 307.1 (M+l) at 1.582 min. 'H NMR (400 MHz, CDCh) 57.75-7.68 (m, 2H), 7.18 (dd, J = 10.0, 8.4 Hz, 1H), 1.34 (s, 12H) ppm.
[00326] Step 2. Synthesis of [4-fluoro-3-(trifluoromethoxy) phenyl] boronic acid.
Figure imgf000092_0001
[00327] To a stirred solution of 2-[4-fluoro-3-(trifluoromethoxy)phenyl]-4,4,5,5-tetramethyl- 1,3,2-dioxaborolane (850 mg, 2.78 mmol) in tetrahydrofuran (20 mL) and water (5 mL) was added a Sodium periodate (1.78 g, 8.33 mmol). After stirring at room temperature for 30 min, HC1 aqueous (2.78 mL, 2.78 mmol) was added and stirred for an additional 16h. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (50 mL*3). The organic layers were washed with brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product [4-fluoro-3 -(trifluoromethoxy) phenyl] boronic acid (576 mg, yield 85.6%) as yellow solid. LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 92.47% (214 nm) Mass: no ms peak, at 1.876 min.
[00328] Step 3. Synthesis of tert-butyl 4-[l-[4-fluoro-3-(trifluoromethoxy) phenyl]-5-
Figure imgf000093_0001
[00329] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine- 1 -carboxylate (100 mg, 0.375 mmol) in chloroform (20 mL) was added [4-fluoro-3 -(trifluoromethoxy) phenyl] boronic acid (109 mg, 0.488 mmol), anhydrous copper acetate (136 mg, 0.751 mmol), pyridine (0.15 mL, 1.88 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[l-[4-fluoro-3 -(trifluoromethoxy) phenyl]-5-methyl-pyrazol-3- yl] piperazine- 1 -carboxylate (160 mg, yield 91.1%) as a yellow solid.
[00330] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 95.04% (214 nm) Mass: found peak 445.3 (M+l) at 1.527 min. 'H NMR (400 MHz, CDCh) 5 7.44-7.40 (m, 1H), 7.39-7.34 (m, 1H), 7.28-7.22 (m, 1H), 5.71 (s, 1H), 3.55 (t, J = 5.2 Hz, 4H), 3.20 (t, J = 5.2 Hz, 4H), 2.31 (s, 3H), 1.49 (s, 9H) ppm.
[00331] Step 4. Synthesis of l-[l-[4-fluoro-3-(trifluoromethoxy) phenyl]-5-methyl-pyrazol- 3-yl] piperazine.
Figure imgf000093_0002
[00332] To a solution of tert-butyl 4-[l-[4-fluoro-3 -(trifluoromethoxy) phenyl]-5-methyl- pyrazol-3-yl] piperazine- 1 -carboxylate (160 mg, 0.36 mmol) in di chloromethane (10 mL) was added TFA (2 mL, 26.9 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 1 -[ 1 -[4- fluoro-3 -(trifluoromethoxy) phenyl]-5-methyl-pyrazol-3-yl] piperazine (120 mg, yield 92.2%) as a yellow oil.
[00333] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95.22% (214 nm) Mass: found peak 345.1 (M+l) at 1.989 min.
[00334] Step 5. Synthesis of 4-[2-[4-[l-[4-fluoro-3-(trifluoromethoxy) phenyl]-5-methyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 6).
Figure imgf000094_0001
[00335] To a solution of l-[l-[4-fluoro-3 -(trifluoromethoxy) phenyl]-5-methyl-pyrazol-3-yl] piperazine (120 mg, 0.349 mmol), potassium carbonate (193 mg, 1.39 mmol) and KI (58 mg, 0.349 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (97 mg, 0.523 mmol). The reaction was stirred at 90°C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4-fluoro-3- (trifluoromethoxy) phenyl]-5-methyl-pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (134.7 mg, yield 84.5%) as a yellow solid.
[00336] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 458.1 (M+l) at 2.029 min. 'H NMR (400 MHz, CD3OD) 5 7.60-7.55 (m, 1H), 7.54-7.44 (m, 2H), 5.87 (s, 1H), 3.72 (t, J = 4.8 Hz, 4H), 3.26 (t, J = 4.8 Hz, 4H), 2.66 (t, J = 4.8 Hz, 4H), 2.63-2.58 (m, 4H), 2.54 (t, J = 4.8 Hz, 4H), 2.31 (s, 3H) ppm.
Example S7. Synthesis of 4-[2-[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] pyrrolidin-3-yl] oxyethyl] morpholine (Compound 7). [00337] Compound 7 was prepared as outlined below.
Figure imgf000095_0001
[00338] Step 1. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] pyr rolidin-3-ol.
Figure imgf000095_0002
[00339] Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole 500 mg, 1.56 mmol), tert-butyl 2,6-diazaspiro-[3.3]-heptane-2-carboxylate (203 mg, 2.34 mmol), tBuXPhos Pd G3 (124 mg, 0.156 mmol) and sodium tert-butoxide (449 mg, 4.67 mmol) in 1,4-dioxane (20 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-80% ethyl acetate in petroleum ether) to afford the desired product l-[5-methyl-l- [4-(trifluoromethoxy) phenyl]pyrazol-3-yl]pyrrolidin-3-ol (97 mg, yield 17.9%) as a yellow oil. [00340] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 93.81% (214 nm) Mass: found peak 328.2 (M+l) at 1.164 min.
[00341] Step 2. Synthesis of 4-[2-[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] pyrrolidin-3-yl] oxyethyl] morpholine (Compound 7).
Figure imgf000095_0003
[00342] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]pyrrolidin-3- ol (97 mg, 0.296 mmol) in dry DMF (9 mL) was added sodium hydride (60%, 237 mg, 5.93 mmol). The reaction was stirred at 60°C for 2h. The reaction was cooled and 4-(2- chloroethyl)morpholine hydrochloride (165 mg, 0.889 mmol) was added and stirred at 80 °C for 16h. The reaction was cooled to room temperature, quenched with water (5 mL), extracted with DCM (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] pyrrolidin-3-yl] oxyethyl] morpholine (53.9 mg, yield 41.3%) as a yellow oil.
[00343] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 441.1 (M+l) at 2.059 min. 'H NMR (400 MHz, CD3OD) 5 7.59-7.54(m, 2H), 7.41 (d, J = 8.4 Hz, 2H), 5.68 (s, 1H), 4.23-4.18 (m, 1H), 3.72-3.63 (m, 6H), 3.50-3.34 (m, 4H), 2.62 (t, J = 5.6 Hz, 2H), 2.54 (t, J = 4.4 Hz, 4H), 2.30 (s, 3H), 2.15-2.07 (m, 2H) ppm.
Example S8. Synthesis of 4-[(7S,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-yl]morpholine (Compound 8).
[00344] Compound 8 was prepared as outlined below.
Figure imgf000096_0001
[00345] Step 1. Synthesis of methyl (2S,4R)-l-[2-(tert-butoxycarbonylamino)acetyl]-4- hydroxy-pyrrolidine-2-carboxylate.
Figure imgf000096_0002
[00346] To a mixture of N-(tert-butoxycarbonyl)glycine (2.89 g, 16.5 mmol), DIEA (8.21 mL, 49.6 mmol) and HBTU (6.26 g, 16.5 mmol) in 75 mL DMF at 0°C, L-4-hydroxyproline methyl ester hydrochloride (3 g, 16.5 mmol) was added and the suspension thus obtained was stirred at room temperature for 16h. The reaction mixture was diluted with EtOAc (750 mL), washed with water (100 mL) and brine (100 mL*4), dried over Sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @75 mL/min, eluting with 0-50% acetone in petroleum ether) to afford the desired methyl (2S,4R)-1- [2-(tert-butoxycarbonylamino)acetyl]-4-hydroxy-pyrrolidine-2-carboxylate (4.0 g, yield 79.6%) as a yellow solid.
[00347] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.42% (214 nm) Mass: found peak 325.1 (M+23)+ at 1.567 min. 'HNMR (400 MHz, DMSO-d6) 5 6.86 (t, J = 5.6 Hz, IH), 3.48-3.39 (m, IH), 4.30 (t, J = 8.0 Hz, IH), 3.84-3.72 (m, IH), 3.67-3.55 (m, 5H), 3.52-3.40 (m, 2H), 2.13-2.05 (m, IH), 1.92-1.84 (m, IH), 1.38 (s, 9H) ppm.
[00348] Step 2. Synthesis of (7R,8aS)-7-hydroxy-2,3,6,7,8,8a-hexahydropyrrolo[l,2- a]pyrazine-l, 4-dione.
Figure imgf000097_0001
[00349] To a solution of methyl (2S,4R)-l-[2-(tert-butoxycarbonylamino)acetyl]-4-hydroxy- pyrrolidine-2-carboxylate (700 mg, 2.32 mmol) in methanol (10 mL), was added a solution of HC1 in dioxane (4 M, 8 mL, 32 mmol). The reaction was stirred at room temperature for Ih and concentrated in vacuo under reduced pressure below 40 °C. The residue was dissolved in MeOH (40 mL), treated slowly dropwise with DIPEA until the pH was stable at 9-10. Another DIPEA (0.575 mL, 3.47 mmol) was added and stirred at room temperature for 16h. The reaction was concentrated in vacuo. The residue was diluted with chloroform (30 mL) and stirred at reflux for 2h, then cooled to room temperature for 2h and overnight at 4°C. The solid was filtered, washed with cold chloroform and Et2O to afford desired product (7R,8aS)-7-hydroxy-2,3,6,7,8,8a- hexahydropyrrolo[l,2-a]pyrazine- 1,4-dione (0.3 g, yield 76.1%) as a white solid.
[00350] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 171 (M+l) at 0.349 min. ‘HNMR (400 MHz, DMSO-d6) 5 8.08 (d, J = 2.4 Hz, IH), 5.11 (d, J = 2.8 Hz, IH), 4.36-4.26 (m, 2H), 4.04 (dd, J = 16.8, 1.2 Hz, IH), 3.56-3.46 (m, 2H), 3.22 (d, J = 12.4 Hz, IH), 2.08-2.01 (m, IH), 1.93-1.85 (m, IH) ppm.
[00351] Step 3. Synthesis of (7R,8aS)-l,2,3,4,6,7,8,8a-octahydropyrrolo[l,2-a]pyrazin-7-ol.
Figure imgf000098_0001
[00352] To a solution of (7R,8aS)-7-hydroxy-2,3,6,7,8,8a-hexahydropyrrolo[l,2-a]pyrazine-
I, 4-dione (1.45 g, 8.52 mmol) in tetrahydrofuran (50 mL), was added a solution of LiAIT in tetrahydrofuran (1.0 M, 50 mL, 50 mmol). The reaction was stirred at 60 °C for 3h, then stirred at room temperature for 16h. The reaction was cooled to 0 °C, quenched with water (1.9 g), 15% NaOH aqueous (1.9 g) and water (5.7 g), diluted with tetrahydrofuran (50 mL), stirred for 15 min, filtered, and concentrated in vacuo to afford the desired product (7R,8aS)-l,2,3,4,6,7,8,8a- octahydropyrrolo[l,2-a]pyrazin-7-ol (680 mg, yield 56.1%) as a yellow oil.
[00353] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B with 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; Mass: found peak 143 (M+l) at 0.367 min. ‘H NMR (500 MHz, DMSO-d6) 5 4.70 (s, 1H), 4.16-4.09 (m, 1H), 3.22 (dd, J = 9.0, 7.0 Hz, 1H), 2.88 (dd, J = 11.5, 2.5 Hz, 1H), 2.78-2.72 (m, 2H), 2.56-2.50 (m, 1H), 2.17 (dd, J =
I I.5, 10.0 Hz, 1H), 2.09-1.98 (m, 2H), 1.88 (dd, J = 9.0, 5.5 Hz, 1H), 1.50-1.41 (m, 2H) ppm.
[00354] Step 4. Synthesis of (7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-ol.
Figure imgf000098_0002
[00355] A mixture of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, 0.934 mmol), (7R,8aS)-l,2,3,4,6,7,8,8a-octahydropyrrolo[l,2-a]pyrazin-7-ol (159 mg, 1.12 mmol), tBuXPhos Pd G3 (74 mg, 0.0934 mmol) and sodium tert-butoxide (269 mg, 2.8 mmol) in 1,4-di oxane (15 mL) was stirred 100 °C for 16h under argon atmosphere. The reaction was cooled to room temperature and directly purified by flash chromatography (Biotage, 40 g silica gel column @75 mL/min, eluting with 0-20% MeOH in DCM) to afford the desired product (7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,4,6,7,8,8a-hexahydro-lH- pyrrolo[l,2-a]pyrazin-7-ol (84 mg, yield 23.9%) as a brown solid.
[00356] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90.87% (214 nm) Mass: found peak 383.1 (M+l) at 1.885 min.
[00357] Step 5. Synthesis of [(7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-yl] 4-methylbenzenesulfonate.
Figure imgf000099_0001
[00358] To a solution of (7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-ol (84 mg, 0.22 mmol), DMAP (81 mg, 0.66 mmol) and triethylamine (67 mg, 0.66 mmol) in dichloromethane (15 mL) was added 4- methylbenzenesulfonyl chloride (126 mg, 0.66 mmol). The reaction was stirred at room temperature for 16h. The reaction was diluted with di chloromethane (50 mL), washed with water (10 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0- 20% MeOH in DCM) to afford the desired product [(7R,8aS)-2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-yl] 4-methylbenzenesulfonate (90 mg, yield 72.3%) as a brown solid.
[00359] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94.66% (214 nm) Mass: found peak 537.1 (M+l) at 2.262 min.
[00360] Step 6. Synthesis of 4-[(7S,8aS)-2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin- 7-yl]morpholine (Compound 8).
Figure imgf000099_0002
[00361] To a solution of [(7R,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,4,6,7,8,8a-hexahydro-lH-pyrrolo[l,2-a]pyrazin-7-yl] 4-methylbenzenesulfonate (85 mg, 0.158 mmol) and DIPEA (102 mg, 0.792 mmol) in l-methylpyrrolidin-2-one (3 mL) was added morpholine (41.4 mg, 0.475 mmol). The reaction was treated with microwave reactor and stirred at 120 °C for Ih. The reaction was cooled to room temperature and directly purified by prep- HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [(7S,8aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,4,6,7,8,8a-hexahydro-lH- pyrrolo[l,2-a]pyrazin-7-yl]morpholine (44.3 mg, yield 61.9%) as a yellow solid.
[00362] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 452.1 (M+l) at 2.054 min. ‘H NMR (400 MHz, CD3OD) 5 7.59-7.54 (m, 2H), 7.41 (d, J = 8.4 Hz, 2H), 5.86 (s, 1H), 3.81 (dd, J = 12.0, 2.0 Hz, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.67-3.61 (m, 1H), 3.21 (dd, J = 10.4, 2.4 Hz, 1H), 3.07-2.98 (m, 2H), 2.92 (qd, J = 11.6, 3.2 Hz, 1H), 2.62 (dd, J = 11.2, 10.8 Hz, 1H), 2.54 (t, J = 4.0 Hz, 4H), 2.40-2.27 (m, 6H), 2.10- 2.02 (m, 1H), 1.59-1.49 (m, 1H) ppm.
Example S9. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]- 3, 6-diazabicyclo [3.1.1] heptan-6-yl] ethyl] morpholine (Compound 9).
[00363] Compound 9 was prepared as outlined below.
Figure imgf000100_0001
[00364] Step 1. Synthesis of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo [3.1.1] heptane-6-carboxylate.
Figure imgf000100_0002
[00365] To a solution of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 0.31mmol), sodium t-butoxide (90 mg, 0.93 mmol), methanesulfonato (2-di-t-butylphosphino- 2',4',6'-tri-i-propyl-l,l'-biphenyl)(2'-amino-l,l'-biphenyl-2-yl)palladium(II) (25 mg, 0.031 mmol) in 1,4-dioxane (6 mL) was added tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (93 mg, 0.47 mmol). The reaction was stirred at 100 °C in tube for 16h. The mixture was then filtered and the filtrate was washed with water (10 mL) and extracted by EtOAc (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo [3.1.1] heptane-6-carboxylate (75 mg, 54.9% yield) as a yellow oil.
[00366] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 76 % (214 nm); Mass: found peak 439.3 (M + H) at 2.16 min.
[00367] Step 2. Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo-[3.1.1]-heptane.
Figure imgf000101_0001
[00368] To a solution of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]- 3, 6- diazabicyclo-[3.1.1]-heptane-6-carboxylate (75 mg, 0.171 mmol) in dichloromethane (6 mL) was added TFA (2.5mL). The reaction mixture was stirred at room temperature for Ih. The mixture was neutralized with saturated potassium carbonate solution (5 mL) and extracted with dichloromethane (10 mL X 3). Combined DCM layers were dried over sodium sulfate and filtered. The filtrate was concentrated to dryness to give 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo-[3.1.1]-heptane (57 mg, crude) as a yellow oil.
[00369] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 78 % (214 nm); Mass: found peak 339.2 (M + H) at 1.55 min.
[00370] Step 3. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-3, 6-diazabicyclo [3.1.1] heptan-6-yl] ethyl] morpholine (Compound 9).
Figure imgf000102_0001
[00371] To a solution of 3-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-3, 6- diazabicyclo [3.1.1] heptane (57 mg, 0.168 mmol), potassium carbonate (70 mg, 0.505 mmol) and KI (28 mg, 0.168 mmol) in 95% ethanol / water (5 mL / 0.5 mL) was added 4-(2- chloroethyl) morpholine (38 mg, 0.253 mmol). The reaction mixture was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[3-[5-methyl-l-[4- (trifluorom ethoxy) phenyl] pyrazol-3-yl]-3, 6-diazabicyclo [3.1.1] heptan-6-yl] ethyl] morpholine (47.8 mg, yield: 62.8%) as a yellow oil.
[00372] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 452.2 (M + H) at 2.00 min.
[00373] 'H NMR (400 MHz, CD3OD): 5 7.60-7.56 (m, 2H), 7.40 (d, J = 8.4 Hz, 2H), 5.76 (s, 1H), 3.76 (d, J = 5.6 Hz, 2H), 3.69-3.61 (m, 6H), 3.39 (d, J = 11.6 Hz, 2H), 2.65 (t, J = 7.2 Hz, 3H), 2.51-2.42 (m, 6H), 2.30 (s, 3H), 1.73 (d, J = 8.4 Hz, 1H) ppm.
Example S10. Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]- 2, 5-diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine (Compound 10).
[00374] Compound 10 was prepared as outlined below.
Figure imgf000102_0002
[00375] Step 1. Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] -2, 5-diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine.
Figure imgf000103_0001
[00376] To a solution of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 0.31mmol), sodium t-butoxide (90 mg, 0.93 mmol), methanesulfonato(2-di-t-butylphosphino- 2',4',6'-tri-i-propyl-l,r-biphenyl)(2'-amino-l,r-biphenyl-2-yl)palladium(II) (25 mg, 0.031 mmol) in 1,4-dioxane (4 mL) was added tert-butyl 3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (93 mg, 0.47 mmol). The reaction was stirred at 100 °C in tube for 16h. The reaction mixture was filtered, and the filtrate was washed with water (10 mL) and extracted by EtOAc (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2,5- diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine (120 mg, 85.3% yield
[00377] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100 % (214 nm); Mass: found peak 439.3 (M + H) at 2.15 min.
[00378] Step 2. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2, 5-diazabicyclo [2.2.1] heptane.
Figure imgf000103_0002
[00379] To a solution of tert-butyl 5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]- 2, 5-diazabicyclo [2.2.1] heptane-2-carboxylate (120 mg, 0.274 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for Ih. The mixture was neutralized with saturated potassium carbonate solution (5 mL) and extracted with dichloromethane (10 mL X 3). Combined DCM layers were dried over sodium sulfate and filtered. The filtrate was concentrated to dryness to give 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2, 5-diazabicyclo [2.2.1] heptane (92 mg, crude). The crude is used directly in the next step.
[00380] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93 % (214 nm);
Mass: found peak 339.1 (M + H) at 1.94 min.
[00381] Step 3. Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-2, 5-diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine (Compound 10).
Figure imgf000104_0001
[00382] To a solution of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2, 5- diazabicyclo [2.2.1] heptane (92 mg, 0.27 mmol), potassium carbonate (113 mg, 0.816 mmol) and KI (45 mg, 0.27 mmol) in 95% ethanol / water (5 mL / 0.5 mL) was added 4-(2-chloroethyl) morpholine (61 mg, 0.408 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate / water / acetonitrile) to afford the desired product 4-[2-[5-[5-methyl-l-[4-(trifhioromethoxy) phenyl] pyrazol-3-yl]-2, 5- diazabicyclo [2.2.1] heptan-2-yl] ethyl] morpholine (83.5 mg, yield 68%) as a yellow oil.
[00383] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 452.1 (M + H) at 1.92 min.
[00384] 'H NMR (400 MHz, CD3OD): 5 7.54-7.58 (m, 2H), 7.41 (d, J = 8.0 Hz, 2H), 5.72 (s, 1H), 4.18 (s, 1H), 3.69 (t, J = 4.4 Hz, 5H), 3.51 (d, J = 10.0 Hz, 1H), 3.35 (d, J = 2.0 Hz, 1H), 2.96 (t, J = 8.4 Hz, 1H), 2.77 (t, J = 7.6 Hz, 3H), 2.48-2.52 (m, 6H), 2.30 (s, 3H), 1.94 (dd, J = 20 Hz, 9.6 Hz, 2H) ppm.
Example Sil. Synthesis of 4-[2-[4-[l-(2, 2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 11).
[00385] Compound 11 was prepared as outlined below.
Figure imgf000105_0001
[00386] Step 1. Synthesis of 3,5-dibromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl) pyrazole.
Figure imgf000105_0002
[00387] To a solution of 3,5-dibromo-lH-pyrazole (0.2 g, 0.885 mmol) in dichloromethane (10 mL) was added (2,2-difluoro-l,3-benzodioxol-5-yl) boronic acid (215 mg, 1.06 mmol), anhydrous copper acetate (482 mg, 2.66 mmol), pyridine (0.356 mL, 4.43 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether: ethyl acetate = 10 / 1) to afford the desired product 3, 5-dibromo-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl) pyrazole (338 mg, yield: 93%) as a yellow oil.
[00388] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 93 % (214 nm); Mass: found peak 382.9 (M + H) at 1.48 min.
[00389] Step 2. Synthesis of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropenyl- pyrazole.
Figure imgf000106_0001
[00390] To a solution of 3,5-dibromo-l-(2,2-difluoro-l,3-benzodioxol-5-yl) pyrazole (338 mg, 0.82 mmol) and 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (124 mg, 0.741 mmol) in 1,4-dioxane (5 mL) was added Sodium carbonate (174 mg, 1.65 mmol) and PdC12(dppf) dichloromethane complex (67.2 mg, 0.082 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)- 5-isopropenyl-pyrazole (91 mg, yield: 29.6%) as a colorless oil.
[00391] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 92 % (214 nm); Mass: found peak 343.0 (M + H) at 2.25 min.
[00392] Step 3. Synthesis of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl- pyrazole.
Figure imgf000106_0002
[00393] To a solution of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropenyl- pyrazole (91 mg, 0.24 mmol) in 2 mL THF was added platinum dioxide (10 mg) in one portion. The mixture was stirred at room temperature under H2 atmosphere for Ih. The mixture was filtered, and the filtrate was concentrated to dryness to give 3 -bromo- 1 -(2, 2-difluoro-l, 3- benzodioxol-5-yl)-5-isopropyl-pyrazole (91 mg, crude). The crude was used directly in the next step without purification.
[00394] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 85 % (214 nm); Mass: found peak 345.0(M + H) at 2.27 min. [00395] Step 4. Synthesis of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5- isopropyl-pyrazol-3-yl] piperazine-l-carboxylate.
Figure imgf000107_0001
[00396] Under argon atmosphere, a mixture of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)- 5-isopropyl-pyrazole (91 mg, 0.26 mmol), tert-butyl piperazine- 1 -carboxylate (98 mg, 0.53 mmol), Pd PEPPSI-IPENT (10 mg, 0.01 mmol) and sodipm; 2-methylpropan-2-olate (76 mg, 0.79 mmol) in THF (2 mL) in a sealed tube was stirred at 60 °C for 16h. Then concentrated and purified by SGC (petroleum ether: ethyl acetate = 1 : 1) to afford the product tert-butyl 4-[l-(2,
2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl-pyrazol-3-yl] piperazine- 1 -carboxylate (61 mg, 43.7%) as a yellow solid.
[00397] LCMS method: Column: HALO C18 2.7pm 4.6*30mm. Mobile phase: Water (0.01%TFA) (A) / ACN (0.01% TFA) (B). Elution program: Gradient from 5 to 95% of B in l.Omin at 2.2 mL/min. Temperature: 40°C. LC purity: 85% (214 nm), Mass: found peak 451.3 (M+l) at 1.55 min.
[00398] Step 5. Synthesis of l-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl-pyrazol-
3-yl] piperazine.
Figure imgf000107_0002
[00399] To a solution of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl] piperazine-l-carboxylate (104 mg, 0.22 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for Ih. The mixture was neutralized with saturated potassium carbonate solution (5 mL) and extracted with dichloromethane (10 mL X 3). Combined DCM layers were dried over sodium sulfate and filtered. The filtrate was concentrated to dryness to give 1 -[ 1 -(2, 2-difluoro-l, 3-benzodioxol-5- yl)-5-isopropyl-pyrazol-3-yl] piperazine (78 mg, crude). The crude was used directly in the next step. [00400] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89 % (214 nm);
Mass: found peak 351.1 (M + H) at 2.11 min.
[00401] Step 6. Synthesis of 4-[2-[4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 11).
Figure imgf000108_0001
[00402] To a solution of 1 -[ l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-isopropyl-pyrazol-3-yl] piperazine (78 mg, 0.22 mmol), potassium carbonate (154 mg, 1.11 mmol) and KI (37 mg, 0.22 mmol) in ethanol / water (5 mL / 0.5 mL) was added 4-(2-chloroethyl) morpholine (50 mg, 0.33 mmol). The reaction was stirred at 90 °C for 16h, then cooled down and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water/acetonitrile) to afford the desired product 4-[2-[4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)- 5-isopropyl-pyrazol-3-yl] piperazin-l-yl] ethyl]morpholine (63 mg, yield 61.1%) as a yellow solid.
[00403] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 464.1 (M + H) at 2.05 min.
[00404] 'H NMR (400 MHz, CD3OD): 5 7.34 (dd, J = 4.0 Hz, 1.6 Hz, 2H), 7.22 (dd, J = 8.8 Hz, 2.4 Hz, 1H), 5.88 (s, 1H), 3.72 (t, J = 4.4 Hz, 4H), 3.26 (t, J = 4.4 Hz, 4H), 2.91-2.98 (m, 1H), 2.69 (t, J = 4.8 Hz, 4H), 2.62-2.65 (m, 4H), 2.56 (s, 4H), 1.20 (s, 3H), 1.18 (s, 3H) ppm.
Example S12. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 12).
Synthesis 1
[00405] Compound 12 was prepared as outlined below.
Figure imgf000109_0001
[00406] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol- 3-yl] piperazine-l-carboxylate.
Boc
Figure imgf000109_0002
[00407] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine-l-carboxylate (0.2 g, 0.75 mmol) in chloroform (10 mL) was added [4-(trifluoromethyl) phenyl] boronic acid (214 mg, 1.13 mmol), anhydrous copper acetate (273 mg, 1.5 mmol), pyridine (0.3 mL, 3.75 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether: ethyl acetate = 4 / 1) to afford the desired product tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine-l-carboxylate (1.39 g, yield: 75.2%) as a yellow solid.
[00408] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 411.1 (M + H) at 2.27 min.
[00409] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine.
Figure imgf000109_0003
[00410] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine-l-carboxylate (1.39 g, 3.39 mmol) in dichloromethane (30 mL) was added 2, 2, 2- trifluoroacetic acid (10 mL). The reaction mixture was stirred at room temperature for Ih. The mixture was neutralized with saturated potassium carbonate solution (5 mL) and extracted with dichloromethane (10 mL X 3). Combined DCM layers were dried over sodium sulfate and filtered. The filtrate was concentrated to dryness to give l-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine (1.05 g, crude). The crude was used directly in the next step. [00411] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 84% (214 nm) Mass: found peak 311.3 (M+l) + at 0.99 min.
[00412] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 12).
Figure imgf000110_0001
[00413] To a solution of l-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine (1.05 g, 3.38 mmol), potassium carbonate (1.4 g, 10.2 mmol) and KI (0.562 g, 3.38 mmol) in 95% ethanol / water (90 mL /10 mL) was added 4-(2-chloroethyl) morpholine (1.01 g, 6.77 mmol). The reaction was stirred at 95°C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate / water / acetonitrile) to afford the desired product 4-[2-[4-[5- methyl-l-[4-(trifluorom ethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (1.04 g, yield: 72%) as a yellow solid.
[00414] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 424.2 (M + H) at 2.01 min.
[00415] 'H NMR (400 MHz, CD3OD): 5 7.79 (d, J = 8.8 Hz, 2H), 7.69 (d, J = 8.8 Hz, 2H), 5.91 (s, 1H), 3.72 (t, J = 4.8 Hz, 4H), 3.29 (t, J = 4.8 Hz, 4H), 2.70 (t, J = 4.8 Hz, 4H), 2.63-2.65 (m, 4H), 2.56 (s, 4H), 2.38 (s, 3H) ppm.
Synthesis 2
[00416] Compound 12 was prepared as outlined below.
Figure imgf000110_0002
[00417] Step 1. Synthesis of tert-butyl 4-(3-oxobutanoyl)piperazine-l-carboxylate.
Figure imgf000111_0001
[00418] To a solution of tert-butyl 3-oxobutanoate (11.2 g, 70.8 mmol) in toluene (150 mL) was added tert-butyl piperazine- 1 -carboxylate (12.0g, 64.4 mmol) and the mixture was stirred at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @100mL/min, eluting with 0-35% MeOH in DCM) to afford the desired product tert-butyl 4-(3- oxobutanoyl)piperazine-l -carboxylate (13.9 g, yield 72.6%) as a transparent oil.
[00419] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 215.1 (M-56)+ at 1.30 min.
[00420] Step 2. Synthesis of tert-butyl 4-(3-oxobutanethioyl)piperazine-l-carboxylate.
Lawesson's reagent
(0.5 eq) toluene, 75°c 16h
Figure imgf000111_0002
Figure imgf000111_0003
[00421] To a solution of tert-butyl 4-(3-oxobutanoyl)piperazine-l -carboxylate (16.9 g, 60.6 mmol) in toluene (200 mL) was added Lawesson's reagent (12.3 g, 30.3 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tertbutyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (9.1g, yield 38.4%) as a brown oil.
[00422] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.32% (254 nm) Mass: found peak 287.0 (M+l) at 1.51 min.
[00423] Step 3. Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000111_0004
,
[00424] To a solution of tert-butyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (9.1 g, 23.3 mmol) in toluene (200 mL) was added hydrazine monohydrate (4.65 mL, 95.9 mmol) and the mixture was stirred at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-6% MeOH in DCM) to afford the desired product tertbutyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (4.35 g, yield 69.0%) as a yellow solid.
[00425] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.32% (254 nm) Mass: found peak 287.0 (M+l) at 1.51 min. LC purity: 98.33% (254 nm) Mass: found peak 267.1 (M+l) at 1.33 min.
[00426] Step 4. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000112_0001
[00427] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (5 g, 18.8 mmol) in chloroform (500 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (7.28 g, 37.5 mmol), anhydrous copper acetate (6.82 g, 37.5 mmol), pyridine (7.42 g, 93.9 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (4.1 g, yield 53.2%) as a white solid.
[00428] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 411.2 (M + 1) + at 2.258 min.
[00429] Step 5. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000112_0002
I l l [00430] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (2.6 g, 6.33 mmol) in dichloromethane (20 mL) was added 2,2,2- trifluoroacetic acid (10 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (2.1 g, crude). The crude product was used directely in the next step.
[00431] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 311.2 (M + 1) + at 1.663 min.
[00432] Step 6. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 12).
Figure imgf000113_0001
[00433] To a solution of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (3.4 g, 11 mmol), potassium carbonate (7.57 g, 54.8 mmol) and KI (1.82 g, 11 mmol) in 95% ethanol (30 mL) was added 4-(2-chloroethyl)morpholine (2.4 g, 16.4 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature, filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (2.16 g, yield: 46.6%) as a white solid.
[00434] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.56 % (214 nm); Mass: found peak 424.2 (M + H) at 1.790 min.
[00435] 'H NMR (400 MHz, CDCh) 5 7.67 (d, J = 8.5 Hz, 2H), 7.58 (d, J = 8.5 Hz, 2H), 5.73 (s, 1H), 3.77-3.54 (m, 4H), 3.36-3.19 (m, 4H), 2.66-2.59 (m, 4H), 2.57 (s, 4H), 2.50 (s, 4H), 2.36 (s, 3H) ppm.
Example S13. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] cyclohexoxy] ethyl] morpholine (Compound 13).
[00436] Compound 13 was prepared as outlined below.
Figure imgf000114_0001
[00437] Step 1. Synthesis of tert-butyl 4-(2-morpholinoethoxy) piperidine-l-carboxylate.
Figure imgf000114_0002
Boc
[00438] To a solution of tert-butyl 4-hydroxypiperidine-l -carboxylate (1.0 g, 5 mmol) in THF (20 mL) was added sodium hydride (1.19 g, 50 mmol). The reaction mixture was stirred at room temperature for 2h. Then N-(2-Chloroethyl) morpholine hydrochloride (925 mg, 5 mmol) was added. The mixture was stirred at reflux overnight. The mixture was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product tert-butyl 4-(2- morpholinoethoxy) piperidine-l-carboxylate (500 mg, yield: 32%) as a white solid.
[00439] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95% (214 nm), Mass: found peak 315.2 (M+l) at 1.861 min.
[00440] Step 2. Synthesis of 4- [2-(4-piperidyloxy)ethyl] morpholine.
Figure imgf000114_0003
[00441] To a solution of tert-butyl 4-(2-morpholinoethoxy) piperidine-l-carboxylate (500 mg, 1.59 mmol) in 1,4-dioxane (10 mL) was added HC1 (8 mL, 4 mol/L in dioxane). The mixture was stirred for 3h at room temperature. The residue was concentrated to afford the desired product 4-[2-(4-piperidyloxy)ethyl]morpholine (300 mg, yield 88%) as a yellow oil.
[00442] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 99% (214 nm), Mass: found peak 215.1 (M+l) at 0.179 min. [00443] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] cyclohexoxy] ethyl] morpholine (Compound 13).
Figure imgf000115_0001
[00444] To a solution of 3-iodo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (50 mg, 0.13 mmol) in dry 1,4-dioxane (3 mL) was added 4-[2-(4-piperidyloxy)ethyl]morpholine (58 mg, 0.27 mmol), Pd-PEPPSI-ipent (10.7 mg, 0.01 mmol), sodium tert-butoxide (39 mg, 0.4 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]cyclohexoxy] ethyl]morpholine (11.7 mg, yield 19%) as a colorless oil.
[00445] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98% (214 nm), Mass: found peak 455.2 (M+l) at 2.108 min.
[00446] 'H NMR (400 MHz, MeOD-d4) 5 7.59-7.50 (m, 2H), 7.42-7.35 (m, 2H), 5.83 (d, J = 0.4 Hz, 1H), 3.68 (dt, J = 9.6, 5.2 Hz, 6H), 3.60-3.44 (m, 3H), 2.95 (ddd, J = 12.8, 9.6, 3.2 Hz, 2H), 2.60 (t, J = 5.6 Hz, 2H), 2.56 (d, J = 4.2 Hz, 4H), 2.33-2.22 (m, 3H), 2.03-1.90 (m, 2H), 1.69-1.55 (m, 2H) ppm.
Example S14. Synthesis of l-[5-methyl-l-[4 (trifluoromethoxy) phenyl]pyrazol-3-yl]-4-(l- tetrahydropyran-4-ylazetidin-3-yl)oxy-piperidine (Compound 14).
[00447] Compound 14 was prepared as outlined below.
Figure imgf000116_0001
[00448] Step 1. Synthesis of tert-butyl 3-(4-pyridyloxy)azetidine-l-carboxylate.
Figure imgf000116_0002
,
[00449] A mixture of pyridin-4-ol (0.824 g, 8.66 mmol), tert-butyl 3-hydroxyazetidine-l- carboxylate (1.0 g 5.77 mmol), triphenylphosphine (1.82 g, 6.93 mmol) and diisopropylazodicarboxylate (1.4 g, 6.93 mmol) in THF (20 mL) was degassed and purged with nitrogen 3 times, and then the mixture was stirred at 50°C for 16h under nitrogen atmosphere. The reaction mixture was quenched by the addition of water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a reside, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate=20/l to 5/1) to afford the desired product tertbutyl 3 -(4-pyridyloxy)azetidine-l -carboxylate (500 mg, 34.6%).
[00450] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 70% (214 nm), Mass: found peak 251.2 (M+l) at 0.871 min.
[00451] Step 2. Synthesis of tert-butyl 3-(4-piperidyloxy)azetidine-l-carboxylate.
Figure imgf000116_0003
[00452] Under hydrogen atmosphere, a mixture of tert-butyl 3-(4-pyridyloxy)azetidine-l- carboxylate (500 mg, 2 mmol), platinum dioxide (200 mg, 40%w/w), p-toluenesulfonic acid (688 mg, 4 mmol) in ethanol (30 mL) was stirred overnight. Then the mixture was filtered, and concentrated to afford the desired product tert-butyl 3 -(4-piperidyloxy)azetidine-l -carboxylate (400 mg, 78.1%) as a yellow oil.
[00453] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 50% (214 nm), Mass: found peak 257.4 (M+l) at 0.87 min.
[00454] Step 3. Synthesis of tert-butyl 3-[[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4-piperidyl] oxy] azetidine-l-carboxylate.
Figure imgf000117_0001
[00455] Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazole (300 mg, 0.92 mmol), tert-butyl 3-(4- piperidyloxy)azetidine-l -carboxylate (471 mg, 1.84 mmol), Pd-PEPPSI-IPent catalyst (26.8 mg, 2.76e-5 mol) and sodium tert-butoxide (442 mg, 4.6 mmol) in anhydrous THF (10 mL) in a sealed tube was stirred at 60 °C for 16h. Then the mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate =2: 1) to afford the desired product (160 mg, 35.1%) [00456] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98% (214 nm), Mass: found peak 497.3 (M+l) at 2.293 min.
[00457] Step 4. Synthesis of 4-(azetidin-3-yloxy)-l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperidine.
Figure imgf000117_0002
[00458] To a solution of tert-butyl 3-[[l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-4-piperidyl] oxy] azetidine- 1 -carboxylate (160 mg, 0.3 mmol) in dichloromethane (10 mL) was added TFA (3 mL). The reaction mixture was stirred at room temperature for 2h, then concentrated to remove solvent, and potassium carbonate was added to pH=10. The product was extracted with DCM and dried over sodium sulfate. The organic layer was concentrated to afford the desired product 4-(azetidin-3-yloxy)-l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperidine (127 mg, yield 99.4 %) as a yellow oil.
[00459] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 86% (214 nm), Mass: found peak 397.2 (M+l) at 1.075 min.
[00460] Step 5. Synthesis of l-[5-methyl-l-[4 (trifluoromethoxy) phenyl]pyrazol-3-yl]-4-(l- tetrahydropyran-4-ylazetidin-3-yl)oxy-piperidine (Compound 14).
Figure imgf000118_0001
[00461] To a solution of 4-(azetidin-3-yloxy)-l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperidine (127 mg, 0.32 mmol) in methanol (10 mL) was added tetrahydropyran- 4-one (64.1 mg, 0.64 mmol), sodium triacetoxyborohydride (204 mg, 0.96 mmol), acetic acid (0.2 mL, 0.22 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product l-[5-methyl-l-[4 (trifluoromethoxy) phenyl]pyrazol-3-yl]-4-(l-tetrahydropyran-4-ylazetidin-3-yl)oxy-piperidine (96.5 mg, yield 62.7 %) as a yellow oil.
[00462] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm), Mass: found peak 481.2 (M+l) at 2.096 min.
[00463] 'H NMR (400 MHz, MeOD-d4) 5 7.56 (d, J = 9.2 Hz, 2H), 7.41 (d, J = 8.3 Hz, 2H), 5.85 (s, 1H), 4.34-4.24 (m, 1H), 3.95 (d, J = 9.6 Hz, 2H), 3.67 (dd, J = 8.4, 6.4 Hz, 2H), 3.57 (d, J = 14.2 Hz, 3H), 3.43-3.37 (m, 2H), 3.03-2.91 (m, 4H), 2.39-2.27 (m, 4H), 1.92 (s, 2H), 1.73 (d, J = 11.2 Hz, 2H), 1.62 (d, J = 9.5 Hz, 2H), 1.37-1.19 (m, 2H) ppm. Example S15. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2- tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (Compound 15).
[00464] Compound 15 was prepared as outlined below.
Figure imgf000119_0001
[00465] Step 1. Synthesis of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2,7-diazaspiro [3.5] nonane-2-carboxylate.
Figure imgf000119_0002
[00466] To a solution of 3-iodo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole(100 mg, 0.27 mmol) in dry 1,4-dioxane (3 mL) was added tert-butyl 2,7-diazaspiro[3.5]nonane-2- carboxylate (61.5 mg, 0.27 mmol), methanesulfonate (2-di-t-butylphosphino-2',4',6'-tri-i-propyl- l,T-biphenyl)(2'-amino-l,r-biphenyl-2-yl)palladium(II) (21.6 mg, 0.027 mmol), and sodium tert-butoxide (78 mg, 0.81 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The reaction mixture was purified by SGC (PE: EA= 1 : 1) to afford the desired product tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-2,7-diazaspiro [3.5] nonane-2-carboxylate (80 mg, yield: 63.1%) as a yellow oil. LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 76% (214 nm), Mass: found peak 467.3 (M+l) at 1.510 min. [00467] Step 2. Synthesis of tert-butyl 2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000120_0001
[00468] A solution of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2,7- diazaspiro [3.5] nonane-2-carboxylate (80 mg, 0.27 mmol) in dichloromethane (10 mL) was stirred for 2h. The mixture was concentrated to remove solvent, and potassium carbonate was added to pH=10. It was extracted with DCM and dried over sodium sulfate. It was concentrated in vacuo to afford the desired product tert-butyl 2,2-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy) phenyl] pyrazol-3-yl]piperazine-l -carboxylate (60 mg, yield 92.8 %) as a yellow oil.
[00469] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 66% (214 nm), Mass: found peak 367.3 (M+l) at 1.077 min.
[00470] Step 3. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2- tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (Compound 15).
[00471] To a solution of 7-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-2,7- diazaspiro [3.5] nonane (80 mg, 0.22mmol) in 1,2-di chloroethane (5 mL) was added tetrahydropyran-4-one (22 mg, 0.22 mmol), sodium triacetoxyborohydride (139 mg, 0.65 mmol), acetic acid (0.01 mL, 0.22 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by prep- HPLC (ammonium hydrogen carbonate) to afford the desired product 7-[5-methyl-l-[4- (trifluoromethoxy) phenyl] pyrazol-3-yl]-2-tetrahydropyran-4-yl-2,7-diazaspiro [3.5] nonane (33.5 mg, yield 34.1%) as a yellow solid.
[00472] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93% (214 nm), Mass: found peak 451.2 (M+l) at 2.144 min.
[00473] 'H NMR (400 MHz, MeOD-d4) 5 7.54 (d, J = 8.8 Hz, 2H), 7.39 (d, J = 8.4 Hz, 2H),
5.84 (s, 1H), 3.97 (d, J = 9.6 Hz, 2H), 3.38 (t, J = 11.6 Hz, 4H), 3.17 (d, J = 4.8 Hz, 6H), 2.67 (s,
1H), 2.28 (s, 3H), 1.90-1.75 (m, 6H), 1.32 (d, J = 12.0 Hz, 2H) ppm. Example S16. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 16).
[00474] Compound 16 was prepared as outlined below.
Figure imgf000121_0001
[00475] Step 1. Synthesis of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl] pyrazole.
Figure imgf000121_0002
CHCI3, RT, o/n
[00476] Under argon atmosphere, a mixture of 3,5-dibromo-lH-pyrazole (200 mg, 0.88 mmol), [4-(trifluoromethoxy)phenyl]boronic acid (365 mg, 1.77 mmol), copper acetate (322 mg, 1.77 mmol) and Pyridine (280 mg, 3.54 mmol) in chloroform (10 mL) was stirred at RT for 16h.
Then the reaction mixture was concentrated and purified by SGC (PE:DCM = 2: 1) to afford the desired product 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl] pyrazole (150 mg, 44%) as a colorless oil.
[00477] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 95% (214 nm); Mass: found peak 387.0 (M+l) at 1.493 min.
[00478] Step 2. Synthesis of 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole.
Figure imgf000122_0001
dioxane/H2O(10:1 )
[00479] Under argon atmosphere, a mixture of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl] pyrazole (150 mg, 0.382 mmol), 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (64.3 mg, 0.382 mmol), l,l'-bis(diphenylphosphino) ferrocene-Palladium(II)dichloride dichloromethane complex (31.2 mg, 0.038 mmol) and sodium carbonate (38.5 mg, 0.4 mmol) in anhydrous dioxane/water (10/2 mL) was stirred at 80 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum : ethyl acetate=l : 1) to afford the desired product 3- bromo-5-isopropenyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 71.3%) as a colorless oil.
[00480] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 67% (214 nm); Mass: found peak 347.1 (M+l) at 1.511 min.
[00481] Step 3. Synthesis of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazole.
Figure imgf000122_0002
[00482] Under hydrogen atmosphere, a mixture of 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (90 mg, 0.01 mmol), PtO2 (30 mg, i/ i, 30%) in THF (5 mL) was stirred at RT for 2h. Then the reaction mixture was filtered and concentrated to afford the desired product 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazole (80 mg, 88%). [00483] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 66% (214 nm) Mass: found peak 349.1 (M+l) at 2.089 min.
[00484] Step 4. Synthesis of tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate. Boc
Figure imgf000123_0001
THF, 60°c, o/n
[00485] Under argon atmosphere, a mixture of 3-bromo-5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (80 mg, 0.22 mmol), tert-butyl 2,2-dimethylpiperazine-l- carboxylate(84 mg, 0.45 mmol), Pd PEPPSI IPENT (5 mg, 0.006 mmol) and sodium tert- butoxide (65 mg, 0.6 mmol) in anhydrous THF (2 mL) in a sealed tube was stirred at 60 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the product tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (50 mg, 46.2%) as a yellow solid.
[00486] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 455.3 (M+l) at 1.562 min.
[00487] Step 5. Synthesis of tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000123_0002
[00488] To a solution of tert-butyl 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (40 mg, 0.088 mmol) in di chloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the reaction mixture was concentrated to remove solvent, and potassium carbonate was added to pH=10. The product was extracted with DCM and dried over sodium sulfate. The mixture was concentrated to afford the desired product tert-butyl 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (35 mg, yield 70 %) as a yellow oil.
[00489] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 81% (214 nm), Mass: found peak 355.3 (M+l) at 1.072 min.
[00490] Step 6. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 16).
Figure imgf000124_0001
[00491] To a solution of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (30 mg, 0.084 mmol), potassium carbonate (58 mg, 0.42 mmol) and KI (14 mg, 0.084 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (23 mg, 0.127 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-isopropyl- l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (9.0 mg, yield 22.5%) as a yellow solid.
[00492] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm), Mass: found peak 468.2 (M+l) at 2.115 min.
[00493] 'H NMR (400 MHz, MeOD-d4) 5 7.56-7.51 (m, 2H), 7.44 (d, J = 8.8 Hz, 2H), 5.90 (s, 1H), 3.78-3.66 (m, 4H), 3.28-3.20 (m, 4H), 2.98 (dt, J = 13.6, 6.8 Hz, 1H), 2.70-2.64 (m, 4H), 2.63-2.58 (m, 4H), 2.55 (s, 4H), 1.20 (d, J = 6.8 Hz, 6H) ppm.
Example S17. Synthesis of l-imino-4-[2-[4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] - 1 ,4-t h iazina ne 1 -oxide (Compound 17).
[00494] Compound 17 was prepared as outlined below.
Figure imgf000125_0001
[00495] Step 1. Synthesis of tert-butyl thiomorpholine-4-carboxylate.
Figure imgf000125_0002
[00496] To a solution of thiomorpholine (500 mg, 4.85 mmol) and TEA (1.47 g, 14.6 mmol) in DCM (40 mL) was added di -tert-butyl di carb onate (1.27 g, 5.82 mmol). The reaction mixture was stirred at room temperature for 16h. The mixture was directly purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 3-45% ethyl acetate in petroleum ether) to afford the desired product tert-butyl thiomorpholine-4-carboxylate (927 mg, yield 94%) as a white solid.
[00497] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min 95%B for 1.7min at 2.0 mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50 °C; LC purity: 97% (214 nm) Mass: found peak 226.3 (M+23) at 1.266min.
[00498] Step 2. Synthesis of tert-butyl l-oxo-l,4-thiazinane-4-carboxylate.
Figure imgf000125_0003
[00499] At 0 °C, to a solution of tert-butyl thiomorpholine-4-carboxylate (827 mg, 4.07 mmol) in 50 mL EtOAc, 25 mL MeOH and 25 mL water was added sodium periodate (870 mg, 4.07 mmol) portion wise. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was filtered, and the filtrate was diluted with brine, extracted with DCM (4*50 mL), dried over sodium sulfate and concentrated in vacuo. The residue was purified via silica gel chromatography to afford the desired product tert-butyl l-oxo-l,4-thiazinane-4-carboxylate (800 mg, yield 89.7%) as a white solid. LCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.3min 95%B for 1.7min at 2.0mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 242.2 (M+23)+ at 0.934 min.
[00500] Step 3. Synthesis of tert-butyl 1-benzyloxy-carbonylimino- l-oxo-l,4-thiazinane-4- carboxylate.
Figure imgf000126_0001
benzyl carbamate(2.5eq) rhodium(ll) acetate dimer(0.05eq
Figure imgf000126_0003
MgO(4eq), DCM
Figure imgf000126_0002
RT 48 h
[00501] A mixture of tert-butyl l-oxo-l,4-thiazinane-4-carboxylate (700 mg, 3.19 mmol), benzyl carbamate (1.21 g, 7.98 mmol), phenyl-13 -iodanediyl diacetate (2.06 g, 6.38 mmol), MgO (515 mg, 12.8 mmol), and [Rh(OAc)2]2 (70 mg, 0.160 mmol) in DCM (10 mL) was stirred at room temperature for 4 days. The reaction mixture was filtered, and concentrated in vacuo. The residue was purified via silica gel chromatography to afford the desired product tert-butyl 1- benzyloxy-carbonylimino- l-oxo-l,4-thiazinane-4-carboxylate (620 mg, yield 52.7%) as a yellow solid.
[00502] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3min 95%B for 1.7min at 2.0mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 369.2(M+1) at 1.250 min.
[00503] Step 4. Synthesis of benzyl N-(l-oxo-l,4-thiazinan-l-ylidene) carbamate.
O N-Cbz
Figure imgf000126_0004
[00504] To a solution of tert-butyl 1 -benzyl oxy carbonylimino- 1 -oxo- l,4-thiazinane-4- carboxylate (620 mg, 1.68 mmol) in dichloromethane (20 mL) was added TFA (3 mL, 40.4 mmol). The reaction was stirred at room temperature for 4h. The reaction mixture was concentrated in vacuo to afford the desired product benzyl N-(l-oxo-l,4-thiazinan-l-ylidene) carbamate as TFA salt (660 mg, yield 100%) as a white solid.
[00505] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 97.91% (214 nm), Mass: found peak 269.2 (M+l)+, at 0.867 min.
[00506] Step 5. Synthesis of benzyl N-[4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l-yl]ethyl]-l-oxo-l,4-thiazinan-l-ylidene]carbamate.
Figure imgf000127_0001
[00507] A mixture of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (150 mg, 0.386 mmol), benzyl N-(l-oxo-l,4-thiazinan-l-ylidene)carbamate TFA salt (221 mg, 0.579 mmol) and DIPEA (249mg, 1.93 mmol) in NMP (3 mL) was treated with microwave reactor and stirred at 160°C for 3h. The reaction was cooled to room temperature and directly purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product benzyl N-[4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazin-l-yl]ethyl]-l-oxo-l,4-thiazinan-l-ylidene]carbamate (110 mg, yield 45.3%) as a white solid.
[00508] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.68% (214 nm) Mass: found peak 621.3 (M+l) at 1.148 min.
[00509] Step 6. Synthesis of l-imino-4-[2-[4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] - 1 ,4-t h iazina ne 1 -oxide (Compo
Figure imgf000127_0002
[00510] Under H2 atmosphere, a mixture of benzyl N-[4-[2-[4-[5-methyl-l-[4- (trifluorom ethoxy) phenyl]pyrazol-3-yl]piperazin- 1 -yl]ethyl]- 1 -oxo- 1 ,4-thiazinan- 1 - ylidene]carbamate (100 mg, 0.068 mmol) and Pd/C (10%, 20 mg) in methanol (50 mL) was stirred at room temperature for 3h. The reaction was filtered. The filtrate was concentrated in vacuo to afford the desired product l-imino-4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1-oxide (67.3 mg, yield 85.9%) as a yellow solid. [00511] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 487.1 (M+l) at 1.861 min.
[00512] 'H NMR (400 MHz, CD3OD) 5 7.57-7.52 (m, 2H), 7.39 (d, J = 8.8 Hz, 2H), 5.84 (s, 1H), 3.25 (t, J = 4.8 Hz, 4H), 3.20-2.94 (m, 8H), 2.74 (t, J = 6.4 Hz, 2H), 2.67 (t, J = 4.4 Hz, 4H), 2.61 (t, J = 6.4 Hz, 2H), 2.29 (s, 3H) ppm.
Example S18. Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3- tetrahydropyran-4-yl-3-azaspiro[5.5] undecane (Compound 18).
[00513] Compound 18 was prepared as outlined below.
Figure imgf000128_0001
[00514] Step 1. Synthesis of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl] oxy-3-azaspiro [5.5] undecane-3-carboxylate.
Figure imgf000128_0002
[00515] To a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol (100 mg, 0.387 mmol), tert-butyl 9-hydroxy-3-azaspiro[5.5]undecane-3-carboxylate (209 mg, 0.775 mmol) and triphenylphosphane (203 mg, 0.775 mmol) in THF (15 mL) were add DIAD (157 mg, 0.775 mmol) at 0 °C. The reaction was stirred at RT under the protection of Argon for 16h. LCMS showed the reaction was complete. The mixture was concentrated and purified by flash chromatography (Biotage, 50 g silica gel column @80mL/min, eluting with 50%-100% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 9-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3-azaspiro[5.5]undecane-3-carboxylate (150 mg, 75.1%) as a colorless oil.
[00516] LCMS method: Column: X Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B in 1.5 min at 1.5 mL/minTemperature: 50 °C; LC purity: 98.8% (214 nm) Mass: found peak 510.2 (M+l) at 2.441 min.
[00517] Step 2. Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy- 3-azaspiro [5.5] undecane.
Figure imgf000129_0001
[00518] To a solution of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]oxy-3-azaspiro[5.5]undecane-3-carboxylate (30 mg, 0.0589 mmol) in dichloromethane (5 mL) was added TFA (336 mg, 1.18 mmol), The reaction mixture was stirred at RT for 2h. The mixture was concentrated and was diluted with DCM (50 mL). The organic mixture was washed with potassium carbonate (aq) and then aq NaCl solution. The organic layer was dried and concentrated to afford 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3- azaspiro[5.5]undecane (100 mg, 69.2%) as a colorless oil.
[00519] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: from 5% increase to 95%B within 1.4 min, 95%B for 1.6 min. Flow Rate: 1.8 mL/min; Column: X Bridge C18, 4.6*50mm, 3.5pm; Column Temperature: 45 °C; LC purity: 83.4% (214 nm) Mass: found peak 409.9 (M+l) at 1.845 min.
[00520] Step 3. Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy- 3-tetrahydropyran-4-yl-3-azaspiro[5.5] undecane (Compound 18).
Figure imgf000129_0002
[00521] To a solution of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3- azaspiro[5.5]undecane (85 mg, 0.208 mmol), tetrahydropyran-4-one (41.6 mg, 0.415 mmol) and sodium triacetoxyborohydride (132 mg, 0.623 mmol) in 1,2-di chloroethane (10 mL) were add acetic acid (125 mg, 2.08 mmol). The reaction was stirred at RT for 16h. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to afford 9-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3-tetrahydropyran-4-yl-3-azaspiro[5.5] undecane (16.5 mg, 14.6%) as a colorless oil.
[00522] LCMS method: Column: Sun fire, 50*4.6mm, 3.5pm; Mobile phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Elution program: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min Flow Rate: 2 mL/min; Temperature: 50°C; LC purity: 91.0% (214 nm) Mass: found peak 494.0 (M+l) at 1.650 min.
[00523] 'H NMR (400 MHz, CDCh) 5 7.46 (d, J = 9.2 Hz, 2H), 7.28 (d, J = 8.0 Hz, 2H), 5.65 (s, 1H), 4.53 (t, J = 4.4 Hz, 1H), 4.01 (t, J = 8.0, 2H), 3.37 (t, J = 11.2 Hz, 2H), 2.48-2.56 (m, 4H), 2.30 (s, 3H), 1.91 (d, J = 5.6 Hz, 2H), 1.81 (d, J = 11.2 Hz, 2H), 1.60-1.71 (m, 8H), 1.49 (s, 2H), 1.26 (d, J = 11.2 Hz, 3H).
Example S19. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-2- tetrahydropyran-4-yl-2-azaspiro[3.5] nonane (Compound 19).
[00524] Compound 19 was prepared as outlined below.
Figure imgf000130_0001
[00526] To a round bottom flask were add isopropyl 3-oxobutanoate (2.0 g, 13.9 mmol), hydrazine hydrochloride (998 mg, 14.6 mmol) and propan-2-ol (25 mL). The mixture was stirred at 90 °C for 16h, and LCMS showed the reaction was complete. The solution was concentrated and EtOAc (200 mL) was added to the mixture. The organic phase was washed with IN solution of sodium hydrogen carbonate, dried over sodium sulfate and concentrated to dryness to afford 3-isopropoxy-5-methyl-lH-pyrazole (850mg, 38.5%) as a colorless oil.
[00527] LCMS method: Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Gradient: 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 88.0% (214 nm) Mass: found peak 141.3 (M+l) at 0.963 min.
[00528] Step 2. Synthesis of 3-isopropoxy-5-methyl-l-[4-(trifluoromethoxy)phenyl].
Figure imgf000131_0001
[00529] A suspension 3-isopropoxy-5-methyl-lH-pyrazole (800 mg, 5.71 mmol), [4- (trifluoromethoxy)phenyl]boronic acid (3.53 g, 17.1 mmol), pyridine (2.26 g, 28.5 mmol), copper(II) acetate (3.11 mg, 17.1 mmol) in di chloromethane (50 mL) was stirred at RT for 48h under the protection of Argon. LCMS showed the reaction was complete, the mixture was filtered, and the filter cake was washed with di chloromethane (30 mL). The DCM solution was concentrated, and purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 10%-60% ethyl acetate in petroleum ether for 30 min) to afford 3- isopropoxy-5-methyl-l-[4-(trifluoromethoxy)phenyl] (700 mg, 40.0%) as a colorless oil. [00530] LCMS method: Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 5% to 95% of B ini.5 min at 1.5 mL/min; Column: X BRIGE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 98.0% (214 nm) Mass: found peak 301.1 (M+l) at 2.237 min.
[00531] Step 3. Synthesis of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol.
Figure imgf000131_0002
[00532] To a screw-cap vial were add 3-isopropoxy-5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazole (670 mg, 2.23 mmol), hydrogen bromide (3.61 g, 17.9 mmol) and acetic acid (3 mL). The reaction was stirred at 140 °C under the protection of Argon for 3h. LCMS showed the reaction was complete. The mixture was concentrated and diluted with ethyl acetate (100 mL), which was washed with sodium bicarbonate solution and aq NaCl solution. Then the organic solution was concentrated and purified by flash chromatography (Biotage, 50 g silica gel column @90mL/min, eluting with 10%-30% EtOAc in PE for 30 min) to afford 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol (430 mg, 73.4%) as awhite solid. [00533] LCMS method: Column: HALO C18 2.7pm 4.6*30mm; Mobile phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Elution program: Gradient from 5% to 95% of B in 1.0 min at 2.2 mL/min; Temperature: 40°C; LC purity: 98.4% (214 nm) Mass: found peak 259.2 (M+l) at 1.204 min.
[00534] Step 4. Synthesis of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] oxy-3-azaspiro [5.5] undecane-3-carboxylate.
Figure imgf000132_0001
[00535] To a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol (30 mg, 0.116 mmol), tert-butyl 7-hydroxy-2-azaspiro[3.5]nonane-2-carboxylate (56.1 mg, 0.232 mmol) and triphenylphosphane (61 mg, 0.232 mmol) in THF (5 mL) were added DIAD (47 mg, 0.232 mmol) at 0 °C. The reaction was stirred at RT under the protection of Argon for 16h. LCMS showed the reaction was complete. The mixture was concentrated and which was purified by flash chromatography (Biotage, 50 g silica gel column @80mL/min to afford tert-butyl 9-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-3-azaspiro[5.5]undecane-3-carboxylate (30 mg, 52.9%) as a colorless oil.
[00536] LCMS method: Column: X Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B ini.5 min at 1.5 mL/min. Temperature: 50 °C; LC purity: 98.7% (214 nm) Mass: found peak 426.1 (M-56+l)+ at 2.364 min.
[00537] Step 5. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy- 2-azaspiro [3.5] nonane.
Figure imgf000133_0001
[00538] To a solution of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]oxy-2-azaspiro[3.5]nonane-2-carboxylate (180 mg, 0.374 mmol) in dichloromethane (5 mL) was added TFA (852 mg, 0.748 mmol), The reaction mixture was stirred at RT for 2h. The mixture was concentrated, diluted with DCM (50mL) and washed with aq potassium carbonate. Then the solution was washed with aq NaCl solution, dried over sodium sulfate and concentrated to afford 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-2- azaspiro[3.5]nonane (110 mg, 69.6%) as a colorless oil.
[00539] LCMS method: Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 10% to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 87.3% (214 nm) Mass: found peak 382.1 (M+l) at 1.907 min.
[00540] Step 6. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-
2-tetrahydropyran-4-yl-2-azaspiro[3.5] nonane (Compound 19).
Figure imgf000133_0002
[00541] To a suspension of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-2- azaspiro[3.5]nonane (110 mg, 0.288 mmol), tetrahydropyran-4-one (57.8 mg, 0.577 mmol) and sodium triacetoxyborohydride (183 mg, 0.865 mmol) in 1,2-di chloroethane (10 mL) was added acetic acid (173 mg, 2.88 mmol). The reaction was stirred at RT for 16h. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to afford 7-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-2-tetrahydropyran-4-yl-2-azaspiro[3.5] nonane (28.3 mg, 20.4%) as a colorless oil.
[00542] LCMS method: Column: Sun fire, 50*4.6mm, 3.5pm; Mobile phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Elution program: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min Flow Rate : 2 mL/min; Temperature: 50°C; LC purity: 97.0% (214 nm) Mass: found peak 466.0 (M+l) at 1.610 min. [00543] 'H NMR (500 MHz, CDCh) 5 7.43-7.46 (m, 2H), 7.26-7.28 (m, 2H), 5.64 (s, 1H), 4.53 (d, J = 7.0 Hz, 1H), 3.94-3.97 (m, 2H), 3.33-3.38 (m, 2H), 2.97 (d, J = 19.5 Hz, 4H), 2.30 (s, 3H), 2.22 (d, J = 9.0 Hz, 1H), 1.96 (d, J = 10.0 Hz, 4H), 1.55-1.65 (m, 6H), 1.30-1.37 (m, 2H).
Example S20. Synthesis of 2-[5-methyl-l-[4-(trifluoro-methoxy)phenyl]pyrazol-3-yl]oxy-7- tetrahydropyran-4-yl-7-azaspiro[3.5]nonane (Compound 20).
[00544] Compound 20 was prepared as outlined below.
Figure imgf000134_0001
[00545] Step 1. Synthesis of tert-butyl 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]oxy-7-azaspiro[3.5]nonane-7-carboxylate.
Figure imgf000134_0002
[00546] To a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-ol (50 mg, 0.194 mmol), tert-butyl 2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylate (93.5 mg, 0.387 mmol) and triphenylphosphine (102 mg, 0.387 mmol) in THF (8 mL) was added DIAD (78.3 mg, 0.387 mmol) at 0 °C. The reaction was stirred at RT under the protection of Argon for 16h. LCMS showed the reaction was complete. The mixture was concentrated and purified by flash chromatography (Biotage, 50 g silica gel column @70mL/min, eluting with 10%-60% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-7-azaspiro[3.5]nonane-7-carboxylate (30 mg, 14.8%) as a colorless oil. [00547] LCMS method: Column: X-Bridge C18 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10% to 95% of B in 1.5 min at 1.8 mL/min; Temperature: 50°C; LC purity: 92.3% (214 nm) Mass: found peak 426.1 (M-56+l)+ at 2.374 min.
[00548] Step 2. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]oxy- 7-azaspiro [3.5] nonane.
Figure imgf000135_0001
[00549] To a solution of tert-butyl 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]oxy-7-azaspiro[3.5]nonane-7-carboxylate (40 mg, 0.0831 mmol) in dichloromethane (3 mL) was added TFA (189 mg, 1.66 mmol). The reaction mixture was stirred at RT for 2h. The mixture was concentrated, diluted with DCM (50 mL), washed with aq potassium carbonate. Then the solution was washed with aq NaCl solution, dried and concentrated to afford 2-[5- methyl-l-[4-(trifluorom ethoxy) phenyl]pyrazol-3-yl]oxy-7-azaspiro[3.5]nonane (20 mg, 32.8%) as a colorless oil.
[00550] LCMS method: Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 5% to 95% of B in 1.5 min at 1.5 mL/min; Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 52.0% (214 nm) Mass: found peak 382.1 (M+l) at 2.086 min.
[00551] Step 3. Synthesis of 2-[5-methyl-l-[4-(trifluoro-methoxy)phenyl]pyrazol-3-yl]oxy-
7-tetrahydropyran-4-yl-7-azaspiro [3.5] nonane (Compound 20).
Figure imgf000135_0002
[00552] To a suspension of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]oxy-7- azaspiro[3.5]nonane (20 mg, 0.0524 mmol), tetrahydropyran-4-one (10.5 mg, 0.105 mmol) and sodium triacetoxyborohydride (33.3 mg, 0.157 mmol) in 1,2-dichloroethane (5 mL) was add acetic acid (173 mg, 2.88 mmol). The reaction was stirred at RT for 48h. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to afford 2-[5- methyl- l-[4-(trifluoro-methoxy)phenyl]pyrazol-3-yl]oxy-7-tetrahydropyran-4-yl-7- azaspiro[3.5]nonane (17.5 mg, 71.7%) as a colorless oil.
[00553] LCMS method: Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10% to 95% of B in 1.5 min at 1.8 mL/min; Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 466.1 (M+l) at 2.146 min.
[00554] 'H NMR (400 MHz, CD3OD) 5 7.54-7.56 (m, 2H), 7.43 (d, J = 8.4 Hz, 2H), 5.74 (s, 1H), 4.84 (t, J = 6.8 Hz, 1H), 3.98-4.02 (m, 2H), 3.40 (t, J = 7.6 Hz, 2H), 2.48-2.58 (m, 4H), 2.38-2.45 (m, 3H), 2.31 (s, 3H), 1.82-1.92 (m, 4H), 1.69 -1.72(m, 4H), 1.55-1.60 (m, 2H).
Example S21. Synthesis of 4-[2-[4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 21).
[00555] Compound 21 was prepared as outlined below.
Figure imgf000136_0001
[00556] Step 1. Synthesis of tert-butyl 4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5- methyl-pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000137_0001
[00557] A suspension of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (200 mg, 0.751 mmol), [3-(difluoromethoxy)-4-fluoro-phenyl]boronic acid (309 mg, 1.50 mmol), pyridine (297 mg, 3.75 mmol), copper(II) acetate (409 mg, 2.25 mmol) in chloroform (25 mL) was stirred at 40°C for 16h under the atmosphere of O2. LCMS showed the reaction was complete, the mixture was filtered, and the filter cake was washed with dichloromethane (50 mL), The filtrate was concentrated, and purified by flash chromatography (Biotage, 50 g silica gel column @80mL/min, eluting with 0%-40% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (190 mg, 59.3%) as a white solid.
[00558] LCMS method: Mobile Phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Gradient: from 5% to 95% of B ini.5 min at 1.5 mL/min, Column: X Bridge C18 (4.6x 50 mm, 3.5pm), Column Temperature: 50 °C, LC purity: 100% (254 nm) Mass: found peak 427.1 (M+l) at 2.179 min.
[00559] Step 2. Synthesis of l-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl-pyrazol- 3-yl] piperazine.
Figure imgf000137_0002
[00560] To a solution of tert-butyl 4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazine- 1-carboxylate (190 mg, 0.446 mmol) in dichloromethane (8 mL) was added TFA (508 mg, 4.46 mmol). The reaction mixture was stirred at RT for 2h. The mixture concentrated, diluted with DCM (50 mL) and washed with aq. potassium carbonate. Then the organic solution was washed with aq. NaCl solution, dried and concentrated to afford 1 -[ 1 -[3- (difluoromethoxy)-4-fluoro-phenyl]-5-methyl-pyrazol-3-yl]piperazine (110 mg, 56.4%) as a colorless oil.
[00561] LCMS method: Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B), Gradient: from 5% to 95% of B in 1.0 min at 2.2 mL/min, Column: HALO C18 2.7pm 4.6*30mm, Column Temperature: 40 °C, LC purity: 74.6% (214 nm) Mass: found peak 327.5 (M+l) at 0.954 min.
[00562] Step 3. Synthesis of 4-[2-[4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 21).
Figure imgf000138_0001
[00563] To a solution of l-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]-5-methyl-pyrazol-3- yl]piperazine (110 mg, 0.337 mmol) in ethanol / water (10 mL/0.5 mL) were added 4-(2- chloroethyl)morpholine;hydrochloride (94.1 mg, 0.506 mmol), KI (56.0 mg, 0.337 mmol) and potassium carbonate (140 mg, 1.01 mmol). The reaction mixture was stirred at 90 °C for 16h under the protection of Argon, then the mixture was concentrated and diluted with methanol. The mixture was filtered, and the filter cake washed with MeOH (5 mL), then the filtrate was concentrated and purified by HPLC to afford 4-[2-[4-[l-[3-(difluoromethoxy)-4-fluoro-phenyl]- 5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (40.3 mg, yield 27.2% ) as a white solid. [00564] LCMS method: Mobile Phase: WATER (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Gradient: from 5% to 95% of B ini.5 min at 1.5 mL/min, Column: X Bridge C18 (4.6x 50 mm, 3.5pm), Column Temperature: 50 °C, LC purity: 100% (214 nm) Mass: found peak 440.2 (M+l) at 1.900 min.
[00565] 'H NMR (400 MHz, CDCh) 5 7.35 (dt, J = 2.4 Hz, 7.2 Hz, 1H), 7.28 (t, J = 1.2 Hz, 1H), 7.23 (dt, J = 9.6 Hz, 18.8 Hz, 1H), 6.40-6.76 (m, 1H), 3.69-3.74 (m, 8H), 5.68 (s, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.25 (t, J = 5.2 Hz, 4H), 2.61 (t, J = 4.8 Hz, 4H), 2.56 (dt, J = 4.8 Hz, 10.8 Hz, 4H), 2.50 (t, J = 4.0 Hz, 4H), 2.28 (s, 3H).
Example S22. Synthesis of 5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethoxy)phenyl] pyrazole-3-carbonitrile (Compound 22) and 5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethoxy)phenyl]pyrazole-3-carboxamide (Compound a3).
[00566] Compounds 22 and a3 were prepared as outlined below.
Figure imgf000139_0001
[00567] Step 1. Synthesis of trimethyl-[2-(pyrazol-l-ylmethoxy)ethyl]silane.
Figure imgf000139_0002
THF 0 C-RT, 16 h
[00568] Sodium hydride (1.29 g, 32.3 mmol) was added portion-wise to IH-pyrazole (2 g, 29.4 mmol) in THF (50 mL) at 21 °C over a period of 15 minutes. The resulting solution was stirred at 21 °C for 1 h. The mixture was cooled to 0 °C and (2-(chloromethoxy)ethyl)trimethylsilane (6.7 mL, 32.3 mmol) was added and the mixture was stirred for 16 h at room temperature. The reaction mixture was quenched by the addition of brine (150 mL) and extracted with EtOAc (3 x 100 mL). The organic phases were combined, dried over sodium sulfate and the volatiles removed under reduced pressure. Purification by flash chromatography 2-30% acetone in petroleum ether to afford trimethyl-[2-(pyrazol-l-ylmethoxy)ethyl]silane (360 mg, 61.0% yield) as a yellow solid.
[00569] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.47% (214 nm) Mass: found peak 199.2 (M+l) at 1.305 min. [00570] Step 2. Synthesis of l-2-(2-trimethylsilylethoxymethyl)pyrazole-3-carbonitrile.
Figure imgf000140_0001
[00571] Lithium magnesium 2,2,6,6-tetramethylpiperidin-l-ide dichloride (13.1 mL, 13.1 mmol) was added dropwise to trimethyl-[2-(pyrazol-l-ylmethoxy)ethyl]silane (2 g, 10.1 mmol) in THF (30 mL) at 21 °C under nitrogen. The resulting suspension was stirred at 21 °C for 1 h and then cooled to 5 °C. p-tolylsulfonyl-formonitrile (2.38 g, 13.1 mmol) was added and the mixture stirred for 45 minutes. The reaction was quenched by the addition of brine and extracted with EtOAc (200 mL). The organic layer was dried and concentrated in vacuo to crude material. The crude product was purified by flash silica chromatography, elution gradient 5 to 40% acetone in petroleum ether to afford l-2-(2-trimethylsilylethoxymethyl)pyrazole-3-carbonitrile (1.5 g, 52.8%) as a colorless liquid.
[00572] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 66.12% (254 nm) Mass: found peak 199.0 (M+l) at 2.018 min.
[00573] Step 3. Synthesis of 5-iodo-2-(2-trimethylsilylethoxymethyl)pyrazole-3- carbonitrile. M (2 eq) ( SEM h _ ^NC^_N
Figure imgf000140_0002
CgH18CI2MgN.Li (1 .5 eq) THF, -16 °c, 2 h
[00574] Lithium magnesium 2,2,6,6-tetramethylpiperidin-l-ide dichloride (12.1 mL, 12.1 mmol) was added dropwise over 15 minutes to 2-(2 -trimethylsilylethoxymethyl) pyrazole-3- carbonitrile (1.8 g, 8.06 mmol) in THF (40 mL) at -16°C under nitrogen. The resulting solution was stirred at
-16°C for 1 h. Iodine (818 mg, 3.22 mmol) was added and the mixture stirred for 1 h. The reaction was quenched by the addition of brine, extracted with EtOAc (2 x 100 mL), dried and concentrated in vacuo to crude material. The crude product was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in isohexane. Pure fractions were concentrated in vacuo to dryness to afford ethyl 5-iodo-2-(2- trimethylsilylethoxymethyl)pyrazole-3-carbonitrile (1.8 g, 47.9%) as a colorless liquid.
[00575] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 61.34% (214 nm) Mass: found peak 470.1 (M+l) at 2.013 min.
[00576] Step 4. Synthesis of 3-iodo-lH-pyrazole-5-carbonitrile.
Figure imgf000141_0001
[00577] Under argon atmosphere, a mixture of 5-iodo-2-(2 -trimethylsilylethoxymethyl) pyrazole-3 -carbonitrile (2 g, 5.73 mmol) in anhydrous ethanol (15 mL) was treated with IN HC1 (15 mL). The mixture was stirred at 90 °C for 3h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with sodium bicarbonate to pH=8, extracted with EtOAc (100 mL*3), dried over sodium sulfate, filtered, and concentrated, then purified by flash chromatography 2-20% MeOH in DCM to afford 3-iodo-lH-pyrazole-5- carbonitrile (1.2 g, 77.5% yield) as a yellow solid.
[00578] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 80.97% (214 nm), Mass: found peak 220.0 (M+l) at 1.638 min.
[00579] Step 5. Synthesis of 5-iodo-2-[4-(trifluoromethoxy)phenyl]pyrazole-3-carbonitrile.
Figure imgf000141_0002
[00580] To a solution of 3-iodo-lH-pyrazole-5-carbonitrile (300 mg, 1.37 mmol) in dichloromethane (20 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (564 mg, 2.74 mmol), pyridine (542 mg, 6.85 mmol) and Copper (II) acetate (498 mg, 2.74 mmol). The reaction mixture was stirred at RT for 16h. The mixture was concentrated, finally purified by flash chromatography (Biotage, 40 g silica gel column @70 mL/min, eluting with 0-40% dichloromethane in petroleum ether for 30 min) to afford 5-iodo-2-[4- (trifluoromethoxy)phenyl]pyrazole-3-carbonitrile (310 mg, 50.6%) as a white solid.
[00581] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 84.72% (214 nm) Mass: found peak 380.0 (M+l) at 2.160 min. [00582] Step 6. Synthesis of tert-butyl 4-[5-cyano-l-[4-(trifluoromethoxy)phenyl]indazol- 3-yl] piperazine- 1-carboxylate. p . (0.1 eq) Pd2(dba)3 Xantphosjf^Seq) Cs2CO3 dioxane, 80uc, in tube
Figure imgf000142_0002
Figure imgf000142_0001
[00583] Under argon atmosphere, a mixture of 3-iodo-l-[4-(trifluoromethoxy)phenyl] indazole- 5-carbonitrile (50 mg, 0.132 mmol), tert-butyl piperazine- 1 -carboxylate (37 mg, 0.2 mmol), xantphos (15 mg, 0.026 mmol), Pd2(dba)3 (12 mg, 0.013 mmol) and cesium carbonate (130 mg, 0.4 mmol) in anhydrous 1,4-di oxane (3 mL) in a sealed tube was stirred at 80 °C for 16h, filtered, and concentrated, then purified by flash chromatography 0-20% ethyl acetate in petroleum ether to afford tert-butyl 4-[5-cyano-l-[4-(trifluoromethoxy)phenyl]indazol-3- yl]piperazine-l -carboxylate (40 mg, 55.7% yield) as a yellow oil.
[00584] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 80.28% (214 nm) Mass: found peak 382.2 (M-55)+ at 2.297 min.
[00585] Step 7. Synthesis of 5-piperazin-l-yl-2-[4-(trifluoromethoxy)phenyl] pyrazole-3- carbonitrile.
Figure imgf000142_0003
[00586] To a solution of tert-butyl 4-[5-cyano-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (40 mg, 0.09 mmol) in di chloromethane (3 mL) was added TFA (0.3 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 5-piperazin-l-yl-2-[4-(trifluoromethoxy)phenyl] pyrazole-3 -carbonitrile (30 mg, yield 77.5%) as a yellow oil. [00587] Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 79.68% (214 nm) Mass: found peak 338.2 (M+l) at 1.066 min.
[00588] Step 8. Synthesis of 5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethoxy)phenyl] pyrazole-3-carbonitrile (Compound 22) and 5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethoxy)phenyl]pyrazole-3-carboxamide (Compound a3).
Figure imgf000143_0001
[00589] To a solution of 5-piperazin-l-yl-2-[4-(trifluoromethoxy)phenyl]pyrazole-3- carbonitrile (25 mg, 0.074 mmol), potassium carbonate (52 mg, 0.37 mmol) and KI (12 mg, 0.074 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (21 mg, 0.111 mmol). The reaction was stirred at 90 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethoxy)phenyl] pyrazole-3 -carbonitrile (6.6 mg, yield 19.6%) as a white solid and 5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethoxy)phenyl]pyrazole-3-carboxamide (6.6 mg, yield 11.9%) as a yellow oil.
[00590] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C.
[00591] Compound 22: LC purity: 99.07% (214 nm) Mass: found peak 451.1 (M+l) at 2.065 min. 'H NMR (400 MHz, MeOD-d4) 5 7.84-7.78 (m, 2H), 7.46 (d, J = 8.4 Hz, 2H), 6.75 (s, 1H), 3.70 (t, J = 4.8 Hz, 4H), 3.32 (t, J = 4.8 Hz, 4H), 2.65 (J = 4.8 Hz, 4H), 2.61-2.57 (m, 4H), 2.58 (t, J = 4.8 Hz, 4H) ppm.
[00592] Compound a3: LC purity: 93.82% (214 nm) Mass: found peak 468.9 (M+l) at 1.555 min. XH NMR (400 MHz, MeOD-d4) 5 7.54-7.50 (m, 2H), 7.35 (d, J= 8.5 Hz, 2H), 6.45 (s, 1H), 3.72(t, J=4.8 Hz, 4H), 3.32 (t, J=4.8 Hz, 4H), 2.68 (t, J = 4.8 Hz, 4H), 3.34-3.29 (m, 4H), 2.60 (t, J=4.8 Hz, 4H) ppm. Example S23. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol- 3-yl]piperazin-l-yl]ethyl]morpholine (Compound 23).
Synthesis 1
[00593] Compound 23 was prepared as outlined below.
Figure imgf000144_0001
[00594] Step 1. Synthesis of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000144_0002
[00595] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.2 g, 0.751 mmol), anhydrous copper acetate (273 mg, 1.5 mmol), and pyridine (0.302 mL, 3.75 mmol) in dichloromethane (5 mL) was added (2,2-difluorobenzo [d][l,3]dioxol-5-yl)boronic acid (227 mg, 1.13 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether / ethyl acetate = 4 / 1) to afford the desired product tert-butyl 4-[ 1 -(2,2-difluoro- 1 ,3 -benzodi oxol-5-yl)-5-methyl-pyrazol-3 -yl]piperazine- 1 -carboxylate (206 mg, yield 64.9%) as a yellow solid.
[00596] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 99 % (214 nm); Mass: found peak 423.3 (M + H) at 1.49 min.
[00597] Step 2. Synthesis of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl] piperazine.
Figure imgf000145_0001
[00598] To a solution of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol- 3 -yl]piperazine-l -carboxylate (206 mg, 0.488 mmol) in 3mL dichloromethane was added TFA (1 mL). The mixture was stirred at room temperature for Ih. The reaction was quenched with saturated potassium carbonate (3mL) and extracted with dichloromethane (10 mL X 3). The organic layer was dried over sodium sulfate, and the organic phase was concentrated to dryness to give l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine (155 mg, crude) as a yellow solid.
[00599] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98 % (214 nm); Mass: found peak 323.2 (M + H) at 1.56 min.
[00600] Step 3. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 23).
Figure imgf000145_0002
[00601] To a solution of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl]piperazine (115 mg, 0.481 mmol), potassium carbonate (199 mg, 1.44 mmol) and KI (79.8 mg, 0.481 mmol) in 95% ethanol / water (lOmL /I mL) was added 4-(2-chloroethyl)morpholine (10.8 mg, 0.721 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water/CFLCN) to afford the desired product 4- [2- [4- [ 1 -(2,2-difluoro- 1 ,3 -benzodi oxol-5-yl)-5-methyl-pyrazol-3 -yl]piperazin- 1 - yl]ethyl]morpholine (141.4 mg, yield 67.4%) as a white solid. [00602] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm);
Mass: found peak 436.2 (M + H) at 1.96 min.
[00603] 'H NMR (400MHz, CDCh): 5 7.22 (d, J = 3.2 Hz, 1H), 7.08-7.13 (m, 2H), 5.71 (s,
1H), 3.73 (t, J = 4.8 Hz, 4H), 3.27 (t, J = 4.8 Hz, 4H), 2.63 (t, J = 4.8 Hz, 4H), 2.56-2.60 (m,
4H), 2.52-2.55 (m, 4H) ppm.
Synthesis 2
[00604] Compound 23 was prepared as outlined below.
Figure imgf000146_0001
[00605] Step 1. Synthesis of tert-butyl 4-(3-oxobutanoyl) piperazine-l-carboxylate.
Figure imgf000146_0002
[00606] A mixture of tert-butyl piperazine-l-carboxylate (30.0 g, 0.161mol) and tert-butyl acetoacetate (28.0 g, 0.177mol) in toluene (300 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-60% acetone in petroleum ether for 8 CV) to afford tert-butyl 4-(3 -oxobutanoyl) piperazine-l- carboxylate (43.0 g, 96.6% yield) as a transparent oil.
[00607] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97.84% (214 nm) Mass: found peak 215.3 (M-55)+ at 1.48 min.
[00608] Step 2. Synthesis of tert-butyl 4-(3-oxobutanethioyl) piperazine-l-carboxylate.
Figure imgf000146_0003
[00609] To a solution of tert-butyl 4-(3-oxobutanoyl)piperazine-l -carboxylate (43.0 g, 0.156mol) in toluene (400 mL) was added Lawesson's reagent (31.5 g, 0.078mol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 4-(3 -oxobutanethioyl) piperazine- 1 -carboxylate (20.1g, yield 39.5%) as a yellow solid.
[00610] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 87.59% (254 nm) Mass: found peak 287.3 (M+l) at 1.65 min.
[00611] Step 3. Synthesis of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000147_0001
[00612] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (3 g, 11.3 mmol) in chloroform (300 mL) was added (2, 2-difluoro-l, 3-benzodioxol-5-yl)boronic acid (4.64 g, 22.5 mmol), anhydrous copper acetate (4.09 g, 22.5 mmol), pyridine (4.45 g, 56.3 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, and concentrated in vacuo The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[l-(2, 2-difluoro- 1, 3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate (3.0 g, yield 63%) as a white solid.
[00613] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97.73% (214 nm) Mass: found peak 422.9 (M + 1)+ at 2.036 min.
[00614] Step 4. Synthesis of l-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl] piperazine.
Figure imgf000147_0002
[00615] To a solution oftert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (3 g, 11.26 mmol) in di chloromethane (20 mL) was added 2,2,2- trifluoroacetic acid (10 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine (2.5 g, crude). The crude product was used directly in the next step.
[00616] LCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 323.2 (M + 1)+ at 1.644 min.
[00617] Step 5. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine.
Figure imgf000148_0001
[00618] To a solution of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (3.4 g, 11 mmol), potassium carbonate (7.57 g, 54.8 mmol) and KI (1.82 g, 11 mmol) in 95% ethanol (30 mL) was added 4-(2-chloroethyl)morpholine (2.4 g, 16.4 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (2.18 g, yield: 64.6%) as a white solid.
[00619] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.56 % (214 nm); Mass: found peak 436.2 (M + H) at 1.758 min.
[00620] 'H NMR (400 MHz, CDCh) 5 7.21 (d, J = 1.3 Hz, 1H), 7.11-7.08 (m, 2H), 5.68 (s, 1H), 3.76-3.66 (m, 4H), 3.35-3.16 (m, 4H), 2.65-2.60 (m, 4H), 2.58 (s, 4H), 2.51 (s, 4H), 2.27 (s, 3H) ppm.
Example S24. Synthesis of 4-[2-[4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 24).
[00621] Compound 24 was prepared as outlined below.
Figure imgf000149_0001
[00622] Step 1. Synthesis of tert-butyl 4-[l-(4-fluoro-3-formyl-phenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000149_0002
[00623] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.1 g, 0.375 mmol), anhydrous copper acetate (0.136 g, 0.751 mmol), and pyridine (0.151 mL, 1.88 mmol) in chloroform (5 mL) were added (4-fluoro-3-formyl-phenyl)boronic acid (94.6 mg, 0.563 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether / ethyl acetate = 4 / 1) to afford the desired product tert-butyl 4-[l-(4-fluoro-3- formyl-phenyl)-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate (100 mg, yield 68.6%) as a yellow solid.
[00624] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: CH3CN (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 95 % (214 nm); Mass: found peak 389.3 (M + H) at 1.36 min. [00625] 'H NMR (400MHz, CDCh): 5 10.39 (s, 1H), 7.90 (dd, J = 6.0, 2.8 Hz, 1H), 7.74-7.79 (m, 1H), 7.28 (d, J = 18.4 Hz, 1H), 5.74 (s, 1H), 3.57 (t, J = 4.8 Hz, 4H), 3.22 (t, J = 5.2 Hz, 4H), 2.34 (s, 3H), 1.50 (s, 9H) ppm.
[00626] Step 2. Synthesis of tert-butyl 4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000150_0001
[00627] At -78 °C, a solution of tert-butyl 4-[l-(4-fluoro-3-formyl-phenyl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (80 mg, 0.206 mmol) in 1 mL DCM was added DAST (0.5 mL, 4.12 mmol) dropwise. The mixture was stirred at -78 °C for 1 h, then warmed to room temperature. The mixture was stirred at room temperature for 16 h. The mixture was diluted with saturated sodium bicarbonate solution (3 mL) and extracted with dichloromethane (3x3 mL). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give tert-butyl 4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (65 mg, 76.9% yield) as a colorless oil.
[00628] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: CH3CN; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X- Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96 % (214 nm); Mass: found peak 411.2 (M + H) at 2.15 min.
[00629] Step 3. Synthesis of l-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-3- yl] piperazine.
Boc
Figure imgf000150_0002
[00630] To a solution of tert-butyl 4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-
3 -yl]piperazine-l -carboxylate (73 mg, 0.178 mmol) in dichloromethane (1.5 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 1 h. The reaction was quenched with saturated potassium carbonate (5 mL). The resulting mixture was extracted with dichloromethane (10 mL* 3). The combined DCM layers were dried over sodium sulfate, filtered, and concentrated to dryness to give l-[l-[3-(difhioromethyl)-4-fluoro-phenyl]-5- methyl-pyrazol-3-yl]piperazine (55 mg, crude) as a yellow solid.
[00631] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95 % (214 nm); Mass: found peak 323.2 (M + H) at 1.49 min.
[00632] Step 4. Synthesis of 4-[2-[4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 24).
Figure imgf000151_0001
[00633] To a solution of l-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-3- yl]piperazine (55 mg, 0.177 mmol), potassium carbonate (73.5 mg, 0.532 mmol) and KI (29.4 mg, 0.177 mmol) in 95% ethanol / water (10 mL /I mL) was added 4-(2-chloroethyl)morpholine (39.8 mg, 0.266 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/CHiCN) to afford the desired product 4- [2-[4-[l-[3-(difluoromethyl)-4-fluoro-phenyl]-5-methyl-pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (28.4 mg, yield 37.8%) as a yellow solid.
[00634] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.5 min, 95%B for 1.5 min, back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column: Sunfire C18 ,4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 423.9 (M + H) at 1.71 min. [00635] 'H NMR (400MHz, CDCh): 5 7.68 (dd, J = 6.0, 2.4 Hz, 1H), 7.62-7.66 (m, 1H), 7.37 (t, J = 9.6 Hz, 1H), 7.05(t, J = 54.4 Hz, 1H), 5.86 (s, 1H), 3.71 (t, J = 4.8 Hz, 4H), 2.66 (t, J = 4.8 Hz, 4H), 2.59-2.62 (m, 4H), 2.54 (s, 4H), 2.29 (s, 3H) ppm
Example S25. Synthesis of 4-[(7R,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido [l,2-a]pyrazin-7-yl]morpholine (Compound 25). [00636] Compound 25 was prepared as outlined below.
Figure imgf000152_0001
[00637] Step 1. Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
Figure imgf000152_0002
[00638] Under argon atmosphere, a mixture of (2S,5S)-5-hydroxypiperidine-2-carboxylic acid (500 mg, 3.44 mmol), thionyl chloride (615 mg, 5.17 mmol) in MeOH (20 mL) was stirred at 65 °C for 2h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate (540 mg, yield 98.5%) as a colorless oil.
[00639] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; Mass: found peak 160 (M + H) at 0.399 mm.
[00640] Step 2. Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
Figure imgf000153_0001
[00641] Under argon atmosphere, a mixture of methyl (2S,5S)-5-hydroxypiperidine-2- carboxylate (540 mg, 3.39 mmol), potassium carbonate (1.41 g, 10.2 mmol), tertbutoxycarbonyl tert-butyl carbonate (830 mg, 3.78 mmol) in THF/water (20 mL/5 mL) was stirred at room temperature for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate (800 mg, yield 90.9%) as a colorless oil.
[00642] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 100% (214 nm); Mass: found peak 160 (M -100) + at 1.061 min.
[00643] Step 3. Synthesis of 1-tert-butyl 2-methyl (2S)-5-oxopiperidine-l,2-dicarboxylate
Figure imgf000153_0002
[00644] To a solution of 01 -tert-butyl O2-methyl (2S, 5S)-5-hydroxypiperidine-l, 2- dicarboxylate (700 mg, 2.7 mmol) in DCM (30 mL) at 0 °C was added DMP (2.29 g, 5.4 mmol). The reaction mixture was stirred at this temperature for Ih. The reaction was diluted with DCM (30 mL), washed with 5% Na2S20s aqueous (20 mL) and 5% sodium bicarbonate aqueous (20 mL), dried over Sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-45% ethyl acetate in petroleum ether) to afford the desired product 1-tert-butyl 2-methyl (2S)-5- oxopiperidine-l,2-dicarboxylate (500 mg, yield 70%) as a yellow oil.
[00645] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak Mass: found peak 158.3 (M- 100)+ at 1.111 min.
[00646] Step 4. Synthesis of 1-tert-butyl 2-methyl (2S)-5-morpholinopiperidine-l,2- dicarboxylate.
Figure imgf000154_0001
[00647] Under argon atmosphere, a mixture of Ol-tert-butyl O2-methyl (2S)-5-oxopiperidine- 1,2-dicarboxylate (50 mg, 0.194 mmol), acetic acid (0.09 mmol, 5.8 mg), morpholine (58.4 mg, 0.389 mmol) and sodium triacetoxyborohydride (124 mg, 0.583 mmol) in DCE (5 mL) was stirred at RT for 16h. The reaction mixture was filtered and diluted with water (5 mL), then extracted with ethyl acetate (20 mL*3), dried over sodium sulfate, filtered, and concentrated, finally purified by flash chromatography 2-30% ethyl acetate in petroleum ether to afford Ol- tert-butyl O2-methyl (2S)-5-morpholinopiperidine-l,2-dicarboxylate (50 mg, 78.3% yield) as a yellow solid.
[00648] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 329.2(M+1) at 1.1850 min.
[00649] Step 5. Synthesis of methyl (2S)-5-morpholinopiperidine-2-carboxylate.
Figure imgf000154_0002
[00650] To a solution of Ol-tert-butyl O2-methyl (2S)-5-morpholinopiperidine-l, 2- dicarboxylate (579 mg, 1.76 mmol) in 1,4-dioxane (10 mL) was added a solution of HCI/dioxane (4 M, 5 mL, 20 mmol). The reaction was stirred at room temperature for 2h. The reaction was concentrated in vacuo. The residue was washed with EtOAc (20 mL), dried in vacuo to afford the desired product methyl (2S)-5-morpholinopiperidine-2-carboxylate (270 mg, 67.1% yield) as a white solid.
[00651] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: 229.1 [M-100]+ at 1.358 min.
[00652] Step 6. Synthesis of methyl l-[2-(l, 3-dioxoisoindolin-2-yl) ethyl]-4-morpholino- piperidine-2-carboxylate.
Figure imgf000155_0001
[00653] To a cooled (0 °C) solution of 2-(2-hydroxyethyl) isoindoline-1, 3-dione (192 mg, 1.0 mmol) in DCM (10 mL) was added trifluoromethanesulfonic anhydride (334 mg, 1.18 mmol) under argon atmosphere. After 10 min 2,6-lutidine (127 mg, 1.18 mmol) and after another lOmin a solution of methyl 4-morpholinopiperidine-2-carboxylate (270 mg, 1.18 mmol) and TEA (120 mg, 1.18 mmol) in DCM (5 mL) were added. The reaction mixture was stirred at room temperature for 16h. The residue was diluted with water (10 mL), extracted with dichloromethane (20 mL x 3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica column chromatography (dichloromethane: methanol = 10 / 1) to give methyl l-[2-(l, 3-dioxoisoindolin-2-yl) ethyl]-4-morpholino-piperidine-2-carboxylate (250 mg, 49.5% yield) as a yellow oil.
[00654] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 94.33% (214 nm); Mass: found peak 402.2 (M + H) at 0.855 min.
[00655] Step 7. Synthesis of (9aS)-7-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2- a]pyrazin-l-one.
Figure imgf000155_0002
[00656] To a solution of methyl (2S)-l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]-5-morpholino- piperidine-2-carboxylate (250 mg, 0.623 mmol) in methanol (15 mL) was added NELNEh water (70 mg , 1.37 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction mixture was concentrated in vacuo. The residue was dissolved with dichloromethane (30 mL), filtered, and concentrated in vacuo to afford (9aS)-7-morpholino-2,3,4,6,7,8,9,9a- octahydropyrido[l,2-a]pyrazin-l-one (130 mg, crude). The crude was used directly in the next step.
[00657] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89% (214 nm); Mass: found peak 240.1 (M + H) at 1.23 min. [00658] Step 8. Synthesis of 4-[(9aS)-2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-
7-yl]morpholine.
Figure imgf000156_0001
[00659] At 0 °C, to a solution of (9aS)-7-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2- a]pyrazin-l-one (130 mg, 1.6 mmol) in THF (20 mL) was added LiAlLL (5.4 mL, 5.4 mmol) under Argon atmosphere. The reaction was stirred at 60 °C for 3h. The reaction mixture was cooled to 0 °C, then 10 drops of water was added. 5 minutes later, 10 drops of 15% NaOH were added to the mixture slowly, then 30 drops of water were added. The solution was dried over sodium sulfate, filtered, and concentrated to dryness to give 4-[(9aS)-2,3,4,6,7,8,9,9a-octahydro- lH-pyrido[l,2-a]pyrazin-7-yl]morpholine (130 mg, crude) as a yellow solid.
[00660] 'H NMR (400 MHz, CDCh): 5 3.73 (t, J = 4.4 Hz, 4H), 2.85-2.99 (m, 4H), 2.78 (td, J = 12.0, 2.4 Hz, 1H), 2.52-2.57 (m, 4H), 2.29-2.36 (m, 1H), 2.07-2.17 (m, 2H), 1.82-1.92 (m, 2H), 1.59-1.68 (m, 2H), 1.18-1.27 (m, 2H) ppm.
[00661] Step 9. Synthesis of 4-[(7R,9aS)-2-[5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido [l,2-a]pyrazin-7- yljmorp
Figure imgf000156_0002
[00662] Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole (247 mg, 0.67 mmol), 4-(2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin- 7-yl)morpholine (162 mg, 0.72 mmol), tBuXPhos Pd G3 (61.1 mg, 0.076 mmol) and sodium tert-butoxide (222 mg, 2.3 mmol) in 1,4-dioxane (16 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 25-P1 4-[(7R,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido [l,2-a]pyrazin-7-yl]morpholine (36.5 mg, yield 10%) as a yellow solid and product 25-P2 4-[(7S,9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7-yl]morpholine (2.3 mg, yield 0.6%) as a yellow solid.
[00663] 25-P1: LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.62% (214 nm) Mass: found peak 465.9 (M+l) at 1.747 min.
[00664] 25-P1: 1 H NMR (400 MHz, CDCh) 5 7.39 (d, J = 8.8 Hz, 2H), 7.20 (d, J = 4.8 Hz, 2H), 5.63 (s, 1H), 3.64 (s, 4H), 3.54 (d, J = 11.8 Hz, 2H), 3.02 (d, J = 10.2 Hz, 1H), 2.88 (t, J = 11.8 Hz, 1H), 2.77 (d, J = 11.2 Hz, 1H), 2.53 (s, 4H), 2.41 (dt, J = 20.7, 11.1 Hz, 3H), 2.23 (s, 3H), 1.95 (t, J = 10.5 Hz, 3H), 1.72 (s, 1H), 1.33-1.15 (m, 2H) ppm.
[00665] 25-P2: LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 465.9 (M+l) at 1.907 min.
[00666] 25-P2: 1 H NMR (400 MHz, CDCh) 5 7.39 (d, J = 8.9 Hz, 2H), 7.20 (d, J = 5.8 Hz, 2H), 5.62 (s, 1H), 3.69 (s, 4H), 3.45 (dd, J = 21.0, 11.8 Hz, 2H), 2.95 (d, J = 10.6 Hz, 2H), 2.73- 2.58 (m, 2H), 2.52 (s, 4H), 2.26 (d, J = 12.7 Hz, 1H), 2.23 (s, 2H), 2.14 (d, J = 7.9 Hz, 2H), 1.99-1.87 (m, 2H), 1.43-1.28 (m, 3H) ppm.
Example S26. Synthesis of 4-[2-[7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 4,7-diazaspiro[2.5]octan-4-yl]ethyl]morpholine (Compound 26).
[00667] Compound 26 was prepared as outlined below.
Figure imgf000157_0001
[00668] Step 1. Synthesis of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-4,7-diazaspiro [2.5] octane-4-carboxylate.
Figure imgf000157_0002
[00669] Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (300 mg, 0.934 mmol), tert-butyl 4,7-diazaspiro[2.5] octane-4-carboxylate (298 mg, 1.4 mmol), tBuXPhos Pd G3 (74 mg, 0.0934 mmol) and sodium tert-butoxide (269 mg, 2.8 mmol) in 1,4-di oxane (18 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0- 10% EA in PE) to afford the desired product tert-butyl 7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7-diazaspiro[2.5]octane-4-carboxylate (160 mg, yield 34.7%) as a yellow solid.
[00670] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 91.72% (214 nm) Mass: found peak 453.2 (M+l) at 2.146 min.
[00671] Step 2. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro [2.5] octane.
Figure imgf000158_0001
[00672] A mixture of tert-butyl 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro[2.5]octane-4-carboxylate (160 mg, 0.354 mmol) and TFA (1.5 mL, 19.6 mmol) in di chloromethane (10 mL) was stirred at room temperature for 2h. The reaction was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7-diazaspiro[2.5] octane (124 mg, yield 91.6%) as a yellow oil.
[00673] Step 3. Synthesis of 4-[2-[7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-4,7-diazaspiro [2.5] octan-4-yl] ethyl] morpholine (Compound 26).
Figure imgf000158_0002
[00674] To a solution of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro [2.5]octane (50 mg, 0.142 mmol), potassium carbonate (98 mg, 0.71 mmol) and KI (24 mg, 0.142 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (40 mg, 0.213 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/Water/acetonitrile) to afford the desired product 4-[2-[7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro[2.5]octan-4-yl]ethyl]morpholine (13.3 mg, yield 20.1%) as a yellow oil.
[00675] LCMS: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 1.7 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C; LC purity: 100% (214 nm) Mass: found peak 466.3 (M+l) at 1.902 min.
[00676] 'H NMR (400 MHz, CDCh) 5 7.48-7.42 (m, 2H), 7.30-7.24 (m, 2H), 5.69 (s, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.23-3.12 (m, 4H), 3.01 (s, 2H), 2.91 (t, J = 7.2 Hz, 2H), 2.53-2.40 (m, 6H), 2.30 (s, 3H), 0.76-0.69 (m, 2H), 0.61-0.56 (m, 2H) ppm.
Example S27. Synthesis of 4-[2-[7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 4,7-diazaspiro[2.5]octan-4-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 27).
[00677] Compound 27 was prepared as outlined below.
Figure imgf000159_0001
[00678] Step 1. Synthesis of 4-(2-chloroethyl)-7-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]-4,7-diazaspiro [2.5] octane.
Figure imgf000159_0002
[00679] A mixture of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro[2.5] octane (124 mg, 0.352 mmol), 2-chloroacetaldehyde (40% aqueous, 138 mg, 0.704 mmol) and acetic acid (10.6 mg, 0.176 mmol) in methanol (8 mL) was stirred at room temperature for 0.5h. The reaction was cooled to 0 °C, sodium cyanoborohydride (44 mg, 0.704 mmol) was added and stirred at room temperature for 8h. The reaction mixture was diluted with water (20 mL), extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75mL/min, eluting with 0-5% methanol in di chloromethane) to afford the desired product 4-(2-chloroethyl)-7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7- diazaspiro[2.5]octane (110 mg, yield 55.6%) as a yellow oil.
[00680] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.78% (214 nm) Mass: found peak 414.9 (M+l) at 2.055 min.
[00681] Step 2. Synthesis of 4-[2-[7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-4,7-diazaspiro[2.5]octan-4-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 27).
Figure imgf000160_0001
[00682] A mixture of 4-(2-chloroethyl)-7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-4,7-diazaspiro[2.5]octane (110 mg, 0.265 mmol), 1,4-thiazinane 1,1-dioxide (72 mg, 0.53 mmol) and N-ethyl-N-isopropyl-propan-2-amine (103 mg, 0.795 mmol) in 1-methylpyrrolidin- 2-one (3 mL) was treated with microwave reactor and stirred at 160 °C for 2h. The reaction was cooled to room temperature, directly purified by prep-HPLC (ammonium hydrogen carbonate/ water/ acetonitrile) to afford the desired product 4-[2-[7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-4,7-diazaspiro[2.5]octan-4-yl]ethyl]-l,4-thiazinane 1,1- dioxide (16.5 mg, yield 12.1%) as a brown solid.
[00683] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 1.7 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C; LC purity: 100% (214 nm) Mass: found peak 514.2 (M+l) at 1.837 min.
[00684] 'H NMR (400 MHz, CDCh) 5 7.48-7.42 (m, 2H), 7.30-7.25 (m, 2H), 5.69 (s, 1H), 3.25- 3.08 (M, 4H), 3.07-2.95 (m, 10H), 2.93-2.82 (m, 2H), 2.66-2.53 (m, 2H), 2.30 (s, 3H), 0.79-0.55 (m, 4H) ppm.
Example S28. Synthesis of 2,2-difluoro-4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound 28). [00685] Compound 28 was prepared as outlined below.
Figure imgf000161_0001
[00686] Step 1. Synthesis of 2-(dibenzylamino)ethanol.
Figure imgf000161_0002
[00687] To a solution of 2-(benzylamino)ethanol (9.39 mL, 66.1 mmol) in acetonitrile (300 mL) and potassium carbonate (18.3 g, 132 mmol) was added benzyl bromide (11.8 mL, 99.2 mmol). The reaction mixture was stirred at 80 °C for 1.5 h. The mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @100mL/min, eluting with 0-10% methanol in di chloromethane) to afford the desired product 2-(dibenzylamino)ethanol (15 g, yield 94%) as a colorless oil.
[00688] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 242.2 (M+l) at 1.233 min.
[00689] Step 2. Synthesis of 2-[2-(dibenzylamino)ethoxy]-2,2-difluoro-acetic acid.
Figure imgf000161_0003
[00690] At 0 °C, to a solution of 2-(dibenzylamino)ethanol (3.0 g, 12.4 mmol) and sodium chloro-2,2-difluoro-acetic acid (1.9 g, 12.4 mmol) in THF (25 mL) was added sodium hydride (60%, 1.24 g, 31.1 mmol). The reaction mixture was stirred at 75 °C for 16h. The mixture was cooled to room temperature and diluted with water (20 mL). The mixture was extracted with diethyl ether (30 mL*2). The mixture solution was separated and the aqueous layer was adjusted to pH 6 with 6 N HC1. The mixture was extracted with EtOAc (50 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[2- (dibenzylamino)ethoxy]-2,2-difluoro-acetic acid (3.5 g, yield 80.9%) as a yellow solid.
[00691] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.37% (214 nm) Mass: found peak 336.2 (M+l) at 1.383 min.
[00692] Step 3. Synthesis of methyl 2-[2-(dibenzylamino)ethoxy]-2,2-difluoro-acetate.
Figure imgf000162_0001
[00693] To a solution of 2-[2-(dibenzylamino)ethoxy]-2,2-difluoro-acetic acid (0.5 g, 1.49 mmol) in toluene (18 mL) and methanol (4 mL) was added (trimethyl silyl) diazomethane (2 M in hexane, 0.82 mL, 1.64 mmol). The reaction mixture was stirred at room temperature for 15 min and quenched with acetic acid (0.5 mL). The mixture was concentrated in vacuo to afford the desired product methyl 2-[2-(dibenzylamino)ethoxy]-2,2-difluoro-acetate (0.5 g, yield 45.6%) as a yellow oil.
[00694] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 45.57% (214 nm) Mass: found peak 350.2 (M+l) at 1.553 min.
[00695] Step 4. Synthesis of 4-benzyl-2, 2-difluoro-morpholin-3-one.
Figure imgf000162_0002
[00696] Under H2 atmosphere, a mixture of methyl 2-[2-(dibenzylamino)ethoxy]-2,2-difluoro- acetate (3.13 g, 8.96 mmol) and Pd/C (10%, 300 mg) in ethanol (150 mL) was stirred at room temperature for 16h. The reaction was filtered and concentrated in vacuo to afford the desired product 4-benzyl-2, 2-difluoro-morpholin-3-one (1.86 g, yield 87.2%) as a white solid.
[00697] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95.45% (214 nm) Mass: found peak 228.1 (M+l) at 1.635 min.
[00698] Step 5. Synthesis of 4-benzyl-2, 2-difluoro-morpholine.
Figure imgf000163_0001
[00699] To a solution of 4-benzyl-2, 2-difluoro-morpholin-3-one (1.8 g, 7.92 mmol) in THF (80 mL) was added a solution of boron dimethyl sulfide complex (2 M in THF, 20 mL, 40 mmol). The reaction was stirred at 55 °C for 6h, then, room temperature for 16h. The reaction was quenched with water slowly, acidified with HC1 aqueous to pH=l and stirred at room temperature for Ih. The mixture was neutralized with sodium bicarbonate, extracted with EtOAc (100 mL*3). The organic layers were washed with brine (50 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the desired product 4-benzyl-2, 2-difluoro-morpholine (700 mg, yield 40.4%) as a yellow oil.
[00700] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: Sunfire C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95.45% (214 nm) Mass: found peak 228.1 (M+l) at 1.635 min. LC purity: 97.48% (214 nm) Mass: found peak 214.2 (M+l) at 1.752 min.
[00701] Step 6. Synthesis of tert-butyl 2,2-difluoromorpholine-4-carboxylate.
Figure imgf000163_0002
Boc
[00702] Under H2 atmosphere, a mixture of 4-benzyl-2, 2-difluoro-morpholine (0.7 g, 3.28 mmol), BOC2O (0.86 g, 3.94 mmol) and Pd(OH)2/C (20%, 450 mg) in ethyl acetate (60 mL) was stirred at room temperature for 16 h. The reaction was filtered and concentrated under reduced pressure to afford the desired product tert-butyl 2,2-difluoromorpholine-4-carboxylate (700 mg, yield 95.5%) as a colorless oil.
[00703] 'H NMR (400 MHz, CDCI3) 54.05 (t, J = 4.8 Hz, 2H), 3.71 (t, J = 8.0 Hz, 2H), 3.54 (t, J = 4.8 Hz, 2H), 1.48 (s, 9H) ppm.
[00704] Step 7. Synthesis of 2,2-difluoromorpholine TFA salt.
Figure imgf000163_0003
[00705] A mixture of tert-butyl 2,2-difluoromorpholine-4-carboxylate (200 mg, 0.896 mmol) and TFA (2 mL, 26.1 mmol) in dichloromethane (10 mL) was stirred at room temperature for 2 h. The reaction was concentrated in vacuo to afford the desired product 2,2-difluoromorpholine TFA salt (160 mg, yield 75.3%) as a brown solid.
[00706] 'H NMR (400 MHz, MeOD-d4) 3 4.27 - 4.20 (m, 2H), 3.58 (t, J= 7.5 Hz, 2H), 3.35 - 3.29 (m, 2H) ppm.
[00707] Step 8. Synthesis of 2,2-difluoro-4-[2-[4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound 28).
Figure imgf000164_0001
[00708] A mixture of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (80 mg, 0.206 mmol), 2,2-difluoromorpholine 2,2,2-trifluoroacetic acid (98 mg, 0.412 mmol) and N-ethyl-N-isopropyl-propan-2-amine (160 mg, 1.23 mmol) in 1- methylpyrrolidin-2-one (3 mL) was treated with microwave reactor and stirred at 160 °C for 2h. The reaction was cooled to room temperature, directly purified by prep-HPLC (ammonium hydrogen carbonate/water/ acetonitrile) to afford the desired product 2,2-difluoro-4-[2-[4-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (36.1 mg, yield 36.9%) as a yellow solid.
[00709] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C; LC purity: 100% (214 nm) Mass: found peak 476.1 (M+l) at 2.031 min.
[00710] 'H NMR (400 MHz, CDC13) 5 7.49-7.43 (m, 2H), 7.27 (d, J = 8.8 Hz, 2H), 5.70 (s, 1H), 4.08-4.03 (m, 2H), 3.26 (t, J = 4.8 Hz, 4H), 2.82 (t, J = 7.2 Hz, 2H), 2.69-2.55 (m, 10H), 2.30 (s, 3H) ppm.
Example S29. Synthesis of 5-[2-[4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro[3,4-c] pyrrole (Compound 29).
[00711] Compound 29 was prepared as outlined below.
Figure imgf000165_0001
[00712] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-
3-yl] piperazine-l-carboxylate.
Figure imgf000165_0002
[00713] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl) piperazine-l-carboxylate (200 mg, 0.75 mmol) in DCM (20 mL) was added [4-(trifluoromethyl) phenyl] boronic acid (285 mg, 1.5 mmol), Copper (II) acetate (409 mg, 2.25 mmol), pyridine (297 mg, 3.75 mmol) and molecular sieves 4A. The reaction mixture was stirred at room temperature for 48h. The mixture was filtered and purified by flash chromatography (PE/DCM=1/1) to afford desired product tertbutyl 4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazine-l-carboxylate (160 mg, yield 52%) as a yellow oil.
[00714] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.4 min; 95% B for 1.6 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C; LC purity: 96% (214 nm) Mass: found peak 411.2 (M+l) at 2.097 min.
[00715] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000165_0003
[00716] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (160 mg, 0.39 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h, then concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with di chloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (130 mg, yield 95.6%) as a yellow oil.
[00717] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 311.2 (M+l) at 1.391 min.
[00718] Step 3. Synthesis of l-(2-chloroethyl)-4-[5-methyl-l-[4-
(trifluoromethyl)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000166_0001
[00719] Under argon atmosphere, a mixture of l-[5-methyl-l-[4-(trifluoromethyl)phenyl] pyrazol-3-yl]piperazine (140 mg, 0.451 mmol), acetic acid (13.5 mg, 0.226 mmol), 2- chloroacetaldehyde (177 mg, 0.902 mmol) and sodium cyanoborohydride (56 mg, 0.91 mmol) in methanol (6 mL) was stirred at RT for 16h. Then the mixture was filtered and concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with sodium bicarbonate to pH = 8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-(2-chloroethyl)-4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (100 mg, yield 59.5%) as a yellow oil.
[00720] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C.
[00721] Step 4. Synthesis of 5-[2-[4-[5-methyl-l-[4-(trifluoromethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro[3,4-c] pyrrole (Compound 29).
Figure imgf000166_0002
[00722] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), 3,3a,4,5,6,6a-hexahydro-lH-furo[3,4-c]pyrrole (30 mg, 0.268 mmol) and DIPEA (87 mg, 0.671 mmol) in NMP (1.5 mL) stirred at 160 °C for 2h in MW. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 5-[2-[4-[5-methyl-l-[4- (trifluoromethyl) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro[3,4- c] pyrrole (30.2 mg, yield: 50.1%) as a brown solid. [00723] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B:
ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 450.3 (M+l) at 1.830 min.
[00724] 'H NMR (400 MHz, CDCh) 5 7.67 (d, J = 8.5 Hz, 2H), 7.58 (d, J = 8.5 Hz, 2H), 5.73 (s, 1H), 3.76 (s, 2H), 3.59 (d, J = 7.4 Hz, 2H), 3.30- 3.25 (m, 4H), 2.79 (s, 4H), 2.64-2.56 (m, 8H), 2.36 (s, 5H) ppm.
Example S30. Synthesis of 5-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro [3,4-c] pyrrole (Compound 30).
[00725] Compound 30 was prepared as outlined below.
Figure imgf000167_0001
[00726] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] piperazine- 1-carboxylate.
Figure imgf000167_0002
[00727] Under Oxygen atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine- 1-carboxylate (1.0 g, 3.75 mmol), [4-(trifluoromethoxy) phenyl ]b or onic acid (1.55 g, 7.51 mmol), copper (II) acetate (1.36 g, 7.51 mmol) and pyridine (1.2 g, 15 mmol) in di chloromethane (20 mL) was stirred at RT for 16h. Then concentrated and purified by flash chromatography (PE: EA = 3: 1) to afford the desired product tert-butyl 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine- 1-carboxylate (800 mg, 46.8%) as a yellow solid.
[00728] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 2.0mL/min Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99.3% (214 nm), Mass: found peak 427.2 (M+l) at 2.090 min.
[00729] Step 2. Synthesis of l-[5-ethyl-l-[4-(trifluoromethyl) phenyl]pyrazol-3- yl] piperazine.
Figure imgf000168_0001
[00730] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (1.5 g, 3.52 mmol) in DCM (30 mL) was added TFA (8 mL). The reaction mixture was stirred at room temperature for 2h. Then the mixture was concentrated and neutralized with a solution of potassium carbonate to pH=10. The resulting mixture was extracted with DCM and dried over sodium sulfate and filtered. The filtrate was concentrated to afford the desired product l-[5-ethyl-l-[4-(trifluoromethyl) phenyl]pyrazol-3-yl]piperazine (1.1 g, yield: 95.8%) as a yellow oil.
[00731] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 2.0mL/min; Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 50 °C. LC purity: 93% (214 nm), Mass: found peak 327.2 (M+l) at 1.406 min.
[00732] Step 3. Synthesis of l-(2-chloroethyl)-4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000168_0002
[00733] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (1.1 g, 3.37 mmol) in methanol (20 mL) was added sodium cyanoborohydride (424 mg, 6.74 mmol), 2-chloroacetaldehyde (530 mg, 6.74 mmol) and acetic acid (100 mg, 1.7 mmol). The reaction mixture was stirred at room temperature for 16h. The mixture was quenched by water (5 mL) and extracted by dichloromethane (10 mL X 3). The combined organic layer was dried over sodium sulfate and filtered. The filtrate was concentrated to afford the desired product l-(2- chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl] piperazine (1.0 g, 75%) as a colorless oil.
[00734] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 2.0mL/min; Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 50 °C. LC purity: 96% (214 nm) Mass: found peak 389.2 (M+l) at 1.454 min.
[00735] Step 4. Synthesis of 5-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperazin-l-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro [3,4-c] pyrrole (Compound 30).
Figure imgf000169_0001
[00736] To a mixture of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol- 3-yl]piperazine (50 mg, 0.13 mmol) in NMP (3 mL) was added 3,3a,4,5,6,6a-hexahydro-lH- furo[3,4-c]pyrrole hydrochloride (25.5 mg, 0.26 mmol) and N-ethyl-N-isopropyl-propan-2- amine (0.11 mL, 0.64 mmol). The reaction mixture was stirred at 140 °C in microwave reactor for 2h. The mixture was purified by prep-HPLC (ammonium hydrogen carbonate/acetonitrile) to afford the desired product 5-[2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl]-l,3,3a,4,6,6a-hexahydrofuro [3,4-c] pyrrole (26.5 mg, 44.3%) as a yellow solid.
[00737] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 45 °C; LC purity: 100% (214 nm) Mass: found peak 466.1 (M+l) at 1.828 min.
[00738] 'H NMR (400 MHz, CDCh) 5 7.46 (d, J = 8.8 Hz, 2H), 7.28 (d, J = 8.8 Hz, 2H), 5.70 (s, 1H), 3.78 - 3.71 (m, 2H), 3.60 (d, J = 8.4 Hz, 2H), 3.30 - 3.22 (m, 4H), 2.84 (s, 4H), 2.69 - 2.57 (m, 8H), 2.36 (d, J = 5.2 Hz, 2H), 2.30 (s, 3H) ppm.
Example S31. Synthesis of 6-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-2-oxa-6-azaspiro[3.3]heptane (Compound 31).
[00739] Compound 31 was prepared as outlined below.
Figure imgf000169_0002
[00740] To a mixture of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol- 3-yl] piperazine (50 mg, 0.13 mmol) in NMP (3 mL) was added 2-oxa-6-azaspiro[3.3]heptane hydrochloride (35 mg, 0.26 mmol) and N-ethyl-N-isopropyl-propan-2-amine (0.11 mL, 0.64 mmol). The reaction mixture was stirred at 140 °C in microwave reactor for 2h. The mixture was purified by prep-HPLC (ammonium hydrogen carbonate/acetonitrile) to afford the desired product 6-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]-2- oxa-6-azaspiro[3.3]heptane (24.7 mg, 42.5%) as a yellow oil.
[00741] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 45 °C; LC purity: 96% (214 nm); Mass: found peak 452.1 (M+l) at 1.743 min.
[00742] 'H NMR (400 MHz, CDCh) 5 7.46 (d, J = 8.8 Hz, 2H), 7.28 (d, J = 8.8 Hz, 2H), 5.70 (s, 1H), 3.78 - 3.71 (m, 2H), 3.60 (d, J = 8.4 Hz, 2H), 3.30 - 3.22 (m, 4H), 2.84 (s, 4H), 2.69 - 2.57 (m, 8H), 2.36 (d, J = 5.2 Hz, 2H), 2.30 (s, 3H) ppm.
Example S32. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-2-(trifluoromethyl)morpholine (Compound 32).
[00743] Compound 32 was prepared as outlined below.
Figure imgf000170_0001
[00744] To a mixture of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol- 3-yl]piperazine (50 mg, 0.13 mmol) in NMP (3 mL) was added 2-(trifluoromethyl)morpholine hydrochloride(49.3 mg, 0.26 mmol) and N-ethyl-N-isopropyl-propan-2-amine (0.11 mL, 0.64 mmol). The reaction mixture was stirred at 160 °C in microwave reactor for 4h. The mixture was purified by prep-HPLC (ammonium hydrogen carbonate/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]-2- (trifhioromethyl)morpholine (7.9 mg, 12.1%) as a yellow oil.
[00745] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 45 °C; LC purity: 98% (214 nm) Mass: found peak 508.1 (M+l) at 2.039 min.
[00746] 'H NMR (400 MHz, CDCh) 5 7.49 - 7.42 (m, 2H), 7.27 (d, J = 6.0 Hz, 2H), 5.70 (s, 1H), 4.03 - 3.87 (m, 2H), 3.71 (td, J = 11.6, 2.4 Hz, 1H), 3.32 - 3.20 (m, 4H), 2.99 (d, J = 11.2 Hz, 1H), 2.77 (d, J = 11.2 Hz, 1H), 2.66-2.54 (m, 8H), 2.33 - 2.26 (m, 4H), 2.21 (t, J = 10.8 Hz, 1H) ppm.
Example S33. Synthesis of 4-[2-[4-[5-(l-methoxyethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl] piperazin-l-yl]ethyl]morpholine (Compound 33). [00747] Compound 33 was prepared as outlined below.
Figure imgf000171_0001
[00748] Step 1. Synthesis of tert-butyl 4-methoxy-3-oxo-pentanoate.
Figure imgf000171_0002
THF, 0 C-rt, 24h
[00749] To a solution of 2-methoxypropanoic acid (1.05 g, 10.1 mmol) in THF (30 mL) at 0°C was added di(imidazol-l-yl) methanone (1.8 g, 11.1 mmol) and the mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (16.6 mL, 33.3 mmol) was added dropwise to a solution of 3-tert-butoxy-3-oxo-propanoic acid (2.42 g, 15.1 mmol) in THF (30 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (100 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-methoxy-3-oxo-pentanoate (1.84 g, yield: 83.9%) as a yellow oil.
[00750] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18,3.5pm,4.6*50mm; Column Temperature: 45 °C. LC purity: 93% (254 nm) Mass: found peak 147.1 (M - 55)+ at 1.628 min.
[00751] Step 2. Synthesis of tert-butyl 4-(4-methoxy-3-oxo-pentanoyl)piperazine-l- carboxylate.
Figure imgf000171_0003
[00752] A mixture of tert-butyl 4-m ethoxy-3 -oxo-pentanoate (1.9 g, 9.4 mmol) and tert-butyl piperazine- 1 -carboxylate (1.92 g, 10.3 mmol) in toluene (40 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(4-methoxy-3-oxo-pentanoyl)piperazine-l -carboxylate (2.6 g, 78.5% yield) as a yellow oil.
[00753] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X bridge C18,3.5pm,4.6*50mm; Column Temperature:45 °C; LC purity: 92.86% (254 nm) Mass: found peak 315.2 (M -55)+ at 1.452 min.
[00754] Step 3. Synthesis of tert-butyl 4-(6-methyl-3-oxo-heptanethioyl)piperazine-l- carboxylate. Lawesson's reagent
_ (° 5 eq) _ toluene, 75°c 16h
Figure imgf000172_0001
Figure imgf000172_0002
[00755] To a solution of tert-butyl 4-(6-methyl-3-oxo-heptanoyl)piperazine-l -carboxylate (2.6 g, 7.65 mmol) in toluene (40 mL) was added Lawesson's reagent (1.55 g, 3.82 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(6-methyl-3-oxo-heptanethioyl)piperazine- 1-carboxylate (1.7 g, yield 46.2%) as a yellow oil.
[00756] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18,3.5pm,4.6*50mm; Column Temperature:45 °C; LC purity: 71.68% (214 nm) Mass: found peak 343.2 (M + H) at 2.015 min.
[00757] Step 4. Synthesis of tert-butyl 4-[5-(l-methoxyethyl)-lH-pyrazol-3-yl]piperazine- 1-carboxylate. Boc
Figure imgf000172_0003
[00758] To a solution of tert-butyl 4-(4-methoxy-3-oxo-pentanethioyl)piperazine-l -carboxylate (1.2 g, 3.63 mmol) in toluene (50 mL) was added NH2NH2 water (560 mg, 11 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-[5-(l-methoxyethyl)-lH-pyrazol-3-yl]piperazine-l-carboxylate (1g, yield 88.7%) as a white solid. [00759] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.5 min, 95%B for 1.5 min. Flow Rate: 2.0mL/min; Column: Poroshell 120 EC-C18,4.6*50mm, 4pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 311.1 (M+H) + at 1.566 min.
[00760] Step 5. Synthesis of tert-butyl 4-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- y 1] piperazine-l-carboxylate.
Figure imgf000173_0001
[00761] To a solution of tert-butyl 4-[5-(l-methoxyethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (1.0 g, 3.2 mmol) in chloroform (50 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (1.35 g, 6.44 mmol), anhydrous copper acetate (1.17 g, 6.44 mmol), pyridine (1.3 mL, 16.1 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (600 mg, yield 36.8%) as a white solid.
[00762] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.5 min, 95%B for 1.5 min; Flow Rate: 2.0 mL/min;
Column: Poroshell 120 EC-C18, 4.6*50mm, 4pm; Column Temperature: 50 °C. LC purity: 97.76% (214 nm) Mass: found peak 471.0 (M + 1) + at 2.179 min.
[00763] Step 6. Synthesis of l-[5-(l-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] piperazine.
Figure imgf000173_0002
[00764] To a solution of tert-butyl 4-[5-(l-methoxyethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 0.213 mmol) in dichloromethane (5 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-(l-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (78 mg, crude). The crude product was used directly in the next step.
[00765] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2 min at 2.2mL/min; Column: HALO Cl 8, 2.7pm, 4.6*30mm; Column Temperature: 40 °C. LC purity: 86% (214nm); Mass: 409.3 (M+l) at 1.043 min.
[00766] Step 7. Synthesis of 4-[2-[4-[5-(l-methoxyethyl)-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl] piperazin-l-yl]ethyl]morpholine (Compound 33).
Figure imgf000174_0001
[00767] To a solution of l-[5-(l-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (100 mg, 0.27 mmol), potassium carbonate (188 mg, 1.36 mmol) and KI (45 mg, 0.27 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine (80.8 mg, 0.54 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-( 1 - methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl] piperazin- l-yl]ethyl]morpholine (51.7 mg, yield: 39.6%) as a white solid.
[00768] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.09 % (214 nm); Mass: found peak 484.2 (M + H) at 1.837 min.
[00769] ‘H NMR (400 MHz, CDCh) 5 7.52 (d, J = 2.1 Hz, 1H), 7.51 (d, J = 2.2 Hz, 1H), 7.29 (s, 2H), 5.92 (s, 1H), 4.37 (q, J = 6.5 Hz, 1H), 3.76 - 3.68 (m, 4H), 3.29 (s, 4H), 3.24 (s, 3H), 2.68 - 2.46 (m, 12H), 1.45 (d, J = 6.5 Hz, 3H) ppm.
Example S34. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]methyl]morpholine (Compound 34). [00770] Compound 34 was prepared as outlined below.
Figure imgf000175_0001
[00771] Step 1. Synthesis of dimethyl pyridine-2,4-dicarboxylate.
Figure imgf000175_0002
[00772] To a solution of pyridine-2,4-dicarboxylic acid (2.0 g, 12 mmol) in MeOH (50 mL) was added thionyl chloride (4.3 mL, 60 mmol) at 0 °C. Then the mixture was stirred at 60 °C for 16h. The mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the desired product dimethyl pyridine-2,4-dicarboxylate (1.4 g , yield 59.9%) as a yellow solid.
[00773] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 95% (214 nm) Mass: found peak 196.2 (M+l) at 0.947 min.
[00774] Step 2. Synthesis of dimethyl piperidine-2,4-dicarboxylate.
Figure imgf000175_0003
[00775] Under hydrogen, to a solution of dimethyl pyridine-2,4-dicarboxylate (1.4 g, 0.5 mmol) in acetic acid (50 mL) was added platinum dioxide (210 mg, 15%w/w). The reaction mixture was stirred at rt for 16h. The mixture was filtered. The filtrate was concentrated to afford the desired product dimethyl piperidine-2,4-dicarboxylate (1.44 g, yield 99.8%) as a colorless oil. [00776] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 202.1 (M+l) at 0.198 min.
[00777] Step 3. Synthesis of dimethyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,5- dicarboxylate.
Figure imgf000176_0001
[00778] A stirred mixture of DCM (20 mL), N-(2-hydroxyethyl)phthalimide (2.0 g, 10.4 mmol) and 2,6-lutidine (1.62 mL, 13.9 mmol ) was cooled to 0 °C. Maintaining the temperature below 15 °C, trifluoromethanesulfonicanhydride (2.34 mL, 13.9 mmol) was added slowly over Ih. The resulting mixture was stirred at rt for 2h, then washed sequentially with water (10 mL), 2N HC1 (10 mL) and water (10 mL) to yield a solution of 2-(l,3-dioxoisoindolin-2-yl)ethyl trifluoromethanesulfonate. At 20-25 °C, a separate reaction vessel was charged with DCM (10 mL), water and sodium carbonate (3.69 g, 34.8 mmol). After stirring for 15 minutes, dimethyl piperidine-2,5-dicarboxylate (1.4 g, 6.96 mmol) in DCM (10 mL) was added, and the mixture was stirred for 16h. The organic layer was separated and crystallized by DCM to afford dimethyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,5-dicarboxylate (2.6 g , yield 99.8%) as a yellow solid.
[00779] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm) Mass: found peak 375.3 (M+l) at 1.17 min.
[00780] Step 4. Synthesis of methyl l-oxo-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine- 8-carboxylate.
Figure imgf000176_0002
[00781] A solution of dimethyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,4- dicarboxylate (2.6 g, 6.94 mmol) and hydrazine hydrate solution (0.7 mL, 13.9 mmol) in MeOH (20 mL) was stirred for 16h at rt. The organic layer was separated and crystallized by DCM (10 mL). The filtrate was concentrated to afford the desired product methyl l-oxo-2,3,4,6,7,8,9,9a- octahydropyrido[l,2-a]pyrazine-8-carboxylate (1.3 g , yield 88.2%) as a yellow solid. [00782] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; Mass: found peak 213.1 (M+l) at 0.1 min.
[00783] Step 5. Synthesis of 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-8- ylmethanol.
Figure imgf000177_0001
[00784] At 0 °C, to a solution of methyl l-oxo-2,3,4,6,7,8,9,9a-octahydropyrido[l,2- a]pyrazine-8-carboxylate (383 mg, 1.6 mmol) in tetrahydrofuran (20 mL) was added LiAlH4 (18.8 mL, 18.8 mmol) under Ar atomosphere. The reaction was stirred at 60 °C for 3h, then were added 20 drops of water and 20 drops of 15% NaOH to the mixture slowly. Then were added 60 drops of water, the organic phase was separated and dried over sodium sulfate, filtered, and concentrated to dryness to afford 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-8- ylmethanol (800 mg, crude) as a yellow oil.
[00785] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; Mass: found peak 171.3 (M + H) at 0.233 min.
[00786] Step 6. Synthesis of [2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]methanol.
Figure imgf000177_0002
[00787] To a solution of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 0.31 mmol) in dry 1,4-dioxane (3 mL) was added 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2- a]pyrazin-8-ylmethanol (106 mg, 0.62 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy- palladipm;dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (26 mg, 0.01 mmol), and sodium tert-butoxide (90 mg, 0.9 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The filtrate was purified by SGC (DCM:MeOH=20: l) to afford the desired product [2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8- yl]methanol (50 mg, yield 39%) as a colorless oil.
[00788] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 92% (214 nm) Mass: found peak 411.3 (M+l) at 1.13 min.
[00789] Step 7. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-8-carbaldehyde.
Figure imgf000178_0001
[00790] To a solution of [2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]methanol (50 mg, 0.12 mmol) and DIPEA (78 mg, 0.61 mmol) in DCM (5 mL) at 0 °C was added dropwise pyridine sulfur trioxide (96 mg, 0.61 mmol) in DMSO (1 mL). The reaction mixture was stirred at this temperature for 16h. The reaction was diluted with DCM (30 mL), washed with 5% Na2SOs aqueous (20 mL) and 5% sodium bicarbonate aqueous (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Then the crude product was diluted with EtOAc (20 mL), washed with saturated sodium chloride (lOmL x 3) dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a- octahydropyrido[l,2-a]pyrazine-8-carbaldehyde (40 mg, yield 78.1%) as a yellow solid.
[00791] LCMS method: Column: HALO C18 2.7pm 4.6*30mm Mobile phase: water (0.01%TFA) (A) / ACN (0.01%TFA) (B) Elution program: Gradient from 5 to 95% of B in l.Omin at 2.2.mL/min Temperature: 40°C LC purity: 40% (214nm); Mass: 409.3 [M + 1]+ at 1.011 min.
[00792] Step 8. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]methyl]morpholine (Compound 34).
Figure imgf000178_0002
[00793] To a solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-8-carbaldehyde (40 mg, 0.0979 mmol) in methanol (5 mL) was added sodium triacetoxyborohydride (62.3 mg, 0.294 mmol), morpholine (17.1 mg, 0.196 mmol), a drop of acetic acid and 4A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched with water (5 mL) and extracted with dichloromethane (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[[2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8- yl]methyl]morpholine (4.9 mg, yield 10.2%) as a colorless oil.
[00794] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 480.2 (M+l) at 2.121 min.
[00795] 'H NMR (400 MHz, CDCh) 5 7.49 - 7.43 (m, 2H), 7.28 (s, 2H), 5.70 (s, 1H), 3.79 - 3.67 (m, 4H), 3.61 (dd, J = 20.0, 12.4 Hz, 2H), 3.06 - 2.80 (m, 3H), 2.58 (m, 1H), 2.40 (s, 5H), 2.30 (s, 3H), 2.18 (d, J = 7.2 Hz, 2H), 2.14 - 1.99 (m, 2H), 1.77 (t, J = 14.8 Hz, 2H), 1.29 (d, J = 26.4 Hz, 2H).
Example S35. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 35).
[00796] Compound 35 was prepared as outlined below.
Figure imgf000179_0001
[00797] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] piperazine- 1-carboxylate. Boc
Figure imgf000180_0001
[00798] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (150 mg, 0.563 mmol), [4-(trifluoromethyl)phenyl]boronic acid (232 mg, 1.13 mmol) and Cu(OAc)2 (224 mg, 1.13 mmol) in DCM (10 mL) was added dry pyridine (89 mg, 1.13 mmol). The mixture was stirred at rt for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=3/1) to afford the desired product tertbutyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (157 mg, yield 62.8%) as a yellow solid.
[00799] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire,
4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 96.4% (214 nm), Mass: found peak 427.2(M+H) at 1.526 min.
[00800] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoro-methoxy)phenyl] pyrazol-3- yl] piperazine.
Boc
Figure imgf000180_0002
[00801] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (157 mg, 0.368 mmol) and TFA (420 mg, 3.68 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[4-(trifluoro-methoxy)phenyl] pyrazol-3-yl]piperazine (110 mg, yield 61.1%) as a yellow oil.
[00802] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm), Mass: found peak 327.2(M+H) at 1.036 min.
[00803] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 35).
Figure imgf000181_0001
[00804] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (118 mg, 0.362 mmol), potassium carbonate (150 mg, 1.08 mmol), and KI (60 mg, 0.362 mmol) in 95% EtOH (10 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (101 mg, 0.542 mmol). The reaction was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (76.7 mg, yield 48.3%) as a white solid.
[00805] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 100.0% (214 nm), Mass: found peak 440.2(M+H) at 2.006 min.
[00806] 'H NMR (400 MHz, DMSO-d6) 5 7.60 (d, J = 12.0 Hz, 2H), 7.44 (d, J = 8.0 Hz, 2H), 5.87 (s, 1H), 3.55 (t, J = 12.0 Hz, 4H), 3.33 (s, 4H), 3.12 (t, J = 12.0 Hz, 4H), 2.50 (t, J = 4.0 Hz, 4H), 2.42 (t, J = 4.0 Hz, 4H), 2.31 (s, 3H) ppm.
Example S36. Synthesis of 4-[2-[4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperazin-l- yl] ethyl] morpholine (Compound 36).
[00807] Compound 36 was prepared as outlined below.
Figure imgf000182_0001
[00808] Step 1. Synthesis of tert-butyl 4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000182_0002
[00809] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (150 mg, 0.563 mmol), (4-chlorophenyl)boronic acid (176 mg, 1.13 mmol), and Cu(OAc)2 (224 mg, 1.13 mmol) in DCM (10 mL) was added dry pyridine (89 mg, 2.26 mmol). The mixture was stirred at rt for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=3/1) to afford the desired product tert-butyl 4-[l-(4- chlorophenyl)-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate (181 mg, yield 85.3%) as a yellow oil.
[00810] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm), Mass: found peak 377.2(M+H) at 1.497 min.
[00811] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000183_0001
[00812] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (182 mg, 0.443 mmol) and TFA (506 mg, 4.43 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (130 mg, yield 94.5%) as a yellow oil.
[00813] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 50 °C; LC purity: 97.4% (214 nm), Mass: found peak 277.2(M+H) at 0.997 min.
[00814] Step 3. Synthesis of 4-[2-[4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl]morpholine (Compound 36).
Figure imgf000183_0002
[00815] To a solution of l-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperazine (130 mg, 0.470 mmol), potassium carbonate (195 mg, 1.41 mmol), and KI (156 mg, 0.939 mmol) in 95% EtOH (10 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (87 mg, 0.47 mmol). The reaction was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(4- chlorophenyl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (84.6 mg, yield 46.2%) as a white solid.
[00816] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min; Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 390.1(M+H) at 1.928 min.
[00817] 'H NMR (400 MHz, DMSO-d6) 5 7.50 (s, 4H), 5.87 (s, 1H), 3.57 (t, J = 4.4 Hz, 4H),
3.33-3.31 (m, 4H), 3.13 (t, J = 4.4 Hz, 4H), 2.49-2.42 (m, 8H), 2.30 (s, 3H) ppm.
Example S37. Synthesis of 4-[2-[4-(5-methyl-l-phenyl-pyrazol-3-yl)-l- piperidyljethyljmorpholine (Compound 37).
[00818] Compound 37 was prepared as outlined below.
Figure imgf000184_0001
[00819] To a solution of 4-(5-methyl-l-phenyl-pyrazol-3-yl)piperidine (90 mg, 0.366 mmol), potassium carbonate (253 mg, 1.83 mmol) and KI (61 mg, 0.336 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (102 mg, 0.55 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature, filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-(5-methyl-l-phenyl-pyrazol- 3-yl)-l-piperidyl]ethyl]morpholine (34.0 mg, yield 26.2%) as a yellow oil.
[00820] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column
50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 355.2 (M+l) at 1.824 min.
[00821] XH NMR (400 MHz, CD3OD) 5 7.57-7.51 (m, 2H), 7.48-7.43 (m, 3H), 6.16 (s, 1H), 3.72 (t, J = 4.4 Hz, 4H), 3.12-3.06 (m, 2H), 2.75-2.57 (m, 5H), 2.57-2.50 (m, 4H), 2.29 (s, 3H), 2.27-2.19 (m, 2H), 2.01-1.94 (m, 2H), 1.87-1.74 (m, 2H) ppm.
Example S38. Synthesis of 4-[2-[4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]-
1-piperidyl] ethyl] morpholine (Compound 38).
[00822] Compound 38 was prepared as outlined below.
Figure imgf000184_0002
[00823] Step 1. Synthesis of tert-butyl 4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol- 3-yl]piperidine-l-carboxylate.
Figure imgf000185_0001
[00824] To a solution of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (200 mg, 0.582 mmol) in anhydrous dichloromethane (10 mL) was added [4-
(difluorom ethoxy )phenyl]boronic acid (220 mg, 1.16 mmol), anhydrous copper acetate (211 mg, 1.16 mmol), dry pyridine (230 mg, 2.91 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 48 h. The mixture was filtered, and the residue was purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[l-[4- (difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperidine-l-carboxylate (280 mg, yield 82.8%) as a yellow solid.
[00825] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93.27% (214 nm), Mass: found peak 352.1 (M— 55)+ at 2.209 min.
[00826] Step 2. Synthesis of 4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl] piperidine.
Figure imgf000185_0002
[00827] To a solution of tert-butyl 4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl]piperidine-l -carboxylate (190 mg, 0.435 mmol) in dichloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[l-[4- (difluoromethoxy )phenyl]-5-methyl-pyrazol-3-yl]piperidine (150 mg, yield 84.4%) as a yellow oil.
[00828] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 76.38% (214 nm), Mass: found peak 308.1 (M+l) at 1.764 min.
[00829] Step 3. Synthesis of 4-[2-[4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl]-l-piperidyl]ethyl]morpholine (Compound 38).
Figure imgf000186_0001
[00830] To a solution of 4-[l-[4-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperidine (150 mg, 0.367 mmol), potassium carbonate (254 mg, 1.83 mmol) and KI (61 mg, 0.367 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (102 mg, 0.55 mmol). The reaction was stirred at 90 °Cfor 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4- (difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (117 mg, yield 75.8%) as a white solid.
[00831] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 421.2 (M+l) at 1.868 min.
[00832] 'H NMR (400 MHz, DMSO-d6) 5 7.54 (d, J = 8.8 Hz, 2H), 7.30 (s, 2H), 7.27 (t, J = 64.8 Hz, 1H), 6.13 (s, 1H), 3.55 (t, J = 4.4 Hz, 4H), 2.95-2.88 (m, 2H), 2.57-2.52 (m, 1H), 2.42- 2.40 (m, 4H), 2.38 (t, J = 4.4 Hz, 4H), 2.29 (s, 3H), 2.02 (t, J = 11.2 Hz, 2H), 1.87-1.79 (m, 2H), 1.60 (qd, J = 12.4, 3.2 Hz, 2H) ppm.
Example S39. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 1-piperidyl] ethyl] morpholine (Compound 39).
[00833] Compound 39 was prepared as outlined below.
Figure imgf000187_0001
[00834] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine-l-carboxylate.
Figure imgf000187_0002
[00835] To a solution of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (300 mg, 1.13 mmol) in anhydrous di chloromethane (20 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (466 mg, 2.26 mmol), anhydrous copper acetate (821 mg, 4.52 mmol), dry pyridine (358 mg, 4.52 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 48 h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @750mL/min, eluting with 0- 20% EA in petroleum ether) to afford the crude product. Then the product was purified by prep- HPLC (acetonitrile/water/ammonium hydrogen carbonate)) to afford the desired product tertbutyl 4-[5-methyl- 1 -[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (210 mg, yield 43.7%) as a yellow solid.
[00836] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 426.1 (M+l) at 2.324 min.
[00837] Step 2. Synthesis of 4-(5-methyl-l-(4-(trifluoro methoxy)phenyl)-lH-pyrazol-3- yl)piperidine.
Figure imgf000188_0001
[00838] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine-l -carboxylate (210 mg, 0.49 mmol) in dichloromethane (10 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-(5-methyl-l-(4-(trifluoro methoxy)phenyl)-lH-pyrazol-3-yl)piperidine (160 mg, yield 93.3%) as an orange oil.
[00839] LCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93.6% (214 nm), Mass: found peak 326.2 (M+l) at 1.492 min.
[00840] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]ethyl]morpholine (Compound 39).
Figure imgf000188_0002
[00841] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (80 mg, 0.49 mmol), potassium carbonate (340 mg, 2.46 mmol) and KI (81.6 mg, 0.49 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (137 mg, 0.73 mmol). The reaction was stirred at 90 °C overnight. After cooled to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (43.9 mg, yield 20.4 %) as a yellow solid.
[00842] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 439.2 (M+l) at 2.013 min.
[00843] 'H NMR (500 MHz, DMSO-d6) 5 7.64 (d, J = 9.0 Hz, 2H), 7.49 (d, J = 8.5 Hz, 2H), 6.16 (s, 1H), 3.55 (t, J = 4.5 Hz, 4H), 2.93 (s, 2H), 2.49-2.34 (m, 10H), 2.32 (s, 3H), 2.05 (s, 1H), 1.85 (d, J = 12.0 Hz, 2H), 1.66-1.56 (m, 2H) ppm.
Example S40. Synthesis of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-[2- (l-piperidyl)ethyl] piperidine (Compound 40).
[00844] Compound 40 was prepared as outlined below.
Figure imgf000189_0001
[00845] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (100 mg, 0.307 mmol), potassium carbonate (127 mg, 0.922 mmol), and KI (51 mg, 0.307 mmol) in 95% EtOH (10 mL) was added l-(2-chloroethyl)piperidine hydrochloride (84 mg, 0.461 mmol). The reaction was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-[2-(l-piperidyl)ethyl]piperidine (43.4 mg, yield 32.3%) as a white solid.
[00846] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 40 °C; LC purity: 100.0 % (214 nm), Mass: found peak 437.3 (M+l) at 1.009 min.
[00847] 'H NMR (400 MHz, DMSO-d6) 5 7.66-7.63 (m, 2H), 7.49 (d, J = 8.0 Hz, 2H), 6.16 (s, 1H), 3.35 (s, 1H), 2.92 (d, J = 8.0 Hz, 2H), 2.68-2.56 (m, 2H), 2.40 (d, J = 8.0 Hz, 6H), 2.32 (s, 3H), 2.04 (t, J = 8.0 Hz, 2H), 1.84 (d, J = 12.0 Hz, 2H), 1.60 (qd, J = 12.0, 4.0 Hz, 2H), 1.48 (t, J = 8.0 Hz, 2H), 1.37 (qt, J = 12.0, 4.0 Hz, 2H) ppm. Example S41. Synthesis of 4-[2-[4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]-l- piperidyl] ethyl] morpholine (Compound 41).
[00848] Compound 41 was prepared as outlined below.
Figure imgf000190_0001
[00849] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine-l-carboxylate.
Figure imgf000190_0002
[00850] To a solution of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (300 mg, 1.13 mmol) in anhydrous di chloromethane (20 mL) was added (4-chlorophenyl)boronic acid (354 mg, 2.26 mmol), anhydrous copper acetate (821 mg, 4.52 mmol) and dry pyridine (358 mg, 4.52 mmol). The reaction mixture was stirred at room temperature for 16 h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @750mL/min, eluting with 0-20% EA in petroleum ether) to afford the crude product. The crude was further purified by prep-HPLC (acetonitrile/water/ammonium hydrogen carbonate) to afford the desired product tert-butyl 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (210 mg, yield 49.4%) as a white solid.
[00851] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89% (214 nm), Mass: found peak 376.1 (M+l) at 2.300 min.
[00852] Step 2. Synthesis of 4- [l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl] piperidine.
Figure imgf000191_0001
[00853] To a solution of tert-butyl 4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperidine-l- carboxylate (210 mg, 0.49 mmol) in dichloromethane (10 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3- yl]piperidine (120 mg, yield 75.6%) as an orange oil.
[00854] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 97.0% (214 nm), Mass: found peak 275.9 (M+l) at 1.602 min.
[00855] Step 3. Synthesis of 4-[2-[4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]-l- piperidyl] ethyl] morpholine (Compound 41).
Figure imgf000191_0002
[00856] To a solution of 4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperidine (120 mg, 0.43 mmol), potassium carbonate (301 mg, 2.18 mmol) and KI (72.2 mg, 0.43 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (121mg, 0.65 mmol). The reaction was stirred at 90 °C overnight. After cooled to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(4- chlorophenyl)-5-methyl-pyrazol-3-yl]-l-piperidyl]ethyl] morpholine (105.2 mg, yield 62.2 %) as a white solid. [00857] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 389.2 (M+l) at 1.939 min.
[00858] ‘H NMR (400 MHz, DMSO-d6) 5 7.54 (s, 4H), 6.15 (s, 1H), 3.57-3.52 (t, J = 4.8 Hz, 4H), 2.91 (d, J = 12.0 Hz, 2H), 2.52 (s, 1H) 2.41 (s, 4H), 2.39-2.36 (m, 4H), 2.31 (s, 3H), 2.02 (t, J = 12.0 Hz, 1H), 1.83 (d, J = 12.0 Hz, 2H), 1.64-1.53 (m, 2H) ppm.
Example S42. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-l- piperidyl]ethyl]morpholine (Compound 42).
[00859] Compound 42 was prepared as outlined below.
Figure imgf000192_0001
[00860] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine-l-carboxylate.
Figure imgf000192_0002
[00861] To a solution of tert-butyl 4-(3-methyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (300 mg, 1.13 mmol) in anhydrous di chloromethane (20 mL) was added [4- (trifluoromethyl)phenyl]boronic acid (429 mg, 2.26 mmol), anhydrous copper acetate (821 mg, 4.52 mmol) and dry pyridine (358 mg, 4.52 mmol). The reaction mixture was stirred at room temperature for 16 h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @750mL/min, eluting with 0-20% EA in petroleum ether) to afford the crude product. The crude was further purified by prep-HPLC (ACN/water (ammonium hydrogen carbonate)) to afford the desired product tert-butyl 4-[5- m ethyl- l-[4-(trifluorom ethoxy )phenyl]pyrazol-3-yl]piperi dine- 1 -carboxylate (210 mg, yield 45.4%) as a white solid. [00862] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 354.1 (M-55)+ at 2.324 min.
[00863] Step 2. Synthesis of 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperidine.
Figure imgf000193_0001
[00864] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperidine-l -carboxylate (210 mg, 0.51 mmol) in dichloromethane (10 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperidine (120 mg, yield 75.6%) as an orange oil.
[00865] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100.0% (214 nm) Mass: found peak 310.1 (M+l) at 1.817 min.
[00866] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]- 1-piperidyl] ethyl] morpholine (Compound 42).
Figure imgf000193_0002
[00867] To a solution of 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperidine (150 mg, 0.48 mmol), potassium carbonate (335 mg, 2.42 mmol) and KI (80 mg, 0.48 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (135 mg, 0.727 mmol). The reaction was stirred at 90 °C overnight. After cooled to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (117.9 mg, yield 57.5 %) as a white solid.
[00868] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 389.2 (M+l) at 1.939 min.
[00869] 'H NMR (400 MHz, DMSO-d6) 5 7.85 (d, J = 8.8 Hz, 2H), 7.76 (d, J = 8.4 Hz, 2H), 6.21 (s, 1H), 3.55 (t, J = 4.8 Hz, 4H), 2.92 (d, J = 10.4 Hz, 2H), 2.60-2.55 (m, 1H), 2.42-2.35 (m, 11H), 2.03 (t, J = 11.6 Hz, 2H), 1.85 (d, J = 11.2 Hz, 2H), 1.68-1.55 (m, 2H) ppm.
Example S43. Synthesis of 4-[2-[4-[5-tert-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 43).
[00870] Compound 43 was prepared as outlined below.
Figure imgf000194_0001
[00871] Step 1. Synthesis of tert-butyl 4,4-dimethyl-3-oxo-pentanoate.
Figure imgf000195_0001
[00872] To a solution of 2,2-dimethylpropanoic acid (2.04 g, 20 mmol) in THF (50 mL) at 0°C was added CDI (4.39 g, 27.1 mmol) and the reaction mixture was stirred at RT for 3h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (4.8 g, 30 mmol) in THF (50 mL) at 0 °C, and the reaction mixture was stirred at RT for 3h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to RT and stirred overnight. The mixture was quenched by addition of 10 % aqueous citric acid (200 mi) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by flash column chromatography on silica gel (cyclohexane/EtOAc 1/0 to 7/3) to afford tert-butyl 4,4-dimethyl-3-oxo-pentanoate (2.0 g, yield 43%) as a yellow oil.
[00873] 'H NMR (400 MHz, CDCh) 5 3.45 (s, 2H), 1.47 (s, 9H), 1.17 (s, 9H) ppm.
[00874] Step 2. Synthesis of tert-butyl 4-(4,4-dimethyl-3-oxo-pentanoyl)piperazine-l- carboxylate.
Figure imgf000195_0002
[00875] A mixture of tert-butyl 4,4-dimethyl-3-oxo-pentanoate (2.0 g, 10 mmol) and tert-butyl piperazine- 1 -carboxylate (2.05g, 11 mmol) in toluene (50 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-20% acetone in petroleum ether to afford tertbutyl 4-(4,4-dimethyl-3-oxo-pentanoyl)piperazine-l -carboxylate (2.9 g, 89% yield) as a yellow oil.
[00876] LCMS method: LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 95.60% (214 nm) Mass: found peak 257.0 (M-55)+ at 1.64 min. [00877] Step 3. Synthesis of tert-butyl 4-(4,4-dimethyl-3-oxo-pentanethioyl)piperazine-l- carboxylate.
(0.5 eq) Lawesson's reagent toluene, 75°c, 16h
Figure imgf000196_0001
Figure imgf000196_0002
[00878] To a solution of tert-butyl 4-(4,4-dimethyl-3-oxo-pentanoyl)piperazine-l -carboxylate (2.9 g, 9.28 mmol) in toluene (80 mL) was added Lawesson's reagent (1.88 g, 4.64 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(4,4-dimethyl-3-oxo- pentanethioyl)piperazine-l -carboxylate (1.4 g, yield 39.1%) as a yellow oil.
[00879] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 82.17% (214 nm) Mass: found peak 273.2 (M-55)+ at 2.210 min.
[00880] Step 4. Synthesis of tert-butyl 4-(5-tert-butyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000196_0003
[00881] To a solution of tert-butyl 4-(4,4-dimethyl-3-oxo-pentanethioyl)piperazine-l- carboxylate (1.4 g, 4.26 mmol) in toluene (10 mL) was added NH2NH2 water (653 mg, 12.8 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 4-(5-tert-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (820 mg, yield 60.1%) as a white solid.
[00882] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.41% (214 nm) Mass: found peak 309.2 (M+l) at 1.981 min.
[00883] Step 5. Synthesis of tert-butyl 4-[5-tert-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000197_0002
[00884] To a solution of tert-butyl 4-(5-tert-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (700 mg, 2.27 mmol) in chloroform (30 mL) was added [3-(trifluoromethoxy)phenyl]boronic acid (954 mg, 4.54 mmol), anhydrous copper acetate (824 mg, 4.54 mmol), pyridine (900 mg, 11.3 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-tert-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (60 mg, yield 4.54%) as a yellow solid.
[00885] Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 83.75% (214 nm) Mass: found peak 469.3 (M+l) at 2.610 min.
[00886] Step 6. Synthesis of l-[5-tert-butyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperazine.
Figure imgf000197_0001
[00887] To a solution of tert-butyl 4-[5-tert-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (60 mg, 0.128 mmol) in dichloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-tert-butyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]piperazine (40 mg, yield 69.5%) as a yellow oil.
[00888] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 81.88% (214 nm) Mass: found peak 369.3 (M+l) at 1.127 min.
[00889] Step 7. Synthesis of 4-[2-[4-[5-tert-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 43).
Figure imgf000198_0001
[00890] To a solution of l-[5-tert-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (40 mg, 0.11 mmol), potassium carbonate (75 mg, 0.54 mmol) and KI (18 mg, 0.11 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (30 mg, 0.14 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-tert-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (27.9 mg, yield 52.6%) as a yellow oil.
[00891] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.63% (214 nm) Mass: found peak 482.2 (M+l) at 2.135 min.
[00892] 'H NMR (400 MHz, CDCh) 5 7.45-7.39 (m, 2H), 7.27-7.24 (m, 2H), 5.67 (s, 1H), 3.73 (t, J = 4.8 Hz, 4H), 3.26 (t, J = 4.8 Hz, 4H), 2.61 (t, J = 4.8 Hz, 4H)„ 2.58-2.55 (m, 4H), 2.51 (t, J = 4.8 Hz, 4H), 1.15 (s, 9H) ppm.
Example S44. Synthesis of 4-[2-[4-[l-(3, 4-difluorophenyl)-5-methyl-pyrazol-3-yl] piperazin-l-yl] ethyl] morpholine (Compound 44).
[00893] Compound 44 was prepared as outlined below.
Figure imgf000198_0002
[00894] Step 1. Synthesis of tert-butyl 4-[l-(3, 4-difluorophenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000199_0001
[00895] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.2 g, 0.751 mmol) in chloroform (50 mL) was added (3,4-difluorophenyl) boronic acid (119 mg, 0.751 mmol), anhydrous copper acetate (273 mg, 1.5 mmol), pyridine (0.3 mL, 3.75 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether: ethyl acetate = 4 / 1) to afford the desired product tert-butyl 4-[ 1 -(3 , 4-difluorophenyl)-5-methyl- pyrazol-3-yl]piperazine-l -carboxylate (224 mg, yield: 78.8%) as a yellow solid.
[00896] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100 % (214 nm); Mass: found peak 379.3 (M + H) at 1.44 min.
[00897] Step 2. Synthesis of l-[l-(3,4-difluorophenyl)-5-methyl-pyrazol-3-yl] piperazine.
Figure imgf000199_0002
[00898] To a solution of tert-butyl 4-[ 1 -(3, 4-difluorophenyl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (224 mg, 0.592 mmol) in dichloromethane (3 mL) was added TFA (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH = 8, extracted with dichloromethane (10 mL X 3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-(3,4-difluorophenyl)-5-methyl-pyrazol-3- yl]piperazine (165 mg, crude). The crude product was used directly in the next step.
[00899] LCMS method: Mobile Phase: A: water (10 mMammonium hydrogen carbonate) B: ACN; Gradient: 5%-95% B in 1.6min; Flow Rate: 1.6 mL / min; Column: YMC Cl 8 Plus, 4.6*50mm, 3 pm, 12nm Oven Temperature: 50 °C. LC purity: 100 % (214 nm); Mass: found peak 279.1 (M + H) at 1.70 min. [00900] Step 3. Synthesis of 4- [2-[4-[l-(3, 4-difluorophenyl)-5-methyl-pyrazol-3-yl] piperazin-l-yl] ethyljmorpholine (Compound 44).
Figure imgf000200_0001
[00901] To a solution of 1 -[ l-(3, 4-difluorophenyl)-5-methyl-pyrazol-3-yl] piperazine (164 mg, 0.589 mmol), potassium carbonate (407 mg, 2.95 mmol) and KI (97.8 mg, 0.589 mmol) in 95% ethanol / water (20mL /2 mL) was added 4-(2-chloroethyl) morpholine (176 mg, 1.18 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature, filtered, concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(3, 4-difluorophenyl)-5- methyl-pyrazol-3-yl] piperazin-l-yl] ethyl morpholine (179.7 mg, yield: 77.9%) as a white solid.
[00902] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 392.1 (M + H) at 1.86 min.
[00903] 'H NMR (400 MHz, CD3OD): 5 7.36-7.47 (m, 2H), 7.26-7.30 (m, 1H), 5.85 (s, 1H), 3.72 (t, J = 4.8 Hz, 4H), 3.26 (t, J = 4.8 Hz, 4H), 2.67 (t, J = 4.8 Hz, 4H), 2.59-2.64 (m, 4H), 2.55-2.58 (m, 4H), 2.30 (s, 3H) ppm.
Example S45. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 45).
[00904] Compound 45 was prepared as outlined below.
Figure imgf000200_0002
[00905] Step 1. Synthesis of l-(4-bromophenyl)-3-[l-(2-chloroethyl)-4-piperidyl]-5- methoxy-pyrazolo [3, 4-c] pyridine.
Figure imgf000201_0001
[00906] A mixture of l-(4-bromophenyl)-5-methoxy-3-(4-piperidyl)pyrazolo[3,4-c]pyridine (110 mg, 0.284 mmol), 2-chloroacetaldehyde (40% aqueous, 11 mg, 0.568 mmol), sodium cyanoborohydride (35.7 mg, 0.568 mmol) and acetic acid (34 mg, 0.568 mmol) in methanol (10 mL) was stirred at room temperature for 16h. The reaction mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 0-10% methanol in dichloromethane) to afford the desired product 1- (4-bromophenyl)-3-[l-(2-chloroethyl)-4-piperidyl]-5-methoxy-pyrazolo [3, 4-c] pyridine (72 mg, yield: 18.6%) as a yellow oil.
[00907] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 33% (214 nm) Mass: found peak 413.1 (M + 1) at 2.25 min.
[00908] Step 2. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 45).
Figure imgf000201_0002
[00909] A mixture of l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-3-yl]piperazine (72 mg, 0.0575 mmol), 1,4-thiazinane 1,1-dioxide (15.6 mg, 0.115 mmol) and N-ethyl-N-isopropyl-propan-2-amine (37.2 mg, 0.288 mmol) in NMP (2 mL) was treated with microwave reactor and stirred at 160 °C for 2h. The reaction was cooled to room temperature and directly purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-isopropyl-pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (6.6 mg, yield: 22%) as a yellow solid.
[00910] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 512.1 (M + H) at 2.01 min.
[00911] 'H NMR (400 MHz, CD3OD): 5 7.33-7.36 (m, 2H), 7.22 (dd, J = 4.4 , 2.0 Hz, 1H), 5.88 (s, 1H), 3.26 (t, J = 4.4 Hz, 4H), 3.07-3.13 (m, 8H), 2.93-2.98 (m, 1H), 2.77 (t, J = 6.4 Hz, 2H), 2.71 (s, 4H), 2.65 (t, J = 6.4 Hz, 2H), 1.19 (d, J = 6.8 Hz, 6H) ppm.
Example S46. Synthesis of 4-[2-[4-[5-cyclopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-
3-yl] piperazin-l-yl] ethyl] morpholine (Compound 46).
[00912] Compound 46 was prepared as outlined below.
Figure imgf000202_0001
[00913] Step 1. Synthesis of tert-butyl 3-cyclopropyl-3-oxo-propanoate.
Figure imgf000202_0002
[00914] To a solution of cyclopropanecarboxylic acid (5 g, 58.1 mmol) in THF (200 mL) at 0°C was added di (imidazol-l-yl)m ethanone (10.4 g, 63.9 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (96 mL, 192 mmol) was added slowly to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (14 g, 87 mmol) in THF (100 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. This solution was added to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred for 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted with EA (100 mL X 2). The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 3-cyclopropyl-3-oxo-propanoate (9.39 g, yield: 87.8%) as a yellow oil.
[00915] 'H NMR (400 MHz, CDCh): 5 3.48 (s, 2H), 2.02-2.07 (m, 1H), 1.49 (s, 9H), 1.09-1.13 (m, 2H), 0.93-0.98 (m, 2H) ppm.
[00916] Step 2. Synthesis of tert-butyl 4-(3-cyclopropyl-3-oxo-propanoyl) piperazine-1- carboxylate.
Figure imgf000203_0001
[00917] A mixture of tert-butyl 3-cyclopropyl-3-oxo-propanoate (0.5 g, 2.5 mmol) and tertbutyl piperazine- 1 -carboxylate (0.511 g, 2.75 mmol) in toluene (50 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(3-cyclopropyl-3-oxo-propanoyl) piperazine- 1 -carboxylate (0.75 g, 92.3% yield) as a yellow oil.
[00918] LCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X-B RIDGE Cl 8 (4.6x 50 mm, 3.5pm). Temperature: 50°C. LC purity: 92.3% (214 nm) Mass: found peak 319.2 (M + Na) + at 1.88 min.
[00919] Step 3. Synthesis of tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl) piperazine- 1-carboxylate. Lawesson's reagent (0 5 eq) toluene, 75°c, 16h
Figure imgf000203_0002
Figure imgf000203_0003
[00920] To a solution of tert-butyl 4-(3-cyclopropyl-3-oxo-propanoyl)piperazine-l -carboxylate (0.75 g, 2.43 mmol) in toluene (40 mL) was added Lawesson's reagent (0.491 g, 1.21 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl) piperazine- 1 -carboxylate (0.65 g, yield: 46.2%) as a yellow oil. [00921] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 54% (214 nm) Mass: found peak 313.2 (M + H) at 1.22 min.
[00922] Step 4. Synthesis of tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl) piperazine-l- carboxylate.
Figure imgf000204_0001
[00923] To a solution of tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl) piperazine-1- carboxylate (0.65 g, 2.08 mmol) in toluene (50 mL) was added NH2NH2 water (0.3 mL, 6.24 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl) piperazine- 1 -carboxylate (350 mg, yield 48.3%) as a yellow oil.
[00924] LCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X-B RIDGE Cl 8 (4.6x 50 mm, 3.5pm). Temperature: 50°C. LC purity: 84% (214 nm) Mass: found peak 293.3 (M + H) at 1.85 min.
[00925] Step 5. Synthesis of tert-butyl 4-[5-cyclopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperazine-l-carboxylate.
Figure imgf000204_0002
[00926] To a solution of tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, 0.342 mmol) in chloroform (15 mL) was added [4-(trifluoromethoxy) phenyl] boronic acid (144 mg, 0.684 mmol), anhydrous copper acetate (124 mg, 0.684 mmol), pyridine (0.14 mL, 1.73 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-cyclopropyl-l- [4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperazine- 1 -carboxylate (150 mg, yield: 84.3%) as a yellow solid.
[00927] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 87% (214 nm) Mass: found peak 453.3 (M + 1) + at 1.54 min.
[00928] Step 6. Synthesis of l-[5-cyclopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000205_0001
[00929] To a solution of tert-butyl 4-[5-cyclopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperazine- 1 -carboxylate (150 mg, 0.288 mmol) in dichloromethane (3 mL) was added 2, 2, 2-trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5mL X 3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-cyclopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (120 mg, crude). The crude product was used directly in the next step.
[00930] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 82% (214 nm) Mass: found peak 353.3 (M + 1) + at 1.08 min.
[00931] Step 7. Synthesis of 4-[2-[4-[5-cyclopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl]piperazin-l-yl]ethyl]morpholine (Compound 46).
Figure imgf000205_0002
[00932] To a solution of l-[5-cyclopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (110 mg, 0.312 mmol), potassium carbonate (216 mg, 1.56 mmol) and KI (52 mg, 0.312 mmol) in 95% ethanol (20mL) was added 4-(2-chloroethyl)morpholine (93 mg, 0.624 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- cyclopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl] ethyl]morpholine (85.7 mg, yield: 58.9%) as a white solid.
[00933] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.84 % (214 nm); Mass: found peak 466.1 (M + H) at 2.094 min.
[00934] 'H NMR (400 MHz, CD3OD): 5 7.71-7.66 (m, 2H), 7.39 (d, J = 8.4 Hz, 2H), 5.66 (s, 1H), 3.69 (t, J = 4.8 Hz, 4H), 3.23 (t, J = 4.8 Hz, 4H), 2.65 (t, J = 4.8 Hz, 4H), 2.62-2.56 (m, 4H), 2.53 (t, J = 4.8 Hz, 4H), 1.81-1.73 (m, 1H), 1.02-0.96 (m, 2H), 0.78-0.73 (m, 2H) ppm.
Example S47. Synthesis of 4-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]morpholine (4-(2,3,4,6,7,8,9,9a- octahydro-lH-pyrido[l,2-a]pyrazin-8-yl)morpholine (Compound 47).
[00935] Compound 47 was prepared as outlined below.
Figure imgf000206_0001
[00936] Step 1. Synthesis of Ol-tert-butyl O2-methyl 4-morpholinopiperidine-l, 2- dicarboxylate.
Figure imgf000206_0002
[00937] To a solution of Ol-tert-butyl O2-methyl 4-oxopiperidine-l, 2-dicarboxylate (1 g, 3.89 mmol) in 1,2-dichloroethane (50 mL) was added sodium triacetoxyborohydride (1.65 g, 7.77 mmol), morpholine (440 mg, 5.05 mmol), one drop of acetic acid and 4 A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched by water (30 mL) and extracted by dichloromethane (50 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (dichloromethane: methanol = 10 / 1) to give Ol-tert-butyl O2-methyl 4-morpholinopiperidine-l, 2-dicarboxylate (1.069 g, 83.7% yield) as a yellow oil.
[00938] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm); Mass: found peak 329.3 (M + H) at 0.84 min.
[00939] Step 2. Synthesis of methyl 4-morpholinopiperidine-2-carboxylate.
Figure imgf000207_0001
[00940] To a solution of Ol-tert-butyl O2-methyl 4-morpholinopiperidine-l, 2-dicarboxylate (1.069 g, 3.26 mmol) in methanol (20 mL) was added HC1 (4M in dioxane, 8.14 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH = 8, extracted with dichloromethane (20 mL X 3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford methyl 4-morpholinopiperidine-2-carboxylate (615 mg, crude). The crude product was used directly in the next step.
[00941] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 78% (214 nm) Mass: found peak 229.1 (M+l) + at 1.30 min.
[00942] Step 3. Synthesis of l-[2-(l, 3-dioxoisoindolin-2-yl) ethyl]-4-morpholino- piperidine-2-carboxylate.
Figure imgf000207_0002
[00943] To a cooled (0 °C) solution of 2-(2-hydroxyethyl) isoindoline-1, 3-dione (695 mg, 3.04 mmol) in DCM (20 mL) was added trifluoromethanesulfonic anhydride (0.512 mL, 3.04 mmol) under argon atmosphere. After 10 min, 2,6-lutidine (355 pL, 3.04 mmol) and after another lOmin, a solution of methyl 4-morpholinopiperidine-2-carboxylate (695 mg, 3.04 mmol) and TEA (424 pL, 3.04 mmol) in DCM (5 mL) were added. The reaction mixture was stirred at room temperature for 16h. The residue was diluted with water (10 mL), extracted with dichloromethane (20 mL X 3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica column chromatography (dichloromethane: methanol = 10 / 1) to give methyl l-[2-(l, 3-dioxoisoindolin-2-yl) ethyl]-4-morpholino-piperidine-2-carboxylate (680 mg, 52.3% yield) as a yellow oil.
[00944] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 94% (214 nm); Mass: found peak 402.2 (M + H) at 0.85 min.
[00945] Step 4. Synthesis of 8-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-
1-one.
Figure imgf000208_0001
[00946] To a solution of methyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]-4-morpholino-piperidine-
2-carboxylate (580 mg, 1.44 mmol) in methanol (30 mL) was added hydrazine hydrate (0.21 mL, 4.33 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction mixture was concentrated in vacuo. The residue was dissolved with dichloromethane (30 mL), filtered, and concentrated in vacuo to afford 8-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2- a]pyrazin-l-one (328 mg, crude). The crude product was used directly in the next step.
[00947] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89% (214 nm); Mass: found peak 240.1 (M + H) at 1.23 min.
[00948] Step 5. Synthesis of 4-(2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-8- yl)morpholine.
Figure imgf000208_0002
[00949] At 0 °C, to a solution of 8-morpholino-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin- 1-one (383 mg, 1.6 mmol) in tetrahydrofuran (20 mL) was added Li Al FL (16 mL, 16 mmol) under Argon atmosphere. The reaction was stirred at 60 °C for 3h. The reaction mixture was cooled to 0 °C, then 20 drops of water was added. After 5 minutes, 20 drops of 15% NaOH was added to the mixture slowly, then were added 60 drops of water. The solution was dried over sodium sulfate, filtered, and concentrated to dryness to give 4-(2,3,4,6,7,8,9,9a-octahydro-lH- pyrido[l,2-a]pyrazin-8-yl)morpholine (340 mg, crude) as a yellow solid.
[00950] 'H NMR (400 MHz, CDCI3): 5 3.73 (t, J = 4.4 Hz, 4H), 2.85-2.99 (m, 4H), 2.78 (td, J = 12.0, 2.4 Hz, 1H), 2.52-2.57 (m, 5H), 2.29-2.36 (m, 1H), 2.07-2.17 (m, 2H), 1.82-1.92 (m, 2H), 1.59-1.68 (m, 2H), 1.18-1.27 (m, 2H) ppm.
[00951] Step 6. Synthesis of 4-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-8-yl]morpholine (4-(2,3,4,6,7,8,9,9a- octahydro-lH-pyrido[l,2-a]pyrazin-8-yl)morpholine (Compound 47).
Figure imgf000209_0001
C, 16h
[00952] To a solution of 4-(2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-8- yl)morpholine (149 mg, 0.0888 mmol), 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazole (142 mg, 0.442mmol), and [2-(2-aminophenyl)phenyl]-methylsulfonyloxy- palladium;ditert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (35 mg, 0.0442 mmol) in 1,4-dioxane (10 mL) was added sodium tert-butoxide (170 mg, 1.77 mmol). The reaction was stirred at 80 °C in tube for 16h. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[5- methyl- 1 -[4-(trifluorom ethoxy )phenyl]pyrazol-3 -y 1 ] - 1 ,3 ,4,6,7, 8,9,9a-octahydropyrido[ 1 ,2- a]pyrazin-8-yl]morpholine (4-(2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-8- yl)morpholine (149 mg, 0.0888 mmol), 92 mg, yield: 44.7%) as a white solid.
[00953] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 466.1 (M + 1) at 2.00 min.
[00954] 'H NMR (400 MHz, CD3OD): 5 7.55-7.59 (m, 2H), 7.40-7.43 (m, 2H), 5.87 (s, 1H), 3.73 (t, J = 4.4 Hz, 4H), 3.61-3.68 (m, 2H), 3.01 (td, J = 8.00, 3.2 Hz, 1H), 2.88-2.97 (m, 2H), 2.64 (t, J = 4.4 Hz, 4H), 2.56-2.59 (m, 1H), 2.36-2.45 (m, 2H), 2.31 (s, 3H), 2.19-2.25 (m, 2H), 1.95-2.02 (m, 2H), 1.61 (dq, J = 12.0, 4.0 Hz, 1H), 1.28 (q, 4H) ppm.
Example S48. Synthesis of l-[2-(3-methylsulfonylazetidin-l-yl)ethyl]-4-[5-methyl-l-[4- (trifluoromethyl)phenyl] pyrazol-3-yl]piperazine (Compound 48).
[00955] Compound 48 was prepared as outlined below.
Figure imgf000210_0001
[00956] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), K2CO3 (92 mg, 0.671 mmol) and KI (22 mg, 0.134 mmol) in 95% ethanol (2 mL) was added 3-methylsulfonylazetidine; hydrochloride (46 mg, 0.268 mmol). The reaction was stirred at 90 °C for 16 h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by prep-HPLC (NH4HCO3/H2O/MeCN) to afford the desired product l-[2-(3- methylsulfonylazetidin-l-yl)ethyl]-4-[5-methyl-l-[4-(trifluoromethyl)phenyl] pyrazol-3- yl]piperazine (54.1 mg, yield 81.4%) as a yellow oil.
[00957] LCMS method: Mobile Phase: A: water (10 mmol NH4HCO3) B: MeCN; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 ml/min; Column: X-Bridge C18, 50*4.6mm, 3.5um; Column Temperature: 50°C; LC purity: 100% (214 nm) Mass: find peak 472.1 (M+l)+ at 1.696 min.
[00958] ‘H NMR (400 MHz, CDC13) 5 5.73 (s, 1H), 3.92 (s, 1H), 3.68 (t, J = 8.2 Hz, 2H), 3.55 (dd, J = 8.4, 6.8 Hz, 2H), 3.26 (dd, J = 11.3, 6.3 Hz, 4H), 2.89 (s, 3H), 2.68 (t, J = 6.6 Hz, 2H), 2.63 - 2.55 (m, 4H), 2.41 (t, J = 6.5 Hz, 2H), 2.36 (d, J = 2.0 Hz, 3H) ppm.
Example S49. Synthesis of 4-[2-[4-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 49).
[00959] Compound 49 was prepared as outlined below.
Figure imgf000211_0001
[00960] Step 1. Synthesis of tert-butyl 6-methyl-3-oxo-heptanoate.
Figure imgf000211_0002
[00961] To a solution of 4-methylpentanoic acid (1.2 g, 10.3 mmol) in THF (30 mL) at 0°C was added di(imidazol-l-yl)methanone (1.84 g, 11.4 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (17 mL, 34.1 mmol) was added dropwise to a solution of 3-tert-butoxy-3-oxo-propanoic acid (2.48 g, 15.5 mmol) in THF (30 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (100 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 6-methyl-3-oxo-heptanoate (1.9 g, yield: 77.2%) as a yellow oil.
[00962] ‘H NMR (400 MHz, CDCh) 5 3.35 (s, 2H), 2.58 - 2.49 (m, 2H), 1.53 - 1.45 (m, 12H), 0.90 (dd, J = 6.4, 3.2 Hz, 6H) ppm.
[00963] Step 2. Synthesis of tert-butyl 4-(6-methyl-3-oxo-heptanoyl)piperazine-l- carboxylate.
Figure imgf000212_0001
[00964] A mixture of tert-butyl 6-methyl-3-oxo-heptanoate (1.9 g, 8.87 mmol) and tert-butyl piperazine- 1 -carboxylate (1.82 g, 9.75 mmol) in toluene (40 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(6-methyl-3-oxo-heptanoyl)piperazine-l -carboxylate (2.6 g, 80.1% yield) as a yellow oil.
[00965] LCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95% B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm), Mass: found peak: 271.2 (M -55) + at 1.809 min.
[00966] Step 3. Synthesis of tert-butyl 4-(6-methyl-3-oxo-heptanethioyl)piperazine-l- carboxylate.
Figure imgf000212_0002
[00967] To a solution of tert-butyl 4-(6-methyl-3-oxo-heptanoyl)piperazine-l -carboxylate (2.6 g, 7.65 mmol) in toluene (40 mL) was added Lawesson's reagent (1.55 g, 3.82 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(6-methyl-3-oxo-heptanethioyl)piperazine- 1-carboxylate (1.7 g, yield 46.2%) as a yellow oil.
[00968] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate); B: Acetonitrile Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18,3.5pm,4.6*50mm; Column Temperature:45 °C; LC purity: 71.68% (214 nm) Mass: found peak 343.2 (M + H) at 2.015 min.
[00969] Step 4. Synthesis of tert-butyl 4-(5-isopentyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000212_0003
[00970] To a solution of tert tert-butyl 4-(6-methyl-3-oxo-heptanethioyl)piperazine-l- carboxylate (1.7 g, 4.96 mmol) in toluene (40 mL) was added NH2NH2' water (745 mg, 14.9 mmol) and the mixture was heated at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 4-(5-isopentyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.93 g, yield 58%) as a white solid.
[00971] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 323.2 (M+l) at 1.667 min.
[00972] Step 5. Synthesis of tert-butyl 4-[5-isopentyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000213_0001
[00973] To a solution of tert-butyl tert-butyl 4-(5-isopentyl-lH-pyrazol-3-yl)piperazine-l- carboxylate (0.93 g, 0.29 mmol) was added [4-(trifluoromethoxy) phenyl]boronic acid (1.19 g, 0.58 mmol), anhydrous copper acetate (1.57 g, 8.67 mmol), pyridine (1.14 g, 14.5 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-isopentyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (750 mg, yield 53.8%) as a yellow oil.
[00974] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 44.2% (254 nm) Mass: found peak 483.3 (M+l) at 2.300 min.
[00975] Step 6. Synthesis of l-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000214_0001
[00976] To a solution of tert-butyl 4-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.207 mmol) in dichloromethane (6 mL) was added 2,2,2- trifluoroacetic acid (2.0 mL, 26.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude desired product l-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine, which was used directly in the next step.
[00977] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93.8% (214 nm) Mass: found peak 383.2 (M+l) at 1.564 min.
[00978] Step 7. Synthesis of 4-[2-[4-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 49).
Figure imgf000214_0002
[00979] To a solution of l-[5-isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (80 mg, 0.209 mmol), potassium carbonate (116 mg, 0.836 mmol) and KI (35 mg, 0.209 mmol) in 95% ethanol (2 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (78 mg, 0.418 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- isopentyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (69.1 mg, yield 66.7%) as a white solid.
[00980] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 478.1 (M+l) at 2.101 min. [00981] 'H NMR (400 MHz, CDCI3) 5 7.44 (d, J = 2.1 Hz, 1H), 7.43 (d, J = 2.2 Hz, 1H), 7.28 (s, 2H), 5.70 (s, 1H), 3.75 - 3.68 (m, 4H), 3.30 - 3.23 (m, 4H), 2.66 - 2.46 (m, 14H), 1.61 - 1.53 (m, 1H), 1.47 (dd, J = 8.8, 7.2 Hz, 2H), 0.86 (d, J = 6.5 Hz, 6H) ppm.
Example S50. Synthesis of l-[2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]piperidin-4-ol (Compound 50).
[00982] Compound 50 was prepared as outlined below.
Figure imgf000215_0001
[00983] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), potassium carbonate (92 mg, 0.671 mmol) and KI (22 mg, 0.134 mmol) in 95% ethanol (2 mL) was added 3-(trifluoromethoxy)azetidine hydrochloride (48 mg, 0.268 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 1- [2-[4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]piperidin-4-ol (23.5 mg, yield 40.1%) as a yellow oil.
[00984] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm) Mass: found peak 438.2 (M+l) at 1.661 min.
[00985] 'H NMR (400 MHz, CDCI3) 5 7.67 (d, J = 8.6 Hz, 2H), 7.58 (d, J = 8.5 Hz, 2H), 5.74 (d, J = 4.8 Hz, 1H), 3.69 (ddd, J = 17.8, 9.6, 4.9 Hz, 1H), 3.32 - 3.23 (m, 4H), 2.94 - 2.75 (m, 2H), 2.60 (dd, J = 14.0, 9.0 Hz, 8H), 2.36 (s, 3H), 2.22 (t, J = 9.7 Hz, 2H), 1.91 (d, J = 9.8 Hz, 3H), 1.64 - 1.56 (m, 2H) ppm.
Example S51. Synthesis of 4-[2-[4-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl] morpholine (Compound 51).
[00986] Compound 51 was prepared as outlined below.
Figure imgf000215_0002
[00987] Step 1. Synthesis of tert-butyl 4-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000216_0001
[00988] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.2 g, 0.751 mmol) in DCM (20 mL) was added (3, 5 -difluorophenyl) boronic acid (237 mg, 1.5 mmol), anhydrous copper acetate (409 mg, 2.25 mmol), pyridine (0.35 mL, 2.97 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 5-30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[l-(3,5-difhiorophenyl)-5-methyl- pyrazol-3-yl]piperazine-l -carboxylate (120 mg, 40% yield) as a yellow solid.
[00989] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 50% (214nm); Mass: found peak 379.2 (M + H) at 2.065 min.
[00990] Step 2. Synthesis of l-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3-yl] piperazine.
Figure imgf000216_0002
[00991] To a solution of tert-butyl 4-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (120 mg, 0.317 mmol) in dichloromethane (8 mL) was added 2,2,2-trifluoroacetic acid (1.5 mL, 19.6 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[l-(3,5- difluorophenyl)-5-methyl-pyrazol-3-yl]piperazine (110 mg, 87% yield) as a yellow oil.
[00992] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 91.31% (214nm); Mass: found peak 279.1 (M + H) at 1.005 min. [00993] Step 3. Synthesis of 4-[2-[4-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl] morpholine (Compound 51).
Figure imgf000217_0001
[00994] To a solution of l-[l-(3,5-difluorophenyl)-5-methyl-pyrazol-3-yl]piperazine (50 mg, 0.180 mmol), potassium carbonate (99 mg, 0.719 mmol) and KI (29 mg, 0.18 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (67 mg, 0.359 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(3,5- difluorophenyl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (39.4 mg, yield 56%) as a red solid.
[00995] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 392.2 (M+l) at 1.737 min.
[00996] 'H NMR (400 MHz, CDCh) 5 7.04 (dd, J = 8.3, 2.1 Hz, 2H), 6.70 (tt, J = 8.9, 2.3 Hz, 1H), 5.71 (s, 1H), 3.75 - 3.70 (m, 4H), 3.31 - 3.25 (m, 4H), 2.63 (dd, J = 14.0, 9.3 Hz, 8H), 2.52 (s, 4H), 2.37 (s, 3H) ppm.
Example S52. Synthesis of 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]-l-piperidyl]ethyl]morpholine (Compound 52).
[00997] Compound 52 was prepared as outlined below.
Figure imgf000217_0002
[00998] Step 1. Synthesis of tert-butyl 4-[5-isopropenyl-3-[4- (trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l-carboxylate.
Figure imgf000218_0001
[00999] To a solution of tert-butyl 4-(3-bromo-5-isopropenyl-pyrazol-l-yl)piperidine-l- carboxylate (100 mg, 0.270 mmol) and [4-(trifluoromethoxy)phenyl] boronic acid (50 mg, 0.243 mmol) in 1,4-dioxane (6 mL) and water (1.2 mL) was added potassium carbonate aqueous (57 mg, 0.54 mmol) and PdC12(dppf) dichloromethane complex (19.8 mg, 0.027 mmol) under argon atmosphere. The reaction mixture was stirred at room temperature for 16h. The mixture was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0- 5% MeOH in DCM) to afford the desired crude product tert-butyl 4-[5-isopropenyl-3-[4- (trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l -carboxylate (140 mg, yield 90%) as a colorless oil.
[001000JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 27.73% (214 nm), Mass: found peak 452.2 (M+l)+ at 2.261 min.
[001001] Step 2. Synthesis of tert-butyl 4-[5-isopropyl-3-[4- (trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l-carboxylate.
Figure imgf000218_0002
[001002] Tert-butyl 4-[5-isopropenyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l- carboxylate (140 mg, 0.28 mmol) was dissolved in THF (3 mL) and the mixture was hydrogenated with PdO2 (15 mg) at H2 pressure and room temperature for 30min. The reaction mixture was filtered and concentrated in vacuo to afford the desired product tert-butyl 4-[5- isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l-carboxylate (120 mg, yield 93%) as a yellow oil.
[001003JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 30.89% (214 nm), Mass: found peak 454.3 (M+l)+ at 2.448 min. [001004] Step 3. Synthesis of 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl] piperidine.
Figure imgf000219_0001
[001005] To a solution of tert-butyl 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]piperidine-l -carboxylate (50 mg, 0.11 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 26.9 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[5-isopropyl- 3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine (45 mg, yield 92%) as a yellow oil. [001006] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO Cl 8,
2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 71.35 % (214 nm), Mass: found peak 354.3 (M+l) at 1.703 min.
[001007] Step 4. Synthesis of 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]-l-piperidyl]ethyl]morpholine (Compound 52).
Figure imgf000219_0002
[001008] To a solution of 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine (40 mg, about 0.113 mmol), potassium carbonate (63 mg, 0.453 mmol) and KI (19 mg, 0.11 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (32 mg, 0.17 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]-l-piperidyl]ethyl]morpholine (16.1 mg, yield 28.4%) as a yellow oil.
[001009JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93% (214 nm) Mass: found peak 467.2 (M+l) at 2.059 min. [001010] 'H NMR (400 MHz, CDCh) 5 7.83 - 7.74 (m, 2H), 7.20 (d, J = 8.1 Hz, 2H), 6.28 (s, 1H), 4.03 (d, J = 11.0 Hz, 1H), 3.77 - 3.67 (m, 4H), 3.13 (d, J = 10.9 Hz, 2H), 2.98 (dt, J = 13.6, 6.8 Hz, 1H), 2.63 - 2.48 (m, 8H), 2.47 - 2.36 (m, 2H), 2.26 - 2.13 (m, 2H), 1.89 (d, J = 11.8 Hz,
2H), 1.30 (d, J = 6.8 Hz, 6H) ppm.
Example S53. Synthesis of 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 53).
[001011] Compound 53 was prepared as outlined below.
Figure imgf000220_0001
[001012] To a solution of 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine (80 mg, about 0.226 mmol), potassium carbonate (126 mg, 0.91 mmol) and KI (38 mg, 0.22 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)-l,4-thiazinane 1,1- dioxide;hydrochloride (80 mg, 0.35 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]-l- piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (28.3 mg, yield 22.6%) as a yellow oil.
[001013JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm) Mass: found peak 515.0 (M+l) at 1.993 min.
[001014] 'H NMR (400 MHz, CDCh) 5 7.79 (d, J = 8.8 Hz, 2H), 7.20 (d, J = 8.2 Hz, 2H), 6.28 (s, 1H), 4.00 (s, 1H), 3.08 (s, 10H), 3.00 - 2.94 (m, 1H), 2.72 (t, J = 6.6 Hz, 2H), 2.55 (t, J = 6.6 Hz, 2H), 2.41 (d, J = 9.3 Hz, 2H), 2.15 (t, J = 11.1 Hz, 2H), 1.87 (d, J = 13.3 Hz, 2H), 1.30 (d, J = 6.8 Hz, 6H) ppm.
Example S54. Synthesis of 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy] pyrazol-1- yl]-l-piperidyl]ethyl]morpholine (Compound 54).
[001015] Compound 54 was prepared as outlined below.
Figure imgf000221_0001
[001016] Step 1. Synthesis of tert-butyl 4-(3,5-dibromopyrazol-l-yl)piperidine-l- carboxylate.
Figure imgf000221_0002
[001017] A mixture of 3,5-dibromo-lH-pyrazole (500 mg, 2.21 mmol), cesium carbonate (1.44 g, 4.43 mmol) and tert-butyl 4-methylsulfonyloxypiperidine-l -carboxylate (0.93 g, 3.32 mmol) in DMF (15 mL) was stirred at 80 °C for 16h. After complete conversion of the starting material, water (40 mL) and EtOAc (40 mL) were added. The precipitate was filtered and the aqueous layer was extracted with EtOAc (100 mL*2). The organic layers were combined, dried over sodium sulfate, and concentrated in vacuo. The residue was directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 0-20% acetone in petroleum ether) to afford the desired product tert-butyl 4-(3,5-dibromopyrazol-l-yl)piperidine- 1 -carboxylate (900 mg, yield 93%) as a colorless solid.
[001018JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: found peak 353.9 (M-55)+ at 2.109 min.
[001019] Step 2. Synthesis of tert-butyl 4-(3-bromo-5-isopropenyl-pyrazol-l-yl)piperidine-
1-carboxylate.
Figure imgf000221_0003
[001020]Under argon atmosphere, a mixture of tert-butyl 4-(3,5-dibromopyrazol-l- yl)piperidine-l -carboxylate (800 mg, 1.96 mmol), 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2- dioxaborolane (296 mg, 1.76 mmol), l,l'-bis(diphenylphosphino) ferrocene- palladium(II)dichloride dichloromethane complex (143 mg, 0.19 mmol) and sodium carbonate (415 mg, 3.9 mmol) in anhydrous dioxane/water (10/1 mL) was stirred at 80 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : l) to afford the product tert-butyl 4-(3-bromo-5-isopropenyl-pyrazol-l-yl)piperidine-l-carboxylate (400 mg, 45%) as a white solid.
[001021JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 57% (214 nm), Mass: found peak 314.0 (M-55)+ at 2.118 min.
[001022] Step 3. Synthesis of tert-butyl 4-(3-bromo-5-isopropyl-pyrazol-l-yl)piperidine-l- carboxylate.
Figure imgf000222_0001
[001023] Tert-butyl 4-(3-bromo-5-isopropenyl-pyrazol-l-yl)piperidine-l-carboxylate (600 mg, 1.42 mmol) was dissolved in THF (3 mL) and the mixture was hydrogenated with PdO2 (15 mg) at H2 pressure and room temperature for 30 min. The reaction mixture was filtered and concentrated in vacuo to afford the desired product tert-butyl 4-(3-bromo-5-isopropyl-pyrazol-l- yl)piperidine- 1-carboxylate (300 mg, yield 53%) as a yellow oil.
[001024JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 95% (214 nm), Mass: found peak 317.8 (M-55)+ at 2.018 min.
[001025] Step 4. Synthesis of tert-butyl 4-[5-isopropyl-3-[4-
(tr ifluoromethoxy)phenoxy] pyrazol- 1-yl] piperidine- 1-carboxylate.
Figure imgf000222_0002
[001026]Under argon atmosphere, a mixture of tert-butyl 4-(3-bromo-5-isopropyl-pyrazol-l- yl)piperidine- 1-carboxylate (250 mg, 0.672 mmol), 4-(trifluoromethoxy) phenol (120 mg, 0.672 mmol), copper(I) iodide (12.8 mg, 0.067mmol), 2-(dimethylamino)acetic acid (20.8 mg, 0.201 mmol) and potassium carbonate (186 mg, 1.34 mmol) in dry DMSO (10 mL) was stirred at 120 °C in tube for 16h. The reaction mixture was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine-l -carboxylate (150 mg, yield 44.7%) as a yellow solid.
[001027] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 18.96% (214nm); Mass: 414.1 [M - 55]+ at 2.286 min.
[001028] Step 5. Synthesis of 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl] piperidine.
Figure imgf000223_0001
[001029] To a solution of tert-butyl 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine-l -carboxylate (150 mg, 0.319 mmol) in dichloromethane (8 mL) was added 2,2,2- trifluoroacetic acid (1.0 mL, 13.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude desired product 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine (130 mg, 90% yield) as a yellow oil.
[001030] LCMS method: Mobile Phase: 3.5pm; Column A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm Temperature: 50 °C; LC purity: 57.36% (214 nm); Mass: 370.1 [M + 1]+ at 2.120 min.
[001031] Step 6. Synthesis of 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy] pyrazol- l-yl]-l-piperidyl]ethyl]morpholine (Compound 54).
Figure imgf000223_0002
[001032] To a solution of 4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine (60 mg, 0.162 mmol), potassium carbonate (90 mg, 0.65 mmol) and KI (27 mg, 0.162 mmol) in 95% ethanol (2 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (45.3 mg, 0.244 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-isopropyl-3-[4-(trifluoromethoxy)phenoxy] pyrazol-l-yl]-l- piperidyl]ethyl]morpholine (26.8 mg, yield 33.2%) as a white solid.
[001033JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97% (214 nm) Mass: found peak 483.1 (M+l) at 2.036 min.
[001034] 'H NMR (400 MHz, CDCh) 5 7.14 (s, 4H), 5.54 (s, 1H), 4.04 (s, 0.5H), 3.92 (t, J = 11.8 Hz, 1H), 3.75 - 3.67 (m, 5.5H), 3.07 (d, J = 11.9 Hz, 2H), 3.00 - 2.89 (m, 1H), 2.52 (dd, J = 10.1, 4.2 Hz, 6H), 2.30 (d, J = 8.8 Hz, 2H), 2.12 (t, J = 12.2 Hz, 2H), 1.81 (d, J = 10.8 Hz, 2H), 1.25 (d, J = 6.8 Hz, 6H) ppm.
Example S55. Synthesis of 4-[2-[4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-l-yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 55).
[001035] Compound 55 was prepared as outlined below.
Figure imgf000224_0001
[001036] Step 1. Synthesis of tert-butyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5- isopropenyl-pyrazol-l-yl]piperidine-l-carboxylate.
Figure imgf000224_0002
[001037]Under argon atmosphere, a mixture of tert-butyl 4-(3-bromo-5-isopropenyl-pyrazol-l- yl)piperidine-l -carboxylate (200 mg, 0.54 mmol), (2,2-difluoro-l,3-benzodioxol-5-yl)boronic acid (98 mg, 0.49 mmol), l,l'-bis(diphenylphosphino) ferrocene-palladium(II)dichloride dichloromethane complex (39.5 mg, 0.05 mmol) and sodium carbonate (114 mg, 1.2 mmol) in anhydrous dioxane/water (10/1 mL) was stirred at 80 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the product tertbutyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropenyl-pyrazol-l-yl]piperidine-l- carboxylate (150 mg, 60%) as a yellow solid.
[001038JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 43.5% (214 nm), Mass: found peak 392.2 (M-55)+ at 2.454 min.
[001039] Step 2. tert-butyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl-pyrazol-l- yl]piperidine-l-carboxylate.
Figure imgf000225_0001
[001040] Tert-butyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropenyl-pyrazol-l-yl] piperidine- 1 -carboxylate (150 mg, 0.29 mmol) was dissolved in THF (3 mL) and the mixture was hydrogenated with PdO2 (20 mg) at H2 pressure and room temperature for 30min. The reaction mixture was filtered and concentrated in vacuo to afford the desired product tert-butyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl-pyrazol-l-yl]piperidine-l-carboxylate (120 mg, yield 85%) as a yellow oil.
[001041JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 80% (214 nm), Mass: found peak 450.1 (M+l)+ at 2.407 min.
[001042] Step 3. Synthesis of 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl-pyrazol-l- yl] piperidine.
Figure imgf000225_0002
[001043] To a solution of tert-butyl 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-l-yl]piperidine-l -carboxylate (120 mg, 0.267 mmol) in dichloromethane (8 mL) was added 2,2,2-trifluoroacetic acid (1.5 mL, 19.6 mmol). The reaction mixture was stirred at room temperature. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5- isopropyl-pyrazol-l-yl]piperidine (110 mg, 99% yield) as a yellow oil.
[001044JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 80% (214 nm); Mass: found peak 350.1 (M + H) at 1.597 min.
[001045] Step 4. Synthesis of 4-[2-[4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl- pyrazol-l-yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 55).
Figure imgf000226_0001
[001046] To a solution of 4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl-pyrazol-l- yl]piperidine (60 mg, 0.172 mmol), potassium carbonate (95 mg, 0.687 mmol) and KI (28.5 mg, 0.172 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)-l,4-thiazinane 1,1- dioxide;hydrochloride (60.3 mg, 0.258 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[3-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-isopropyl-pyrazol-l- yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (10.3 mg, yield 11.7%) as a yellow oil.
[001047] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 511.1 (M+l) at 1.935 min.
[001048] XH NMR (400 MHz, CDCh) 5 7.53 (d, J = 1.4 Hz, 1H), 7.47 (dd, J = 8.3, 1.6 Hz, 1H), 7.03 (d, J = 8.3 Hz, 1H), 6.24 (s, 1H), 4.04 - 3.93 (m, 1H), 3.08 (s, 10H), 3.01 - 2.94 (m, 1H), 2.72 (t, J = 6.7 Hz, 2H), 2.55 (t, J = 6.6 Hz, 2H), 2.39 (tt, J = 12.6, 6.3 Hz, 2H), 2.15 (t, J = 11.7 Hz, 2H), 1.87 (d, J = 12.2 Hz, 2H), 1.30 (d, J = 6.8 Hz, 6H) ppm.
Example S56. 4-[2-[4-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l- yl] ethyl] morpholine (Compound 56).
[001049] Compound 56 was prepared as outlined below.
Figure imgf000227_0001
[001050] Step 1. Synthesis of tert-butyl 4-methyl-3-oxohexanoate.
Figure imgf000227_0002
[001051] To a solution of 2-methylbutanoic acid (2 g, 20 mmol) in THF (200 mL) at 0°C was added di(imidazol-l-yl)methanone (3.56 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-tert-butoxy-3-oxo-propanoic acid (4.8 g, 30 mmol) in THF (40 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-methyl-3 -oxohexanoate (3.1 g, yield: 78.3%) as a yellow oil.
[001052] ‘H NMR (400 MHz, CDCh) 5 3.41 (s, 2H), 2.37 - 2.35 (m, 1H), 1.68 - 1.66 (m, 2H), 1.42 (s, 9H), 1.04 - 1.03 (m, 3H), 0.95 - 0.93 (m, 3H).
[001053] Step 2. Synthesis of tert-butyl 4-(4-methyl-3-oxo-hexanoyl) piperazine-1- carboxylate.
Figure imgf000228_0001
[001054] A mixture of tert-butyl 4-methyl-3 -oxo-hexanoate (1.5 g, 7.6 mmol) and tert-butyl piperazine- 1 -carboxylate (1.55 g, 8.32 mmol) in toluene (75 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(4-methyl-3-oxo-hexanoyl) piperazine- 1 -carboxylate (1.4 g, 55.4% yield) as a yellow oil.
[001055] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (254 nm) Mass: found peak 335 (M + Na) + at 1.66 min.
[001056] Step 3. Synthesis of tert-butyl 4-(4-methyl-3-oxo-hexanethioyl)piperazine-l- carboxylate.
Lawesson's reagent
Figure imgf000228_0002
toluene, 75°c, 16h
Figure imgf000228_0003
Figure imgf000228_0004
[001057] To a solution of tert-butyl 4-(4-methyl-3-oxo-hexanoyl) piperazine- 1 -carboxylate (0.75 g, 2.43 mmol) in toluene (40 mL) was added Lawesson's reagent (0.491 g, 1.21 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(4-methyl-3-oxo-hexanethioyl)piperazine- 1-carboxylate (0.65 g, yield 46.2%) as a yellow oil.
[001058JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 56% (214 nm) Mass: found peak 351.1 (M + Na) + at 1.81 min.
[001059] Step 4. Synthesis of tert-butyl 4-(5-sec-butyl-lH-pyrazol-3-yl) piperazine-1- carboxylate.
Figure imgf000228_0005
c, 16h
[001060] To a solution of tert-butyl 4-(4-methyl-3-oxo-hexanethioyl) piperazine- 1 -carboxylate (0.65 g, 2.08 mmol) in toluene (50 mL) was added NELNEU water (0.3 mL, 6.24 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-sec-butyl-lH-pyrazol-3-yl) piperazine- 1 -carboxylate (540 mg, yield 75.1%) as a yellow oil.
[001061JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99% (254 nm) Mass: found peak 309.2 (M + H) at 1.48 min.
[001062] Step 5. Synthesis of tert-butyl 4-[5-sec-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000229_0001
[001063] To a solution of tert-butyl 4-(5-sec-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, 0.342 mmol) in chloroform (15 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (144 mg, 0.684 mmol), anhydrous copper acetate (124 mg, 0.684 mmol), pyridine (0.14 mL, 1.73 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-sec-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (150 mg, yield 84.3%) as a yellow solid.
[001064JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 95% (254 nm) Mass: found peak 469.2 (M + 1)+ at 2.14 min.
[001065] Step 6. Synthesis of l-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000230_0001
[001066] To a solution of tert-butyl 4-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.186 mmol) in dichloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-sec-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (70 mg, crude). The crude product was used directely in the next step.
[001067] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97% (254 nm) Mass: found peak 369.1 (M + H) at 1.55min.
[001068] Step 7. Synthesis of 4-[2-[4-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 56).
Figure imgf000230_0002
[001069] To a solution of l-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (80 mg, 0.218 mmol), potassium carbonate (151 mg, 1.09 mmol) and KI (36.2 mg, 0.218 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (39.2 mg, 0.262 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature, filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-sec-butyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (29.8 mg, yield: 28.5%) as a white solid.
[001070JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 482.4 (M + H) at 1.95 min.
[001071] 'H NMR (400 MHz, CDCh) 5 7.42 - 7.40 (m, 2H), 7.29 - 7.27 (m, 2H), 5.69 (s, 1H),
3.73 - 3.71 (m, 4H), 3.29 - 3.26 (m, 4H), 2.73 - 2.72 (m, 1H), 2.71 - 2.70 (m, 4H), 2.38 - 2.34 (m, 4H), 2.55 - 2.52 (m, 4H), 1.59 - 1.50 (m, 2H), 1.20 - 1.18 (m, 3H), 0.82 - 0.79 (m, 3H).
Example S57. Synthesis of 4-[2-[4-[5-(2-methoxyethyl)-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound 57).
[001072] Compound 57 was prepared as outlined below.
Figure imgf000231_0001
[001073] Step 1. Synthesis of tert-butyl 5-methoxy-3-oxo-pentanoate.
Figure imgf000231_0002
THF, 0°C-rt, 24h
[001074] To a solution of 3-methoxypropanoic acid (2 g, 20 mmol) in THF (200 mL) at 0°C was added di(imidazol-l-yl)methanone (3.56 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-tert-butoxy-3-oxo-propanoic acid (4.8 g, 30 mmol) in THF (40 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 5-methoxy-3-oxo-pentanoate (3.1 g, yield: 78.3%) as a yellow oil.
[001075] 'H NMR (400 MHz, CDCh) 5 3.65 - 3.60 (m, 2H), 3.41 (s, 2H), 3.23 (s, 3H), 3.12 - 3.06 (m, 2H), 1.42 (s, 9H).
[001076] Step 2. Synthesis of tert-butyl 4-(5-methoxy-3-oxo-pentanoyl)piperazine-l- carboxylate.
Figure imgf000232_0001
[001077] A mixture of tert-butyl 5 -m ethoxy-3 -oxo-pentanoate (1.5 g, 7.6 mmol) and tert-butyl piperazine- 1 -carboxylate (1.55 g, 8.32 mmol) in toluene (75 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(5-methoxy-3-oxo-pentanoyl)piperazine-l-carboxylate (1.4 g, 55.4% yield) as a yellow oil.
[001078JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97% (254 nm) Mass: found peak 315.1 (M + H) at 1.48 min.
[001079] Step 3. Synthesis of tert-butyl 4-(5-methoxy-3-oxo-pentanethioyl) piperazine-1- carboxylate.
Figure imgf000232_0002
[001080] To a solution of tert-butyl 4-(5-methoxy-3-oxopentanoyl)piperazine-l-carboxylate (0.75 g, 2.43 mmol) in toluene (40 mL) was added Lawesson's reagent (0.491 g, 1.21 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(5-methoxy-3-oxo-pentanethioyl) piperazine- 1 -carboxylate (0.65 g, yield 46.2%) as a yellow oil. [001081JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97% (254 nm) Mass: found peak 331.2 (M + H) at 1 ,66min.
[001082] Step 4. Synthesis of tert-butyl 4-(5-sec-butyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000233_0001
[001083] To a solution of tert-butyl 4-(5-methoxy-3-oxopentanethioyl)piperazine-l-carboxylate (0.65 g, 2.08 mmol) in toluene (50 mL) was added NFENFU water (0.3 mL, 6.24 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-sec-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (540 mg, yield 75.1%) as a yellow oil.
[001084JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97% (254 nm) Mass: found peak 311.2 (M + H) at 1 ,36min.
[001085] Step 5. Synthesis of tert-butyl 4-[5-(2-methoxyethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000233_0002
[001086] To a solution of tert-butyl 4-[5-(2-methoxyethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (100 mg, 0.342 mmol) in chloroform (15 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (144 mg, 0.684 mmol), anhydrous copper acetate (124 mg, 0.684 mmol), pyridine (0.14 mL, 1.73 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10- 50% dichloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-(2- methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (150 mg, yield 84.3%) as a yellow solid.
[001087] LCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C. Mass: found peak 471.1 (M + H) at 1.85 min.
[001088] Step 6. Synthesis of l-[5-(2-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl] piperazine.
Figure imgf000234_0001
[001089] To a solution of tert-butyl 4-[5-(2-methoxyethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 0.186 mmol) in dichloromethane (3 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-(2-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (120 mg, crude). The crude product was used directly in the next step.
[001090JLCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C. Mass: found peak 371.1 (M + H) at 1.64 min.
[001091] Step 7. Synthesis of 4-[2-[4-[5-(2-methoxyethyl)-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound 57).
Figure imgf000234_0002
[001092] To a solution of l-[5-(2-methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (80 mg, 0.218 mmol), potassium carbonate (151 mg, 1.09 mmol) and KI (36.2 mg, 0.218 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (39.2 mg, 0.262 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-(2- methoxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (29.8 mg, yield: 28.5%) as a yellow oil.
[001093JLCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C. Mass: found peak 484.2 (M + H) at 1.63 min.
[001094] 'H NMR (400 MHz, CDCI3) 5 7.48 - 7.46 (m, 2H), 7.28 - 7.26 (m, 2H), 5.78 (s, 1H), 3.73 - 3.71 (m, 4H), 3.60 - 3.56 (m, 2H), 3.33 (s, 3H), 3.29 - 3.26 (m, 4H), 2.63 - 2.61 (m, 4H), 2.58 - 2.56 (m, 4H), 2.52 - 2.50 (m, 4H).
Example S58. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 58).
[001095] Compound 58 was prepared as outlined below.
Figure imgf000235_0001
[001096] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000235_0002
[001097] A solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (110 mg, 0.374 mmol), [4-(difluoromethyl)phenyl]boronic acid (128 mg, 0.747 mmol), pyridine (59 mg, 0.747 mmol), and copper(II)acetate (136 mg, 0.747 mmol) in dichloromethane (20 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered and the filter cake washed with di chloromethane (10 mL), the solution concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (150 mg, 95.5%) as a white solid.
[001098JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Flow Rate: 1.6mL/min, Column: Proshell 120 EC-C18 30*3mm, 2.7pm, Oven Temperature: 50 °C; LC purity: 100.00 % (214 nm), Mass: found peak 421.0 (M+l) at 1.880 min.
[001099] Step 2. Synthesis of l-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3- yl] piperazine.
Figure imgf000236_0001
[001100] To a solution of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (140 mg, 0.33 mmol) in 2,2,2-trifluoroacetic acid (1 mL) was added DCM (3 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-[4-(difhioromethyl)phenyl]-5- isopropyl-pyrazol-3-yl]piperazine (70 mg, yield 65.6 %) as a yellow oil.
[001101JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: ACN (0.01%TFA); Flow Rate: 1.6mL/min; Column: Proshell 120 EC-C18 30*3mm, 2.7pm; Oven Temperature: 50 °C; LC purity: 100.00 % (214 nm), Mass: found peak 421.0 (M+l) at 1.305 min.
[001102] Step 3. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 58).
Figure imgf000236_0002
[001103] To a solution of l-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (70 mg, 0.21 mmol), potassium carbonate (116 mg, 0.85 mmol) and KI (35 mg, 0.21 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (77 mg, 0.42 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NFLHCCh/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (35 mg, yield 35.1 %) as a white solid. [001104JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate); B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100.00 % (214 nm) Mass: found peak 434.0 (M+l) at 1.604 min.
[001105] 'H NMR (400 MHz, MeOD-d4) 5 7.68 (d, J = 8.3 Hz, 2H), 7.53 (d, J = 8.3 Hz, 2H), 6.84 (t, J = 56.1 Hz, 1H), 5.89 (s, 1H), 3.75 - 3.62 (m, 4H), 3.27 - 3.16 (m, 4H), 3.07 - 2.89 (m, 1H), 2.72 - 2.43 (m, 12H), 1.18 (d, J = 6.8 Hz, 6H).
Example S59. Synthesis of 4-[2-[4-[l-[6-(difluoromethoxy)-3-pyridyl]-5-isopropyl-pyrazol-
3-yl] piperazin-l-yl]ethyl] morpholine (Compound 59).
[001106] Compound 59 was prepared as outlined below.
Figure imgf000237_0001
[001107] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000237_0002
[001108] A solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, 0.340 mmol), [4-(difluoromethyl)phenyl]boronic acid (128 mg, 0.679 mmol), pyridine (54 mg, 0.679 mmol), and copper(II) acetate (123 mg, 0.679 mmol) in dichloromethane (20 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered, the filter cake washed with dichloromethane (10 mL), the solution concentrated, and the mixture was purified by flash chromatography (Biotage, 40 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (140 mg, 92.3%) as a yellow oil. [001109JLCMS method: Mobile Phase: A: Water (0.01%TFA); B: Acetonitrile (0.01%TFA);
Flow Rate: 1.6mL/min; Column: Proshell 120 EC-C18 30*3mm, 2.7pm; Oven Temperature: 50 °C; LC purity: 100.00 % (214 nm), Mass: found peak438.7 (M+l)+ at 1.958 min.
[001110] Step 2. Synthesis of l-[l-[6-(difluoromethoxy)-3-pyridyl]-5-isopropyl-pyrazol-3- yl] piperazine.
Figure imgf000238_0001
[OOllllJTo a solution of tert-butyl 4-[l-[6-(difluoromethoxy)-3-pyridyl]-5-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (140 mg, 0.316 mmol) in 2,2,2-trifluoroacetic acid (1 mL) was added DCM (4 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with di chloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-[6-(difhioromethoxy)-3- pyridyl]-5-isopropyl-pyrazol-3-yl]piperazine (110 mg, yield 94 %) as a yellow oil. [001112JLCMS method: Mobile Phase: A: Water (0.01%TFA); B: Acetonitrile (0.01%TFA); Flow Rate: 1.6mL/min; Column: Proshell 120 EC-C18 30*3mm, 2.7pm; Oven Temperature: 50 °C; LC purity: 100.00 % (214 nm), Mass: found peak 338.7 (M+l) at 1.308 min.
[001113] Step 3. Synthesis of 4-[2-[4-[l-[6-(difluoromethoxy)-3-pyridyl]-5-isopropyl- pyrazol-3-yl] piperazin-l-yl]ethyl] morpholine (Compound 59).
Figure imgf000238_0002
[001114] To a solution of l-[l-[6-(difluoromethoxy)-3-pyridyl]-5-isopropyl-pyrazol-3- yl]piperazine (100 mg, 0.27 mmol), potassium carbonate (149 mg, 1.08 mmol) and KI (45 mg, 0.027mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (100 mg, 0.54 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NHqHCO water/acetonitrile) to afford the desired product 4-[2-[4-[l-[6-(difhioromethoxy)-3- pyridyl]-5-isopropyl-pyrazol-3-yl] piperazin- l-yl]ethyl] morpholine (35 mg, yield 27.9 %) as yellow solid.
[001115] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate); B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100.00 % (214 nm) Mass: found peak 451.7 (M+l) at 1.633 min.
[001116] 'H NMR (400 MHz, MeOD-d4) 5 8.27 (d, J = 2.6 Hz, 1H), 7.91 (dd, J = 8.7, 2.7 Hz, 1H), 7.59 (t, J = 72.7 Hz, 1H), 7.11 (d, J = 8.7 Hz, 1H), 5.90 (s, 1H), 3.74 - 3.60 (m, 4H), 3.27- 3.20 (m, 4H), 2.89 (dt, J = 13.6, 6.8 Hz, 1H), 2.66 - 2.50 (m, 12H), 1.18 (d, J = 6.8 Hz, 6H)
Example S60. Synthesis of 4-[2-[4-[5-isopropyl-l-[6-(trifluoromethoxy)-3-pyridyl]pyrazol- 3-yl]piperazin-l-yl]ethyl]-l,4-oxazepane (Compound 60).
[001117] Compound 60 was prepared as outlined below.
Figure imgf000239_0001
[001118] Step 1. Synthesis of tert-butyl 4-[5-isopropyl-l-[6-(trifluoromethoxy)-3- pyridyl] pyrazol-3-yl] piperazine- 1-car boxylate.
Figure imgf000239_0002
[001119JA solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (70 mg, 0.238 mmol), [6-(trifluoromethoxy)-3-pyridyl]boronic acid (98.4 mg, 0.476 mmol), pyridine (37.6 mg, 0.476 mmol), copper(II)acetate (108 mg, 0.22 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2. LCMS showed the reaction was complete. The mixture was filtered, and the filter cake washed with dichloromethane (10 mL). The solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-isopropyl-l-[6-(trifluoromethoxy)-3-pyridyl]pyrazol-3- yl]piperazine-l -carboxylate (55 mg, 50.9%) as a white solid.
[001120JLCMS method: Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B) Gradient: 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm); Mass: found peak 455.0 (M+l) at 1.80 min. [001121] Step 2. Synthesis of l-[5-isopropyl-l-[6-(trifluoromethoxy)-3-pyridyl]pyrazol-3- yl] piperazine.
Figure imgf000240_0001
[001122] To a solution of tert-butyl 4-[5-isopropyl-l-[6-(trifluoromethoxy)-3-pyridyl]pyrazol-3- yl]piperazine-l -carboxylate (53 mg, 0.121 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-isopropyl-l-[6- (trifluoromethoxy)-3-pyridyl]pyrazol-3-yl]piperazine (40 mg, yield 93.2%) as a yellow oil. [001123JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 355.0 (M+l) at 1.41 min.
[001124] Step 3. Synthesis of l-(2-chloroethyl)-4-[5-isopropyl-l-[6-(trifluoromethoxy)-3- pyridyl] pyrazol-3-yl] piperazine.
Figure imgf000240_0002
[001125]Under argon atmosphere, a mixture of l-[5-isopropyl-l-[6-(trifluoromethoxy)-3- pyridyl]pyrazol-3-yl]piperazine (40 mg, 0.113 mmol), 2-chloroacetaldehyde (44.2 mg, 0.225 mmol), and sodium cyanoborohydride(14.1 mg, 0.225 mmol) in methanol (6 mL) was stirred at rt for 16h. The mixture was quenched by sodium bicarbonate and concentrated in vacuo to remove most of the solvent. The mixture was diluted with DCM (5 mL) and 3 mL water. The product was extracted with DCM (5 x 3 mL), and the organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @75 mL/min, eluting with 0-10% methanol in dichloromethane) to afford the desired product l-(2-chloroethyl)-4-[5-isopropyl-l-[6- (trifluoromethoxy)-3-pyridyl]pyrazol-3-yl] piperazine (30 mg, yield: 63.8%) as a yellow oil. [001126] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 88% (254 nm); Mass: found peak 417(M+1)+ at 1.38 min. [001127] Step 4. Synthesis of 4-[2-[4-[5-isopropyl-l-[6-(trifluoromethoxy)-3- pyridyl]pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-oxazepane (Compound 60).
Figure imgf000241_0001
[001128] A mixture of l-(2-chloroethyl)-4-[5-isopropyl-l-[6-(trifluoromethoxy)-3- pyridyl]pyrazol-3-yl]piperazine (50 mg, 0.0126mmol), 1,4-oxazepane (30 mg, 0.144 mmol) and N-ethyl-N-isopropyl-propan-2-amine (46.4 mg, 0.359 mmol) in NMP (2 mL) was treated with microwave reactor and stirred at 140 °C for 2h. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[5-isopropyl-l-[6-(trifhioromethoxy)-3-pyridyl]pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4- oxazepane (6.1 mg, yield: 17.6%) as a brown solid.
[001129JLCMS method: Mobile Phase: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate); B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100.0 % (214 nm); Mass: found peak 483.3 (M + H) at 1.85 min.
[001130] 'H NMR (400 MHz, CDCh) 5 8.38 (d, J = 2.6 Hz, 1H), 7.89 (dd, J = 8.7, 2.7 Hz, 1H), 7.10 (d, J = 8.7 Hz, 1H), 5.75 (s, 1H), 3.78 (dt, J = 9.0, 5.3 Hz, 4H), 3.33 - 3.21 (m, 4H), 2.96 - 2.89 (m, 1H), 2.87 - 2.73 (m, 6H), 2.66 - 2.58 (m, 6H), 1.97 - 1.90 (m, 2H), 1.21 (d, J = 6.8 Hz, 6H) ppm.
Example S61. Synthesis of 4-[2-[4-[5-methyl-l-[6-(trifluoromethyl)-2-naphthyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 61).
[001131] Compound 61 was prepared as outlined below.
Figure imgf000241_0002
[001132] Step 1. Synthesis of 6-bromonaphthalene-2-boronic acid
Figure imgf000242_0001
[001133] 5.0 g of 2,6-dibromonaphthalene was placed into a 500-mL three-neck flask, and the air in the flask was replaced with nitrogen. To this compound was added 200 mL of THF, and this solution was stirred at -78°C for 20 minutes. Then, 12 mL of a 1.7M hexane solution of n- butyllithium (n-BuLi) was dripped into this mixture solution, followed by stirring at -78°C for 2 h. After the predetermined time had elapsed, 3.5 mL of trimethyl borate was added to the mixture and this solution was stirred for 18 h while the temperature was raised to room temperature. After the predetermined time had elapsed, 80 mL of 1.0M hydrochloric acid was poured into this solution, and the mixture was stirred for 1 h. Then, the mixture solution was separated into an organic layer and an aqueous layer. Organic substances were extracted with ethyl acetate from the obtained aqueous layer. The solution of the extract was combined with the organic layer that had been first obtained, and the mixture was washed with saturated brine and dried by addition of anhydrous magnesium sulfate. Then, the filtrate obtained by gravity filtration was concentrated to give a white solid. The obtained white solid was washed with ethanol, whereby 6-bromonaphthalene-2-boronic acid was obtained with powder (yield 35%). [001134JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C Mass: No peak at LCMS.
[001135] Step 2. Synthesis of tert-butyl 4-[l-(6-bromo-2-naphthyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000242_0002
[001136] A solution of tert-butyl 4-[l-(6-bromo-2-naphthyl)-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (3.18 g, 11.9 mmol), (6-bromo-2-naphthyl)boronic acid (1.5 g, 5.98), pyridine (2.36 g, 29.9 mmol), copper(II)acetate (3.26 g, 17.9 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2. LCMS showed the reaction was complete. The mixture was filtered, and the filter cake was washed with dichloromethane (10 mL). The solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[ 1 -(6-bromo-2-naphthyl)-5-methyl-pyrazol-3-yl]piperazine- 1 -carboxylate (350 mg, 12.4%) as a white oil. [001137] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 97.83% (214nm); LC purity: 74.92% (254 nm) Mass: found peak 471.2 (M+l) at 2.458 min.
[001138] Step 3. Synthesis of tert-butyl 4-[5-methyl-l-[6-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-2-naphthyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000243_0001
[001139] A solution of tert-butyl 4-[l-(6-bromo-2-naphthyl)-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (350 mg, 0.74 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan2yl)l,3,2dioxaborolane (283mg, 1.11 mmol), [l,rbis(diphenylphosphino)ferrocene]dichloropalladium(II) (350mg, 0.0478mmol), and acetoxypotassium (219 mg, 2.23 mmol) in 1,4-di oxane (3 mL) was stirred at 100 °C for 16h. LCMS showed the reaction was complete. The mixture was filtered. The solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-methyl-l-[6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-2- naphthyl]pyrazol-3-yl]piperazine-l -carboxylate (350 mg, 90.9%) as a white solid.
[001140JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 54.82% (214 nm) Mass: found peak 519.4 (M+l) at 2.550 min.
[001141] Step 4. Synthesis of tert-butyl 4-[5-methyl-l-[6-(trifluoromethyl)-2- naphthyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000243_0002
[001142] A 10 mL Schlenk tube containing a stirring bar was charged with tert-butyl 4-[5- methyl-l-[6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-2-naphthyl]pyrazol-3-yl]piperazine- 1-carboxylate (350mg, 0.675 mmol), 3,3-dimethyl-l-(trifhioromethyl)-llambda3,2- benziodoxole (223mg, 0.397 mmol), thiophene-2-carbonyloxycopper (12.9mg, 0.068 mmol), 1,10-phenanthroline (12.2mg, 0.068 mmol), and LiOH (32.3mg., 1.35 mmol). The tube was evacuated and back-filled with argon three times. Anhydrous DCM (10 mL) was added to the reaction mixture. The tube was sealed by a plastic screw cap. The reaction mixture was heated at 45 °C for 6 h. The reaction mixture was cooled to room temperature. LCMS showed the reaction was complete. The solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-methyl-l-[6-(trifluoromethyl)-2- naphthyl]pyrazol-3-yl]piperazine-l -carboxylate (183 mg, 58.9%) as a white oil.
[001143JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min at 2.0 mL/min; Column: SUNFIRE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 30.4% (254 nm) Mass: found peak 461.3 (M+l) at 2.383 min.
[001144] Step 5. Synthesis of l-[5-isopropyl-l-[6-(trifluoromethoxy)-3-pyridyl]pyrazol-3- yl] piperazine.
Figure imgf000244_0001
[001145] To a solution of tert-butyl 4-[5-methyl-l-[6-(trifhioromethyl)-2-naphthyl]pyrazol-3- yl]piperazine-l -carboxylate (183 mg, 0.397 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-isopropyl-l-[6- (trifluoromethoxy)-3-pyridyl]pyrazol-3-yl]piperazine (123 mg, yield 83.8%) as a yellow oil. [001146] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min at 2.0 mL/min; Column: SUNFIRE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 36.77% (214 nm); Mass: found peak 361.3 (M+l) at 1.419 min.
[001147] Step 6. Synthesis of 4-[2-[4-[5-methyl-l-[6-(trifluoromethyl)-2-naphthyl]pyrazol-
3-yl]piperazin-l-yl]ethyl]morpholine (Compound 61).
Figure imgf000244_0002
[001148] To a solution of l-[5-methyl-l-[6-(trifluoromethyl)-2-naphthyl]pyrazol-3-yl]piperazine (120 mg, 0.333 mmol), potassium carbonate (184 mg, 0.333 mmol), and KI (5.53 mg, 0.033 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (35.8 mg, 0.076 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NHqHCO water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[6- (trifluoromethyl)-2-naphthyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (28.7 mg, yield 22.7%) as a white solid.
[001149JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.32 % (214 nm) Mass: found peak 474.3 (M+l) at 1.88 min.
[001150] 'H NMR (400 MHz, CDCh) 5 8.15 (s, 1H), 8.01 - 7.92 (m, 2H), 7.90 (s, 1H), 7.78 (dd, J = 8.8, 2.1 Hz, 1H), 7.67 (dd, J = 8.6, 1.6 Hz, 1H), 5.76 (d, J = 5.0 Hz, 1H), 3.72 (dd, J = 11.4, 6.8 Hz, 4H), 3.38 - 3.22 (m, 4H), 2.66 - 2.48 (m, 12H), 2.43 - 2.36 (m, 3H) ppm.
Example S62. Synthesis of 4-[[4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]methyl]morpholine (Compound 62). [001151] Compound 62 was prepared as outlined below.
Figure imgf000245_0001
[001152] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-
3-yl] piperazine- 1-carboxylate.
Figure imgf000245_0002
[001153] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (II5601-3, 120 mg, 0.45 mmol) in dichloromethane (10 mL) was added [3- (trifluoromethoxy)phenyl]boronic acid (189 mg, 0.901 mmol), anhydrous copper acetate (164 mg, 0.901 mmol), pyridine (0.18 mL, 2.25 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (110 mg, yield 57.3%) as a yellow solid.
[001154JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min at 2.0 mL/min; Column: SUNFIRE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 427.2 (M+l) at 2.316 min.
[001155] 'H NMR (400 MHz, CDCh) 5 7.44 (t, J = 8.0 Hz, 1H), 7.39 (dt, J = 8.0, 1.6 Hz, 1H), 7.34 (s, 1H), 7.13 (d, J = 7.6 Hz, 1H), 5.72 (s, 1H), 3.55 (t, J = 5.2 Hz, 4H), 3.21 (t, J = 5.2 Hz, 4H), 2.34 (s, 3H), 1.48 (s, 9H) ppm.
[001156] Step 2. Synthesis of l-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000246_0001
[001157] To a solution of tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (110 mg, 0.258 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l- [3-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (84 mg, yield 97%) as a yellow oil. [001158JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 97.65% (214 nm), Mass: found peak 327.2 (M+l)+ at 1.040 min.
[001159] Step 3. Synthesis of 4-[[4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]methyl]morpholine (Compound
Figure imgf000246_0003
Figure imgf000246_0002
[001160] To a solution of l-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (84 mg, 0.257 mmol), potassium carbonate (142 mg, 1.03 mmol) and KI (43 mg, 0.257 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (72 mg, 0.386 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[[4-[5- methyl-1 -[3 -(trifluorom ethoxy )phenyl]pyrazol-3-yl]piperazin-l-yl]methyl]morpholine (86.5 mg, yield 76.5%) as a white solid.
[001161JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 440.1 (M+l) at 2.025 min.
[001162] 'H NMR (400 MHz, DMSO-d6) 5 7.61-7.52 (m, 2H), 7.49 (s, 1H), 7.31-7.26 (m, 1H), 5.90 (s, 1H), 3.55 (t, J = 4.4 Hz, 4H), 3.12 (t, J = 4.4 Hz, 4H), 2.53-2.46 (m, 4H), 2.46-2.42 (m, 4H), 2.38 (t, J = 4.4 Hz, 4H, 4H), 2.34 (s, 3H) ppm.
Example S63. Synthesis of 4-[2-[4-[l-(3-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl]morpholine (Compound 63).
[001163] Compound 63 was prepared as outlined below.
Figure imgf000247_0001
[001164] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-
3-yl] piperazine- 1-carboxylate.
Figure imgf000247_0002
[001165] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (120 mg, 0.45 mmol) in dichloromethane (10 mL) was added (3-methoxyphenyl)boronic acid (140 mg, 0.901 mmol), anhydrous copper acetate (164 mg, 0.901 mmol), pyridine (0.18 mL, 2.25 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (115 mg, yield 66.5%) as a yellow solid.
[001166] LCMS method: Mobile Phase: Water (0.01%TFA) (A) / Acetonitrile (0.01%TFA) (B); Gradient: 5 % - 95 % B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 97.38% (214 nm) Mass: found peak 373.3 (M+l) at 1.403 min.
[001167] Step 2. Synthesis of l-[l-(3-methoxyphenyl)-5-methyl-pyrazol-3-yl] piperazine.
Figure imgf000248_0001
[001168] To a solution of tert-butyl 4-[l-(3-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (115 mg, 0.309 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[l-(3-methoxyphenyl)-5- methyl-pyrazol-3-yl]piperazine (84 mg, yield 92%) as a yellow oil.
[001169JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 92.14% (214 nm), Mass: found peak 273.3 (M+l)+ at 0.951 min.
[001170] Step 3. Synthesis of 4-[2-[4-[l-(3-methoxyphenyl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 63).
Figure imgf000248_0002
[001171] To a solution of l-[l-(3-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazine (84 mg, 0.308 mmol), potassium carbonate (171 mg, 1.23 mmol) and KI (51 mg, 0.308 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (86 mg, 0.463 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(3- methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (72.1 mg, yield 60.6%) as a white solid.
[001172JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1. 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 386.2 (M+l) at 1.817 min. [001173] 'H NMR (400 MHz, DMSO-d6) 5 7.35 (t, J = 8.0 Hz, 1H), 7.05-6.99 (m, 2H), 6.90- 6.85 (m, 1H), 5.83 (s, 1H), 3.79 (s, 3H), 3.55 (t, J = 4.4 Hz, 4H), 3.10 (t, J = 4.4 Hz, 4H), 2.53- 2.46 (m, 4H), 2.46-2.41 (m, 4H), 2.38 (t, J = 4.4 Hz, 4H), 2.29 (s, 3H) ppm.
Example S64. Synthesis of cis-4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclobutoxy] ethyl] morpholine (Compound 64).
[001174] Compound 64 was prepared as outlined below.
Figure imgf000249_0001
[001175] Step 1. Synthesis of 5-methyl-l-[4-(trifluoromethoxy)phenyl]-3-vinyl-pyrazole.
Figure imgf000249_0002
[001176]Under Ar, a mixture of 3-iodo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (0.63 g, 1.71 mmol), 4,4,5,5-tetramethyl-2-vinyl-l,3,2-dioxaborolane (0.395 g, 2.57 mmol), Pd(dppf)C12-DCM (139 mg, 0.171 mmol) and potassium carbonate (0.71 g, 5.13 mmol) in dioxane/water (10mL/2 mL) was stirred at 80 °C for 16h. The reaction was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100 mL/min, eluting with 0-10% ethyl acetate in petroleum ether) to afford the desired product 5-methyl-l-[4-(trifluoromethoxy)phenyl]-3-vinyl-pyrazole (393 mg, yield 82.2%) as a colorless oil.
[001177] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96% (214 nm) Mass: found peak 269.2 (M+l) at 1.405 min.
[001178] Step 2. Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclobutanone.
Figure imgf000250_0001
[001179] To a solution of N,N-dimethylacetamide (0.223 mL, 2.4 mmol) in 1,2-dichloroethane (16 mL) under argon atmosphere, trifluoromethyl sulfonyl trifluoromethanesulfonate (0.471 mL, 2.8 mmol) was added dropwise around -15 °C. The mixture was stirred at -15 °C for 10 min, then a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]-3-vinyl-pyrazole (215 mg, 0.8 mmol) and 2,4,6-trimethylpyridine (0.423 mL, 3.2 mmol) in 1,2-dichloroethane (4 mL) was added dropwise. After completion of the addition, the cooling bath was removed and the mixture was warmed to room temperature. The reaction was allowed to proceed for another 15h at 90 °C. The reaction mixture was cooled to room temperature and water (10 mL) was added. The mixture was stirred at 100 °C for 2h. After cooling to room temperature, the mixture was neutralized with sat. sodium bicarbonate aqueous, extracted with DCM (50 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 0-25% ethyl acetate in petroleum ether) to afford the desired product 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]cyclobutanone (48 mg, yield 19.3%) as a yellow solid.
[001180JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 311.2 (M+l) at 1.331 min. [00118111 H NMR (400 MHz, CDCI3) 5 7.51-7.46 (m, 2H), 7.32 (d, J = 8.4 Hz, 2H), 6.13 (s, 1H), 3.73-3.64 (m, 1H), 3.52-3.34 (m, 4H), 2.35 (s, 3H) ppm.
[001182] Step 3. Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclobutanol.
Figure imgf000251_0001
[001183] To a solution of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]cyclobutanone (95 mg, 0.306 mmol) in tetrahydrofuran (20 mL) and ethanol (3 mL) was added NaBH4 (23.2 mg, 0.612 mmol). The reaction was stirred at room temperature for 2h. The reaction was directly purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 12-100% ethyl acetate in petroleum ether) to afford the desired product 3- [5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclobutanol (79 mg, yield 82.6%) as a yellow oil.
[001184JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 313.3 (M+l)+ at 1.238 min.
[001185] Step 4. Synthesis of cis-4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] cyclobutoxy] ethyl] morpholine (Compound 64).
Figure imgf000251_0002
[001186] At 0 °C, to a solution of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]cyclobutanol (79 mg, 0.253 mmol) in dry DMF (10 mL) was added sodium hydride (60%, 202 mg, 5.06 mmol). The reaction was warmed to room temperature and stirred for Ih, then, 60 °C for Ih. The reaction was cooled to room temperature and 4-(2-chloroethyl)morpholine hydrochloride (94.1 mg, 0.506 mmol) was added and stirred at 80 °C for 16h. The reaction was cooled to room temperature, quenched with water, and filtered. The filtrate was purified by prep- HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product cis-4-[2- [3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclobutoxy]ethyl]morpholine (19.1 mg, yield 17.5%) as a yellow oil.
[001187] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.74% (214 nm) Mass: found peak 426.1 (M+l) at 2.073 min.
[001188] ‘H NMR (400 MHz, CDCh) 5 7.51-7.44 (m, 2H), 7.29 (d, J = 8.4 Hz, 2H), 6.13 (s, 1H), 3.99-3.90 (m, 1H), 3.74 (t, J = 4.4 Hz, 4H), 3.53 (t, J = 6.0 Hz, 2H), 3.10-2.98 (m, 1H), 2.73-2.64 (m, 2H), 2.60 (t, J = 6.0 Hz, 2H), 2.52(t, J = 4.4 Hz, 4H), 2.32 (s, 3H), 2.18-2.07 (m, 2H) ppm.
Example S65. Synthesis of l-[l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4- piperidyl]-4-tetrahydropyran-4-yl-piperazine (Compound 65).
[001189] Compound 65 was prepared as outlined below.
Figure imgf000252_0001
[001190] Step 1. Synthesis of l,4-dioxa-8-azaspiro[4.5]decane.
Figure imgf000252_0002
[001191] A mixture of l,4-dioxa-8-azaspiro[4.5]decane (3.0 g, 21 mmol) and tert-butyl acetoacetate (3.65 g, 23 mmol) in toluene (80 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product l,4-dioxa-8-azaspiro[4.5]decane (4.7 g, yield 98.7%) as a yellow solid.
[001192JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 228.3 (M+l)+ at 0.794 min.
[001193] Step 2. Synthesis of 4-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-thioxo-butan-2-one.
Lawesson's reagent (0.5 eq)
Figure imgf000253_0001
toluene, 75°c, 16h
Figure imgf000253_0002
[001194] To a solution of l-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)butane-l, 3-dione (4.7 g, 20.7 mmol) in toluene (100 mL) was added Lawesson's reagent (4.18 g, 10.3 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether) to afford the desired product 4-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-thioxo-butan-2-one (3.0 g, yield 45.2%) as a brown oil.
[001195] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm4.6*30mm; Column Temperature: 40 °C; LC purity: 75.87% (214 nm), Mass: found peak 244.3 (M+l)+ at 0.968 min.
[001196] Step 3. Synthesis of 8-(5-methyl-lH-pyrazol-3-yl)-l,4-dioxa-8-azaspiro[4.5]decane.
Figure imgf000253_0003
[001197] To a solution of 4-(l,4-dioxa-8-azaspiro[4.5]decan-8-yl)-4-thioxo-butan-2-one (2.3 g, 9.45 mmol) in toluene (100 mL) was added hydrazine monohydrate (1.4 mL, 28.4 mmol) and the mixture was stirred at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-6% MeOH in DCM) to afford the desired product 8- (5-methyl-lH-pyrazol-3-yl)-l,4-dioxa-8-azaspiro[4.5]decane (0.83 g, yield 38.2%) as a yellow solid.
[001198JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.22% (214 nm), Mass: found peak 224.1 (M+l)+ at 1.584 min.
[001199] 'H NMR (400 MHz, CDCh) 5 5.51 (s, 1H), 3.98 (s, 4H), 3.35-3.30 (m, 4H), 2.23 (s, 3H), 1.83-1.77(m, 4H) ppm. [001200] Step 4. Synthesis of 8-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4- dioxa-8-azaspiro [4.5] decane.
Figure imgf000254_0001
[001201] To a solution of 8-(5-methyl-lH-pyrazol-3-yl)-l,4-dioxa-8-azaspiro[4.5]decane (0.83 g, 3.72 mmol) in chloroform (50 mL) was added [4-(trifluoromethyl) phenyl]boronic acid (1.53 g, 7.43 mmol), anhydrous copper(II) acetate (1.35 g, 7.43 mmol), pyridine (1.5 mL, 18.6 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 48h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 120g silica gel column @100mL/min, eluting with 0-38% dichloromethane in petroleum ether) to afford the desired product 8-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-dioxa-8- azaspiro[4.5]decane (1.02 g, yield 70.4%) as a yellow solid.
[001202JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.34% (214 nm), Mass: found peak 284.1 (M+l)+ at 2.155 min.
[001203] 'H NMR (400 MHz, CDCh) 5 7.50-7.43 (m, 2H), 7.30-7.23 (m, 2H), 5.71 (s, 1H), 3.99 (s, 4H), 3.42-3.35 (m, 4H), 2.30 (s, 3H), 1.85-1.78 (m, 4H) ppm.
[001204] Step 5. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperidin-4-one.
Figure imgf000254_0002
[001205] To a solution of 8-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-dioxa-8- azaspiro[4.5]decane (600 mg, 1.57 mmol) in tetrahydrofuran (20 mL) was added HC1 aqueous (5 mol/L, 5.0 mL, 25 mmol). The reaction mixture was stirred at room temperature for 4h. The mixture was added to cold sodium bicarbonate aqueous, extracted with DCM, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @75 mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product l-[5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3- yl]piperidin-4-one (467 mg, yield 85.9%) as a yellow solid.
[001206] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 97.63% (214 nm) Mass: found peak 340.1 (M+l) at 2.069 min.
[001207] Step 6. Synthesis of l-[l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4- piperidyl]-4-tetrahydropyran-4-yl-piperazine (Compound 65).
Figure imgf000255_0001
[001208] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidin-4- one (100 mg, 0.28 mmol), l-tetrahydropyran-4-ylpiperazine (72 mg, 0.42 mmol), acetic acid (25 mg, 0.42 mmol) and molecular sieves 4 A in di chloroethane (15 mL) was added sodium triacetoxyborohydride (119 mg, 0.56 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product l-[l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4-piperidyl]-4-tetrahydropyran-4- yl-piperazine (62.1 mg, yield 44.6%) as a white solid.
[001209JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.22% (214 nm) Mass: found peak 494.2 (M+l) at 2.025 min.
[001210] 'H NMR (400 MHz, DMSO-d6) 5 7.61 (d, J = 8.4 Hz, 2H), 7.45 (d, J = 8.0 Hz, 2H), 5.88 (s, 1H), 3.86 (d, J = 9.6 Hz, 2H), 3.71 (d, J = 11.6 Hz, 2H), 3.25 (t, J = 11.6 Hz, 2H), 2.63 (t, J = 11.6 Hz, 2H), 2.50-2.36 (m, 7H), 2.36-2.22 (m, 6H), 1.79 (d, J = 11.2 Hz, 2H), 1.69 (d, J = 11.6 Hz, 2H), 1.50-1.26 (m, 4H) ppm.
Example S66. Synthesis of 4-[l-[l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 4-piperidyl]pyrrolidin-3-yl] morpholine (Compound 66).
[001211] Compound 66 was prepared as outlined below.
Figure imgf000256_0001
[001212] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidin-4- one (100 mg, 0.28 mmol), 4-pyrrolidin-3-ylmorpholine (66 mg, 0.42 mmol), acetic acid (25 mg, 0.42 mmol) and molecular sieves 4 A in 1,2-di chloroethane (15 mL) was added sodium triacetoxyborohydride (119 mg, 0.56 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[l-[l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4-piperidyl]pyrrolidin-3- yl]morpholine (87.3 mg, yield 64.6%) as a yellow solid.
[001213] LCMS method: Mobile Phase: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.41% (214 nm) Mass: found peak 480.2 (M+l) at 2.036 min.
[001214] 'H NMR (400 MHz, DMSO-d6) 5 7.60 (d, J = 8.8 Hz, 2H), 7.45 (d, J = 8.8 Hz, 2H), 5.88 (s, 1H), 3.60 (d, J = 13.6 Hz, 2H), 3.55 (t, J = 4.0 Hz, 4H), 2.81-2.60 (m, 6H), 2.36-2.24 (m, 8H), 2.13-2.03 (m, 1H), 1.90-1.78 (m, 3H), 1.63-1.53 (m, 1H), 1.48-1.34 (m, 2H) ppm.
Example S67. Synthesis of 4-[2-[4-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl]morpholine (Compound 67).
[001215] Compound 67 was prepared as outlined below.
Figure imgf000256_0002
[001216] Step 1. Synthesis of tert-butyl 4-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000257_0001
[001217] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (107 mg, 0.402 mmol) in dry di chloromethane (10 mL) was added (4-methoxyphenyl)boronic acid (125 mg, 0.804 mmol), anhydrous copper acetate (146 mg, 0.804 mmol) and dry pyridine (0.162 mL, 2.01 mmol). The reaction mixture was stirred at room temperature for 16h. The mixture was filtered, and the filtrate was purified by silica gel chromatography (PE : EA = 3 : 1) to afford the desired product tert-butyl 4-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazine-l- carboxylate (115 mg, yield 74.5%) as a yellow oil.
[001218JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: from 10 to 95% of B within 1.5min; Flow Rate: 1.5mL/min; Column: X-
Bridge C18, 3.5 gm, 50*4.6mm; Column Temperature: 50 °C; LC purity: 97% (214 nm), Mass: found peak 317.1 (M-55)+ at 2.149 min.
[001219] Step 2. Synthesis of l-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3-yl] piperazine.
Figure imgf000257_0002
[001220] To a solution of tert-butyl 4-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazine- 1-carboxylate (115 mg, 0.299 mmol) in dichloromethane (8 mL) was added TFA (0.667 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[l-(4-methoxyphenyl)-5- methyl-pyrazol-3-yl]piperazine (80 mg, yield 90%) as a yellow oil.
[001221JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 91.72% (254 nm), Mass: found peak 273.1 (M+l) at 1.685 min.
[001222] Step 3. Synthesis of 4-[2-[4-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 67).
Figure imgf000258_0001
[001223] To a solution of l-[l-(4-methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazine (80 mg, 0.269 mmol), potassium carbonate (149 mg, 1.08 mmol) and KI (44.7 mg, 0.269 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (65.2 mg, 0.39 mmol). The reaction was stirred at 90 °C overnight. After cooled to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(4- methoxyphenyl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (53.8 mg, yield 51.7%) as a white solid.
[001224JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.73% (214 nm), Mass: found peak 386.1 (M+l) at 1.783 min.
[001225] 'H NMR (400 MHz, DMSO-d6) 5 7.35 (d, J = 9.2 Hz, 2H), 6.99 (d, J = 9.2 Hz, 2H), 5.77 (s, 1H), 3.78 (s, 3H), 3.55 (t, J = 4.4 Hz, 4H), 3.08 (t, J = 4.4 Hz, 4H), 2.51-2.38 (m, 12H), 2.20 (s, 3H) ppm.
Example S68. Synthesis of 4-[2-[4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 68).
[001226] Compound 68 was prepared as outlined below.
Figure imgf000258_0002
[001227] Step 1. Synthesis of tert-butyl 4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-
3-yl] piperazine- 1-carboxylate.
Boc
Figure imgf000259_0001
[001228] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (0.12 g, 0.451 mmol) in dichloromethane (5 mL) was added (3-boronophenyl) (0.109 g, 0.901 mmol), anhydrous copper acetate (0.164 g, 0.901 mmol), pyridine (71.3 mg, 0.901 mmol) and molecular sieves 4 A. The reaction mixture was stirred at rt for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether: ethyl acetate=4/l) to afford the desired product tert-butyl 4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperazine- 1 -carboxylate (154 mg, yield 83.7%) as a yellow oil.
[001229JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column :Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 98% (214 nm); Mass: found peak 409.3 (M + H) at 1.40 min.
[001230] Step 2. Synthesis of l-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl] piperazine.
Boc
Figure imgf000259_0002
[001231] To a solution of tert-butyl 4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl]piperazine- 1-carboxylate (154 mg, 0.377 mmol) was added HC1 (3 mol/L, 0.628 mL). The mixture was stirred at rt for Ih. The mixture was concentrated to dryness to give 1 -[ 1 -[3- (difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperazine (120 mg, crude) as a white solid. The crude product was used directly in the next step without purification.
[001232JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 84% (214 nm); Mass: found peak 309.2 (M + H) at 0.95 min. [001233] Step 3. Synthesis of 4-[2-[4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 68).
Figure imgf000260_0001
[001234] To a solution of l-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperazine (116 mg, 0.376 mmol), potassium carbonate (260 mg, 1.88 mmol) and KI (62.5 mg, 0.376 mmol) in 95% ethanol / water (10 mL /0.5 mL) was added 4-(2-chloroethyl)morpholine (84.4 mg, 0.564 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[l-[3-(difluoromethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (110 mg, yield 69.4%) as a white solid.
[001235] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.5min, 95%B for 1.5min, back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column: Sunfire C18, 4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 421.9 (M + H) at 1.66 min. [001236] 'H NMR (400MHz, CDCh): 5 7.41 (d, J =8.0Hz, 1H), 7.29-7.32 (m, 2H), 7.04 (dd, J =8.0Hz, 1.6Hz, 1H), 6.38-6.74 (m, 1H), 5.72 (s, 1H), 3.74 (t, J =4.8Hz, 4H), 3.28 (t, J =4.8Hz, 4H), 2.64 (t, J =4.8Hz, 4H), 2.59 (t, J =6.0Hz, 4H), 2.52 (t, J =4.4Hz, 4H), 2.34 (s, 3H) ppm.
Example S69. Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3- tetrahydropyran-4-yl-3,9-diazaspiro [5.5] undecane (Compound 69).
[001237] Compound 69 was prepared as outlined below.
Figure imgf000261_0001
[001238] Step 1. Synthesis of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-3,9-diazaspiro [5.5] undecane-3-carboxylate.
Boc
Figure imgf000261_0002
[001239] To a solution of 3-iodo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (60 mg, 0.163 mmol), tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (82.9 mg, 0.326 mmol) and PEPPSI-Ipent (6.47 mg, 0.00815 mmol) in 1,4-di oxane (0.5 mL) was added sodium tert- butoxide (47 mg, 0.489 mmol). The reaction was stirred at 100 °C in tube for 16h. Wash the organic layer with saturated sodium bicarbonate (2 mL), extracted. The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (PE/EA=4/1) to give tert-butyl 9-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3,9-diazaspiro[5.5]undecane-3-carboxylate (36 mg, 44.7% yield) as a yellow solid.
[001240JLCMS Method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 100 % (214 nm); Mass: found peak 495.4 (M + H) at 1.52 min.
[001241] Step 2. Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,9- diazaspiro [5.5] undecane.
Figure imgf000262_0001
[001242] To a solution of tert-butyl 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,9-diazaspiro[5.5]undecane-3-carboxylate (36 mg, 0.0728 mmol) was added HC1 (3 mol/L, 0.121 mL). The mixture was stirred at rt for Ih. The mixture was concentrated to dryness to give 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,9-diazaspiro[5.5]undecane (28 mg, crude) as a yellow oil. The crude product was used directly in the next step without purification. [001243JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min;
Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 100% (214 nm); Mass: found peak 395.3 (M+l) at 1.07 min.
[001244] Step 3. Synthesis of 9-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3- tetrahydropyran-4-yl-3,9-diazaspiro [5.5] undecane (Compound 69).
Figure imgf000262_0002
[001245] To a solution of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,9- diazaspiro[5.5]undecane (28.7 mg, 0.0728 mmol) in 1,2-di chloroethane (0.5 mL) was added sodium triacetoxyborohydride (30.8 mg, 0.146 mmol), tetrahydropyran-4-one (10.9 mg, 0.109 mmol) and acetic acid (one drop). The reaction mixture was stirred at room temperature for 16h.The mixture was washed with water (10 mL) and extracted by EA (10 mLx2). The combined organic layer was dried over Sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (DCM:MeOH=10/l) to give 9-[5- methyl-l-[4-(tri fluoromethoxy )phenyl]pyrazol-3-yl]-3-tetrahydropyran-4-yl-3, 9- diazaspiro[5.5]undecane (17.6 mg, 50.5% yield) as a yellow solid.
[001246] LCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B for 0.2 min, increase to 95%B within 1.5min, 95%B for 1.5min, back within O.Olmin. Flow Rate: 2mL/min; Column: Sunfire, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99 % (214 nm); Mass: found peak 479.1 (M + H) at 1.50 min.
[001247] 'H NMR (400MHz, CDCh): 5 7.54-7.58 (m, 2H), 7.41 (d, J =8.4Hz, 2H), 5.85 (s, 1H), 4.01 (dd, J =11.6Hz, 4.0Hz, 2H), 3.42 (t, J =10.8Hz, 2H), 3.21 (t, J =5.6Hz, 4H), 2.63 (t, J =4.4Hz, 4H), 2.47-2.54 (m, 1H), 2.30 (s,3H), 1.87 (d, J =10.8Hz, 2H), 1.52-1.63 (m, 10H) ppm. Example S70. Synthesis of l-[2-(3-methyl-l-piperidyl)ethyl]-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl] piperidine (Compound 70).
[001248] Compound 70 was prepared as outlined below.
Figure imgf000263_0001
[001249] Step 1. Synthesis of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperidine.
Figure imgf000263_0002
[001250] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine-l -carboxylate (600 mg, 1.41 mmol) in DCM (10 mL) was added TFA (1.61 g, 14.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (450 mg, yield 98.9%) as a yellow oil. [001251JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100% (214 nm); Mass: found peak 326.2(M+H) at 1.057 min.
[001252] Step 2. Synthesis of 2-chloro-l-[4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] - 1-piperidyl] ethenone.
Figure imgf000264_0001
[001253] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (260 mg, 0.799 mmol) and TEA (162 mg, 1.6 mmol) in DCM (10 mL) was added chloroacetyl chloride (903 mg, 7.99 mmol). The reaction mixture was stirred at 0 °C for Ih. Then the reaction mixture was concentrated in vacuo. The residue was purified by flash chromatography (PE/EA= 1/1) to afford the desired product 2-chloro-l-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethanone (290 mg, yield 90.3%) as a yellow oil.
[001254JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 100.0% (214 nm); Mass: found peak 402.0(M+H) at 2.083 min.
[001255] Step 3. Synthesis of 2-(3-methyl-l-piperidyl)-l-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]-l-piperidyl]ethenone.
Figure imgf000264_0002
[001256] To a solution of 2-chloro-l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l-piperidyl]ethanone (120 mg, 0.299 mmol) and TEA (90 mg, 0.896 mmol) in DCM (10 mL) was added 3 -methylpiperidine (44 mg, 0.448 mmol). The reaction mixture was stirred at rt for 16h. Then concentrated in vacuo. The residue was purified by flash chromatography (PEZEA= 1 : 1) to afford the desired product 2-(3-methyl-l-piperidyl)-l-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]-l-piperidyl]ethanone (125 mg, yield 69.4%) as a yellow oil.
[001257] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.3min; Flow Rate: 2.0 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 77.4% (214 nm); Mass: found peak 465.3(M+H) at 1.665 min.
[001258] Step 4. Synthesis of l-[2-(3-methyl-l-piperidyl)ethyl]-4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl]piperidine (Compound 70).
Figure imgf000265_0001
[001259] A solution of 2-(3-methyl-l-piperidyl)-l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-l-piperidyl]ethanone (130 mg, 0.280 mmol) in BH3 THF (10 mL) was stirred at 90 °C for 3h. After cooling to 0 °C, the reaction mixture was treated with MeOH dropwise and then 3N HC1 (15 mL). The resulting mixture was stirred under reflux for 6h. After cooling to 0 °C, the mixture was neutralized with sodium bicarbonate to pH=9 and extracted with DCM (20 mL*3). Combined DCM layers were dried over sodium sulfate and concentrated. The residue was purified by Prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-[2-(3-methyl-l-piperidyl)ethyl]-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl] piperidine (26.8 mg, yield 21.3%) as a white solid.
[001260JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 451.4(M+H) at 1.034 min.
[001261] 'H NMR (400 MHz, DMSO) 5 7.65 (t, J = 8.0 Hz, 2H), 7.49 (d, J = 8.0 Hz, 2H), 6.16 (s, 1H), 2.92 (d, J = 12.0 Hz, 2H), 2.77 (s, 2H), 2.54 (s, 1H), 2.41 (s, 6H), 2.32 (s, 3H), 2.03 (t, J = 12.0 Hz, 2H), 1.84 (d, J = 12.0 Hz, 2H), 1.60 (dd, J = 24.0, 12.0 Hz, 6H), 1.45-1.32 (m, 1H), 0.82 (d, J = 8.0 Hz, 3H) ppm.
Example S71. Synthesis of 4,4-difluoro-l-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]piperidine (Compound 71). [001262] Compound 71 was prepared as outlined below.
Figure imgf000266_0001
[001263] Step 1. Synthesis of 2-(4,4-difluoro-l-piperidyl)-l-[4-[5-methyl-l-[4- (tr ifluoromethoxy)phenyl] pyrazol-3-yl] - 1-piperidyl] ethenone.
Figure imgf000266_0002
[001264] To a solution of 2-chloro-l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l-piperidyl]ethanone (150 mg, 0.373 mmol) was added 4,4-difluoropiperidine (67 mg, 0.560 mmol) and potassium carbonate (155 mg, 1.12 mmol) in DMF (10 mL). The reaction mixture was stirred at 50 °C for 2h. Then 80 mL of water and 120 mL of EtOAc were added and the organic layer was separated and washed with brine (40 mL x 4), dried over sodium sulfate, filtered, and concentrated in vacuo in vacuo to afford the desired product 2-(4,4-difluoro-l- piperidyl)-l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethanone (135 mg, yield 49.1%) as a yellow oil.
[001265] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 66.4% (214 nm); Mass: found peak 487.3(M+H) at 1.161min.
[001266] Step 2. Synthesis of 4,4-difluoro-l-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]piperidine (Compound 71).
Figure imgf000267_0001
[001267] To a solution of 2-(4,4-difhioro-l-piperidyl)-l-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethanone (135 mg, 0.278 mmol) was added BH3-THF (10 mL) in THF (10 mL). The reaction mixture was stirred at 90 °C for 3h. The reaction mixture was then cooled to 0 °C and MeOH was added dropwise, concentrated, then was added 3N HC1 (15mL) and the mixture refluxed for 6h. The reaction mixture was cooled to 0 °C and neutralized with sodium bicarbonate to pH=9. The product was extracted with DCM (20mL*3), dried over sodium sulfate, and concentrated to afford a residue. The residue was purified by Prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4,4-difluoro- 1 -[2-[4-[5-methyl- 1 -[4-(trifluoromethoxy)phenyl]pyrazol-3 -yl ] - 1 - piperidyl]ethyl]piperidine (10.6 mg, yield 8.08%) as a colorless oil.
[001268JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire,
4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 473.3(M+H) at 1.142 min.
[001269] 'H NMR (400 MHz, MeOD-d4) 5 7.59 (d, J = 8.0 Hz, 2H), 7.46 (d, J = 8.0 Hz, 2H), 6.19 (s, 1H), 3.50 (s, 1H), 3.15 (s, 2H), 2.70-2.16 (m, 10H), 2.33 (s, 3H), 1.95 (dt, J = 56.0, 12.0 Hz, 8H) ppm.
Example S72. Synthesis of 3-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l-piperidyl]ethyl]-8-oxa-3-azabicyclo[3.2.1]octane (Compound 72).
[001270] Compound 72 was prepared as outlined below.
Figure imgf000267_0002
[001271] Step 1. Synthesis of l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l- piperidyl]-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)ethenone.
Figure imgf000268_0001
[001272] To a solution of 2-chloro-l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l-piperidyl]ethanone (150 mg, 0.373 mmol) was added 8-oxa-3- azabicyclo[3.2.1]octane;hydrochloride (83 mg, 0.560 mmol) and potassium carbonate (155 mg, 1.12 mmol) in DMF (10 mL). The reaction mixture was stirred at 50 °C for 2h. Then 80 mL of water and 120 mL of EtOAc were added. The organic layer was separated, washed with brine (40 mL x 4), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]-2-(8-oxa-3- azabicyclo[3.2.1]octan-3-yl)ethanone (120 mg, yield 52.4%) as a yellow oil.
[001273JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 78.7% (214 nm); Mass: found peak 479.1(M+H) at 2.084 min.
[001274] Step 2. Synthesis of 3-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]ethyl]-8-oxa-3-azabicyclo[3.2.1]octane (Compound 72).
Figure imgf000268_0002
[001275] A solution of l-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l- piperidyl]-2-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)ethanone (120 mg, 0.251 mmol) and BEL THF (10 mL) was stirred at 90 °C for 3h. After cooling to 0 °C, the reaction mixture was treated with MeOH dropwise and then 3N HC1 (15 mL). The resulting mixture was stirred under reflux for 6h. After cooling to 0 °C, the mixture was neutralized with sodium bicarbonate to pH=9, and extracted with DCM (20 mL*3). The combined DCM layers were dried over sodium sulfate and concentrated in vacuo. The residue was purified by Prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 3-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]-8-oxa-3-azabicyclo[3.2.1] octane (37.1 mg, yield 31.8%) as a white solid.
[001276] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B with in l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 465.3(M+H) at 1.046 min.
[001277] 'H NMR (400 MHz, DMSO) 5 7.64 (d, J = 8.0 Hz, 2H), 7.49 (d, J = 8.6 Hz, 2H), 6.16 (s, 1H), 4.18 (s, 2H), 2.93 (d, J = 12.0 Hz, 2H), 2.59 (s, 1H), 2.56 (d, J = 8.5 Hz, 2H), 2.38 (s, 4H), 2.32 (s, 3H), 2.15 (d, J = 8.0 Hz, 2H), 2.04 (t, J = 8.0 Hz, 2H), 1.81 (dd, J = 16.0, 12.0 Hz, 4H), 1.71-1.53 (m, 4H) ppm.
Example S73. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclohexoxy] ethyl] morpholine (Compound 73).
[001278] Compound 73 was prepared as outlined below.
Figure imgf000269_0001
[001279] Step 1. Synthesis of 3-(l,4-dioxaspiro[4.5]dec-7-en-8-yl)-5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000269_0002
[001280] To a solution of 3-iodo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, 0.815 mmol) and 2-(l,4-dioxaspiro[4.5]dec-7-en-8-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (332 mg, 1.22 mmol) in 1,4-dioxane (8 mL) was added potassium carbonate aqueous (338 mg, 2.45 mmol) in water (2 mL) and PdC12(dppf) dichloromethane complex (66 mg, 0.081 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was purified by flash chromatography (PE/EA=3/1) to afford the desired product 3 -(1 ,4- dioxaspiro[4.5]dec-7-en-8-yl)-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, yield 91.9%) as a colorless oil.
[001281JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 LCMS; LC purity: 100.0% (214 nm); Mass: found peak 381.2(M+H) at 1.446 min.
[001282] Step 2. Synthesis of 3-(l,4-dioxaspiro[4.5]decan-8-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole.
Figure imgf000270_0001
[001283] Under H2 atmosphere, a mixture of 3-(l,4-dioxaspiro[4.5]dec-7-en-8-yl)-5-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazole (280 mg, 0.736 mmol) and Pd/C (10%, 150 mg) in ethyl acetate (18 mL) was stirred at room temperature for 2h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=5/1) to afford the desired product 3-(l,4-dioxaspiro[4.5]decan-8-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (270 mg, yield 78.7%) as a colorless oil.
[001284JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 82.2% (214 nm); Mass: found peak 383.2(M+H) at 1.439 min.
[001285] Step 3. Synthesis of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclohexanone.
Figure imgf000271_0001
[001286] To a solution of 3-(l,4-dioxaspiro[4.5]decan-8-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (210 mg, 0.549 mmol) in tetrahydrofuran (15 mL) was added HC1 aqueous (5 mol/L, 0.8 mL, 4.2 mmol). The reaction mixture was stirred at room temperature for 4h. The mixture was diluted with EtOAc (60 mL), washed with water (20 mL) and brine (20 mL*2), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=8/1) to afford the desired product 4-[5-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclohexanone (170 mg, yield 91%) as a colorless oil. [001287] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 339.2(M+H) at 1.357 min.
[001288] Step 4. Synthesis of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclohexanol.
Figure imgf000271_0002
[001289] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]cyclohexanone (170 mg, 0.502 mmol) in methanol (10 mL) was added NaBEL (38 mg, 1.00 mmol) portion-wise. The reaction mixture was stirred at room temperature for Ih. The mixture was directly purified by flash chromatography (PE/EA=10/l) to afford the desired product 4-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclohexanol (160 mg, yield 86.1%) as a colorless oil.
[001290JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 92.0% (214 nm); Mass: found peak 341.3(M+H) at 1.291 min.
[001291] Step 5. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] cyclohexoxy] ethyl] morpholine (Compound 73).
Figure imgf000272_0001
[001292] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclohexanol (215 mg, 0.632 mmol) in dry DMF (15 mL) at 0 °C was added sodium hydride (60%, 505 mg, 12.6 mmol). The reaction was warmed to room temperature and stirred for Ih, then 60 °C for Ih. The reaction was cooled to room temperature and 4-(2-chloroethyl)morpholine hydrochloride (235 mg, 1.26 mmol) was added and stirred at 80 °C for 16h. The reaction was cooled to room temperature, quenched with water, and filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]cyclohexoxy]ethyl]morpholine (11.1 mg, yield 3.53%) as a yellow solid.
[001293JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 91.0% (214 nm); Mass: found peak 454.1(M+H) at 2.165 min.
[001294] 'H NMR (400 MHz, DMSO) 5 7.70-7.58 (dd, J = 8.0, 4.0 Hz, 2H), 7.49 (d, J = 8.0 Hz, 2H), 6.14 (s, IH), 3.54 (dd, J = 8.0, 4.0 Hz, 6H), 3.25-3.24 (m, IH), 2.51-2.50 (m, IH), 2.50 (t, J = 4.0 Hz, 2H), 2.45 (t, J = 8.0 Hz, 4H), 2.32 (s, 3H), 2.03 (d, J = 12.0 Hz, 2H), 1.95 (d, J = 12.0 Hz, 2H), 1.42 (dd, J = 16.0, 8.0 Hz, 2H), 1.25 (dd, J = 12.0, 8.0 Hz, 2H) ppm.
Example S74. Synthesis of 4-[2-[4-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 74).
[001295] Compound 74 was prepared as outlined below.
Figure imgf000272_0002
[001296] Step 1. Synthesis of (4,4-difluorocyclohexyl) 4-methylbenzenesulfonate.
Figure imgf000273_0001
[001297] To a solution of 4,4-difhiorocyclohexanol (300 mg, 2.2 mmol) in pyridine (5 mL) at 0 °C was added 4-methylbenzenesulfonyl chloride (630 mg, 3.31 mmol). The reaction mixture was stirred at RT for 16h. The mixture was directly purified by flash chromatography (PE/EA=3/1) to afford the desired product (4,4-difluorocyclohexyl) 4-methylbenzenesulfonate (534 mg, yield 83.5%) as a white solid.
[001298JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 98.7% (214 nm); Mass: found peak 313.1[M+23]+ at 1.35 min.
[001299] Step 2. Synthesis of tert-butyl 4-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000273_0002
[001300] A mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l -carboxylate (300 mg, 1.13 mmol), (4,4-difluorocyclohexyl) 4-methylbenzenesulfonate (491 mg, 1.69 mmol) and cesium carbonate (1.1 g, 3.38 mmol) in N,N-dimethylformamide (10 mL) was stirred at 90 °C for 48h. The reaction mixture was cooled to room temperature and diluted with EtOAc (150 mL), washed with water (15 mL), brine (15 mL*4), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=2/1) to afford the desired product tert-butyl 4-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3-yl]piperazine-l- carboxylate (135 mg, yield 31.3%) as a white solid.
[001301JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 385.3(M+H) at 1.397 min.
[001302] Step 3. Synthesis of l-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3- yl] piperazine.
Figure imgf000274_0001
[001303] A solution of tert-butyl 4-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (130 mg, 0.338 mmol) and TFA (385 mg, 3.38 mmol) in DCM (10 mL) stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3-yl]piperazine (95 mg, yield 92.9%) as a yellow oil.
[001304JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 94.1% (214 nm); Mass: found peak 285.3(M+H) at 0.935 min.
[001305] Step 4. Synthesis of 4-[2-[4-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 74).
Figure imgf000274_0002
[001306] A solution of l-[l-(4,4-difluorocyclohexyl)-5-methyl-pyrazol-3-yl]piperazine (95 mg, 0.334 mmol), 4-(2-chloroethyl)morpholine;hydrochloride (93 mg, 0.501 mmol), potassium carbonate (139 mg, 1.0 mmol) and KI (55 mg, 0.334 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h, then concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(4,4- difluorocyclohexyl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (68.6 mg, yield 49.6%) as a white solid.
[001307] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 96.7% (214 nm); Mass: found peak 398.2(M+H) at 1.830 min.
[001308] 'H NMR (400 MHz, DMSO) 5 5.46 (s, IH), 4.16 (s, IH), 3.55 (t, J = 4.0 Hz, 4H), 2.99 (d, J = 12.0 Hz, 4H), 2.51 (s, 4H), 2.41 (d, J = 12.0 Hz, 4H), 2.36 (d, J = 24.0 Hz, 4H), 2.17 (s, 3H), 2.11 (d, J = 8.0 Hz, 2H), 2.06-1.91 (m, 4H), 1.80 (s, 2H) ppm. Example S75. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 75).
[001309] Compound 75 was prepared as outlined below.
Figure imgf000275_0001
[001310] Step 1. Synthesis of [4-(trifluoromethyl)cyclohexyl] 4-methylbenzenesulfonate.
Figure imgf000275_0002
[001311] To a solution of 4-(trifluoromethyl)cyclohexanol (400 mg, 2.38 mmol) in pyridine (5 mL) at 0 °C was added 4-methylbenzenesulfonyl chloride (680 mg, 3.57 mmol). The reaction mixture was stirred at rt for 16h. The mixture was directly purified by flash chromatography (PE/EA=3/1) to afford the desired product [4-(trifluoromethyl)cyclohexyl] 4- methylbenzenesulfonate (657 mg, yield 82.3%) as a colorless oil.
[001312JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 70.9% (214 nm); Mass: found peak 345.2[M+23]+ at 1.417 min.
[001313] Step 2. Synthesis of tert-butyl 4-[5-methyl-l-[4- (trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000275_0003
[001314] A mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l -carboxylate (400 mg, 1.5 mmol), [4-(trifluoromethyl)cyclohexyl] 4-methylbenzenesulfonate (726 mg, 2.25 mmol) and cesium carbonate (1.47 g, 4.51 mmol) in N,N-dimethylformamide (10 mL) was stirred at 90 °C for 48h. The reaction mixture was cooled to room temperature and diluted with EtOAc (150 mL), washed with water (15 mL), brine (15 mL*4), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=2/1) to afford the desired product tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazine-l -carboxylate (150 mg, yield 12.3%) as a colorless oil.
[001315] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire,
4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 417.3 (M+H) at 2.220 min.
[001316] Step 3. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3- yl] piperazine.
Figure imgf000276_0001
[001317] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazine-l -carboxylate (150 mg, 0.360 mmol) and TFA (411 mg, 3.60 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine (110 mg, yield 71.4%) as a colorless oil.
[001318JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 74.2% (214 nm); Mass: found peak 317.3(M+H) at 1.041 min.
[001319] Step 4. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 75).
Figure imgf000276_0002
[001320] A suspension of l-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine (106 mg, 0.334 mmol), 4-(2-chloroethyl)morpholine hydrochloride (93 mg, 0.501 mmol), potassium carbonate (139 mg, 1.0 mmol) and KI (55 mg, 0.334 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h. Then the mixture was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (17.4 mg, yield 11.4%) as a colorless oil.
[001321JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 93.8% (214 nm); Mass: found peak 430.2(M+H) at 2.000 min.
[001322] XH NMR (400 MHz, MeOD-d4) 5 5.56 (s, 1H), 4.25 (dd, J = 12.0, 4.0 Hz, 1H), 3.71 (dd, J = 12.0, 8.0 Hz, 4H), 3.25-3.11 (m, 4H), 2.66 (dd, J = 8.0, 4.0 Hz, 4H), 2.62-2.58 (m, 4H), 2.54 (s, 4H), 2.28 (dd, J = 12.0, 4.0 Hz, 2H), 2.23 (s, 3H), 2.11-2.02 (m, 2H), 1.80 (ddd, J = 28.0, 12.0, 8.0 Hz, 4H) ppm.
Example S76. Synthesis of 4-[2-[4-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 76).
[001323] Compound 76 was prepared as outlined below.
Figure imgf000277_0001
[001324] Step 1. Synthesis of [3-(trifluoromethyl)cyclohexyl] 4-methylbenzenesulfonate.
Figure imgf000277_0002
[001325] To a solution of 3-(trifluoromethyl)cyclohexanol (400 mg, 2.38 mmol) in pyridine (5 mL) at 0 °C was added 4-methylbenzenesulfonyl chloride (680 mg, 3.57 mmol). The reaction mixture was stirred at rt for 16h. The mixture was directly purified by flash chromatography (PE/EA=2/1) to afford the desired product [3-(trifluoromethyl)cyclohexyl] 4- methylbenzenesulfonate (645 mg, yield 84.1%) as a white solid.
[001326] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100% (214 nm); Mass: found peak 345.1 (M+H) at 1.408 min.
[001327] Step 2. Synthesis of tert-butyl 4-[5-methyl-l-[3-
(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000278_0001
[001328] A mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l -carboxylate (300 mg, 1.13 mmol), [3-(trifluoromethyl)cyclohexyl] 4-methylbenzenesulfonate (545 mg, 1.69 mmol) and cesium carbonate (1.1 g, 1.38 mmol) in N,N-dimethylformamide (10 mL) was stirred at 90 °C for 48h. The reaction mixture was cooled to room temperature and diluted with EtOAc (60 mL), washed with water (15 mL), brine (15 mL*4), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (PE/EA=2/1) to afford the desired product tert-butyl 4-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazine-l -carboxylate (170 mg, yield 33.2%) as a colorless oil.
[001329JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 46.8% (214 nm); Mass: found peak 417.3 (M+H) at 1.541 min.
[001330] Step 3. Synthesis of l-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3- yl] piperazine.
Figure imgf000278_0002
[001331] A solution of tert-butyl 4-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazine-l -carboxylate (120 mg, 0.288 mmol) and TFA (328 mg, 2.88 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine (85 mg, yield 52.2%) as a colorless oil.
[001332JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 56.8% (214 nm); Mass: found peak 317.3(M+H) at 1.024 min.
[001333] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 76).
Figure imgf000279_0001
[001334] A solution of l-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazine (85 mg, 0.269 mmol), 4-(2-chloroethyl)morpholine;hydrochloride (75 mg, 0.403 mmol), potassium carbonate (111 mg, 0.806 mmol) and KI (44 mg, 0.269 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h. Then the mixture was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[3-(trifluoromethyl)cyclohexyl]pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (33.1 mg, yield 27.8%) as a white solid.
[001335] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 97.5% (214 nm); Mass: found peak 451.4(M+H) at 1.034 min.
[001336] 'H NMR (400 MHz, MeOD-d4) 5 5.55 (s, 1H), 4.41(t, J = 8.0 Hz, 1H), 3.69 (t, J = 4.0 Hz, 4H), 3.30 (t, J = 4.0 Hz, 5H), 2.64 (t, J = 8.0 Hz, 4H), 2.61 (t, J = 8.0 Hz, 4H), 2.52 (t, J = 8.0 Hz, 4H), 2.20 (s, 3H), 2.11 (dd, J = 12.0, 8.0 Hz, 1H), 2.03-1.94 (m, 1H), 1.92-1.71 (m, 4H), 1.70-1.49 (m, 2H) ppm.
Example S77. Synthesis of N-ethyl-N-[2-[4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] cyclohexanamine (Compound 77).
[001337] Compound 77 was prepared as outlined below.
Figure imgf000280_0001
[001338] Step 1. Synthesis of tert-butyl 4-(3-oxobutanoyl)piperazine-l-carboxylate.
Figure imgf000280_0002
[001339] A mixture of tert-butyl piperazine- 1 -carboxylate (10.0 g, 53.7 mmol) and tert-butyl acetoacetate (9.34 g, 59.1 mmol) in toluene (150 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 4-(3-oxobutanoyl)piperazine-l- carboxylate (14.3 g, 51.7% yield) as a yellow oil.
[001340JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 97.8% (214 nm); Mass: found peak 293.2[M+23]+ at 1.033 min.
[001341] Step 2. Synthesis of tert-butyl 4-(3-oxobutanethioyl)piperazine-l-carboxylate.
Lawesson's reagent
(0.5 eq) toluene, 75°c, 16h
Figure imgf000280_0003
Figure imgf000280_0004
[001342] To a solution of tert-butyl 4-(3-oxobutanoyl)piperazine-l -carboxylate (14.3 g, 52.9 mmol) in toluene (200 mL) was added Lawesson's reagent (10.7 g, 26.4 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tertbutyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (8.6 g, 54.3% yield) as a brown oil. [001343JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.3min; Flow Rate:2.0 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 50 °C; LC purity: 72.5% (214 nm); Mass: found peak 287.2(M+H) at 1.847 min.
[001344] Step 3. Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Boc
Figure imgf000281_0001
[001345] To a solution of tert-butyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (8.6 g, 30.0 mmol) in toluene (150 mL) was added hydrazine monohydrate (4.46 mL, 90.1 mmol) and the mixture was stirred at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-6% MeOH in DCM) to afford the desired product tertbutyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (6.80 g, yield 62.1%) as a yellow solid.
[001346] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 73.0% (214 nm); Mass: found peak 267.3(M+H) at 1.033 min.
[001347] Step 4. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl] piperazine- 1-carboxylate.
Figure imgf000281_0002
[001348] To a solution of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (380 mg, 1.43 mmol) was added [4-(trifluoromethoxy)phenyl]boronic acid (588 mg, 2.85 mmol), Copper(II) acetate (518 mg, 2.85 mmol), pyridine (564 mg, 7.13 mmol) and molecular sieves 4 . The reaction mixture was stirred at room temperature for 48h. The mixture was filtered and purified by flash chromatography (PE/DCM=1/1) to afford the desired product tert-butyl 4-[5- methyl- l-[4-(trifluorom ethoxy )phenyl]pyrazol-3-yl]piperazine-l -carboxylate (410 mg, yield 64%) as a yellow oil.
[001349JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA);
Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire,
4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 95.4% (214 nm); Mass: found peak 427.3(M+H) at 1.516 min.
[001350] Step 5. Synthesis of ll-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl] piperazine.
Figure imgf000282_0001
[001351] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (410 mg, 0.961 mmol) and TFA (1.1 g, 9.61 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with di chloromethane (20 mL*3), and dried over sodium sulfate to afford the desired product 11-[5- methyl-l-[4-(trifluorom ethoxy) phenyl] pyrazol-3-yl]piperazine (266 mg, yield 77.2%) as a yellow oil.
[001352JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 91.2% (214 nm); Mass: found peak 327.3(M+H) at 1.045 min.
[001353] Step 6. Synthesis of l-(2-chloroethyl)-4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000283_0001
[001354] Under argon atmosphere, a mixture of l-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (246 mg, 0.754 mmol), 2-chloroacetaldehyde (88 mg, 1.13 mmol) and sodium cyanoborohydride (94 mg, 1.51 mmol) in methanol (10 mL) was stirred at rt for 16h. Then the mixture was filtered and concentrated in vacuo. The residue was purified by flash to afford l-(2-chloroethyl)-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (170 mg, 56.3% yield) as a white solid. [001355] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 97.1% (214 nm); Mass: found peak 389.3(M+H) at 1.065 min.
[001356] Step 7. Synthesis of N-ethyl-N-[2-[4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin-l-yl] ethyl] cyclohexanamine (Compound
Figure imgf000283_0002
[001357] A solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazine (80 mg, 0.206 mmol), N-ethylcyclohexanamine (39 mg, 0.309 mmol) and DIPEA (133 mg, 1.03 mmol) in NMP (2 mL) was stirred at 160 °C for 2h in MW. Then the reaction mixture was concentrated in vacuo. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product N-ethyl-N-[2-[4-[5-methyl- l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]cyclohexanamine (46.9 mg, yield 46.1%) as a yellow oil. [001358JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA;
Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 97.2% (214 nm); Mass: found peak 480.4(M+H) at 1.104 min.
[001359] 'H NMR (500 MHz, MeOD-d4) 5 7.61 (t, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 5.85 (s, 1H), 3.30 (t, J = 12.0 Hz, 6H), 3.25 (d, J = 8.0 Hz, 4H), 2.70 (s, 1H), 2.66 (t, J =12.0 Hz, 4H), 2.29 (s, 3H), 1.91 (dd, J = 8.0, 4.0 Hz, 4H), 1.71 (d, J = 12.0 Hz, 1H), 1.56-1.21 (m, 8H) ppm.
Example S78. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-oxazepane (Compound 78).
[001360] Compound 78 was prepared as outlined below.
Figure imgf000284_0001
[001361] A solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (70 mg, 0.180 mmol), l,4-oxazepane;hydrochloride (37 mg, 0.270 mmol), potassium carbonate (74 mg, 0.540 mmol) and KI (29 mg, 0.180 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h. Then concentrated in vacuo. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- methyl- 1 -[4-(trifluorom ethoxy )phenyl]pyrazol-3 -yl]piperazin- 1 -yl]ethyl]- 1 ,4-oxazepane (39.4 mg, yield 47.3%) as a white solid.
[001362JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 98.9% (214 nm); Mass: found peak 454.2 (M+H) at 2.029 min.
[001363] 'H NMR (400 MHz, MeOD-d4) 5 7.57 (tt, J = 8.0, 4.0 Hz, 2H), 7.42 (t, J = 8.0 Hz, 2H), 5.86 (d, J = 4.0 Hz, 1H), 3.78 (ddd, J = 12.0, 8.0, 4.0 Hz, 4H), 3.29 (t, J = 8.0 Hz, 4H), 2.80 (ddd, J = 12.0, 8.0, 4.0 Hz, 6H), 2.59 (ddd, J = 12.0, 8.0, 4.0 Hz, 6H), 2.31 (s, 3H), 1.94 (dt, J = 12.0, 4.0 Hz, 2H) ppm.
Example S79. Synthesis of N-ethyl-N-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l-yl]ethyl]tetrahydropyran-4-amine (Compound 79).
[001364] Compound 79 was prepared as outlined below.
Figure imgf000285_0001
[001365] A solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazine (60 mg, 0.154 mmol), N-ethyltetrahydropyran-4-amine (29 mg, 0.231 mmol) and DIPEA (99 mg, 0.772 mmol) in NMP (2 mL) was stirred at 160 °C for 2h in MW, then concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product N-ethyl-N-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]piperazin-l-yl]ethyl]tetrahydropyran-4-amine (28 mg, yield 36%) as a yellow oil.
[001366] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 96.1% (214 nm); Mass: found peak 482.2 (M+H) at 2.15 min.
[001367] 'H NMR (400 MHz, MeOD-d4) 5 7.55 (d, J = 8.0 Hz, 2H), 7.39 (d, J = 8.0 Hz, 2H), 5.84 (s, 1H), 3.99 (d, J = 8.0 Hz, 2H), 3.42 (dd, J = 24.0, 12.0 Hz, 2H), 3.23 (d, J = 4.0 Hz, 4H), 2.90 (s, 1H), 2.77 (d, J = 28.0 Hz, 4H), 2.65 (s, 4H), 2.56 (d, J = 8.0 Hz, 2H), 2.29 (s, 3H), 1.78 (d, J = 12.0 Hz, 2H), 1.61 (dd, J = 8.0, 4.0 Hz, 2H), 1.12 (t, J = 8.0 Hz, 3H) ppm.
Example S80. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4-(l- tetrahydropyran-4-yl-4-piperidyl)piperazine (Compound 80).
[001368] Compound 80 was prepared as outlined below.
Figure imgf000286_0001
[001369] Step 1. Synthesis of tert-butyl 4-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]piperidine-l-carboxylate.
Boc
Figure imgf000286_0002
[001370] Under argon atmosphere, a mixture of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine (450 mg, 1.38 mmol) and tert-butyl 4-oxopiperidine-l -carboxylate (412 mg, 2.07 mmol) was stirred in THF (5mL). Titanium (IV) isopropoxide (470 mg, 1.65 mmol) was added under nitrogen atmosphere and the mixture was heated at 45°C for 3.5h. 6 mL of distilled ethanol and sodium cyanoborohydride (173 mg, 2.76 mmol) were added and the mixture was further stirred at 45°C for 3h. After one night at room temperature, the mixture was poured onto 300 mL of water and stirred for Ih. The reaction mixture was filtered through a plug of Celite and washed with dichloromethane. The aqueous layer was then extracted with dichloromethane and the organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The mixture was directly purified by flash chromatography to afford the desired product tert-butyl 4-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l- yl]piperidine-l -carboxylate (75 mg, yield 9.29%) as a colorless oil.
[001371JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 87.5% (214 nm); Mass: found peak 510.4(M+H) at 1.165 min.
[001372] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4-(4- piperidyl)piperazine.
Figure imgf000287_0001
[001373] A solution of tert-butyl 4-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]piperidine-l -carboxylate (75 mg, 0.147 mmol) and TFA (168 mg, 1.47 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 4-(4-piperidyl)piperazine (55 mg, yield 76.7%) as a yellow oil.
[001374JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.3min; Flow Rate:2.0 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 50 °C; LC purity: 84.2% (214 nm); Mass: found peak 410.3(M+H) at 0.998 min.
[001375] Step 3. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4- (l-tetrahydropyran-4-yl-4-piperidyl)piperazine (Compound 80).
Figure imgf000287_0002
[001376] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-4-(4- piperidyl)piperazine (59 mg, 0.144 mmol) in 1,2-dichloroethane (5 mL) was added sodium triacetoxyborohydride(61 mg, 0.288 mmol), tetrahydropyran-4-one (21 mg, 0.216 mmol) and acetic acid (one drop). The reaction mixture was stirred at room temperature for 16h. Then the reaction mixture was concentrated in vacuo. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/ water/acetonitrile) to afford the desired product l-[5-methyl-l- [4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4-(l-tetrahydropyran-4-yl-4-piperidyl)piperazine (10.6 mg, yield 14.9%) as a white solid. [001377] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 100.0% (214 nm); Mass: found peak 494.1(M+H) at 2.01 min.
[001378] 'H NMR (400 MHz, MeOD-d4) 5 7.55 (d, J = 8.0 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 5.85 (s, 1H), 4.01 (d, J = 8.0 Hz, 2H), 3.40 (t, J = 12.0 Hz, 2H), 3.25 (t, J = 8.0 Hz, 8H), 2.74 (t, J = 8.0 Hz, 4H), 2.41 (s, 2H), 2.29 (s, 3H), 2.04 (d, J = 12.0 Hz, 2H), 1.88 (d, J = 12.0 Hz, 2H), 1.61 (t, J = 12.0 Hz, 4H) ppm.
Example S81. Synthesis of 4-[2-[4-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]- 1-piperidyl] ethyl] morpholine (Compound 81).
[001379] Compound 81 was prepared as outlined below.
Figure imgf000288_0001
[001380] Step 1. Synthesis of tert-butyl 4-(3-iodo-5-methyl-pyrazol-l-yl)piperidine-l- carboxylate.
Figure imgf000288_0002
[001381] To a solution of 3-iodo-5-methyl-lH-pyrazole (300 mg, 1.44 mmol) in N,N- dimethylformamide (25 mL) was added tert-butyl 4-methylsulfonyloxy- piperidine- 1- carboxylate (604mg, 2.16 mmol), cesium carbonate (1.17 g, 3.61 mmol), and the reaction mixture was stirred at 90 °C for 16h under the protection of N2. The mixture poured into icewater and partitioned with EtOAc (300 mL). The organic layers were washed with water three times, then NaCl solution once. The organic phase was separated, dried (Na2SO4) and concentrated in vacuo. The residue was flash chromatographed (Biotage, 80 g silica gel column @80mL/min, eluting with 5%-50% ethyl acetate in petroleum ether for 30 min) to afford tert- butyl 4-(3-iodo-5-methyl-pyrazol-l-yl)piperidine-l-carboxylate (210 mg, 35.9%) as a white solid.
[001382JLCMS method: Mobile Phase: WATER (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Gradient: from 10% to 95% of B ini.5 min at 1.8 mL/min; Column: X BRIGE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 96.6% (214 nm) Mass: found peak 335.9 (M-56+l)+ at 2.11 min.
[001383] Step 2. Synthesis of tert-butyl 4-[3-methyl-5-[4-(trifluoromethoxy)phenyl]pyrazol- l-yl]piperidine-l-carboxylate.
Figure imgf000289_0001
, ,
[001384] To a solution of tert-butyl 4-(5-iodo-3-methyl-pyrazol-l-yl)piperidine-l-carboxylate (50 mg, 0.128 mmol), [4-(trifluoromethoxy)phenyl]boronic acid (39.5 mg, 0.192 mmol), and potassium carbonate (35.3 mg, 0.256 mmol) in 1,4-dioxane/water (10 mL/1 mL) was added [1'1- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (9.35 mg, 0.013 mmol). The reaction was stirred at 80 °C under the protection of N2 for 16h. The solution was filtered, and the precipitate was washed with MeOH (15 mL), the solution was concentrated in vacuo to give a crude product, which was purified by flash chromatography (Biotage, 50 g silica gel column @70mL/min, eluting with 30%-80% ethyl acetate in petroleum ether for 30 min) to afford tertbutyl 4-[3-methyl-5-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine-l-carboxylate (60 mg, 79.6%) as a white solid.
[001385] LCMS method: Column: X BRIGE C18 (4.6x 50 mm, 3.5pm); Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B ini.5 min at 1.5 mL/min Temperature: 50°C; LC purity: 72.2% (214 nm) Mass: found peak 370.1 (M-56+l)+ at 2.306 min.
[001386] Step 3. Synthesis of 4-[5-methyl-3-[4-(trifluoromethoxy) phenyl]pyrazol-l- yl] piperidine.
Figure imgf000289_0002
[001387] To a solution of tert-butyl 4-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]piperidine-l -carboxylate (110 mg, 0.259 mmol) in dichloromethane (5 mL) was added TFA (2 mL, 26.9 mmol), The reaction mixture was stirred at RT for 2h. The mixture was concentrated, diluted with DCM (100 mL) and washed with aq potassium carbonate. The solution washed with aq NaCl solution, the solution concentrated in vacuo to afford 4-[5- methyl-3-[4-(trifluorom ethoxy) phenyl]pyrazol-l-yl]piperidine (80 mg, 80.5%) as a colorless oil.
[001388JLCMS method: Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Gradient: from 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO Cl 8 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 84.6% (214 nm) Mass: found peak 326.2 (M+l) at 1.075 min.
[001389] Step 4. Synthesis of 4-[2-[4-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]-l-piperidyl]ethyl]morpholine (Compound 81).
Figure imgf000290_0001
[001390] To a solution of 4-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]piperidine (80 mg, 0.246 mmol) in ethanol / water (10 mL/0.5 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (68.8 mg, 0.369 mmol), KI (40.8 mg, 0.246 mmol) and potassium carbonate (136 mg, 0.984 mmol). The reaction mixture was stirred at 90 °C for 16h under the protection of argon. The mixture was concentrated, diluted with 50 mL MeOH and filtered. The filter cake was washed with MeOH (5 mL), the filtrate was concentrated and the residue was purified by HPLC to afford 4-[2-[4-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l-yl]-l- piperidyl]ethyl]morpholine (27.1 mg, yield 24.8%) as a white solid.
[001391JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.4 min, 95%B for 1.6 min. Flow Rate: 1.8 mL/min; Column: X Bridge C18, 4.6*50mm, 3.5pm; Column Temperature: 45 °C; LC purity: 98.8% (214 nm) Mass: found peak 439.3 (M+l) at 1.95 min.
[001392] 'H NMR (400 MHz, DMSO-d6) 5 7.85 (d, J = 8.4Hz, 2H), 7.36 (d, J = 8.4Hz, 2H), 6.48 (s, 1H), 4.06-4.10 (m, 1H), 3.56 (t, J = 4.4 Hz, 4H), 3.00 (d, J = 4.4 Hz, 2H), 2.34-2.48 (m, 8H), 2.30 (s, 3H), 2.04-2.14 (m, 4H), 1.82 (d, J = 10.0 Hz, 2H).
Example S82. Synthesis of 4-[2-[3-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]pyrrolidin-l-yl]ethyl]morpholine (Compound 82).
[001393] Compound 82 was prepared as outlined below.
Figure imgf000291_0001
[001394] Step 1. Synthesis of tert-butyl 3-(3-iodo-5-methyl-pyrazol-l-yl)pyrrolidine-l- carboxylate.
B
Figure imgf000291_0002
[001395] To a solution of 3-iodo-5-methyl-lH-pyrazole (400 mg, 1.92 mmol) in N,N- dimethylformamide (20 mL) was added tert-butyl 3-methylsulfonyloxy-pyrrolidine-l- carboxylate (765mg, 2.88 mmol), and cesium carbonate (1.57 g, 4.81 mmol). The reaction mixture was stirred at 90 °C for 16h under the protection of N2, and then LCMS showed the reaction was complete. The mixture was poured into ice-water, EtOAc (300 mL) was added to the solution, the organic layers washed with water three times, then NaCl solution once, the solution concentrated and the mixture purified by flash chromatography (Biotage, 80 g silica gel column @80mL/min, eluting with 5%-50% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 3-(3-iodo-5-methyl-pyrazol-l-yl)pyrrolidine-l-carboxylate (350 mg, 45.4%) as a white solid.
[001396] LCMS method: Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B);
Gradient: from 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO Cl 8 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 94.1% (214 nm) Mass: found peak 322.1 (M-56+l)+ at 1.365 min.
[001397] Step 2. Synthesis of tert-butyl 3-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol- l-yljpyrrolidine-l-carboxylate.
Figure imgf000291_0003
[001398] To a solution of tert-butyl 3-(3-iodo-5-methyl-pyrazol-l-yl)pyrrolidine-l-carboxylate (100 mg, 0.265 mmol), [4-(trifluoromethoxy)phenyl]boronic acid (81.9 mg, 0.398 mmol), potassium carbonate (73.3 mg, 0.530 mmol) in 1,4-dioxane/water (5 mL/0.5 mL) was added [Tl-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (19.4 mg, 0.027 mmol). The reaction was stirred at 80 °C under the protection of N2 for 16h. LCMS showed the reaction was complete, the solution was filtered and the filter-cake washed with MeOH (10 mL), the solution concentrated in vacuo to give a crude product, which was purified by flash chromatography (Biotage, 50 g silica gel column @70mL/min, eluting with 30%-80% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 3-[5-methyl-3-[4-(trifluoromethoxy) phenyl]pyrazol-l- yl]pyrrolidine-l -carboxylate (110 mg, 92.1%) as a white solid.
[001399] LCMS method: Column: X Bridge Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: WATER (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 5% to 95% of B in 1.5 min at 1.5 mL/min; Temperature: 50 °C; LC purity: 91.3% (214 nm) Mass: found peak 412.1 (M+l) at 2.339 min.
[001400] Step 3. Synthesis of 5-methyl-l-pyrrolidin-3-yl-3-[4- (trifluoromethoxy)phenyl]pyrazole.
Figure imgf000292_0001
[001401] To a solution of tert-butyl 3-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]pyrrolidine-l -carboxylate (110 mg, 0.267 mmol) in dichloromethane (5 mL) was added TFA (2 mL, 26.9 mmol). The reaction mixture was stirred at RT for 2h. The mixture was concentrated, DCM (100 mL) was added to the mixture, the solution washed with potassium carbonate (aq), the solution washed with NaCl solution (aq), the organic layer was separated, and concentrated in vacuo to afford 5-methyl-l-pyrrolidin-3-yl-3-[4- (trifluoromethoxy)phenyl]pyrazole (80 mg, 86.1%) as a colorless oil.
[001402JLCMS method: Mobile Phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B); Gradient: from 5% to 95% of B in 1.0 min at 2.2 mL/min; Column: HALO Cl 8 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 89.6% (214 nm) Mass: found peak 312.3 (M+l) at 1.093 min.
[001403] Step 4. Synthesis of 4-[2-[3-[5-methyl-3-[4-(trifluoromethoxy)phenyl]pyrazol-l- yl]pyrrolidin-l-yl]ethyl]morpholine (Compound 82).
Figure imgf000293_0001
[001404] To a solution of 5-methyl-l-pyrrolidin-3-yl-3-[4-(trifluoromethoxy)phenyl]pyrazole (80 mg, 0.257 mmol) in ethanol / water (10 mL/0.5 mL) were added 4-(2- chloroethyl)morpholine;hydrochloride (71.7 mg, 0.385 mmol), KI (42.7 mg,0.257mmol) and potassium carbonate (142 mg, 1.03 mmol). The reaction mixture was stirred at 90 °C for 16h under the protection of argon, the mixture was concentrated, MeOH was added to the mixture, the solution filtered, and the filter cake washed with MeOH (5mL). The filtrate was concentrated and the residue was purified by HPLC to afford 4-[2-[3-[5-methyl-3-[4- (trifluoromethoxy)phenyl]pyrazol-l-yl]pyrrolidin-l-yl]ethyl]morpholine (70.9 mg, yield 61.9%) as a colorless oil.
[001405] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min Flow Rate: 1.8 mL/min; Column: X bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 95.3% (214 nm) Mass: found peak 424.9 (M+l) at 1.911 min.
[001406] 'H NMR (400 MHz, DMSO-d6) 5 7.85 (d, J = 8.8Hz, 2H), 7.36 (d, J = 8.4Hz, 2H), 6.50 (s, 1H), 4.84 (t, J = 6.8 Hz, 1H), 3.54 (t, J = 4.4 Hz, 4H), 3.35 (s, 1H), 3.10 (dt, J = 16.8, 8.0 Hz, 1H), 2.52-2.83 (m, 4H), 2.38-2.43 (m, 6H), 2.33 (s, 3H), 2.20-2.26 (m, 2H) ppm.
Example S83. Synthesis of 4-[2-[4-[5-(difluoromethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l piperidyl]ethyl]morpholine (Compound 83). [001407] Compound 83 was prepared as outlined below.
Figure imgf000293_0002
[001408] Step 1. Synthesis of 4, 4-difluoro-l-(4-pyridyl)butane-l, 3-dione.
Figure imgf000294_0001
tBuOMe
[001409] A mixture of l-(4-pyridyl)ethanone (970mg, 8.01 mmol), methyl 2,2- difluoroacetate (970 mg, 8.81 mmol) and sodium methoxide (30% w/w in MeOH, 1.7 g, 9.61 mmol ) in methyl-tert-butyl ether (5 mL) was stirred at rt for 16 h. To the mixture was added a 10% citric acid aqueous solution until the reaction solution was pH 3~4. The precipitate was collected by filtration, washed with water and dried to obtain the title compound 4,4-difluoro-l- (4-pyridyl)butane-l,3-dione (800 mg, 48.9% yield) as a yellow solid.
[001410JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.51% (214 nm), Mass: 200.0 found peak (M+l) at 1.140 min.
[001411] Step 2. Synthesis of 4-[3-(difluoromethyl)-lH-pyrazol-5-yl]pyridine.
Figure imgf000294_0002
[001412] 4,4 -difluoro- l-(4-pyridyl)butane- 1,3 -di one (300 mg, 1.47 mmol) was suspended in ethanol (10 mL), hydrazinium hydroxide (110 mg, 2.2 mmol) was added and the mixture was stirred at 85°C for 5 h, then were added molecular sieves (50 mg), and the reaction mixture was stirred another 5 h at 85 °C. The mixture was filtered, the filtrate was concentrated in vacuo and the residue was purified by flash chromatography eluting with 0-20% dichloromethane in methanol to afford the desired product 4-[3-(difluoromethyl)-lH-pyrazol-5-yl]pyridine (180 mg, yield 54.7%) as a yellow solid
[001413] LCMS Method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C;. LC purity: 87.05% (214 nm), Mass: found peak 196.3 (M+l) at 0.696 min.
[001414] Step 3. Synthesis of 4-[5-(difluoromethyl)-lH-pyrazol-3-yl]piperidine.
Figure imgf000294_0003
[001415] 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]pyridine (320 mg, 1.38 mmol) was dissolved in ethanol (3 mL). Hydrochloric acid solution (1 M, 5.53 mL; 5.53 mmol) was added and the mixture was hydrogenated with platinum oxide hydrate (80% Pt, 60 mg) at atmospheric pressure and room temperature for 16 h. The reaction mixture was filtered and concentrated in vacuo to give the product 4-[5-(difluoromethyl)-lH-pyrazol-3-yl]piperidine (280 mg, 92.4%) as a yellow solid.
[001416] Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm;
Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 201.9 (M+l) at 0.359 min.
[001417] Step 4. Synthesis of tert-butyl 4-[5-(difluoromethyl)-lH-pyrazol-3-yl]piperidine-l- carboxylate.
Figure imgf000295_0001
[001418] 4-[5-(difluoromethyl)-lH-pyrazol-3-yl]piperidine (160 mg; 0.795 mmol) was dissolved in water (2.0 mL). Sodium bicarbonate (200 mg, 2.39 mmol) and tert-butoxycarbonyl tert-butyl carbonate (260 mg, 1.19 mmol) dissolved in dioxane (4.0 mL) were added and the mixture was stirred for 2 h at room temperature. The reaction mixture was diluted with water and extracted with dichloromethane (20 mL * 3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography 5- 40% acetone in petroleum ether to afford tert-butyl 4-[5-(difluoromethyl)-lH-pyrazol-3- yl]piperidine-l -carboxylate (180 mg, 75.1% yield) as a white solid.
[001419JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity:85.98% (214 nm) Mass: found peak 246.2 (M-55)+ at 1.917 min.
[001420] Step 5. Synthesis of tert-butyl 4-[5-(difluoromethyl)-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl] piperidine- 1-carboxylate.
Figure imgf000295_0002
[001421] To a solution of tert-butyl 4-[5-(difluoromethyl)-lH-pyrazol-3-yl]piperidine-l- carboxylate (180 mg, 0.597 mmol) in dichloromethane (10 mL) were added [4- (trifluoromethoxy)phenyl]boronic acid (246 mg, 1.19 mmol), anhydrous copper acetate (217 mg, 1.19 mmol), pyridine (236 mg, 2.99 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 48 h. The mixture was filtered, then concentrated in vacuo. The residue was purified by prep HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product tert-butyl 4-[5-(difluoromethyl)-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]piperidine-l -carboxylate (100 mg, yield 36.3%) as a yellow oil.
[001422JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 461.9 (M+l) at 2.047 min.
[001423] Step 6. Synthesis of 4-[5-(difluoromethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine.
Figure imgf000296_0001
[001424] To a solution of tert-butyl 4-[5-(difhioromethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (100 mg, 0.217 mmol) in dichloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h, then concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 4-[5-(difhioromethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (70 mg, yield 80.3%) as a yellow oil.
[001425] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity:89.78% (214 nm), Mass: found peak 362.1 (M+l) at 1.585 min.
[001426] Step 7. Synthesis of 4-[2-[4-[5-(difluoromethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l piperidyl]ethyl]morpholine (Compound 83).
Figure imgf000297_0001
[001427] To a solution of 4-[5-(difluoromethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine (70 mg, 0.174 mmol), potassium carbonate (120 mg, 0.87 mmol) and KI (29 mg, 0.261 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (49 mg, 0.261 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered, then concentrated in vacuo. The residue was purified by prep-
HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-(difluoromethyl)- l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l piperidyl]ethyl]morpholine (48.6 mg, yield 54.2%) as a yellow oil.
[001428JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B:
ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C. LC purity: 92.54% (214 nm) Mass: found peak 475.0 (M+l) at 2.004 min.
[001429] 'H NMR (400 MHz, MeOD-d4) 5 7.66-7.58 (m, 2H), 7.54-7.49 (m 2H), 6.92-6.61 (m, 1H), 6.59 (s, 1H), 3.70 (t, J= 4.4 Hz, 4H), 3.13-2.99 (m, 2H), 2.80-2.67 (m, 1H), 2.58-2.47 (m, 8H), 2.09-2.00 (m, 2H), 1.91-1.67 (m, 4H) ppm.
Example S84. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 84). [001430] Compound 84 was prepared as outlined below.
Figure imgf000297_0002
[001431] Step 1. Synthesis of 4,4,4-trifluoro-l-(4-pyridyl)butane-l,3-dione.
Figure imgf000298_0001
[001432] A mixture of l-(4-pyridyl)ethanone (2.6 g, 21.5 mmol), ethyl 2,2,2-trifluoroacetate (3.35 g, 23.6 mmol) and NaOMe (30% w/w in MeOH, 4.6 g, 25.8 mmol ) in methyl-tert-butyl ether (5 mL) was stirred at rt for 16 h. Next, 10% citric acid aqueous solution was added until the reaction solution became about pH=4. The precipitate was collected by filtration, washed with water, and dried to obtain the title compound 4,4,4-trifluoro-l-(4-pyridyl)butane-l,3-dione (4.0 g, 73.9% yield) as a yellow solid.
[001433JLCMS method: Mobile Phase: A: water (10 mmol NH4HCO3) B: MeCN; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 ml/min; Column: X-Bridge C18, 50*4.6mm, 3.5um; Column Temperature: 50 °C; LC purity: 86.11% (214 nm) Mass: 218.0 (M+l)+ at 1.417 min.
[001434] Step 2. Synthesis of 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]pyridine.
Figure imgf000298_0002
[001435] 4,4,4 -trifluoro- l-(4-pyridyl)butane-l, 3-dione (1.0 g; 3.97 mmol) was suspended in ethanol (20 mL), hydrazinium hydroxide (298 mg, 5.95 mmol) and molecular sieves (100 mg) were added, and the mixture was stirred at 85°C for 24 h. The reaction mixture was filtered, concentrated in vacuo and the residue purified by flash chromatography 20-75% ethyl acetate in petroleum ether to afford 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]pyridine (900 mg, 98.0% yield) as a white solid.
[001436] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 92.06% (214 nm) Mass: found peak 214.0 (M+l) at 1.744 min.
[001437] Step 3. Synthesis of 4-(3-Methyl-lH-pyrazol-5-yl)piperidine.
Figure imgf000298_0003
[001438] 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]pyridine (320 mg; 1.38 mmol) was dissolved in ethanol (3 mL). Hydrochloric acid solution (1 M, 5.53 mL; 5.53 mmol) was added and the mixture was hydrogenated with platinum oxide hydrate (80% Pt, 60 mg) at atmospheric pressure and room temperature for 16 h. The reaction mixture was filtered and concentrated in vacuo to give the product (280 mg, 92.4%) as a yellow solid.
[001439JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 220.1 (M+l) at 1.468 min.
[001440] Step 4. Synthesis of tert-butyl 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]piperidine- 1-carboxylate. Boc
Figure imgf000299_0001
[001441] 4-(3-Methyl-lH-pyrazol-5-yl)piperidine (230 mg; 0.933 mmol) was dissolved in water (3.0 mL). Sodium bicarbonate (235 mg; 2.8 mmol) and tert-butoxy carbonyl tert-butyl carbonate (305 mg; 1.4 mmol) dissolved in dioxane (6.0 mL) were added and the mixture was stirred for 2 h at room temperature. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography 0-27% acetone in petroleum ether to afford tert-butyl 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]piperidine-l- carboxylate (300 mg, 86.6% yield) as a white solid.
[001442JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 85.98%(214 nm), Mass: found peak 264.2 (M-55)+ at 1.326 min.
[001443] Step 5. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]piperidine-l-carboxylate.
Figure imgf000299_0002
[001444] To a solution of tert-butyl 4-[3-(trifluoromethyl)-lH-pyrazol-5-yl]piperidine-l- carboxylate (300 mg, 0.808 mmol) in dichloromethane (10 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (333 mg, 1.62 mmol), anhydrous copper acetate (293 mg, 1.62 mmol), pyridine (320 mg, 4.04 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16 h. The mixture was filtered, then concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100 mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol-3-yl]piperidine- 1-carboxylate (250 mg, yield 60.8%) as a colorless oil.
[001445] LCMS method: Mobile Phase: A:Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C. LC purity: 94.13% (214 nm), Mass: found peak 480.1 (M+l) at 2.347 min.
[001446] Step 6. Synthesis of 4-[l-[4-(trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol- 3-yl]piperidine.
Figure imgf000300_0001
[001447] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]piperidine-l -carboxylate (120 mg, 0.236 mmol) in dichloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH = 8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 4-[l-[4-(trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol-3-yl]piperidine (100 mg, yield 92.1%) as a yellow oil.
[001448JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 82.33% (214 nm), Mass: found peak 380.2 (M+l) at 1.122 min.
[001449] Step 7. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 84).
Figure imgf000301_0001
[001450] To a solution of 4-[l-[4-(trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol-3- yl]piperidine (100 mg, 0.217 mmol), potassium carbonate (150 mg, 1.09 mmol) and KI (36 mg, 0.217 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (61 mg, 0.326 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[l-[4- (trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (76.4 mg, yield 75.8%) as a yellow oil.
[001451JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.95% (214 nm) Mass: found peak 493.1 (M+l) at 2.121 min.
[001452] 'H NMR (400 MHz, DMSO-t/e) 5 7.74-7.68 (m, 2H), 7.62-7.56 (m, 2H), 6.89 (s, 1H), 3.53 (t, J= 4.4 Hz, 4H), 2.91-2.85 (m, 2H), 2.65-2.55 (m, 1H), 2.39-2.30 (m, 8H), 1.85 (t, J= 10.8 Hz, 2H), 1.76-1.68 (m, 2H), 1.66-1.54 (m, 2H) ppm.
Example S85. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 85).
[001453] Compound 85 was prepared as outlined below.
Figure imgf000301_0002
[001454] Step 1. Synthesis of tert-butyl 2-methyl-4-(trifluoromethylsulfonyloxy)-3,6- dihydro-2H-pyridine-l-carboxylate.
Figure imgf000302_0001
-78°C - 0 °C, 5 h
[001455] To a solution of tert-butyl 2-methyl-4-oxo-piperidine-l -carboxylate (427 mg, 2 mmol) in THF (6 mL) was added LiHMDS (IM, 2.2 mL, 2.2 mmol) dropwise at -78 °C, After 1 h. l,l,l-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (858 mg, 2.4 mmol) in THF (3 mL) was added dropwise, the temperature was gradually increased to rt and the resulting mixture was stirred for 5 h. The reaction mixture was concentrated in vacuo (below 40°C) and extracted with EtOAc (20 mL*3), dried over sodium sulfate, filtered, and purified by flash chromatography 2-20% ethyl acetate in petroleum ether to afford tert-butyl 2-methyl-4- (trifluoromethylsulfonyloxy)-3,6-dihydro-2H-pyridine-l-carboxylate (600 mg, 54.9% yield) as a yellow oil.
[001456] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 63.25% (214 nm) Mass: found peak 290.1 (M-55)+ at 2.220 min.
[001457] Step 2. Synthesis of tert-butyl 2-methyl-4-(trifluoromethylsulfonyloxy)-3,6- dihydro-2H-pyridine-l-carboxylate.
Figure imgf000302_0002
dioxane, 110°c, 16h
[001458] Under argon atmosphere, a mixture of tert-butyl 2-methyl-4-(trifluoromethyl- sulfonyloxy)-3,6-dihydro-2H-pyridine-l-carboxylate (600 mg, 1.1 mmol), 4,4,5,5-tetramethyl-2- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (335 mg, 1.32 mmol), Pd(dppf)C12 (81 mg, 0.11 mmol) and potassium acetate (324 mg, 3.3 mmol) was stirred in 1,4- di oxane (10 mL) at 110 °C for 16 h. The reaction mixture was filtered and purified by flash chromatography 2-20% acetone in petroleum ether to afford tert-butyl 2-methyl-4- (trifluoromethylsulfonyloxy)-3,6-dihydro-2H-pyridine-l-carboxylate (360 mg, 89.3% yield) as a yellow oil. [001459JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 88.13% (214 nm), Mass: found peak 268.3 (M-55)+ at 1.540 min.
[001460] Step 3. Synthesis of tert-butyl 2-methyl-4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000303_0001
[001461] Under argon atmosphere, a mixture of 3-iodo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (232 mg, 0.436 mmol), tert-butyl 2-methyl-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l -carboxylate (160 mg, 0.436 mmol), Pd(dppf)C12 (32 mg, 0.043 mmol) and potassium carbonate (181 mg, 1.13 mmol) was stirred in 1,4-dioxane / water (3 mL / 0.5 mL) at 80 °C for 16 h. The filtrate was purified by flash chromatography (Biotage, 80g silica gel column @100 mL/min, eluting with 10-30% di chloromethane in petroleum ether) to afford the desired product tert-butyl 2-methyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l-carboxylate (90 mg, yield 35.6%) as a yellow solid.
[001462JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 75.59% (214 nm), Mass: found peak 438.1 (M+l) at 2.370 min.
[001463] Step 4. Synthesis of tert-butyl 2-methyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate.
Figure imgf000303_0002
[001464] Tert-butyl 2-methyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,6- dihydro-2H-pyridine-l -carboxylate (90 mg, 0.156 mmol) was dissolved in methanol (3 mL), and the mixture was hydrogenated with Pd/C (10%, 50 mg) at atmospheric pressure and room temperature for 0.5 h. The mixture was filtered to afford the desired product tert-butyl 2-methyl- 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate (95 mg, yield 95.4%) as a yellow oil.
[001465] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 68.64% (214 nm), Mass: found peak 440.3 (M+l) at 1.594 min.
[001466] Step 5. Synthesis of 2-methyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine.
Figure imgf000304_0001
[001467] To a solution of tert-butyl 2-methyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (50 mg, 0.113 mmol) in dichloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 2-methyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (50 mg, yield 75.9%) as a yellow oil.
[001468JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity:76.46% (214 nm) Mass: found peak 340.3 (M+l) at 1.090 min.
[001469] Step 6. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5- (trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 85).
Figure imgf000304_0002
[001470] To a solution of 2-methyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine (50 mg, 0.11 mmol), potassium carbonate (78 mg, 0.56 mmol) and KI (19 mg, 0.11 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (32 mg, 0.17 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo, and the residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[l-[4- (trifluoromethoxy)phenyl]-5-(trifluoromethyl)pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (17 mg, yield 33.3%) as a yellow oil.
[001471JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 452.9 (M+l) at 1.854 min.
[001472] 'H NMR (400 MHz, MeOD-d4) 5 7.48-7.43 (m, 2H), 7.36-7.31 (m, 2H), 6.09-6.04 (m, 1H), 3.63-3.57 (m, 4H), 3.07-2.88 (m, 2H), 2.71-2.56 (m, 1H), 2.55-2.26 (m, 8H), 2.23 (s, 3H), 1.99- 1.50 (m, 4H), 1.25-1.17 (m, 1H), 1.14-1.01 (m, 3H) ppm.
Example S86. Synthesis of 4-[2-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 86). [001473] Compound 86 was prepared as outlined below.
Figure imgf000305_0001
[001474] Step 1. Synthesis of l-benzyl-2,6-dimethyl-piperidin-4-one.
Figure imgf000305_0002
[001475] To a stirring solution of 3 -oxopentanedioic acid (5.0 g, 34.2 mmol) in water (15 mL), acetaldehyde (3.02 g, 68.4 mmol) was added and stirred at 0 °C for 20 min. To this solution, benzylamine (3.67 g, 34.2 mmol) was added slowly. The reaction mixture was stirred at RT for 72 h. The mixture was filtered, and diluted with water (20 mL), then extracted with ethyl acetate (50 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo, the residue was purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-60% ethyl acetate in petroleum ether) to afford the desired product l-benzyl-2,6-dimethyl- piperidin-4-one (350 mg, yield 3.34%) as a yellow oil.
[001476] Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.4 min; 95% B for 1.6 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C; LC purity: 71.02% (214 nm), Mass: found peak 218.3 (M-55)+ at 1.893 min.
[001477] Step 2. Synthesis of tert-butyl 2,6-dimethyl-4-oxo-piperidine-l-carboxylate.
Figure imgf000306_0001
isopropyl alcohol
48 h
[001478] l-benzyl-2,6-dimethyl-piperidin-4-one (350 mg; 1.61 mmol) was dissolved in propan- 2-ol (5.0 mL). Tert-butoxy carbonyl tert-butyl carbonate (884 mg; 4.05 mmol) was added and the mixture was hydrogenated with Pd/C (10%, 200 mg) at atmospheric pressure and room temperature for 48 h. The reaction mixture was filtered and purified by flash chromatography 0- 30% acetone in petroleum ether to afford tert-butyl 2,6-dimethyl-4-oxo-piperidine-l -carboxylate (350 mg, 40.3% yield) as a yellow solid.
[001479JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 42.17% (214 nm) Mass: found peak 172.3 (M-55)+ at 1.211 min.
[001480] Step 3. Synthesis of tert-butyl 2,6-dimethyl-4-(trifluoromethylsulfonyloxy)-3,6- dihydro-2H-pyridine-l-carboxylate.
Figure imgf000306_0002
[001481] To a solution of tert-butyl 2,6-dimethyl-4-oxo-piperidine-l -carboxylate (600 mg, 2.64 mmol) in THF (6 mL) was added LHMDS (2.9 mL, 2.9 mmol) dropwise at -78 °C. After 1 h, l,l,l-trifhioro-N-phenyl-N-(trifhroromethylsulfonyl) methanesulfonamide (1.13 g, 3.17 mmol) in THF (3 mL) was added dropwise to the reaction mixture, the temperature was gradually increased to 0 °C and the resulting mixture was stirred for 4 h. The reaction mixture was concentrated in vacuo (below 40 °C) and extracted with EtOAc (20 mL*3), dried over sodium sulfate, filtered, and purified by flash chromatography 0-10% ethyl acetate in petroleum ethe to afford tert-butyl 2,6-dimethyl-4-(trifluoromethylsulfonyloxy)-3,6-dihydro-2H-pyridine-l- carboxylate (800 mg, 36.6% yield) as a yellow oil.
[001482JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in l.Omin; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 43.40% (214 nm) Mass: found peak 304.1 (M-55)+ at 1.503 min.
[001483] Step 4. Synthesis of tert-butyl 2,6-dimethyl-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000307_0001
dioxane, 110 c, 16h
[001484] Under argon atmosphere, a mixture of tert-butyl 2,6-dimethyl-4- (trifluoromethylsulfonyloxy)-3,6-dihydro-2H-pyridine-l-carboxylate (500 mg, 1.39 mmol), 4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (424 mg, 1.67 mmol), Pd(dppf)C12 (102 mg, 0.139 mmol) and potassium acetate (410 mg, 4.17 mmol) was stirred in 1,4-di oxane (10 mL) at 110 °C for 16 h. The mixture was filtered, concentrated in vacuo and the residue was purified by flash chromatography 2-20% acetone in petroleum ether to afford tert-butyl 2,6-dimethyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-3,6-dihydro-2H-pyridine-l-carboxylate (410 mg, 70.6% yield) as a yellow oil.
[001485] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in l.Omin; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 80.81% (214 nm) Mass: found peak 282.3 (M-55)+ at 1.577 min.
[001486] Step 5. Synthesis of tert-butyl 2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000308_0001
[001487] Under argon atmosphere, a mixture of tert-butyl 2,6-dimethyl-4-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l-carboxylate (280 mg, 0.83 mmol), 3-iodo- 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (336 mg, 0.913 mmol), Pd(dppf)C12 (62 mg, 0.083 mmol) and potassium carbonate (229 mg, 1.66 mmol) was stirred in 1,4-dioxane / water ( 10 mL / 1 mL) at 80 °C for 16 h. The filtrate was concentrated in vacuo, and the residue was purified by flash chromatography (Biotage, 40g silica gel column @60 mL/min, eluting with 0- 30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 2,6-dimethyl-4-[5- methyl-l-[4-(tri fluoromethoxy )phenyl]pyrazol-3-yl]-3,6-dihydro-2H-pyri dine- 1 -carboxylate (110 mg, yield 40.3%) as a yellow solid.
[001488JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in l.Omin; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 91.07% (214 nm), Mass: found peak 452.3 (M+l) at 1.635 min.
[001489] Step 6. Synthesis of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-8-azabicyclo[3.2.1]octane-8-carboxylate.
Figure imgf000308_0002
[001490] Tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo[3.2.1]oct-2-ene-8-carboxylate (110 mg, 0.244 mmol) was dissolved in methanol (5 mL), and the mixture was hydrogenated with Pd/C (10%, 90 mg) at atmospheric pressure and room temperature for 0.5 h. The mixture was filtered, and concentrated in vacuo to afford the desired product tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo[3.2.1]octane-8-carboxylate (85 mg, yield 73.3%) as a yellow oil.
[001491JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.4 min; 95% B for 1.6 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C. LC purity: 95.24% (214 nm), Mass: found peak 454.1 (M+l) at 2.415 min.
[001492] Step 7. Synthesis of 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] - 1-piperidyl] ethenone.
Figure imgf000309_0001
[001493] A mixture of tert-butyl 2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (85 mg, 0.187 mmol) in DCM (3 mL) was added TFA (1.0 mL), and the mixture was stirred overnight at room temperature for 0.5 h. The mixture was concentrated, then dissolved in DCM (3 mL), followed by addition of TEA (190 mg, 1.87 mmol) and 2-chloroacetyl chloride (64 mg, 0.56 mmol) at 0 °C. The mixture was stirred overnight at room temperature. DCM (5 mL) was added and washed with water (3 mL). The organic layer was concentrated and purified by flash chromatography (Biotage, 40g silica gel column @80 mL/min, eluting with 10-30% ethyl acetate in petroleum ether) to afford the desired product 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethanone (70 mg, yield 86.1%) as a yellow solid.
[001494JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 99.09% (214 nm) Mass: found peak 430.0 (M+l) at 2.121 min.
[001495] Step 8. Synthesis of l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]-2-morpholino-ethanone.
Figure imgf000309_0002
[001496] A mixture of 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethanone (60 mg, 0.14 mmol) and morpholine (0.14 mmol) in dichloromethane (4.0 mL) was stirred for 2 h at room temperature. The mixture was concentrated and residue purified by flash chromatography (Biotage, 40g silica gel column @80 mL/min, eluting with 10-10% methanol in di chloromethane) to afford the desired product l-[2,6-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l- piperidyl]-2-morpholino-ethanone (65 mg, yield 95.5%) as a yellow oil.
[001497] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 98.58% (214 nm) Mass: found peak 481.1 (M-55)+ at 2.062 min.
[001498] 'H NMR (400 MHz, MeOD-d4) 5 7.60-7.55 (m, 2H), 7.44 (d, J = 8.6 Hz, 2H), 6.21 (s, 1H), 4.43 (s, 1H), 4.13 (s, 1H), 3.70 (t, J = 4.4 Hz, 4H), 3.35-3.31 (m, 1H), 3.15 (d, J = 14.6 Hz, 1H), 2.60-2.46 (m, 4H), 2.40-2.27 (m, 4H), 2.27-1.91 (m, 4H), 1.44-1.33 (m, 6H) ppm.
[001499] Step 9. Synthesis of 4-[2-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound 86).
Figure imgf000310_0001
[001500] A mixture of l-[2,6-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]-2-morpholino-ethanone (40 mg, 0.083 mmol) in BHrTHF (2 mL) was stirred at 80°C overnight. The reaction mixture was quenched by MeOH (0.5 mL), and the reaction mixture was concentrated. Then the crude was dissolved in HCI (4N, 1 mL), reflux for 4 h. The reaction was cooled to room temperature and filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[2,6-dimethyl-4-[5-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (14.0 mg, yield 35.2%) as yellow oil.
[001501JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.63% (254 nm) Mass: found peak 467.1 (M+l) at 2.163 min. [001502] XH NMR (400 MHz, MeOD-d4) 5 7.60-7.52 (m, 2H), 7.47-7.40 (m, 2H), 6.15 (s, 1H), 3.70 (t, J = 4.4 Hz, 4H), 3.36-3.31 (m, 1H) 3.14-3.01 (m, 1H), 2.99-2.87 (m, 2H), 2.67-2.45 (m, 7H), 2.30 (s, 3H), 1.95 (td, J = 13.0., 4.8 Hz, 1H), 1.87-1.74 (m, 2H), 1.47 (dd, J = 24.8, 12.4 Hz, 1H), 1.17 (m, 6H) ppm.
Example S87. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 8-azabicyclo[3.2.1] octan-8-yl]ethyl]morpholine (Compound 87-P1 and Compound 87-P2).
[001503] Compound 87 (87-P1 and 87-P2) was prepared as outlined below.
Figure imgf000311_0001
[001504] Step 1. Synthesis of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-amine.
Figure imgf000311_0002
[001505] To a solution of 5-methyl-lH-pyrazol-3-amine (4.85 g, 50 mmol) in N,N- dimethylformamide (50 mL) was added l-iodo-4-(trifluoromethoxy)benzene (15.8 g, 54.9 mmol), cesium carbonate (32.6 g, 100 mmol), and copper(I) iodide (951 mg, 5 mmol). The reaction mixture was stirred at 110 °C for 4 h. The mixture was filtered, diluted with water (50 mL), then extracted with ethyl acetate (100 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100 mL/min, eluting with 10-60% ethyl acetate in petroleum ether) to afford the desired product 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-amine (1.1 g, yield 8.24%) as a yellow oil. [001506] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 81.88% (214 nm), Mass: found peak 258.1 (M— 55)+ at 1.790 min.
[001507] Step 2. Synthesis of 3-iodo-5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000312_0001
[001508] To a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-amine (1.1 g; 3.49 mmol) in concentrated HC1 solution (10 mL) was added a solution of sodium nitrite (32 mg;
0.453 mmol) in water (0.5 mL) over 3 min at 0°C. A solution of potassium iodide (83 mg; 0.495 mmol) in water (0.5 mL) was added to the reaction mixture over 5 min, and the reaction was stirred for 15 min. The reaction mixture was diluted with water, extracted with EtOAc (20 mL * 3), and washed with Na2S2Ch two times. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography using 0-20% ethyl acetate in petroleum ether to afford 3-iodo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (800 mg, 43.1% yield) as a yellow oil.
[001509JLCMS method: Mobile phase: A: water (0.1%TFA), B: Acetonitrile (0.1% TFA); Gradient: 10% increase to 90%B within 1.3 min, 90%B for 1.5min; Flow Rate:2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 69.20% (214 nm) Mass: found peak 369.0 (M+l) at 2.251 min.
[001510] Step 3. Synthesis of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-8-azabicyclo[3.2.1]oct-2-ene-8-carboxylate.
Figure imgf000312_0002
[001511]Under argon atmosphere, a mixture of 3-iodo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (220 mg, 0.414 mmol), tert-butyl 4-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l-carboxylate (208 mg, 0.62 mmol),
Pd(dppf)C12 (31 mg, 0.041 mmol) and potassium carbonate (114 mg, 0.83 mmol) was stirred in 1,4-di oxane / water (3 mL / 0.5 mL) at 80 °C for 16 h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-20% ethyl acetate in petroleum ether) to afford the desired product, tert-butyl 3-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-8-azabicyclo[3.2.1]oct-2-ene-8-carboxylate (190 mg, yield 86.9%) as a yellow solid.
[001512JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 85.06% (214 nm), Mass: found peak 393.9 (M-55)+ at 2.150 min.
[001513] Step 4. Synthesis of Pl and P2.
Figure imgf000313_0001
P1 P2
[001514] Tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo[3.2.1]oct-2-ene-8-carboxylate (190 mg, 0.36 mmol) was dissolved in methanol (3 mL), and the mixture was hydrogenated with Pd/C (10%, 100 mg) at atmospheric pressure and room temperature for 0.5 h. The mixture was filtered, concentrated in vacuo, and purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to give the desired product Pl (62 mg, yield 32.6%) and P2 (70 mg, yield 42.0%) as a yellow oil.
[001515] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 85.26% (214 nm), Mass: found peak 452.3 (M+l) at 2.363 min; LC purity: 97.48% (214 nm), Mass: found peak 452.3 (M+l) at 2.437 min.
[001516] Step 5. Synthesis of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo [3.2.1] octane.
Figure imgf000314_0001
[001517] To a solution of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo[3.2.1]octane-8-carboxylate (62 mg, 0.121 mmol) in di chloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 3-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-8-azabicyclo[3.2.1]octane (45 mg, yield 87.8%) as a yellow oil
[001518JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 83% (214 nm) Mass: found peak 352.2 (M+l) at 1.602 min.
[001519] Step 6. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-8-azabicyclo[3.2.1] octan-8-yl] ethyl] morpholine (Compound 87-P1 and 87-P2).
Figure imgf000314_0002
[001520] To a solution of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8- azabicyclo[3.2.1]octane (45 mg, 0.106 mmol), potassium carbonate (74 mg, 0.531 mmol) and KI (18 mg, 0.106 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (30 mg, 0.159 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo and the residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2- [3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-8-azabicyclo[3.2.1] octan-8- yl]ethyl]morpholine (87-P1, 11.2 mg, yield 21.7%) as a yellow oil. [001521] 87-P2 was prepared following the same method as for 87-P1).
[001522JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C.
[001523] 87-P1: LC purity: 95.73% (254 nm) Mass: found peak 465.1 (M+l) at 1.978 min.
[001524] 87-P2: LC purity: 100% (214 nm) Mass: found peak 465.1 (M+l) at 2.094 min.
[001525] 87-P1: 'H NMR (400 MHz, MeOD-d4) 5 7.59-7.55 (m, 2H), 7.48-7.43 (m, 2H), 6.19 (s, 1H), 3.72 (t, J=4.4 Hz, 4H), 3.47-3.40 (m, 2H), 3.13-3.03 (m, 1H), 2.74-2.47 (m, 8H), 2.32 (s, 3H), 2.16-2.06 (m, 3H), 2.00-1.87 (m, 3H), 1.84-1.72 (m, 4H) ppm.
[001526] 87-P2: ‘HNMR (400 MHz, MeOD-d4) 5 7.64-7.57 (m, 2H), 7.48-7.43 (m, 2H), 6.28 (s, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.33-3.32 (m, 2H), 3.09-3.02 (m, 1H), 2.66-2.48 (m, 8H), 2.38- 2.22 (m, 7H), 1.88-1.78 (m, 2H), 1.72-1.63 (m, 2H) ppm.
Example S88. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]ethyl]morpholine (Compound 88).
[001527] Compound 88 was prepared as outlined below.
Figure imgf000315_0001
[001529] To a solution of methyl pyridine-4-carboxylate (1 g, 7.29 mmol) in ether (20 mL) was added sodium methoxide (394 mg, 7.29 mmol). Then a solution of methylbutan-2-one (3.14 g, 36.5 mmol) in ether (20 mL) was added. The suspension was stirred at reflux for 6 h, cooled, and filtered. The isolated solid was washed with ether and dissolved in water (40 mL). The mixture was added glacial acetic acid (5.2 mL) and extracted with chloroform (40 mL x 2). The organic layers were dried over anhydrous sodium sulfate and purified by SGC (petroleum ether: ethyl acetate = 2:1), concentrated to afford the desired product 4-methyl-l-(4- pyridyl)pentane-l, 3-dione (380 mg, 27.3% yield) as a colorless oil.
[001530] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 96% (214 nm) Mass: found peak 192.0 (M+l) at 1.686 min.
[001531] Step 2. Synthesis of 4-(3-isopropyl-lH-pyrazol-5-yl)pyridine.
Figure imgf000316_0001
[001532] To a solution of 4-methyl-l-(4-pyridyl)pentane-l, 3-dione (380 mg, 2 mmol) in ethanol (10 mL) was added hydrazinium hydroxide (150 mg, 3 mmol) and the mixture was stirred at 85°C for 48 h. The reaction mixture was diluted with saturated sodium bicarbonate solution (pH 7-8) and extracted with dichloromethane (20 mL x 2). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-(3- isopropyl-lH-pyrazol-5-yl)pyridine (340 mg, 88.6%) as a beige solid.
[001533JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 188.2 (M+l) at 0.808 min.
[001534] Step 3. Synthesis of 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine.
Figure imgf000316_0002
[001535] To a solution of 4-(3-isopropyl-lH-pyrazol-5-yl)pyridine (340 mg; 1.82 mmol) in ethanol (50 mL) was added hydrochloric acid solution (1 M, 31.4 mL; 31.4 mmol) and platinum oxide hydrate (80% Pt, 375 mg). The mixture was hydrogenated at atmospheric pressure and room temperature for 14 h. The reaction mixture was filtered, and concentrated in vacuo to afford the product 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine (340 mg, 96.9%) as a yellow solid. [001536] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 91% (214 nm), Mass: found peak 194.0(M+l) at 1.467 min. [001537] Step 4. Synthesis of tert-butyl 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine-l- carboxylate.
Figure imgf000317_0002
[001538] 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine (340 mg, 1.76 mmol) was dissolved in water (10.0 mL). Sodium bicarbonate (443 mg; 5.28 mmol) and di -tertbutyl di carb onate (384 mg; 1.76 mmol) dissolved in dioxane (15.0 mL) were added and the reaction mixture was stirred for 2 h at room temperature. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to afford the product tert-butyl 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (480 mg, 93%) as a colorless oil.
[001539JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 238.1 (M-55)+ at 2.016 min.
[001540] Step 5. Synthesis of tert-butyl 4-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate.
Figure imgf000317_0001
[001541] To a solution of tert-butyl 4-(3-isopropyl-lH-pyrazol-5-yl)piperidine-l-carboxylate (180 mg, 0.61 mmol) in dichloromethane (20 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (253 mg, 1.23 mmol), anhydrous copper acetate(446 mg, 2.45 mmol), pyridine (194 mg, 2.45 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 40 h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @750mL/min, eluting with 0-20% EA in petroleum ether) to afford the desired product tert-butyl 4-[5- isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate (140 mg, yield 42.8%) as a yellow solid. [001542JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
[001543] X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 85% (214 nm), Mass: found peak 454.1 (M+l) at 2.418 min.
[001544] Step 6. Synthesis of 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperidine.
Figure imgf000318_0001
[001545] To a solution of tert-butyl 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine-l -carboxylate (114 mg, 0.3 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The mixture was stirred for 2h. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), and dried over sodium sulfate. The combined organic layers were concentrated to afford the product 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (100 mg, 71%) as an orange oil.
[001546] LCMS method: Mobile Phase : A: water (0.01% TFA) B: ACN (0.01% TFA); Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.2min; Flow Rate :2.2mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 79% (214 nm), Mass: found peak 354.3 (M+l) at 1.119 min.
[001547] Step 7. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]ethyl]morpholine (Compound 88).
Figure imgf000318_0002
[001548] To a solution of 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (100 mg, 0.28 mmol), KI (47 mg, 0.28 mmol), and potassium carbonate (117 mg, 0.85 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (79 mg, 0.42 mmol). The reaction was stirred at 90 °C overnight. After the reaction mixture was cooled to room temperature, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l- piperidyl]ethyl]morpholine (58.6 mg, yield 44.4 %) as a yellow solid.
[001549] LCMS method: Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Temperature: 50°C; LC purity: 100% (214 nm), Mass: found peak 467.1(M+1) at 2.125 min.
[001550] XH NMR (400 MHz, DMSO-d6) 5 7.61-7.55 (m, 2H), 7.50 (d, J = 8.4 Hz, 2H), 6.19 (s, 1H), 3.63-3.51 (m, 4H), 3.17 (d, J = 5.2 Hz, 1H), 3.03-3.00 (m, 1H), 2.91 (d, J = 11.6 Hz, 2H), 2.46-2.31 (m, 8H), 2.01 (t, J = 10.8 Hz, 2H), 1.86 (d, J = 12.0 Hz, 2H), 1.60 (qd, J = 12.4, 3.6 Hz, 2H), 1.11 (d, J = 6.8 Hz, 6H) ppm.
Example S89. Synthesis of 4-[2-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound 89). [001551] Compound 89 was prepared as outlined below.
Figure imgf000319_0001
[001552] Step 1. Synthesis of tert-butyl 2,6-dimethyl-4-(3-oxobutanoyl)piperazine-l- carboxylate.
Figure imgf000319_0002
, [001553] A mixture of tert-butyl 2,6-dimethylpiperazine-l -carboxylate (500 mg, 2.33 mmol) and tert-butyl acetoacetate (406 mg, 2.57 mmol) in toluene (50 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 2,6-dimethyl-4-(3- oxobutanoyl)piperazine-l -carboxylate (700 mg, 96.7% yield) as a yellow oil.
[001554JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 96% (214 nm), Mass: found peak 243.3 (M-55)+ at 1.116 min.
[001555] Step 2. Synthesis of tert-butyl 2,6-dimethyl-4-(3-oxobutanethioyl)piperazine-l- carboxylate.
. Lawesson's reagent
6 ^NC^V_BOC toluene^cj eh"
Figure imgf000320_0001
Boc
[001556] To a solution of tert-butyl 2,6-dimethyl-4-(3-oxobutanoyl)piperazine-l-carboxylate (750 mg, 2.51 mmol) in toluene (50 mL) was added Lawesson's reagent (508 mg, 1.26 mmol), and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 2,6-dimethyl-4-(3-oxobutanethioyl)piperazine-l -carboxylate (250 mg, yield 31.6%) as a yellow oil.
[001557] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 56.74% (214 nm), Mass: found peak 259.2 (M-55)+ at 1.261 min. LC purity: 24.56% (214 nm) Mass: found peak 259.2 (M-55)+ at 1.436 min.
[001558] Step 3. Synthesis of tert-butyl 2,6-dimethyl-4-(5-methyl-lH-pyrazol-3- yl)piperazine-l-carboxylate. Boc
Figure imgf000320_0002
[001559] To a solution of tert-butyl 2,6-dimethyl-4-(3-oxobutanethioyl)piperazine-l-carboxylate (250 mg, 0.795 mmol) in toluene (20 mL) was added hydrazine hydrate (122 mg, 2.39 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 2,6-dimethyl-4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (200 mg, yield 85.4%) as a yellow oil.
[001560JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 295.3 (M+l) at 1.083 min.
[001561] Step 4. Synthesis of tert-butyl 2,6-dimethyl-4-(5-methyl-lH-pyrazol-3- yl)piperazine-l-carboxylate.
Figure imgf000321_0001
DCM, RT, 16h
[001562] To a solution of tert-butyl 2,6-dimethyl-4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate (200 mg, 0.68 mmol) in dichloromethane (10 mL) was added [3- (trifluoromethoxy)phenyl]boronic acid (286 mg, 1.36 mmol), anhydrous copper acetate (247 mg, 136 mmol), pyridine (0.28 mL, 3.4 mmol), and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 2,6-dimethyl-4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, yield 51.8%) as a yellow oil.
[001563JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 455.3 (M+l) at 1.578 min.
[001564] Step 5. Synthesis of 3,5-dimethyl-l-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000321_0002
[001565] To a solution of tert-butyl 2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (160 mg, 0.35 mmol) in di chloromethane (10 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h, then concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 3,5- dimethyl-l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (120 mg, yield 93%) as a yellow oil.
[001566] LCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE
C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95% (214 nm), Mass: found peak 355.2 (M+l) at 1.610 min.
[001567] Step 6. Synthesis of 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethanone.
Figure imgf000322_0001
[001568] To a solution of 3,5-dimethyl-l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (90 mg, 0.25 mmol) and TEA (257 mg, 2.54 mmol) in DCM (10 mL) was added 2-chloroacetyl chloride(86 mg, 0.76 mmol) at 0 °C. The mixture was stirred overnight at room temperature, then concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 20 g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM for 8 CV) to afford the desired product 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethenone (100 mg, yield 91.4%) as a yellow oil.
[001569JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm), Mass: found peak 431.1 (M+l) at 2.155 min.
[001570] Step 7. Synthesis of l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]-2-morpholino-ethanone.
Figure imgf000323_0001
[001571] To a solution of 2-chloro-l-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethanone (100 mg, 0.232 mmol) and TEA (235 mg, 2.32 mmol) in DCM (10 mL) was added morpholine (101 mg, 1.16 mmol). The mixture was stirred for 3 Oh at room temperature, then concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 20 g silica gel column @75mL/min, eluting with 0- 10% MeOH in DCM for 8 CV) to afford the crude. Then the product was purified by prep- HPLC (ammonium hydrogen carbonate) to afford the desired product l-[2,6-dimethyl-4-[5- methyl- 1 -[4-(trifluorom ethoxy )phenyl]pyrazol-3 -yl]piperazin- 1 -yl]-2-morpholino-ethanone (80 mg, yield 71.6%) as a white solid.
[001572JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm), Mass: found peak 482.0 (M+l) at 2.100 min.
[001573] 'H NMR (400 MHz, MeOD-d4) 5 7.61-7.51 (m, 2H), 7.40 (d, J = 8.4 Hz, 2H), 5.90 (s, 1H), 4.62 (s, 1H), 4.42 (s, 1H), 3.75-3.68 (m, 4H), 3.61 (d, J = 11.6 Hz, 2H), 3.50-3.39 (m, 1H), 3.25-3.11 (m, 1H), 2.86 (s, 2H), 2.56 (s, 4H), 2.30 (s, 3H), 1.42 (d, J = 42.4 Hz, 6H) ppm.
[001574] Step 8. Synthesis of 4-[2-[2,6-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound 89).
Figure imgf000323_0002
[001575] A solution of l-[4-[5-methoxy-l-[4-(trifluoromethoxy)phenyl]indazol-3-yl]-2- (trifluoromethyl)-l-piperidyl]-2-morpholino-ethanone (60mg, 0.10 mMol) in BH3-THF (10 mL) was stirred at 90 °C overnight. After cooling to 0 °C, the reaction mixture was treated with MeOH (2 mL) dropwise and then HC1 (3 mol/L, 2 mL). The mixture was stirred for Ih at 100 °C. The reaction was allowed to cool to room temp and concentrated in vacuo. The residue was purified by Prep-HPLC to afford crude. The crude was purification by prep-HPLC to afford the desired product 4-[2-[2,6-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl] morpholine (16.5 mg, 24.3%) as a white solid. [001576] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98% (214 nm), Mass: found peak 468.1 (M+l) at 2.081 min.
[001577] 'H NMR (400 MHz, MeOD-d4) 5 7.59-7.50 (m, 2H), 7.39 (d, J = 8.4 Hz, 2H), 5.84 (s, 1H), 3.74-3.64 (m, 4H), 3.57 (d, J = 11.2 Hz, 2H), 3.01-2.92 (m, 2H), 2.87 (s, 2H), 2.60-2.46 (m, 8H), 2.28 (s, 3H), 1.19 (d, J = 6.4 Hz, 6H) ppm.
Example S90. Synthesis of 4-[2-[2,2-dimethyl-4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound 90).
[001578] Compound 90 was prepared as outlined below.
Figure imgf000324_0001
[001579] Step 1. Synthesis of tert-butyl 2,2-dimethyl-4-(3-oxobutanoyl)piperazine-l- carboxylate.
Figure imgf000324_0002
[001580] A mixture of tert-butyl 2,2-dimethylpiperazine-l -carboxylate (250 mg, 1.17 mmol) and tert-butyl 3-oxobutanoate (203 mg, 1.28 mmol) in toluene (10 mL) was heated at 100°C overnight, then the mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 2,2-dimethyl-4-(3- oxobutanoyl)piperazine-l -carboxylate (340 mg, 94% yield) as a yellow oil. [001581JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 94% (214 nm), Mass: found peak 243.3 (M+l) at 1.115 min.
[001582] Step 2. Synthesis of 4-(3,3-dimethylpiperazin-l-yl)-4-thioxo-butan-2-one.
Lawesson's reagent \ /
(0.5 eq) - >
B
Figure imgf000325_0002
Do 0c CT
Figure imgf000325_0001
— ~N— Boc toluene, 75 c, 16h s
[001583] To a solution of 1 -(3, 3-dimethylpiperazin-l-yl)butane-l, 3-dione (400 mg, 2.02 mmol) in toluene (20 mL) was added Lawesson's reagent (408 mg, 1.01 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford 4-(3,3- dimethylpiperazin-l-yl)-4-thioxo-butan-2-one (80 mg, yield 18.5%) as a yellow oil. [001584JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 73% (214 nm), Mass: found peak 259.2 (M-55)+ at 1.262 min. LC purity: 16.7% (214 nm) Mass: found peak 259.2 (M-55)+ at 1.430 min.
[001585] Step 3. Synthesis of tert-butyl 2,2-dimethyl-4-(5-methyl-lH-pyrazol-3- yl)piperazine-l-carboxylate.
Figure imgf000325_0003
, c, 16h — N Boc
[001586] To a solution of tert-butyl 2,2-dimethyl-4-(3-oxobutanethioyl)piperazine-l -carboxylate (90 mg, 0.28 mmol) in toluene (10 mL) was added NH2NH2 water (43.9 mg, 0.86 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 2,2-dimethyl-4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (80 mg, yield 94.9%) as a yellow oil.
[001587] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 87% (214 nm), Mass: found peak 295.3 (M-17)+ at 1.047 min.
[001588] Step 4. Synthesis of tert-butyl 2,2-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl] piperazine-l-carboxylate.
Figure imgf000326_0001
[001589] To a solution of tert-butyl 2,2-dimethyl-4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate (80 mg, 0.27 mmol) in dichloromethane (10 mL) was added [3- (trifluoromethoxy)phenyl]boronic acid (114 mg, 0.54 mmol), anhydrous copper acetate (98.7 mg, 0.54 mmol), pyridine (0.11 mL, 1.71 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl] piperazine- 1 -carboxylate (120 mg, yield 97.2%) as a yellow oil.
[001590JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 87% (214 nm), Mass: found peak 295.3 (M-17)+ at 1.047 min.
[001591] Step 5. Synthesis of 3,3-dimethyl-l-(5-methyl-l-(4-(trifluoromethoxy)phenyl)-lH- pyrazol-3-yl)piperazine.
Figure imgf000326_0002
[001592] To a solution of tert-butyl 2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazine- 1 -carboxylate (130 mg, 0.28 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 3,3- dimethyl-l-(5-methyl-l-(4-(trifluoromethoxy)phenyl)-lH-pyrazol-3-yl)piperazine (90 mg, 96.2%) as a yellow oil.
[001593JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 68% (214 nm), Mass: found peak 355.3 (M-17)+ at 1.074 min.
[001594] Step 6. Synthesis of 2-chloro-l-[2,2-dimethyl-4-[5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethenone.
Figure imgf000327_0001
[001595] To a solution of 3,3-dimethyl-l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (90 mg, 0.25 mmol) and TEA (257 mg, 2.54 mmol) in DCM (10 mL) was added 2-chloroacetyl chloride (86 mg, 0.76 mmol) at 0 °C. The mixture was stirred overnight at room temperature. The residue was purified by flash chromatography (Biotage, 20 g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM for 8 CV) to afford the desired product 2- chl oro- l-[2,2-dimethyl-4-[5-methyl-l-[4-(tri fluoromethoxy )phenyl]pyrazol-3-yl]piperazin- 1- yl]ethenone (70 mg, yield 64%) as a yellow oil.
[001596] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 92% (214 nm), Mass: found peak 431.0 (M+l) at 2.164 min.
[001597] Step 7. Synthesis of l-[2,2-dimethyl-4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]-2-morpholino-ethanone.
Figure imgf000327_0002
[001598] To a solution of 2-chloro-l-[2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- l-yl]ethanone (80 mg, 0.186 mmol) and TEA (188 mg, 1.86 mmol) in DCM (10 mL) was added morpholine (80.9 mg, 0.93 mmol). The mixture was stirred for 30h at room temperature. The residue was purified by flash chromatography (Biotage, 20 g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM for 8 CV) to afford the crude product. Then the crude material was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product l-[2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]-2-morpholino-ethanone (70 mg, yield 78.3%) as a white solid.
[001599JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 94% (214 nm), Mass: found peak 482.3 (M+l) at 1.122 min.
[001600] 'H NMR (400 MHz, MeOD-d4) 5 7.62-7.50 (m, 2H), 7.39 (d, J = 8.4 Hz, 2H), 5.77 (s, 1H), 3.96-3.82 (m, 2H), 3.78-3.64 (m, 4H), 3.48 (t, J = 5.6 Hz, 2H), 3.38 (s, 2H), 3.23 (s, 2H), 2.52 (s, 4H), 2.29 (s, 3H), 1.49 (s, 6H).
[001601] Step 8. Synthesis of 4-[2-[2,2-dimethyl-4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound 90).
Figure imgf000328_0001
[001602] A solution of l-[2,2-dimethyl-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]-2-morpholino-ethanone (50 mg, 0.10 mmol) in BH3 THF (10 mL) was stirred at 90 °C overnight. After cooling to 0 °C, the reaction mixture was treated with MeOH (2 mL) dropwise. Then HC1 (3 mol/L, 2 mL) was added. The mixture was stirred for Ih at 100 °C. The residue was purified by Prep-HPLC to afford crude product. The crude product was purified by prep-HPLC to afford the desired product 4-[2-[2,2-dimethyl-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (16.5 mg, 24.3%) as a yellow solid.
[001603JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 98% (214 nm) Mass: found peak 467.9 (M+l) at 1.942 min.
[001604] 'H NMR (400 MHz, MeOD-d4) 5 7.57-7.49 (m, 2H), 7.39 (d, J = 8.3 Hz, 2H), 5.82 (s, IH), 3.75-3.65 (m, 4H), 3.26-3.17 (m, 2H), 2.96 (s, 2H), 2.82-2.72 (m, 2H), 2.66-2.57 (m, 2H), 2.57-2.44 (m, 6H), 2.28 (s, 3H), 1.13 (s, 6H). Example S91. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-
3,8-diazabicyclo[3.2.1]octan-8-yl]ethyl] morpholine (Compound 91).
[001605] Compound 91 was prepared as outlined below.
Figure imgf000329_0001
[001606] Step 1. Synthesis of tert-butyl 3-(3-oxobutanoyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate.
Figure imgf000329_0002
[001607] A mixture of tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (200 mg, 0.94 mmol) and tert-butyl acetoacetate (164 mg, 1.04 mmol) in toluene (20 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 3-(3-oxobutanoyl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (270 mg, 93% yield) as a yellow oil.
[001608] LCMS method: Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Temperature: 50°C; LC purity: 98% (214 nm), Mass: found peak 279.1(M+l) at 1.702 min.
[001609] Step 2. Synthesis of tert-butyl 3-(3-oxobutanethioyl)-3,8-diazabicyclo[3.2.1]octane-
8-carboxylate.
Lawesson's reagent (0.5 eq)
Figure imgf000329_0004
N— Boc toluene, 75°c, 16h
Figure imgf000329_0003
[001610] To a solution of tert-butyl 3-(3-oxobutanoyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (270 mg, 0.91 mmol) in toluene (20 mL) was added Lawesson's reagent (184 mg, 0.45 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford the desired product tert-butyl 3-(3-oxobutanethioyl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (120 mg, yield 42.2%) as a yellow oil. [001611JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 75% (214 nm), Mass: found peak 257.3 (M-55)+ at 1.228 min. LC purity: 18% (214 nm) Mass: found peak 257.3 (M-55)+ at 1.409 min.
[001612] Step 3. Synthesis of tert-butyl 3-(5-methyl-lH-pyrazol-3-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate.
Figure imgf000330_0001
[001613] To a solution of tert-butyl 3-(3-oxobutanethioyl)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (120 mg, 0.38 mmol) in toluene (10 mL) was added NH2NH2 water (43.9 mg, 0.86 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 3-(5-methyl-lH-pyrazol-3-yl)-3,8-diazabicyclo[3.2.1]octane- 8-carboxylate (100 mg, yield 89%) as a yellow oil.
[001614JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 92% (214 nm), Mass: found peak 293.3 (M+l) at 1.024 min.
[001615] Step 4. Synthesis of tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate.
Figure imgf000330_0002
[001616] To a solution of tert-butyl 3-(5-methyl-lH-pyrazol-3-yl)-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, 0.34 mmol) in dichloromethane (10 mL) was added [3-(trifluoromethoxy)phenyl]boronic acid (144 mg, 0.68 mmol), anhydrous copper acetate (124 mg, 0.68 mmol), pyridine (0.14 mL, 1.71 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,8- diazabicyclo[3.2.1]octane-8-carboxylate (100 mg, yield 64.6%) as a yellow solid.
[001617] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95. Column Temperature: 40 °C; LC purity: 97% (214 nm), Mass: found peak 453.3 (M+l) at 1.566 min.
[001618] Step 5. Synthesis of l-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000331_0001
[001619] To a solution of tert-butyl 4-[5-methyl-l-[3-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (110 mg, 0.258 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l- [3-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (84 mg, yield 97%) as a yellow oil. [001620JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 97% (214 nm), Mass: found peak 353.3 (M+l)+ at 1.076 min.
[001621] Step 6. Synthesis of 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-3,8-diazabicyclo[3.2.1]octan-8-yl]ethyl] morpholine (Compound 91).
Figure imgf000331_0002
[001622] To a solution of 3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,8- diazabicyclo[3.2.1]octane (75 mg, 0.213 mmol), potassium carbonate (118 mg, 0.851 mmol) and KI (35.3 mg, 0.213 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (59.4 mg, 0.319 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water/acetonitrile) to afford the desired product 4-[2-[3-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,8- diazabicyclo[3.2.1]octan-8-yl]ethyl] morpholine (33.8 mg, yield 34.1%) as a yellow solid. [001623JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm) Mass: found peak 466.1 (M+l) at 2.076 min.
[001624] 'H NMR (400 MHz, DMSO-d6) 5 7.59 (d, J = 9.2 Hz, 2H), 7.44 (d, J = 8.4 Hz, 2H), 5.76 (s, 1H), 3.54 (t, J = 4.4 Hz, 4H), 3.24 (d, J = 11.2 Hz, 2H), 2.83 (d, J = 9.6 Hz, 2H), 2.41 (m, 10H), 2.30 (s, 3H), 1.90-1.82 (m, 2H), 1.61 (d, J = 7.2 Hz, 2H).
Example S92. Synthesis of 4-[2-[4-[l-[4-(l,l-difluoroethyl) phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 92).
[001625] Compound 92 was prepared as outlined below.
Figure imgf000332_0001
[001627] At -78°C, to a solution of l-bromo-4-(l, 1 -difluoroethyl) benzene (200 mg, 0.9 mmol) in THF (10 mL) was added n-BuLi (0.2 mL, 0.2 mmol) dropwise. The reaction mixture was stirred at
-78°C for 20 min. Then a solution of trimethyl borate (141 mg, 1.36 mmol) in THF (2 mL) was added dropwise and stirred at -78°C for Ih. The reaction was quenched by sat. NH4Q aqueous, extracted with EtOAc (20 mL), washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product [4-(l,l-difluoroethyl)phenyl]boronic acid (220 mg) as a yellow solid.
[001628] Step 2. Synthesis of tert-butyl 4-[l-[4-(l,l-difluoroethyl)phenyl]-5-isopropyl- pyrazol-3-yl] piperazine-l-carboxylate.
Figure imgf000333_0001
[001629] To a solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, 0.34 mmol) in chloroform (20 mL) was added [4-(trifluoromethoxy) phenyl] boronic acid (129 mg, 0.679 mmol), anhydrous copper acetate (123 mg, 0.679 mmol), pyridine (134 mg, 1.7 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[l-[4-(l,l- difluoroethyl)phenyl]-5-isopropyl-pyrazol-3-yl] piperazine- 1 -carboxylate (94 mg, yield 63.7%) as a white solid.
[001630JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 435.3 (M + 1) + at 1.861 min.
[001631] Step 3. Synthesis of l-[l-[4-(l,l-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3- yl] piperazine.
Figure imgf000333_0002
[001632] To a solution of tert-butyl 4-[l-[4-(l,l-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (94 mg, 0.216 mmol) in di chloromethane (5 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 1 -[ 1 -[4-(l , 1 - difluoroethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (103 mg, crude). The crude product was used directly in the next step.
[001633JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 335.3 (M + 1)+ at 1.161 min. [001634] Step 4. Synthesis of 4-[2-[4-[l-[4-(l,l-difluoroethyl) phenyl]-5-isopropyl-pyrazol- 3-yl]piperazin-l-yl]ethyl]morpholine (Compound 92).
Figure imgf000334_0001
[001635] To a solution of l-[l-[4-(l,l-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (103 mg, 0.3 mmol), potassium carbonate (213 mg, 1.54 mmol) and KI (51 mg, 0.3 mmol) in 95% ethanol (30 mL) was added 4-(2-chloroethyl)morpholine (2.4 g, 16.4 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[ l-[4-(l , 1 - difluoroethyl) phenyl]-5-isopropyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (39.3 mg, yield: 27.9%) as a white solid.
[001636] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.39 % (214 nm); Mass: found peak 448.3 (M + H) at 1.822 min.
[001637] 'H NMR (400 MHz, CDCh) 5 7.57 (d, J = 8.5 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 5.72 (s, 1H), 3.80-3.63 (m, 4H), 3.35-3.15 (m, 4H), 3.00 (dt, J = 13.8, 6.9 Hz, 1H), 2.65-2.60 (m, 4H), 2.59-2.55 (m, 4H), 2.51 (s, 4H), 1.93 (t, J = 18.1 Hz, 3H), 1.18 (d, J = 6.8 Hz, 6H) ppm.
Example S93. Synthesis of 4-[2-[4-[l-[4-(2,2-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 93).
[001638] Compound 93 was prepared as outlined below.
Figure imgf000334_0002
[001639] Step 1. Synthesis of l-bromo-4-(2, 2-difluoroethyl)benzene.
Figure imgf000334_0003
[001640] To a solution of 5-bromo-3-isopropyl-lH-pyrazin-2-one (1.0g, 5.46 mmol) and [phenyl-(2,2,2-trifluoroacetyl)oxy-lambda3-iodanyl] 2,2,2-trifluoroacetate (846mg, 1.97 mmol) in
DCM (10 mL) was added pyridine hydrofluoride (217 g,21.85mmol) at 0°C. The mixture reaction was stirred at rt for 16h. The mixture was quenched with aqueous sodium bicarbonate solution (50mL) and extracted with ethyl acetate (100mL><2). The combined organic phases were dried in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @35mL/min, eluting with petroleum ether in petroleum ether for 5 CV) to afford 1- bromo-4-(2, 2-difluoroethyl)benzene (260 mg, yield 15.9%) as a transparent oil.
[001641] Step 2. Synthesis of [4-(l,l-difluoroethyl)phenyl]boronic acid.
Figure imgf000335_0001
[001642] At -78°C, to a solution of l-bromo-4-(l,l-difluoroethyl)benzene (200 mg, 0.9 mmol) in THF (10 mL) was added n-BuLi (0.2 mL, 0.2 mmol) dropwise. The reaction mixture was stirred at
-78°C for 20 min. Then a solution of trimethyl borate (141 mg, 1.36 mmol) in THF (2 mL) was added dropwise and stirred at -78°C for Ih. The reaction was quenched by sat. NH4Q aqueous, extracted with EtOAc (20 mL), washed with brine (10 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product [4-(l,l-difluoroethyl)phenyl]boronic acid ( 220 mg ) as a yellow solid.
[001643] Step 3. Synthesis of tert-butyl 4-[l-[4-(2,2-difluoroethyl)phenyl]-5-isopropyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000335_0002
[001644] To a solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (165 mg, 0.89 mmol) in chloroform (15 mL) was added [4-(2,2-difluoroethyl)phenyl]boronic acid (213 mg, 0.11 mmol), anhydrous copper acetate (12.3 mg, 0.06 mmol), pyridine (13.4 mg, 1.7 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[l-[4-(2,2-difluoroethyl)phenyl]-5- isopropyl-pyrazol-3-yl]piperazine-l-carboxylate (70 mg, yield 28.7%) as a yellow solid. [001645] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. Mass: found peak 435.1 (M + 1)+ at 2.236 min.
[001646] Step 4. Synthesis of l-[l-[4-(l,l-difluoroethyl) phenyl]-5-isopropyl-pyrazol-3- yl] piperazine.
Figure imgf000336_0001
[001647] To a solution of tert-butyl 4-[l-[4-(l, l-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (94 mg, 0.216 mmol) in di chloromethane (5 mL) was added 2,2,2- trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-[4-(l,l -difluoroethyl) phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (103 mg, crude). The crude product was used directly in the next step.
[001648JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 335.3 (M + 1)+ at 1.161 min.
[001649] Step 5. Synthesis of 4-[2-[4-[l-[4-(2,2-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 93).
Figure imgf000336_0002
[001650] To a solution of l-[l-[4-(2,2-difluoroethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (71 mg, 0.2 mmol), potassium carbonate (147 mg, l.Ommol) and KI (35.2 mg, 0.2 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine (47.6 mg, 3.18 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4-(2,2- difluoroethyl)phenyl]-5-isopropyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (7.5 mg, yield: 8%) as a yellow oil. [001651JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.39 % (214 nm); Mass: found peak 448.2 (M + H) at 1.908 min.
[001652] 'H NMR (400 MHz, CDCh) 5 7.37 (d, J = 8.4 Hz, 2H), 7.31 (d, J = 8.3 Hz, 2H), 5.93 (tt, J = 56.5, 4.5 Hz, 1H), 5.70 (s, 1H), 3.78-3.63 (m, 4H), 3.30- 3.24 (m, 4H), 3.17 (td, J = 17.3, 4.5 Hz, 2H), 2.97 (dt, J = 13.6, 6.8 Hz, 1H), 2.65-2.59 (m, 4H), 2.57 (s, 4H), 2.51 (d, J = 4.0 Hz, 4H), 1.16 (t, J = 5.9 Hz, 6H) ppm.
Example S94. Synthesis of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy-2- tetrahydropyran-4-yl-2-azaspiro[3.5]nonane (Compound 94).
[001653] Compound 94 was prepared as outlined below.
Figure imgf000337_0001
[001654] Step 1. Synthesis of tert-butyl 7-(4-methyl-3-oxo-pentanoyl)oxy-2- azaspiro[3.5]nonane-2-carboxylate.
Figure imgf000337_0002
[001655] A mixture of methyl 4-methyl-3-oxo-pentanoate (500 mg, 3.47 mmol) and tert-butyl 7- hydroxy-2-azaspiro[3.5]nonane-2-carboxylate (837 mg, 3.47 mmol) in toluene (15 mL) was stirred at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 7- (4-methyl-3-oxo-pentanoyl)oxy-2-azaspiro[3.5]nonane-2-carboxylate (870 mg, 63.9% yield) as a white solid.
[001656] LCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm) Mass: found peak 298.3 (M -55)+ at 1.992 min. [001657] Step 2. Synthesis of tert-butyl 7-[(5-isopropyl-lH-pyrazol-3-yl)oxy]-2- azaspiro[3.5]nonane-2-carboxylate.
Figure imgf000338_0001
[001658] To a solution of methyl tert-butyl 7-(4-methyl-3-oxo-pentanoyl)oxy-2- azaspiro[3.5]nonane-2-carboxylate (200 mg, 0.56 mmol) and hydrazine hydrate (58 mg, 0.85 mmol) in EtOH (20 mL) was stirred at 80°C for 2h. The mixture was extracted with ethyl acetate (30mL><2) and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @60mL/min, eluting with 0-10% MeOH in DCM) to afford the desired product tert-butyl 7-[(5-isopropyl-lH-pyrazol-3-yl)oxy]-2-azaspiro[3.5]nonane-2- carboxylate (150 mg, yield 76%) as a yellow solid.
[001659JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 70% (214 nm) Mass: found peak 294.3 (M -55)+ at 1.909 min.
[001660] Step 3. Synthesis of tert-butyl 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl- pyrazol-3-yl] oxy-2-azaspiro [3.5] nonane-2-carboxylate.
Figure imgf000338_0002
[001661] To a solution of tert-butyl 7-[(5-isopropyl-lH-pyrazol-3-yl)oxy]-2- azaspiro[3.5]nonane-2-carboxylate (150 mg, 0.43 mmol) was added [4- (trifluoromethoxy)phenyl]boronic acid (150 mg, 0.85 mmol), anhydrous copper acetate (240 mg, 1.35 mmol), pyridine (172 mg, 2.25 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered and concentrated in vacuo. The filtrate was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 10-50% dichloromethane in petroleum ether) to afford the desired product tert-butyl 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy-2- azaspiro[3.5]nonane-2-carboxylate (160 mg, yield 78.3%) as a yellow oil.
[001662JLCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 87% (254 nm) Mass: found peak 476.3 (M +H)+ at 2.155 min. [001663] Step 4. Synthesis of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy-
2-azaspiro[3.5] nonane.
Figure imgf000339_0001
[001664] To a solution of tert-butyl 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3- yl]oxy-2-azaspiro[3.5]nonane-2-carboxylate (160 mg, 0.336 mmol) in dichloromethane (6 mL) was added 2,2,2-trifluoroacetic acid (2.0 mL, 26.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude desired product 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy- 2-azaspiro[3.5] nonane (160 mg, yield 79%), which was used directly in next step.
[001665] LCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18
2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 60% (254 nm) Mass: found peak
376.2 (M +H)+ at 1.479 min.
[001666] Step 5. Synthesis of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy- 2-tetrahydropyran-4-yl-2-azaspiro [3.5] nonane (Compound 94).
Figure imgf000339_0002
[001667]Under argon atmosphere, a mixture of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl- pyrazol-3-yl]oxy-2-azaspiro[3.5]nonane (80 mg, 0.02 mmol), tetrahydropyran-4-one (32 mg, 0.32 mmol) and sodium triacetoxyborohydride (93 mg, 0.44 mmol) in DCE (25mL) was stirred at rt for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO3/water/acetonitrile) to afford the desired product 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy-2-tetrahydropyran-4-yl-2- azaspiro[3.5]nonane (14.5 mg, yield 14.8%) as a yellow solid.
[001668JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90% (214 nm) Mass: found peak 460.3 (M+l) at 2.172 min.
[001669] 'H NMR (500 MHz, CDCh) 5 7.84 (d, J = 8.5 Hz, 2H), 7.53 (d, J = 8.5 Hz, 2H), 6.65 (t, J = 56.6 Hz, 1H), 5.48 (s, 1H), 4.24 (s, 1H), 3.97 (d, J = 11.3 Hz, 2H), 3.36 (t, J = 10.5 Hz, 2H), 3.04 (s, 4H), 2.96 - 2.90 (m, 1H), 2.28 (s, 1H), 1.92 (d, J = 9.7 Hz, 4H), 1.67 (dt, J = 24.7, 9.0 Hz, 6H), 1.38 (d, J = 10.0 Hz, 2H), 1.28 (d, J = 6.9 Hz, 6H) ppm.
Example S95. Synthesis of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl-pyrazol-3-yl]oxy-2- (oxetan-3-yl)-2-azaspiro [3.5] nonane (Compound 95).
[001670] Compound 95 was prepared as outlined below.
Figure imgf000340_0001
[001671]Under argon atmosphere, a mixture of 7-[l-[4-(difluoromethyl)phenyl]-5-isopropyl- pyrazol-3-yl]oxy-2-azaspiro[3.5]nonane (80 mg, 0.02 mmol), oxetan-3-one (57 mg, 0.32 mmol) and sodium triacetoxyborohydride (93 mg, 0.44 mmol) in DCE (25mL) was stirred at rt for 16h. The reaction was filtered and concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO3/water/acetonitrile) to afford the desired product 7-[l-[4-(difhroromethyl)phenyl]-5- isopropyl-pyrazol-3-yl]oxy-2-(oxetan-3-yl)-2-azaspiro[3.5]nonane (13.7 mg, yield 14.8%) as a yellow solid.
[001672JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90% (214 nm) Mass: found peak 432.2 (M+l) at 2.045 min.
[001673] 'H NMR (400 MHz, CDCh) 5 7.83 (d, J = 8.5 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 6.59 (d, J = 56.5 Hz, 1H), 5.49 (s, 1H), 4.76 (s, 2H), 4.64 - 4.51 (m, 2H), 4.27 (s, 1H), 3.81 (d, J = 13.8 Hz, 1H), 3.25 (s, 2H), 2.98 - 2.89 (m, 1H), 1.92 (d, J = 13.5 Hz, 4H), 1.72 (s, 4H), 1.28 (dd, J = 6.9, 2.1 Hz, 8H) ppm.
Example S96. Synthesis of 4-[2-[4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]-l- piperidyl]ethyl]morpholine (Compound 96).
[001674] Compound 96 was prepared as outlined below.
Figure imgf000340_0002
[001675] Step 1. Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)-3,6-dihydro-2H- pyridine-l-carboxylate.
Figure imgf000341_0001
[001676]Under argon atmosphere, a mixture of 3-bromo-5-methyl-lH-pyrazole (600 mg, 3.73 mmol), tert-butyl 4-(4, 4,5, 5-tetramethyl- 1,3, 2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyri dine- 1- carboxylate (1.04 g, 3.35 mmol), PdC12(dppf) (273 mg, 0.373 mmol) and sodium carbonate (1.18 g, 11.2 mmol) in 1,4-dioxane (15 mL) was stirred at 100 °C for 16h. The reaction mixture was cooled to room temperature, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-20% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-(5-methyl-lH-pyrazol- 3 -yl)-3,6-dihydro-2H-pyri dine- 1 -carboxylate (600 mg, yield 55%) as a white solid.
[001677] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 97% (214 nm) Mass: found peak 286.2 (M+23)+ at 1.71 min.
[001678] Step 2. Synthesis of tert-butyl 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]- 3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000341_0002
[001679] A solution of tert-butyl tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)-3,6-dihydro-2H- pyridine-1 -carboxylate (600 mg, 2.28 mmol), (4-tert-butylphenyl)boronic acid (811 mg, 4.56 mmol), pyridine (360 mg, 4.56 mmol), and copper(II) acetate (828 mg, 4.56 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered, the filter cake was washed with di chloromethane (10 mL), the solution concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[l-(4-tert-butylphenyl)-5-methyl- pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l-carboxylate (500 mg, 55.5%) as a colorless oil. [001680JLCMS method: Mobile phase: Water (0.01% TFA) (A) / Acetonitrile (0.01% TFA) (B); Elution program: Gradient from 5 to 95% of B in 1.4min at 2.2mL/min; Temperature: 50°C; LC purity: 81% (214 nm) Mass: found peak 396.3 (M+l) at 2.293 min.
[001681] Step 3. Synthesis of tert-butyl 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3- yl]piperidine-l-carboxylate.
Figure imgf000342_0001
[001682]Under EE atmosphere, a mixture of tert-butyl 4-[l-(4-tert-butylphenyl)-5-methyl- pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l-carboxylate (500 mg, 1.26 mmol) and Pd/C (10%, 224 mg) in methanol (3 mL) was stirred at room temperature for 16h. The reaction was filtered and concentrated in vacuo to afford the desired product tert-butyl 4-[l-(4-tert-butylphenyl)-5- methyl-pyrazol-3-yl]piperidine-l -carboxylate (300 mg, yield 43.6%) as a yellow oil. [001683JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8mL/min; Column: Sunfire C18,4.6*50mm,3.5pm; Column Temperature: 50 °C; LC purity: 73%; Mass: found peak 398.3 (M+l) at 2.345 min.
[001684] Step 4. Synthesis of 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]piperidine.
Figure imgf000342_0002
[001685] To a solution of tert-butyl 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]piperidine- 1-carboxylate (300 mg, 0.755 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol- 3-yl]piperidine (180 mg, yield 63.4%) as a yellow oil.
[001686] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 1.8mL/min; Column: Sunfire C18,4.6*50mm, 3.5pm; Column Temperature: 50 °C; LC purity: 79% (214 nm) Mass: found peak 298.3 (M+l) at 1.46 min.
[001687] Step 5. Synthesis of 4-[2-[4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]-l- piperidyl]ethyl]morpholine (Compound 96).
Figure imgf000343_0001
[001688] To a solution of 4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]piperidine (100 mg, 0.336 mmol), potassium carbonate (186 mg, 1.34 mmol) and KI (5.58 mg, 0.0034 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (93.8 mg, 0.504 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(4-tert-butylphenyl)-5-methyl-pyrazol-3-yl]-l- piperidyl]ethyl]morpholine (28.7 mg, yield 20.8%) as a yellow oil.
[001689JLCMS method: Mobile phase: Water (0.01%TFA) (A) / Acetonitrile (0.01%TFA) (B); Elution program: Gradient from 5 to 95% of B in 1.4min at 2.2mL/min; Temperature: 50°C; LC purity: 100 % (214 nm) Mass: found peak 411.3 (M+l) at 1.49 min.
[001690] 'H NMR (400 MHz, CDCh) 5 7.46 (d, J = 8.3 Hz, 2H), 7.32 (d, J = 8.3 Hz, 2H), 6.04 (s, 1H), 3.70 (s, 4H), 3.37 (d, J = 11.2 Hz, 2H), 2.88 (d, J = 6.2 Hz, 4H), 2.78 - 2.72 (m, 3H), 2.52 (s, 4H), 2.31 (s, 3H), 2.20 - 1.95 (m, 4H), 1.32 (d, J = 19.7 Hz, 9H).
Example S97. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-(l- methoxyethyl)pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 97). [001691] Compound 97 was prepared as outlined below.
Figure imgf000343_0002
[001692] Step 1. Synthesis of tert-butyl 4-methoxy-3-oxopentanoate.
Figure imgf000343_0003
[001693] To a solution of 2-methoxypropanoic acid (2 g, 19.2 mmol) in THF (200 mL) was added di(imidazol-l-yl)methanone (3.4g,21.1mmol) and stirred at 0 °C for 16h. To a solution of 3-tert-butoxy-3-oxo-propanoic acid (4.6, 28.8mmol) in THF (70 mL) was added 2 mol/L chloro(isopropyl)magnesium (31.7 mL) at 0 °C and stirred at rt for 2h. The 3-tert-butoxy-3-oxo- propanoic acid mixture was added in 2-methoxypropanoic acid mixture under 0°C and stirred at rt for 16h. The reaction mixture was quenched with 10% citric acid aqueous solution (200 mL) and extracted with ethyl acetate (150mLx2).The organic phase was concentrated in vacuo and purified by flash chromatography (Biotage, 40 g silica gel column><2 @60mL/min, eluting with 0-7% ethyl acetate in petroleum ether for 8 CV) to afford the desired product tert-butyl 4- methoxy-3-oxopentanoate (3.6 g, yield 92.7%) as a yellow oil.
[001694] LCMS method: Mobile Phase: A: water (A 10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 5 to 95% of B in 1.4min at 1.8mL/min; Column: HALO C18 3.5pm 4.6*50mm; Column Temperature: 50°C; LC purity: 68.22% (214 nm) Mass: found peak 147.3 (M+l) at 1.68 min.
[001695] Step 2. Synthesis of tert-butyl 4-(4-methoxy-3-oxopentanoyl)piperazine-l- carboxylate.
Figure imgf000344_0001
[001696] To a solution of tert-butyl piperazine- 1 -carboxylate (2.7 g, 14.5 mmol) in toluene (35 mL) was added tert-butyl 4-methoxy-3-oxo-pentanoate (3.2 g, 15.9 mmol) and stirred at 100°C for 16h. The reaction mixture was concentrated in vacuo and purified by flash chromatography (Biotage, 40 g silica gel column><2 @60mL/min, eluting with 0-6% methanol in di chloromethane for 8 CV) to afford the desired product tert-butyl 4-(4-methoxy-3- oxopentanoyl)piperazine-l -carboxylate (4.9 g, yield 95.8%) as a yellow oil.
[001697] LCMS method: Mobile Phase: A: Water (0.01%TFA); B: Acetonitrile (0.01%TFA); Flow Rate: 1.6mL/min; Column: Proshell 120 EC-C18 30*3mm, 2.7pm; Oven Temperature: 50 °C; LC purity: 66.87 % (214 nm); Mass: found peak 315.1 (M+l) at 1.64 min; LC purity: 10.39 % (214 nm), Mass: found peak 315.1 (M+l) at 1.85 min.
[001698] Step 3. Synthesis of tert-butyl 4-(4-methoxy-3-oxopentanethioyl)piperazine-l- carboxylate.
Figure imgf000344_0002
[001699] To a solution of tert-butyl 4-(4-methoxy-3-oxo-pentanoyl) piperazine- 1 -carboxylate (1 g, 2.83 mmol) in toluene (20 mL) was added 2,4-bis(4-methoxyphenyl)-2,4-dithioxo- I,3,21ambda5,41ambda5-dithiadiphosphetane (573 mg, 1.42 mmol). The reaction was stirred at 75 °C for 16h. The reaction mixture was concentrated in vacuo and purified by flash chromatography (Biotage, 25 g silica gel column><2 @60mL/min, eluting with 0-35% ethyl acetate in petroleum ether for 8 CV) to afford the desired product tert-butyl 4-(4-methoxy-3- oxopentanethioyl)piperazine- 1-carboxylate (320 mg, yield 29.9%) as a yellow oil.
[001700JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8mL/min; Column: Sunfire C18,4.6*50mm,3.5pm; Column Temperature: 50°C; Detection: UV (214, 4 nm) and MS (ESI, Pos mode, 50 to 900); LC purity: 33.43 % (214 nm) Mass: found peak 331.2 (M+l) at 1.80 min.
[001701] Step 4. Synthesis of tert-butyl 4-(5-(l-methoxyethyl)-lH-pyrazol-3-yl)piperazine-
1-carboxylate.
Figure imgf000345_0001
, 16h
[001702] To a solution of tert-butyl 4-(4-methoxy-3-oxo-pentanethioyl)piperazine- 1-carboxylate (320 mg, 0.76 mmol) in toluene (5 mL) was added hydrazine monohydrate (114 mg, 2.27mmol) and the mixture was stirred at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @100mL/min, eluting with 0-16% MeOH in DCM) to afford the desired product tert-butyl 4-(5-(l-methoxyethyl)-lH-pyrazol-3-yl)piperazine-l-carboxylate (220 mg, yield 93.8%) as a yellow solid.
[001703JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8mL/min; Column: Sunfire C18,4.6*50mm, 3.5pm; Column Temperature: 50°C; Detection: UV (214, 4nm) and MS (ESI, Pos mode, 50 to 900); LC purity: 100.00% (254 nm) Mass: found peak 311.3 (M+l) at 1.60 min.
[001704] Step 5. Synthesis of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-5-(l- methoxyethyl)pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000345_0002
[001705] A solution of tert-butyl 4-[5-(l-methoxyethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (300 mg, 0.967 mmol), [4-(difluoromethyl)phenyl]boronic acid (332 mg, 1.93 mmol), pyridine (153 mg, 1.93 mmol), and copper(II) acetate (351 mg, 1.93 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered, the filter cake washed with dichloromethane (10 mL), the solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-5-(l- methoxyethyl)pyrazol-3-yl]piperazine-l -carboxylate (200 mg, 47.4%) as a white oil.
[001706] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 1.8mL/min;
Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm) Mass: found peak 437.3 (M+l) at 2.06 min.
[001707] Step 6. Synthesis of l-[l-[4-(difluoromethyl)phenyl]-5-(l-methoxyethyl)pyrazol-3- yl] piperazine.
Figure imgf000346_0001
[001708] To a solution of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-5-(l-methoxyethyl)pyrazol- 3 -yl]piperazine-l -carboxylate (200 mg, 0.458 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 1 -[ 1 -[4- (difluoromethyl)phenyl]-5-(l-methoxyethyl)pyrazol-3-yl]piperazine (170 mg, yield 91.5%) as a yellow oil.
[001709JLCMS method: Mobile phase: Water (0.01%TFA) (A) / Acetonitrile (0.01%TFA) (B); Elution program: Gradient from 5 to 95% of B in 1.4min at 2.2mL/min; Temperature: 50°C; LC purity: 83% (214 nm); Mass: found peak 337.2 (M+l) at 1.42 min.
[001710] Step 7. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-(l- methoxyethyl)pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 97).
Figure imgf000346_0002
[001711] To a solution of l-[l-[4-(difluoromethyl)phenyl]-5-(l-methoxyethyl)pyrazol-3- yl]piperazine (70 mg, 0.208 mmol), potassium carbonate (115 mg, 0.832 mmol) and KI (3.45 mg, 0.0021 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (58.1 mg, 0.312 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4- (difluoromethyl)phenyl]-5-(l-methoxyethyl)pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (52.0 mg, yield 55.6%) as a yellow oil.
[001712JLCMS method: Mobile phase: Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min; Temperature: 50°C; LC purity: 100 % (214 nm) Mass: found peak 450.4 (M+l) at 1.62 min.
[001713] 'H NMR (400 MHz, CDCI3) 5 7.58 (s, 3H), 7.26 (s, 1H), 6.68 (t, J = 56.4 Hz, 1H), 5.94 (s, 1H), 4.41 (d, J = 6.5 Hz, 1H), 3.74 - 3.69 (m, 4H), 3.34 - 3.26 (m, 4H), 3.25 (d, J = 6.9 Hz, 3H), 2.66 - 2.60 (m, 4H), 2.57 (s, 4H), 2.50 (d, J = 4.3 Hz, 4H), 1.46 (d, J = 6.5 Hz, 3H).
Example S98. Synthesis of 4-[2-[4-[l-[3-(difluoromethyl)-5-fluoro-phenyl]-5-isopropyl- pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound 98).
[001714] Compound 98 was prepared as outlined below.
Figure imgf000347_0001
[001715] Step 1. Synthesis of [3-(difluoromethyl)-5-fluoro-phenyl]boronic acid.
Figure imgf000347_0002
[001716] 1.0 g of l-bromo-3-(difluoromethyl)-5-fluoro-benzene was placed into a 100-mL three- neck flask, and the air in the flask was replaced with nitrogen. To this compound was added 10 mL of tetrahydrofuran, and this solution was stirred at -78°C for 20 minutes. Next, 0.27g of a 1.7 M hexane solution of n-butyllithium was dripped into this mixture solution, followed by stirring at
-78°C for 10 minutes. After 30 min, 0.46 g of trimethyl borate was added to the mixture and this solution was stirred for 20 min, while the temperature was warmed to room temperature. The residue was diluted with water (10 mL), extracted with di chloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product [3- (difluoromethyl)-5-fluoro-phenyl]boronic acid as a white solid, yield 59.2%.
[001717] LCMS: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 1.8mL/min; Column: Sunfire C18,4.6*50mm, 3.5pm; Column Temperature: 50 °C; Mass: No peak observed by LCMS.
[001718] Step 2. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000348_0001
[001719] A solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (150 mg, 0.51 mmol), [4-(difluoromethoxy)phenyl]boronic acid (128 mg, 1.02 mmol), pyridine (57.8 mg, 1.02 mmol), and copper(II)acetate (128 mg, 1.02 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2. The mixture was filtered, the filter cake washed with di chloromethane (10 mL), the filtrate was concentrated, and the residue was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tert-butyl 4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 45%) as a white oil. [001720JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min. Flow Rate: 1.8mL/min;
Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm); Mass: found peak 439.3 (M+l) at 2.16 min.
[001721] Step 3. Synthesis of l-[l-[3-(difluoromethyl)-5-fluoro-phenyl]-5-isopropyl- pyrazol-3-yl] piperazine.
Figure imgf000348_0002
[001722] To a solution of tert-butyl 4-[l-[3-(difluoromethyl)-5-fluoro-phenyl]-5-isopropyl- pyrazol-3-yl]piperazine-l -carboxylate (200 mg, 0.457 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 1 -[ 1 -[3- (difluoromethyl)-5-fluoro-phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (100 mg, yield 74.5%) as a yellow oil.
[001723JLCMS method: Mobile phase: Water (0.01%TFA) (A) / ACN (0.01%TFA) (B);
Elution program: Gradient from 5 to 95% of B in 1.4 min at 2.2mL/min; Temperature: 50°C; LC purity: 82.3% (214 nm) Mass: found peak 339.1 (M+l) at 1.51 min.
[001724] Step 4. Synthesis of 4-[2-[4-[l-[3-(difluoromethyl)-5-fluoro-phenyl]-5-isopropyl- pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound 98).
Figure imgf000349_0001
[001725] To a solution of l-[l-[3-(difluoromethyl)-5-fluoro-phenyl]-5-isopropyl-pyrazol-3- yl]piperazine (100 mg, 0.296 mmol), potassium carbonate (163 mg, 1.18 mmol) and KI (4.91 mg, 0.0030 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (82.5 mg, 0.443 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered and concentrated in vacuo. The residue was purified by prep-HPLC (NH-tHCCWwater/acetonitrile) to afford the desired product 4-[2-[4-[l-[3-(difhioromethyl)-5- fluoro-phenyl]-5-isopropyl-pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (80.2 mg, yield 60.1%) as a yellow solid.
[001726] LCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B); Elution program: Gradient from 10 to 95% of B ini.5 min at 1.8mL/min; Temperature: 50°C; LC purity: 100 % (214 nm); Mass: found peak 452.4 (M+l) at 1.79 min.
[001727] XH NMR (400 MHz, CDCh) 5 7.39 (s, 1H), 7.29 (s, 1H), 7.17 (d, J = 8.3 Hz, 1H), 6.65 (t, J = 56.2 Hz, 1H), 5.74 (s, 1H), 3.75 - 3.67 (m, 4H), 3.28 (s, 4H), 3.07 - 2.96 (m, 1H), 2.60 (d, J = 18.6 Hz, 8H), 2.52 (s, 4H), 1.21 (d, J = 6.8 Hz, 6H).
Example S99. Synthesis of 4-[2-[4-[5-methoxy-l-[3-(trifluoromethyl)-l- bicyclo[l.l.l]pentanyl]pyrazolo[3,4-b]pyridin-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 99).
[001728] Compound 99 was prepared as outlined below.
Figure imgf000350_0001
[001729] Step 1. Synthesis of tert-butyl 5-benzyloxy-3-oxo-pentanoate.
Figure imgf000350_0002
[001730] To a solution of 3 -methylbicyclofl.1.1 ]pentane-l -carboxylic acid (3.6 g, 20 mmol) in THF (50 mL) at 0 °C was added CDI (3.56 g, 22 mmol) and the reaction mixture was stirred at RT for 3h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (4.8 g, 30 mmol) in THF (40 mL) at 0°C, and the reaction mixture was stirred at RT for 3h. Next, this solution was added dropwise to the acyl imidazole solution at 0 °C and the resulting mixture was allowed to warm up to RT and stirred overnight. The mixture was quenched by addition of 10 % aqueous citric acid (50 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by flash column chromatography on 7/3) to afford tert-butyl 5-benzyloxy-3-oxo-pentanoate (2.5 g, yield 38.2%) as a yellow oil.
[001731] 'H NMR (500 MHz, CDCh) 5 7.38-7.26 (m, 5H), 4.48-4.44 (m, 2H), 3.64 (t, J = 6.4 Hz, 2H), 3.48 (s, 2H), 2.77 (t, J = 6.4 Hz, 2H), 1.40 (s, 9H) ppm.
[001732] Step 2. Synthesis of tert-butyl 4-(5-benzyloxy-3-oxo-pentanoyl)piperazine-l- carboxylate.
Figure imgf000351_0001
[001733] A mixture of tert-butyl 5-benzyloxy-3-oxo-pentanoate (400 mg, 1.44 mmol) and tertbutyl piperazine- 1 -carboxylate (294 mg, 1.58.8mmol) in toluene (10 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tert-butyl 4-(5-benzyloxy-3-oxo-pentanoyl)piperazine-l-carboxylate (450 mg, 73% yield) as a yellow oil.
[001734JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge : C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 87.72% (214 nm) Mass: found peak 391.2 (M+l) at 1.848 min.
[001735] Step 3. Synthesis of tert-butyl 4-(5-benzyloxy-3-oxo-pentanethioyl)piperazine-l- carboxylate.
Figure imgf000351_0002
[001736] To a solution of tert-butyl 4-(5-benzyloxy-3-oxo-pentanoyl)piperazine-l-carboxylate (2.7 g, 6.64 mmol) in toluene (50 mL) was added Lawesson's reagent (1.34 g, 3.32 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(5-benzyloxy-3-oxo- pentanethioyl)piperazine-l -carboxylate (1.8 g, yield 38.6%) as a yellow oil.
[001737] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 57.9% (214 nm) Mass: found peak 406.9 (M+l) at 1.819 min.
[001738] Step 4. Synthesis of tert-butyl 4-[5-(2-benzyloxyethyl)-lH-pyrazol-3-yl]piperazine- 1-carboxylate.
Figure imgf000352_0001
[001739] To a solution of tert-butyl 4-(5-benzyloxy-3-oxo-pentanethioyl)piperazine-l- carboxylate (1.8 g, 4.43 mmol) in toluene (40 mL) was added NH2NH2 water (680 mg, 13.2 mmol) and the mixture was heated at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 4-[5-(2 -benzyloxy ethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (1.4 g, yield 63%) as a white solid.
[001740JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge : C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 77.04% (214 nm) Mass: found peak 387.2 (M+l) at 2.042 min.
[001741] Step 5. Synthesis of tert-butyl 4-[5-(2-benzyloxyethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000352_0002
[001742] To a solution of tert-butyl 4-[5-(2-benzyloxyethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (700 mg, 1.81 mmol) was added [3 -(trifluoromethoxy) phenyl] boronic acid (761 mg, 3.62 mmol), anhydrous copper acetate (660 mg, 3.62 mmol), pyridine (716 mg, 9.06 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16h. The mixture was filtered, concentrated in vacuo and the residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 10-50% dichloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-(2- benzyloxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (520 mg, yield 49.5%) as a yellow oil.
[001743JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge : C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 94.17% (214 nm) Mass: found peak 547.3 (M+l) at 2.003 min. [001744] Step 6. Synthesis of tert-butyl 4-[5-(2-hydroxyethyl)-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000353_0001
[001745] Tert-butyl 4-[5-(2-benzyloxyethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (300 mg, 0.55 mmol) was dissolved in methanol (12 mL), and the reaction mixture was hydrogenated with Pd/C (10%, 200 mg) at atmospheric pressure and room temperature for 4 h. The reaction mixture was filtered to afford tert-butyl 4-[5-(2-hydroxyethyl)- l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine-l -carboxylate (200 mg, 50% yield) as a yellow solid.
[001746] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.4 min; 95% B for 1.6 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C. LC purity: 62% (214 nm) Mass: found peak 457.3 (M+l) at 1.752 min.
[001747] Step 7. Synthesis of tert-butyl 4-[5-(2-oxoethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000353_0002
[001748] To a solution of tert-butyl 4-[5-(2-hydroxyethyl)-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine-l -carboxylate (50 mg, 0.11 mmol) in DCM (5 mL) at 0 °C was added DMP (93 mg, 0.23 mmol). The reaction mixture was stirred at this temperature for Ih. The reaction was diluted with DCM (30 mL) and filtered to afford the desired product tert-butyl 4- [5-(2-oxoethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (50 mg, crude) as a yellow solid.
[001749JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 21%(214 nm) Mass: found peak 455.3 (M+l) at 1.490 min.
[001750] Step 8. Synthesis of tert-butyl 4-[5-(2,2-difluoroethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000354_0001
[001751] To a solution of tert-butyl 4-[5-(2-oxoethyl)-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazine-l -carboxylate (40 mg, 0.088 mmol) in DCM (6 mL) at 0 °C was added DAST (71 mg, 0.44 mmol). The reaction mixture was stirred at this temperature for Ih. The reaction mixture was filtered and diluted with water (5 mL), then extracted with DCM (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography 2-30% ethyl acetate in petroleum ether to afford tert-butyl 4-[5-(2,2- difluoroethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (34 mg, 75.2% yield) as a yellow solid.
[001752JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 92.76% (214 nm) Mass: found peak 477.3 (M+l) at 1.886 min.
[001753] Step 9. Synthesis of l-[5-(2,2-difluoroethyl)-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000354_0002
[001754] To a solution of tert-butyl 4-[5-(2,2-difluoroethyl)-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine-l -carboxylate (34 mg, 0.07 mmol) in dichloromethane (3 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5- (2,2-difluoroethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (25 mg, 72.4% yield) as a yellow oil.
[001755] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 77.78% (214 nm); Mass: found peak 377.2 (M+H)+ at 1.443 min. [001756] Step 10. Synthesis of 4-[2-[4-[5-methoxy-l-[3-(trifluoromethyl)-l- bicyclo[l.l.l]pentanyl]pyrazolo[3,4-b]pyridin-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 99).
Figure imgf000355_0001
[001757] To a mixture of l-[5-(2,2-difluoroethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (15 mg, 0.04 mmol) and DIPEA (16 mg, 0.12 mmol) in NMP (2 mL) was added 4- (2-chloroethyl)morpholine hydrochloride (9 mg, 0.06 mmol). The reaction was stirred at 120 °C for 16h in microwave. The reaction was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methoxy-l-[3-(trifhioromethyl)-l- bicyclo[ 1.1.1 ]pentanyl]pyrazolo[3 ,4-b]pyri din-3 -yl]piperazin- 1 -yl]ethyl]morpholine (3.5 mg, yield 18%) as a yellow oil.
[001758JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X-
Bridge : C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100 % (214 nm) Mass: found peak 490.1 (M+l) at 1.968 min.
[001759] 'H NMR (400 MHz, CDCh) 5 7.45-7.39 (m, 2H), 7.33-7.27 (m, 2H), 6.11-5.77 (m, 2H), 3.72 (t, J = 4.4 Hz, 4H), 3.29 (t, J = 4.4 Hz, 4H), 3.17 (td, J = 16.2, 4.2 Hz, 2H), 2.68 - 2.43 (m, 12H) ppm.
Example S100. Synthesis of 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol- 3-yl]piperazin-l-yl]ethyl]morpholine (Compound 100).
[001760] Compound 100 was prepared as outlined below.
Figure imgf000355_0002
[001761] Step 1. Synthesis of ethyl 2-(4-bromo-2-fluoro-phenyl)-2,2-difluoro-acetate.
Figure imgf000356_0001
[001762] To a stirred solution of ethyl 2-bromo-2,2-difluoro-acetate (2.53 g, 12.5 mmol) in DMSO (5 mL) was added copper powder (0.792 g, 12.5 mmol) at room temperature. After Ih, 4-bromo-2-fluoro-l -iodo-benzene (1.5 g, 4.99 mmol) was added and the resulting solution heated to 70°C and stirred for an additional 3 h. After complete conversion of the starting material, ice water (40 mL) and EtOAc (40 mL) were added. Precipitates were removed by filtration and the aqueous layer was extracted with EtOAc (40 mL*2). The organic layers were combined, dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0-10% acetone in petroleum ether) to afford the desired product ethyl 2-(4-bromo-2-fluoro-phenyl)-2,2- difluoro-acetate (1.4 g, 93.5% yield) as a colorless oil.
[001763JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 98.87% (214nm); Mass: No peak, at 1.390 min. [001764] XH NMR (500 MHz, CDCh) 5 7.52 (t, J = 8.0 Hz, IH), 7.42(dd, J = 8.0, 1.5 Hz, IH), 7.36 - 7.33 (m, IH), 4.35 (q, J = 7.0 Hz, 2H), 1.32 (t, J = 7.0 Hz, 3H) ppm.
[001765] Step 2. Synthesis of 2-(4-bromo-2-fluoro-phenyl)-2,2-difluoro-ethanol.
Figure imgf000356_0002
[001766] To a solution of ethyl 2-(4-bromo-2-fluoro-phenyl)-2,2-difluoro-acetate (1.4 g, 4.71 mmol) in tetrahydrofuran (80 mL) and ethanol (12 mL) was added NaBH4 (357 mg, 9.43 mmol). The reaction was stirred at room temperature for 2h. The reaction was quenched with water, then directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 4-20% acetone in petroleum ether) to afford the desired product 2-(4-bromo-2- fhioro-phenyl)-2,2-difhioro-ethanol (1.17 g, yield 97.3%) as a yellow solid.
[001767] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: no peak, at 1.184 min. [001768] 'H NMR (500 MHz, CDCh) 5 7.45 (t, J = 8.0 Hz, 1H), 7.39 (dd, J = 8.5, 1.5 Hz, 1H), 7.34 (d, J = 10.0 Hz, 1H), 4.08 (dt, J = 13.5, 6.0 Hz, 2H), 1.94 (t, J = 7.0 Hz, 1H) ppm.
[001769] Step 3. Synthesis of 6-bromo-3,3-difluoro-2H-benzofuran.
Figure imgf000357_0001
[001770] A mixture of 2-(4-bromo-2-fluoro-phenyl)-2,2-difluoro-ethanol (1.17 g, 4.59 mmol), cesium carbonate (4.48 g, 13.8 mmol) and 18-crown-6 (0.606 g, 2.29 mmol) in tetrahydrofuran (15 mL) was stirred at 80 °C for 16h. The reaction was directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 0-20% acetone in petroleum ether) to afford the desired product 6-bromo-3,3-difluoro-2H-benzofuran (620 mg, yield 55.6%) as a yellow solid.
[001771JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 96.62% (214nm); Mass: no peak at 1.488 min. [001772] 'H NMR (500 MHz, CDCh) 5 7.88 (s, 1H), 7.39 (d, J = 8.0 Hz, 1H), 7.36 (dd, J = 8.5, 1.0 Hz,lH), 4.88 (t, J = 11.0 Hz, 2H) ppm.
[001773] Step 4. Synthesis of tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000357_0002
[001774]Under argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (100 mg, 0.375 mmol), 6-bromo-3,3-difluoro-2H-benzofuran (132 mg, 0.563 mmol), copper(I) iodide (7.2 mg, 0.0375 mmol), 2-(dimethylamino)acetic acid (7.7 mg, 0.075 mmol) and potassium carbonate (114 mg, 0.826 mmol) in dry DMSO (10 mL) was stirred at 120 °C in tube for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (27 mg, yield 10.9%) as a yellow solid.
[001775] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 64.02% (214nm); Mass: 365.3 [M - 55]+ at 1.590 min.
[001776] 1 H NMR (500 MHz, CDCh) 57.56 (d, J = 11.0 Hz, 1H), 7.34 (s, 1H), 7.14 (dd, J = 10.5, 2.0 Hz, 1H), 5.74 (s, 1H), 4.69 (t, J = 13.0 Hz, 2H), 3.56 (t, J = 6.5 Hz, 4H), 3.23 (t, J = 6.5 Hz, 4H), 2.38 (s, 3H), 1.48 (s, 9H) ppm.
[001777] Step 5. Synthesis of l-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3- yl] piperazine.
Figure imgf000358_0001
[001778] To a solution of tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (purity: 64%, 27 mg, 0.041 mmol) in dichloromethane (8 mL) was added 2,2,2-trifluoroacetic acid (1.0 mL, 13.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude desired product l-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3- yl]piperazine (30 mg, 68.1% yield) as a yellow solid.
[001779JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 29.88% (214nm); Mass: 321.4 [M + 1]+ at 0.975 min.
[001780] Step 6. Synthesis of 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 100).
Figure imgf000359_0001
[001781] To a solution of l-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3- yl]piperazine (crude, about 0.041 mmol), potassium carbonate (28 mg, 0.205 mmol) and KI (6.8 mg, 0.041 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (11.4 mg, 0.061 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-6-yl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl]morpholine (7.6 mg, yield 42.8%) as a white solid.
[001782JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 434.3 (M+l) at 1.997 min.
[001783] 'H NMR (400 MHz, CDCh) 5 7.52 (d, J = 8.4 Hz, 1H), 7.31 (s, 1H), 7.12 (dd, J = 8.4, 1.2 Hz, 1H), 5.70 (s, 1H), 4.66 (t, J = 10.8 Hz, 2H), 3.70 (t, J = 4.4 Hz, 4H), 3.26 (t, J = 4.8 Hz, 4H), 2.61(t, J = 4.8 Hz, 4H), 2.58 - 2.52 (m, 4H), 2.49 (t, J = 4.4 Hz, 4H), 2.35 (s, 3H) ppm.
Example S101. Synthesis of 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-
3-yl]piperazin-l-yl]ethyl]morpholine (Compound 101).
[001784] Compound 101 was prepared as outlined below.
Figure imgf000359_0002
[001785] Step 1. Synthesis of ethyl 2-(5-bromo-2-fluoro-phenyl)-2,2-difluoro-acetate.
Figure imgf000360_0001
[001786] To a stirred solution of ethyl 2-bromo-2,2-difluoro-acetate (5.06 g, 24.9 mmol) in DMSO (10 mL) was added copper powder (1.58 g, 24.9 mmol) at room temperature. After Ih, 4-bromo-l-fluoro-2-iodo-benzene (3.0 g, 9.97 mmol) was added, and the resulting solution heated to 70 °C and stirred for an additional 3 h. After complete conversion of the starting material, ice water (40 mL) and EtOAc (40 mL) were added. Precipitates were removed by filtration and the aqueous layer was extracted with EtOAc (100 mL*2). The organic layers were combined, dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120g silica gel column @100mL/min, eluting with 0-10% acetone in petroleum ether) to afford the desired product ethyl 2-(5-bromo-2-fluoro-phenyl)-2,2- difluoro-acetate (2.6 g, 87.8% yield) as a colorless oil.
[001787] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: found peak no peak, at 1.386 min.
[001788] 'H NMR (500 MHz, CDCh) 5 7.77 (dd, J = 6.0, 3.0 Hz, IH), 7.63-7.59 (m, IH), 7.04 (t, J = 9.0 Hz, IH), 4.36 (q, J = 7.0 Hz, 2H), 1.33 (t, J = 7.0 Hz, 3H) ppm.
[001789] Step 2. Synthesis of 2-(5-bromo-2-fluoro-phenyl)-2,2-difluoro-ethanol.
Figure imgf000360_0002
[001790] To a solution of ethyl 2-(5-bromo-2-fluoro-phenyl)-2,2-difluoro-acetate (2.6 g, 8.75 mmol) in tetrahydrofuran (150 mL) and ethanol (22 mL) was added NaBH4 (662 mg, 17.5 mmol). The reaction was stirred at room temperature for 2h. The reaction quenched with water, then directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 4-20% acetone in petroleum ether) to afford the desired product 2-(5-bromo-2- fluoro-phenyl)-2,2-difluoro-ethanol (2.2 g, yield 98.6%) as a yellow solid.
[001791JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 97.83% (214nm); Mass: no peak at 1.181 min. [001792] 'H NMR (500 MHz, CDCh) 5 7.70 (dd, J = 6.0, 3.0 Hz, 1H), 7.59-7.55 (m, 1H), 7.04 (t, J = 9.5 Hz, 1H), 4.10 (t, J = 13 Hz, 2H), 1.95 (brs, 1H) ppm.
[001793] Step 3. Synthesis of 5-bromo-3,3-difluoro-2H-benzofuran.
Figure imgf000361_0001
[001794] A mixture of 2-(5-bromo-2-fluoro-phenyl)-2,2-difluoro-ethanol (1.1 g, 4.31 mmol), cesium carbonate (4.22 g, 12.9 mmol) and 18-crown-6 (0.57 g, 2.16 mmol) in tetrahydrofuran (15 mL) was stirred at 80 °C for 16h. The reaction was directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 0-20% acetone in petroleum ether) to afford the desired product 5-bromo-3,3-difluoro-2H-benzofuran (500 mg, yield 49.3%) as a white solid.
[001795] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: no peak at 1.493 min.
[001796] 'H NMR (500 MHz, CDCh) 5 7.64 (d, J = 2.5 Hz, 1H), 7.57 (dd, J = 9.0, 2.0 Hz, 1H), 7.06 (d, J = 9.5 Hz,lH), 4.63 (t, J = 11.0 Hz, 2H) ppm.
[001797] Step 4. Synthesis of tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000361_0002
[001798]Under argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (50 mg, 0.188 mmol), 5-bromo-3,3-difluoro-2H-benzofuran (66 mg, 0.282 mmol), copper(I) iodide (35.8 mg, 0.188 mmol), 2-(dimethylamino)acetic acid (38.7 mg, 0.375 mmol) and potassium carbonate (77.8 mg, 0.563 mmol) in dry DMSO (6 mL) was stirred at 120 °C for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3-yl]piperazine-l- carboxylate (18 mg, yield 15.7%) as a yellow solid. [001799JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 gm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 69.0% (214nm); Mass: found peak 365.3 (M + H) at 2.386 min.
[001800] XH NMR (400 MHz, CDCh) 57.60 (d, J = 2.8 Hz, 1H), 7.51 (dd, J = 8.4, 2.8 Hz, 1H), 7.22 (d, J = 9.2 Hz, 1H), 5.70 (s, 1H), 4.70 (t, J = 11.2 Hz, 2H), 3.55 (t, J = 4.8 Hz, 4H), 3.20 (t, J = 4.8 Hz, 4H), 2.28 (s, 3H), 1.47 (s, 9H) ppm.
[001801] Step 5. Synthesis of l-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3- yl] piperazine.
Figure imgf000362_0001
[001802] To a solution of tert-butyl 4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (purity: 60%, 55 mg, 0.078 mmol) in dichloromethane (10 mL) was added 2,2,2-trifluoroacetic acid (2.0 mL, 26.1 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude desired product l-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3- yl]piperazine, which was used directly in the next step.
[001803JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.3 min at 2.0 mL/min; Column: SUNFIRE C18 (4.6x 50 mm, 3.5gm); Column Temperature: 50 °C; LC purity: 36.98% (214nm); Mass: 321.4 [M + 1]+ at 1.349 min.
[001804] Step 6. Synthesis of 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 101).
Figure imgf000362_0002
[001805] To a solution of l-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3- yl]piperazine (crude, about 0.078 mmol), potassium carbonate (54 mg, 0.39 mmol) and KI (12.9 mg, 0.078 mmol) in 95% ethanol (12 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (21.8 mg, 0.117 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(3,3-difluoro-2H-benzofuran-5-yl)-5-methyl-pyrazol-3-yl]piperazin- l-yl]ethyl]morpholine (6.5 mg, yield 19.2%, 2 steps) as a yellow solid.
[001806] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 434.3 (M+l) at 1.951 min.
[001807] 1 H NMR (400 MHz, CDCh) 5 7.60 (d, J = 2.4 Hz, 1H), 7.51 (dd, J = 8.8, 2.4 Hz, 1H), 7.22 (d, J = 8.8 Hz, 1H), 5.68 (s, 1H), 4.69 (t, J = 10.8 Hz, 2H), 3.73 (t, J = 4.8 Hz, 4H), 3.28 (t, J = 4.8 Hz, 4H), 2.65(t, J = 4.4 Hz, 4H), 2.63-2.55 (m, 4H), 2.52 (t, J = 4.4 Hz, 4H), 2.27 (s, 3H) ppm.
Example S102. Synthesis of 4-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]morpholine (Compound 102).
[001808] Compound 102 was prepared as outlined below.
Figure imgf000363_0001
[001809] Step 1. Synthesis of 2,3,3a,4,7,7a-hexahydro-lH-isoindole.
Figure imgf000363_0002
[001810] To a solution of 3a, 4,7, 7a-tetrahydroisoindole-l, 3-dione (5.0 g, 33.1 mmol) in tetrahydrofuran (100 mL) at 0 °C was added a solution of LiAlH4 in tetrahydrofuran (1.0 M, 132 mL, 132 mmol) dropwise. The reaction was warmed to room temperature and stirred at 40 °C for 16h. The reaction was quenched by water (5.0 g), followed by 15% NaOH aqueous (5.0 g) and water (15 g). The reaction mixture was filtered, and the filtrate was concentrated in vacuo to afford the crude desired product 2,3,3a,4,7,7a-hexahydro- IH-isoindole, which was used directly in next step.
[001811JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak Mass: found peak 124.2 (M+l) at 1.279 min.
[001812] Step 2. Synthesis of tert-butyl l,3,3a,4,7,7a-hexahydroisoindole-2-carboxylate.
Figure imgf000364_0002
Boc
[001813] At 0 °C, to a solution of 2,3,3a,4,7,7a-hexahydro-lH-isoindole (about 33.1 mmol) and EtiN (6.97 mL, 49.6 mmol) in tetrahydrofuran (80 mL) was added BOC2O (7.94 g, 36.4 mmol) dropwise. The reaction was warmed to room temperature and stirred at this temperature for 16h.The reaction was diluted with water (100 mL), extracted with EtOAc (100 mL*3), the organic layers were combined, dried over sodium sulfate and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120g silica gel column @100mL/min, eluting with 0-15% acetone in petroleum ether) to afford the desired product tert-butyl 1, 3, 3a, 4, 7,7a- hexahydroisoindole-2-carboxylate (4.5 g, 48.4% yield (2 steps)) as a brown oil.
[001814JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 79.5% (214 nm) Mass: found peak 246.3 (M+23)+ at 1.383 min.
[001815] XH NMR (500 MHz, CDCI3) 5 5.64 (brs, 2H), 3.40 (brs, 2H), 3.12 (d, J = 8.5 Hz, 2H), 2.29 (brs, 2H), 2.21 (dd, J = 17.0, 6.5 Hz, 2H), 1.90 (dd, J = 16.5, 4.5 Hz, 2H), 1.46 (s, 9H) ppm.
[001816] Step 3. Synthesis of tert-butyl 5-hydroxy-l,3,3a,4,5,6,7,7a-octahydroisoindole-2- carboxylate.
Figure imgf000364_0001
[001817] To a solution of tert-butyl l,3,3a,4,7,7a-hexahydroisoindole-2- (4.1 g, 18.4 mmol) in anhydrous THF (70 mL) at 0 °C was added BH3 DMS (2.0 M in THF, 16.5 mL, 3.3 mmol). The mixture was stirred for 3h at the same temperature. The reaction mixture was then treated with aqueous NaOH (3M, 12.3 mL, 36.9 mmol) and 30% water (9.38 mL, 91.8 mmol), gradually allowed to warm to room temperature, and further stirred for 3h. The reaction mixture was diluted with water (50 mL) and extracted with DCM (2*200 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 5-hydroxy-l,3,3a,4,5,6,7,7a- octahydroisoindole-2-carboxylate (3.54 g, yield 79%) as a yellow solid.
[001818JLCMS method: Mobile Phase: A: water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 264.3 (M+23)+ at 1.090 min.
[001819] 'H NMR (500 MHz, CDCh) 5 3.92-3.86 (m, 1H), 3.40-3.30 (m, 2H), 3.26-3.10 (m, 2H), 2.50-2.42 (m, 1H), 2.14-2.08 (m, 1H), 1.88-1.76 (m, 3H), 1.58-1.52 (m, 1H), 1.46 (s, 10H), 1.40-1.32 (m, 2H) ppm.
[001820] Step 4. Synthesis of 2,3,3a,4,5,6,7,7a-octahydro-lH-isoindol-5-ol hydrochloride.
Figure imgf000365_0001
[001821] To a solution of tert-butyl 5-hydroxy-l,3,3a,4,5,6,7,7a-octahydroisoindole-2- carboxylate (3.5 g, 14.5 mmol) in 1,4-dioxane (20 mL) was added a solution of HCl/dioxane (4 M, 40 mL, 160 mmol). The reaction was stirred at room temperature for 2h. The reaction was concentrated in vacuo. The residue was washed with EtOAc (20 mL), concentrated in vacuo to afford the desired product 2,3,3a,4,5,6,7,7a-octahydro-lH-isoindol-5-ol hydrochloride (2.5 g, 97% yield) as a purple oil.
[001822JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak Mass: found peak 142 (M+l) at 0.418 min.
[001823] XH NMR (400 MHz, CD3OD) 5 3.86-3.78 (m, 1H), 3.37-3.27 (m, 2H), 3.16-3.04 (m, 2H), 2.66-2.56 (m, 1H), 2.38-2.29 (m, 1H), 1.94-1.86 (m, 1H), 1.85-1.77 (m, 2H), 1.67-1.59 (m, 1H), 1.45-1.35 (m, 2H) ppm.
[001824] Step 5. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-ol.
Figure imgf000366_0001
[001825]Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazole (200 mg, 0.623 mmol), 2,3,3a,4,5,6,7,7a-octahydro-lH- isoindol-5-ol hydrochloride (166 mg, 0.934 mmol), tBuXPhos Pd G3 (50 mg, 0.063 mmol) and sodium tert-butoxide (240 mg, 2.5 mmol) in 1,4-di oxane (16 mL) was stirred 100 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was directly purified by flash chromatography (Biotage, 40g silica gel column @75mL/min, eluting with 0- 60% ethyl acetate in petroleum ether) to afford the desired product 2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,3a,4,5,6,7,7a-octahydroisoindol-5-ol (45 mg, yield 18.9%) as a yellow oil.
[001826] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 (4.6x 30 mm, 2.7 pm); Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: 382.3 [M + 1]+ at 1.211 min.
[001827] Step 6. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,3a,4,6,7,7a-hexahydro-lH-isoindol-5-one.
Figure imgf000366_0002
[001828] At 0 °C, to a solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-ol (90 mg, 0.236 mmol) in DCM (10 mL) was added DMP (200 mg, 0.472 mmol). The reaction mixture was stirred at this temperature for Ih. The reaction was diluted with DCM (30 mL), washed with 5% Na2S20s aqueous (20 mL) and 5% sodium bicarbonate aqueous (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-45% ethyl acetate in petroleum ether) to afford the desired product 2-[5-methyl-l -[4-(tri fluoromethoxy )phenyl]pyrazol-3-yl]-3, 3 a, 4,6,7, 7a-hexahydro- IH-isoindol- 5-one (40 mg, yield 43.4%) as a yellow solid. [001829JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.17% (214nm); Mass: 380.1 [M + 1]+ at 1.124 min.
[001830] Step 7. Synthesis of 4-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]morpholine (Compound 102).
Figure imgf000367_0001
[001831] To a solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,3a,4,6,7,7a-hexahydro-lH-isoindol-5-one (68 mg, 0.179 mmol) in 1,2-dichloroethane (10 mL) was added sodium triacetoxyborohydride (114 mg, 0.538 mmol), morpholine (31.2 mg, 0.358 mmol), a drop of acetic acid and 4A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched with water (5 mL) and extracted with dichloromethane (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]morpholine (50.1 mg, yield 61.5%) as a colorless oil.
[001832JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.12% (214 nm) Mass: found peak 451.1 (M+l) at 2.229 min.
[001833] 'H NMR (500 MHz, CDCh) 5 7.49-7.44 (m, 2H), 7.25 (d, J = 8.5 Hz, 2H), 5.51 (s, 1H), 3.77-3.67 (m, 4H), 3.50-3.24 (m, 3H), 3.21-3.15 (m, 1H), 2.67-2.59 (m, 1H), 2.59-2.50 (m, 4H), 2.49-2.39 (m, 1H), 2.30 (s, 3H), 2.25-2.12 (m, 1H), 1.96-1.78 (m, 3H), 1.66-1.57 (m, 1H), 1.43-1.24 (m, 2H) ppm.
Example S103. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]methyl]morpholine (Compound 103).
[001834] Compound 103 was prepared as outlined below.
Figure imgf000368_0001
[001835] Step 1. Synthesis of benzyl 5-hydroxy-l,3,3a,4,5,6,7,7a-octahydroisoindole-2- carboxylate.
Figure imgf000368_0002
[001836] To a solution of 2,3,3a,4,5,6,7,7a-octahydro-lH-isoindol-5-ol hydrochloride (2.0 g, 11.3 mmol) and sodium bicarbonate (2.84 g, 33.8 mmol) in dichloromethane (50 mL) and water (50 mL) was added benzyl carb onochlori date (1.74 mL, 12.4 mmol) dropwise. The reaction was stirred at room temperature for 16h. The reaction was diluted with water (50 mL), extracted with DCM (100 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-60% acetone in petroleum ether) to afford the desired product benzyl 5 -hydroxy - l,3,3a,4,5,6,7,7a-octahydroisoindole-2-carboxylate (2.15 g, 69.4% yield) as a yellow oil. [001837] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 276.3 (M+l) at 1.111 min.
[001838] Step 2. Synthesis of benzyl 6-oxo-3,3a,4,5,7,7a-hexahydro-lH-isoindole-2- carboxylate.
[001839] To a solution of pyridinium chlorochromate (3.57 g, 16.6 mmol) in dichloromethane (100 mL) was added silica gel (3 g). Then a solution of benzyl 5-hydroxy-l,3,3a,4,5,6,7,7a- octahydroisoindole-2-carboxylate (2.28 g, 8.28 mmol) in DCM (30 mL) was added dropwise over the course of 5 min. The reaction mixture was stirred at room temperature for 2h. The reaction was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product benzyl 6-oxo-3, 3a, 4,5,7, 7a-hexahydro-lH- isoindole-2-carboxylate (2.2 g, 95.2% yield) as a yellow oil. [001840JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 97.9% (214 nm) Mass: found peak 274.3 (M+l) at 1.146 min.
[001841] XH NMR (500 MHz, CDCh) 57.40-7.28 (m, 5H), 5.18-5.08 (m, 2H), 3.66-3.56 (m, 2H), 3.47-3.37 (m, 1H), 3.24-3.13 (m, 1H), 2.77-2.67 (m, 1H), 2.57-2.47 (m, 2H), 2.43-2.29 (m, 3H), 2.10-2.01 (m, 1H), 1.92-1.82 (m, 1H) ppm.
[001842] Step 3. Synthesis of benzyl (6Z)-6-(methoxymethylene)-3, 3a, 4,5,7, 7a-hexahydro- lH-isoindole-2-carboxylate.
[001843]At 0 ° C, to a suspension of methoxymethyl(triphenyl)phosphonium chloride (2.51 g, 7.31 mmol) in THF (40 mL) was added tBuOK (903 mg, 8.05 mmol) in portions. The reaction mixture was stirred at 0 °C for 30 min. The hexahydro-lH-isoindole-2-carboxylate (1.0 g, 3.66 mmol) in THF (10 mL) was added dropwise. After warming to room temperature, the mixture was stirred at room temperature for 2h. The reaction was diluted with water (30 mL), extracted with EtOAc (200 mL), washed with sat. NH4Q aqueous (10 mL) and brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product benzyl (6Z)-6-(methoxymethylene)-3,3a,4,5,7,7a- hexahydro-lH-isoindole-2-carboxylate (1.0 g, 90.7% yield) as a yellow oil.
[001844JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 302.3 (M+l) at 1.378 min.
[001845] Step 4. Synthesis of benzyl 5-formyl-l,3,3a,4,5,6,7,7a-octahydroisoindole-2- carboxylate.
Figure imgf000369_0001
[001846] To a solution of benzyl (6Z)-6-(methoxymethylene)-3, 3a, 4,5,7, 7a-hexahydro-lH- isoindole-2-carboxylate (1.0 g, 3.32 mmol) in THF (45 mL) was added HC1 aqueous (2 M, 15 mL, 30 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction was diluted with water (30 mL), extracted with EtOAc (200 mL), washed with sat. sodium bicarbonate (aq) (30 mL) and brine (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product benzyl 5-formyl-l,3,3a,4,5,6,7,7a- octahydroisoindole-2-carboxylate (0.92 g, 96.5% yield) as a yellow oil.
[001847] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm;
Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 288.3 (M+l) at 1.247 min.
[001848] Step 5. Synthesis of benzyl 5-(morpholinomethyl)-l,3,3a,4,5,6,7,7a- octahydroisoindole-2-carboxylate.
Figure imgf000370_0001
[001849] To a solution of benzyl 5-formyl-l,3,3a,4,5,6,7,7a-octahydroisoindole-2-carboxylate (0.92 g, 3.2 mmol) in 1,2-di chloroethane (50 mL) was added sodium triacetoxyborohydride (2.04 g, 9.6 mmol), morpholine (0.558 g, 6.4 mmol), 3 drops of acetic acid and 4A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched with water (5 mL) and extracted with dichloromethane (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0- 10% MeOH in DCM) to afford the desired product benzyl 5-(morpholinomethyl)- l,3,3a,4,5,6,7,7a-octahydroisoindole-2-carboxylate (1.1 g, yield 94.9%) as a yellow oil. [001850JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99% (214 nm) Mass: found peak 359.2 (M+l) at 2.093 min.
[001851] Step 6. Synthesis of 4-(2,3,3a,4,5,6,7,7a-octahydro-lH-isoindol-5-
Figure imgf000370_0002
[001852] Under H2 atmosphere, a mixture of benzyl 5-(morpholinomethyl)-l,3,3a,4,5,6,7,7a- octahydroisoindole-2-carboxylate (0.5 g, 1.39 mmol) and Pd/C (10%, 50 mg) in methanol (20 mL) was stirred at room temperature for 16h. The reaction was filtered, and concentrated in vacuo to afford the desired product 4-(2,3,3a,4,5,6,7,7a-octahydro-lH-isoindol-5- ylmethyl)morpholine (0.35 g, yield about 89.5%) as a yellow oil.
[001853JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak, Mass: found peak 225.2 (M+l) at 1.367 min.
[001854] Step 7. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]methyl]morpholine (Compound 103).
Figure imgf000371_0001
,
[001855]Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazole (150 mg, 0.467 mmol), 4-(2,3,3a,4,5,6,7,7a-octahydro-lH- isoindol-5-ylmethyl)morpholine (210 mg, 0.934 mmol), tBuXPhos Pd G3 (37 mg, 0.0467 mmol) and sodium tert-butoxide (135 mg, 1.4 mmol) in 1,4-dioxane (16 mL) was stirred at 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,3a,4,5,6,7,7a-octahydroisoindol-5-yl]methyl]morpholine (120.9 mg, yield 55.7%) as a white solid.
[001856] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 465.2 (M+l) at 2.340 min.
[001857] 'H NMR (400 MHz, CDCh) 5 7.50-7.44 (m, 2H), 7.25 (d, J = 8.0 Hz, 2H), 5.52 (s, 1H), 3.80-3.63 (m, 4H), 3.48-3.40 (m, 1H), 3.40-3.23 (m, 2H), 3.15 (dd, J = 9.2, 3.6 Hz, 1H), 2.60-2.31 (m, 5H), 2.30 (s, 3H), 2.22-2.05 (m, 3H), 1.93-1.60 (m, 5H), 1.40-1.20 (m, 1H), 1.13- 0.90 (m, 1H) ppm.
Example S104. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,3a,4,5,6,6a-hexahydro-lH-cyclopenta[c]pyrrol-5-yl]methyl]morpholine (Compound 104). [001858] Compound 104 was prepared as outlined below.
Figure imgf000372_0001
[001859] Step 1. Synthesis of tert-butyl 5-hydroxy-3, 3a, 4,5,6, 6a-hexahydro-lH- cyclopenta [c] pyrr ole-2-carboxylate.
Figure imgf000372_0002
[001860] To a solution of tert-butyl 5-oxo-l,3,3a,4,6,6a-hexahydrocyclopenta[c]pyrrole-2- carboxylate (2.0 g, 8.9 mmol) in tetrahydrofuran (150 mL) and ethanol (22 mL) was added NaBH4 (672 mg, 17.8 mmol). The reaction was stirred at room temperature for 2h. The reaction quenched with water (2 mL) and directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 4-20% acetone in petroleum ether) to afford the desired product tert-butyl 5-hydroxy-3,3a,4,5,6,6a-hexahydro-lH-cyclopenta[c]pyrrole-2-carboxylate (1.64 g, yield 81.3%) as a colorless oil.
[001861JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: 250.3 [M+23]+ at 1.073 min.
[001862] 'H NMR (500 MHz, CDCh) 54.33-4.27 (m, 1H), 3.53-3.47 (m, 2H), 3.34 (dd, J = 11.5, 3.5 Hz, 2H), 2.65-2.56 (m, 2H), 2.21-2.14 (m, 2H), 1.54-1.47 (m, 2H), 1.46 (s, 9H) ppm.
[001863] Step 2. Synthesis of l,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrol-5-ol hydrochloride.
Figure imgf000372_0003
[001864] To a solution of tert-butyl 5-hydroxy-3, 3a, 4,5,6, 6a-hexahydro-lH- cyclopenta[c]pyrrole-2-carboxylate (1.6 g, 7.04 mmol) in 1,4-dioxane (10 mL) was added a solution of HCl/di oxane (4 M, 20 mL, 80 mmol). The reaction was stirred at room temperature for 2h. The reaction was concentrated in vacuo. The residue was washed with EtOAc (20 mL), dried in vacuo to afford the desired product l,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrol-5-ol hydrochloride (1.0 g, 86.8% yield) as a white solid.
[001865] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: 250.3 [M+23]+ at 1.073 min. [001866] 'H NMR (500 MHz, CD3OD) 5 4.29-4.25 (m, 1H), 3.30-3.25 (m, 4H), 3.05-2.97 (m, 2H), 2.65-2.56 (m, 2H), 2.04-1.95 (m, 2H), 1.72 (d, J = 13.5 Hz, 2H) ppm.
[001867] Step 3. Synthesis of benzyl 5-hydroxy-3, 3a, 4,5,6, 6a-hexahydro-lH- cyclopenta [c] pyrr ole-2-carboxylate.
Figure imgf000373_0001
[001868] To a solution of l,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrol-5-ol hydrochloride (0.7 g, 4.28 mmol) and sodium bicarbonate (1.08 g, 123.8 mmol) in di chloromethane (20 mL) and water (20 mL) was added benzyl carb onochlori date (0.66 mL, 4.71 mmol) dropwise. The reaction was stirred at room temperature for 16h. The reaction diluted with water (50 mL), extracted with DCM (100 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product benzyl 5-hydroxy-3,3a,4,5,6,6a-hexahydro-lH-cyclopenta[c]pyrrole-2-carboxylate (1.1 g, 98.4% yield) as a colorless oil.
[001869JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 262.3 (M+l) at 1.208 min.
[001870] Step 4. Synthesis of benzyl 5-oxo-l,3,3a,4,6,6a-hexahydrocyclopenta[c]pyrrole-2- carboxylate.
Figure imgf000373_0002
[001871] To a solution of pyridinium chlorochromate (1.81 g, 8.42 mmol) in dichloromethane (50 mL) was added silica gel (2 g). The mixture was stirred at room temperature for 0.5h. Then a solution of benzyl 5-hydroxy-3,3a,4,5,6,6a-hexahydro-lH-cyclopenta[c]pyrrole-2-carboxylate (1.1 g, 4.21 mmol) in DCM (15 mL) was added dropwise over the course of 5 min. The reaction mixture was stirred at room temperature for 2h. The reaction was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product benzyl 5-oxo-l,3,3a,4,6,6a-hexahydrocyclopenta[c]pyrrole-2-carboxylate (1.05 g, 96.2% yield) as a yellow oil.
[001872JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 260.3 (M+l) at 1.218 min.
[001873] Step 5. Synthesis of benzyl 5-(methoxymethylene)-l,3,3a,4,6,6a- hexahydr ocyclopenta [c] pyrrole-2-carboxylate.
Figure imgf000374_0001
[001874] To a suspension of methoxymethyl(triphenyl)phosphonium chloride (2.78 g, 8.1 mmol) in THF (40 mL) at 0 °C was added tBuOK (1.0 g, 8.91 mmol) in portions. The reaction mixture was stirred at 0 °C for 30 min. Then, a solution of benzyl 5-oxo-l,3,3a,4,6,6a- hexahydrocyclopenta[c]pyrrole-2-carboxylate (1.05 g, 4.05 mmol) in THF (10 mL) was added dropwise. After warming to room temperature, the mixture was stirred at room temperature for 2h. The reaction was diluted with water (30 mL), extracted with EtOAc (200 mL), washed with sat. NH4Q aqueous (10 mL) and brine (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product benzyl 5-(methoxymethylene)-l,3,3a,4,6,6a-hexahydrocyclopenta[c]pyrrole-2- carboxylate (0.83 g, 70.1% yield) as a colorless oil.
[001875] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 98.31% (214 nm) Mass: found peak 288.3 (M+l) at 1.344 min. [001876] Step 6. Synthesis of benzyl 5-formyl-3, 3a, 4,5,6, 6a-hexahydro-lH- cyclopenta [c] pyrr ole-2-carboxylate.
Figure imgf000375_0001
[001877] To a solution of benzyl 5-(methoxymethylene)-l,3,3a,4,6,6a-hexahydrocyclopenta[c] pyrrole-2-carboxylate (0.83 g, 2.89 mmol) in THF (30 mL) was added HC1 aqueous (2 M, 15 mL, 30 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction was diluted with water (30 mL), extracted with EtOAc (200 mL), washed with sat. sodium bicarbonate aqueous (30 mL) and brine (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product benzyl 5-formyl-3, 3a, 4,5,6, 6a-hexahydro- lH-cyclopenta[c]pyrrole-2-carboxylate (0.77 g, 94.5% yield) as a yellow oil.
[001878JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.88% (214 nm) Mass: found peak 288.3 (M+l) at 1.255 min.
[001879] Step 7. Synthesis of benzyl 5-formyl-3,3a,4,5,6,6a-hexahydro-lH- cyclopenta [c] pyrr ole-2-carboxylate.
Figure imgf000375_0002
[001880] To a solution of benzyl 5-formyl-3, 3a, 4,5,6, 6a-hexahydro-lH-cyclopenta[c]pyrrole-2- carboxylate (0.77 g, 2.82 mmol) in 1,2-di chloroethane (35 mL) was added sodium triacetoxyborohydride (1.79 g, 8.45 mmol), morpholine (0.491 g, 5.63 mmol), 3 drops of acetic acid and 4A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched with water (5 mL) and extracted with di chloromethane (10 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the desired product benzyl 5-formyl-3,3a,4,5,6,6a- hexahydro-lH-cyclopenta[c]pyrrole-2-carboxylate (0.95 g, yield 97.9%) as a yellow oil.
[001881JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 345.2 (M+l) at 2.008 min.
[001882] Step 8. Synthesis of 4-(l,2,3,3a,4,5,6,6a-octahydrocyclopenta[c]pyrrol-5- ylmethyl)morpholine.
Figure imgf000376_0001
[001883] Under H2 atmosphere, a mixture of benzyl 5-(morpholinomethyl)-3,3a,4,5,6,6a- hexahydro-lH-cyclopenta[c]pyrrole-2-carboxylate (0.95 g, 2.76 mmol) and Pd/C (10%, 100 mg) in methanol (40 mL) was stirred at room temperature for 16h. The reaction mixture was filtered and concentrated in vacuo to afford the desired product 4-(l,2,3,3a,4,5,6,6a- octahydrocyclopenta[c]pyrrol-5-ylmethyl) morpholine (0.58 g, yield 100%) as a yellow oil. [001884JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak; Mass: found peak 211.2 (M+l) at 1.292 min.
[001885] Step 9. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,3a,4,5,6,6a-hexahydro-lH-cyclopenta[c]pyrrol-5-yl]methyl]morpholine (Compound 104).
Figure imgf000376_0002
100°c, 16h F'
[001886]Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl] pyrazole (100 mg, 0.311 mmol), 4-(l,2,3,3a,4,5,6,6a- octahydrocyclopentafc] pyrrol-5-ylmethyl)morpholine (131 mg, 0.623 mmol), tBuXPhos Pd G3 (25 mg, 0.0311 mmol) and sodium tert-butoxide (90 mg, 0.934 mmol) in 1,4-dioxane (16 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[[2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-3, 3a, 4,5,6, 6a-hexahydro-lH-cyclopenta[c]pyrrol-5- yl]methyl]morpholine (110.8 mg, yield 74.2%) as a white solid. [001887] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93.99% (214 nm) Mass: found peak 451.2 (M+l) at 2.276 min.
[001888] 'H NMR (400 MHz, CDCh) 5 7.50-7.44 (m, 2H), 7.26 (d, J = 8.4 Hz, 2H), 5.64-5.58 (m, 1H), 3.77-3.65 (m, 4H), 3.48-2.96 (m, 4H), 3.87-3.78 (m, 2H), 2.50-2.31 (m, 5H), 2.29 (s, 3H), 2.28-2.10 (m, 2H), 1.76-1.63 (m, 2H), 1.581.45 (m, 2H) ppm.
Example S105. Synthesis of 4-[2-[4-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 105).
[001889] Compound 105 was prepared as outlined below.
Figure imgf000377_0001
[001890] Step 2. Synthesis of 3-bromo-l-[4-(trifluoromethyl)phenyl]-5-vinyl-pyrazole.
Figure imgf000377_0002
Na2CO3(3.0 eq) dioxane/H20
[001891]Under argon atmosphere, a mixture of 3,5-dibromo-l-[4- (trifluoromethyl)phenyl]pyrazole (500 mg, 1.3 mmol), potassium ethenyltrifluoroborate (180 mg, 1.3 mmol), l,r-bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (110 mg, 0.13 mmol) and sodium carbonate (280 mg, 2.6 mmol) in anhydrous dioxane/water (10/2 mL) was stirred at 80 °C for 16h.Then concentrated and purified by SGC (petroleum etherethyl acetate=l : l) to afford the product 3-bromo-l-[4-(trifluoromethyl)phenyl]- 5-vinyl-pyrazole (400 mg, 94.9%) as colorless oil.
[001892JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 85% (214 nm), Mass: found peak 318.0 (M+l) at 2.278 min.
[001893] Step 3. Synthesis of 3-bromo-5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazole.
Figure imgf000378_0001
[001894]Under hydrogen atmosphere, a mixture of 3-bromo-l-[4-(trifluoromethyl)phenyl]-5- vinyl-pyrazole (400 mg, 0.01 mmol) and PtCh (15 mg, w/w, 30%) in THF (2 mL) was stirred at RT for Ih. Then the reaction was filtered with Celite. The filtrate was concentrated to afford the desired product 3-bromo-5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazole (400 mg, 99%) as a yellow oil.
[001895] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 82% (214 nm), Mass: found peak 319.1 (M+l) at 1.462 min.
[001896] Step 4. Synthesis of tert-butyl 4-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- y 1] piperazine-l-carboxylate.
Figure imgf000378_0002
[001897]Under argon atmosphere, a mixture of 3-bromo-5-ethyl-l-[4- (trifluoromethyl)phenyl]pyrazole (400 mg, 1.23 mmol), tert-butyl piperazine-l-carboxylate (460 mg, 0.247 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladipm;dicyclohexyl-[2- (2,4,6-triisopropylphenyl)phenyl]phosphane (104 mg, 0.123 mmol) and sodium tert-butoxide (356 mg, 37 mmol) in anhydrous dioxane (10 mL) in a sealed tube was stirred at 100 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the product tert-butyl 4-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (20 mg, 3.62%) as a yellow solid.
[001898JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 2 mL/min; Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 89% (214 nm), Mass: found peak 425.3(M+1) at 2.333 min.
[001899] Step 5. Synthesis of l-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000378_0003
[001900] To a solution of tert-butyl 4-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (20 mg, 0.04 mmol) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the reaction mixture was concentrated to remove solvent, and potassium carbonate was added to give pH=10. The product was extracted with DCM and dried over sodium sulfate. The reaction mixture was concentrated to afford the desired product l-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (15 mg, yield: 98.2 %) as a yellow oil.
[001901JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 77% (254 nm) Mass: found peak 325.3(M+1) at 1.035 min.
[001902] Step 6. Synthesis of 4-[2-[4-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 105).
Figure imgf000379_0001
[001903] To a solution of l-[5-ethyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (15 mg, 0.046 mmol), potassium carbonate (32 mg, 0.2 mmol) and KI (7.7 mg, 0.04 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine (10 mg, 0.07 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-ethyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (9.3 mg, yield 45%) as a white solid.
[001904JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity:92% (214 nm) Mass: found peak 438.1 (M+l) at 2.075 min.
[001905] 'H NMR (400 MHz, CDCh) 5 7.60 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 8.4 Hz, 2H), 5.69 (s, 1H), 3.65 (t, J = 4.8 Hz, 4H), 3.22 (t, J = 4.8 Hz, 4H), 2.62 (q, J = 7.6 Hz, 2H), 2.56 (t, J = 5.2 Hz, 4H), 2.53 - 2.47 (m, 4H), 2.47 - 2.39 (m, 4H), 1.17 (t, J = 7.6 Hz, 3H) ppm. Example S106. Synthesis of 4-[2-[4-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 106).
[001906] Compound 106 was prepared as outlined below.
Figure imgf000380_0001
[001907] Step 1. Synthesis of 3-bromo-5-(2-methylprop-l-enyl)-l-[4-
(trifluoromethoxy)phenyljpyrazole.
Figure imgf000380_0002
80°c, 16h
[001908] Under argon atmosphere, a mixture of 3,5-dibromo-l-[4- (trifluoromethoxy)phenyl]pyrazole (200 mg, 0.510 mmol), 2-methylprop-l-enylboronic acid (46 mg, 0.46 mmol), l,r-bis(diphenylphosphino)ferrocene-palladium(II)di chloride dichloromethane complex (41 mg, 0.051 mmol) and sodium carbonate (54 mg, 51 mmol) in anhydrous dioxane/water (3/0.5 mL) was stirred at 80 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : l) to afford the product 3- bromo-5-(2-methylprop-l-enyl)-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 54.3%) as a colorless oil.
[001909JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 67% (214 nm), Mass: found peak 347.1 (M+l) at 1.511 mm. [001910] Step 2. Synthesis of 3-bromo-5-isobutyl-l-(4-(trifluoromethoxy)phenyl)-lH- pyrazole.
Figure imgf000381_0001
[001911]Under hydrogen atmosphere, a mixture of 3-bromo-5-(2-methylprop-l-en-l-yl)-l-(4- (trifluoromethoxy)phenyl)-lH-pyrazole (100 mg, 0.27 mmol) and PtCh(15 mg, vi/vi, 30%) in THF (2 mL) was stirred at RT for 30 min. Then the reaction was filtered with Celite. The filtrate was concentrated to afford the desired product 3-bromo-5-isobutyl-l-(4- (trifluoromethoxy)phenyl)-lH-pyrazole (80 mg, 80%) as a yellow oil.
[001912JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 51% (214 nm), Mass: found peak 363.0 (M+l) at 1.541 min.
[001913] Step 3. Synthesis of tert-butyl 4-[5-isobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000381_0002
[001914] To a solution of 3-bromo-5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (50 mg, 0.138 mmol) in dry 1,4-dioxane (3 mL) was added tert-butyl piperazine- 1 -carboxylate (51 mg, 0.27 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladipm;dicyclohexyl-[2-(2,4,6- triisopropylphenyl)phenyl]phosphane (11 mg, 0.01 mmol), and sodium tert-butoxide (40 mg, 0.4 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 16h. The mixture was filtered. The filtrate was purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the desired product tert-butyl 4-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine- 1-carboxylate (60 mg, yield 93%) as a colorless oil. [001915] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 88% (214 nm) Mass: found peak 469.3 (M+l) at 1.591 min.
[001916] Step 4. Synthesis of l-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000382_0001
[001917] To a solution of tert-butyl 4-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (50 mg, 0.11 mmol) in dichloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the reaction mixture was concentrated to remove solvent and potassium carbonate was added to give pH=10. The product was extracted with DCM and dried over sodium sulfate. The reaction mixture was concentrated to afford the desired product l-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperazine (35 mg, yield 89 %) as a yellow oil.
[001918JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 80% (214 nm), Mass: found peak 369.1 (M+l) at 2.288 min.
[001919] Step 5. Synthesis of 4-[2-[4-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 106).
Figure imgf000382_0002
[001920] To a solution of l-[5-isobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (50 mg, 0.136 mmol), potassium carbonate (93.8 mg, 0.68 mmol) and KI (22.5 mg, 0.136 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine (30.5 mg, 0.204 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-isobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (10.5 mg, yield 15.9%) as a yellow solid.
[001921JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C.LC purity:98.7% (214 nm) Mass: found peak 482.2 (M+l) at 2.195 min.
[001922] 'H NMR (500 MHz, CDCh) 5 7.43 - 7.10 (m, 2H), 7.27 (d, J = 8.5 Hz, 2H), 5.69 (s, 1H), 3.71 (t, J = 4.5 Hz, 4H), 3.26 (t, J = 4.5 Hz, 4H), 2.62 (t, J = 5.0 Hz, 2H), 2.59-2.54 (m, 4H), 2.50 (s, 4H), 2.46 (d, J = 7.5 Hz, 4H), 1.82 (m, lH),0.88 (d, J = 7.0 Hz, 6H) ppm.
Example S107. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7-yl]methyl]morpholine (Compound 107). [001923] Compound 107 was prepared as outlined below.
Figure imgf000383_0001
[001924] Step 1. Synthesis of dimethyl piperidine-2,5-dicarboxylate
Figure imgf000383_0002
[001925]Under hydrogen, to a solution of dimethyl pyridine-2,5-dicarboxylate (1.0 g, 0.5 mmol) in acetic acid (20 mL) was added platinum dioxide (150 mg, 15%w/w). The reaction mixture was stirred at rt for 16h. The mixture was filtered. The filtrate was concentrated to afford the desired product dimethyl piperidine-2,5-dicarboxylate (1.0 g, yield 97%) as a colorless oil. [001926] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 202.1 (M+l) at 0.186 min.
[001927] Step 2. Synthesis of dimethyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,5- dicarboxylate.
Figure imgf000384_0001
[001928] A stirred mixture of DCM (20 mL), N-(2-hydroxyethyl)phthalimide (1.43 g, 7.45 mmol) and 2,6-lutidine (1.16 mL, 9.94 mmol ) was cooled to 0 °C. Maintaining the temperature below 15 °C, trifluoromethanesulfonicanhydride (1.67 mL, 9.94 mmol) was added slowly over Ih. The resulting mixture was stirred at rt for 2h, then washed sequentially with water (10 mL), 2N HC1 (10 mL) and water (10 mL) to yield a solution of 2-(l,3-dioxoisoindolin-2-yl)ethyl trifluoromethanesulfonate. At 20-25 °C, a separate reaction vessel was charged with DCM (10 mL), water and sodium carbonate (2.63 g, 24.8 mmol). After stirring for 15 minutes, dimethyl piperidine-2,5-dicarboxylate (1 g, 4.97 mmol) in DCM (10 mL) was added, and the mixture was stirred for 16h. The organic layer was separated and crystallized by DCM to afford dimethyl 1- [2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,5-dicarboxylate (800 mg , yield 43%) as a yellow solid.
[001929JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm) Mass: found peak 375.3 (M+l) at 1.17 min.
[001930] Step 3. Synthesis of methyl l-oxo-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine- 7-carboxylate.
Figure imgf000385_0001
[001931] A solution of dimethyl l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]piperidine-2,5- dicarboxylate (800 mg, 2.14 mmol) and hydrazine hydrate solution (0.2 mL, 4 mmol) in MeOH (10 mL) was stirred for 16h at rt. The organic layer was separated and crystallized by DCM (10 mL). The filtrate was concentrated to afford the desired product methyl l-oxo-2,3,4,6,7,8,9,9a- octahydropyrido[l,2-a]pyrazine-7-carboxylate (400 mg , yield 88.2%) as a yellow solid.
[001932JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90% (214 nm) Mass: found peak 213.1 (M+l) at 1.395 min.
[001933] Step 4. Synthesis of 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-7- ylmethanol.
Figure imgf000385_0002
[001934] To a solution of methyl l-oxo-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-7- carboxylate (383 mg, 1.6 mmol) in tetrahydrofuran (20 mL) at 0 °C was added LiAlFL (18.8 mL, 18.8 mmol) under Ar atomosphere. The reaction was stirred at 60 °C for 3h. Next, 20 drops of water and 20 drops of 15% NaOH were added to the mixture, followed by addition of 60 drops of water, then the reaction mixture was dried over sodium sulfate, filtered, and concentrated to dryness to afford 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-7- ylmethanol (400 mg, crude) as a yellow oil.
[001935] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; Mass: found peak 171.3 (M + H) at 0.40 min.
[001936] Step 5. Synthesis of [2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7-yl]methanol.
Figure imgf000386_0001
[001937] To a solution of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (370 mg, 1.15 mmol) in dry 1,4-dioxane (10 mL) was added 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2- a]pyrazin-7-ylmethanol (294 mg, 1.73 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy- palladipm;dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (48 mg, 0.05 mmol), and sodium tert-butoxide (332 mg, 0.34 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The filtrate was purified by SGC (DCM:MeOH= 20: 1) to afford the desired product [2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7- yl]methanol (90 mg, yield 19%) as a yellow oil.
[001938JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 86% (214 nm) Mass: found peak 411.3 (M+l) at 1.041 min.
[001939] Step 6. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-7-carbaldehyde.
Figure imgf000386_0002
[001940] At 0 °C, to a solution of [2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7-yl]methanol (45 mg, 0.11 mmol) and DIPEA (70.9 mg, 0.55 mmol) in DCM (5 mL) was added dropwise pyridine sulfur trioxide (87.3 mg, 0.55 mmol) in DMSO (1 mL). The reaction mixture was stirred at this temperature for Ih. The reaction was diluted with DCM (30 mL), washed with 5% Na2SOs aqueous (20 mL) and 5% sodium bicarbonate aqueous (20 mL), then the organic phase was dried over sodium sulfate, filtered, and concentrated in vacuo. Then the crude product was diluted with EtOAc (20 mL), washed with saturated sodium chloride (lOmL x 3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-7-carbaldehyde (40 mg, yield 43.4%) as a yellow solid.
[001941JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 86% (214nm); Mass: 409.3 [M + 1]+ at 1.043 min.
[001942] Step 7. Synthesis of 4-[[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7-yl]methyl]morpholine (Compound 107).
Figure imgf000387_0001
[001943] To a solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazine-7-carbaldehyde (40 mg, 0.0979 mmol) in methanol (5 mL) was added sodium triacetoxyborohydride (62.3 mg, 0.294 mmol), morpholine (17.1 mg, 0.196 mmol), a drop of acetic acid and 4A molecular sieves. The reaction mixture was stirred at room temperature for 16h. The mixture was quenched with water (5 mL) and extracted by dichloromethane (10 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[[2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-7- yl]methyl]morpholine (21.4 mg, yield 44.7%) as a colorless oil.
[001944JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.63% (214 nm) Mass: found peak 480.2 (M+l) at 2.258 min.
[001945] 'H NMR (400 MHz, CDCh) 5 7.46 (d, J = 8.9 Hz, 2H), 7.28 (s, 2H), 5.69 (s, 1H), 3.70 (s, 4H), 3.53 (dd, J = 18.0, 12.0 Hz, 2H), 2.94 (s, 1H), 2.72 (t, J = 10.0 Hz, 2H), 2.63 - 2.49 (m, 2H), 2.43 (d, J = 21.6 Hz, 5H), 2.30 (s, 4H), 2.21 (d, J = 8.8 Hz, 1H), 2.03 (s, 1H), 1.89 (s, 1H), 1.80 (d, J = 11.6 Hz, 1H), 1.47 (s, 1H), 1.40 (s, 2H) ppm.
Example S108. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,4-diazepan-l-yl]ethyl]morpholine (Compound 108).
[001946] Compound 108 was prepared as outlined below.
Figure imgf000388_0001
[001947] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-l,4-diazepane-l-carboxylate.
Figure imgf000388_0002
[001948]Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (150 mg, 0.460 mmol), sodium tert-butoxide (133 mg, 1.38 mmol), 4-(2-piperazin-l-ylethyl)morpholine (183 mg, 0.919 mmol) and Pd PEPPSI IPENT (36.5 mg, 0.0459 mmol) in anhydrous 1,4-dioxane (10 mL) was stirred at 100 °C for 16h. The reaction was cooled down to room temperature, directly purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 2-30% ethyl acetate in petroleum ether) to afford tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4- diazepane-1 -carboxylate (130 mg, yield 60.8 %) as a yellow solid.
[001949JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 90% (214 nm) Mass: found peak 441.3 (M+l) at 1.488 min.
[001950] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4- diazepane.
Figure imgf000388_0003
[001951] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 1,4-diazepane-l -carboxylate (130 mg, 0.295 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-diazepane (110 mg, yield 95%) as a yellow oil. [001952JLCMS method: Mobile phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA) Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.2min; Flow Rate: 2.2 mL/min;
Column: Chromolith Fast gradient RP-18e, 50*3 mm, Column Temperature: 40 °C; LC purity: 87.02% (214 nm) Mass: found peak 340.8 (M+l) at 0.782 min.
[001953] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l,4-diazepan-l-yl]ethyl]morpholine (Cmpound 108).
Figure imgf000389_0001
[001954] To a solution of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4- diazepane (60 mg, 0.176 mmol), potassium carbonate (97.5 mg, 0.705 mmol) and KI (30 mg, 0.176 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (65 mg, 0.353 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-diazepan-l-yl]ethyl]morpholine (17.5 mg, yield 21.9%) as a yellow oil.
[001955] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 454.1 (M+l) at 2.103 min.
[001956] 'H NMR (400 MHz, CDCh) 5 7.49-7.43 (m, 2H), 7.26-7.23 (m, 2H), 5.57 (s, 1H), 3.70 (t, J = 4.4 Hz, 4H), 3.57-3.51 (m, 2H), 3.48 (t, J = 6.0 Hz, 2H), 2.87-2.77 (m, 2H), 2.74- 2.60 (m, 4H), 2.56-2.40 (m, 6H), 2.30 (s, 3H), 1.97-1.90 (m, 2H) ppm.
Example S109. Synthesis of 4-[2-[4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 109).
[001957] Compound 109 was prepared as outlined below.
Figure imgf000390_0001
[001958] Step 1. Synthesis of 5-bromo-2-isopropoxy-pyrimidine.
Figure imgf000390_0002
p p
[001959] Sodium hydride was added slowly to propan-2-ol (50 mL) at 0 °C. Then the mixture was stirred for 30 min at 60 °C. Then 5-bromo-2-chloro-pyrimidine (1.0 g, 5.17 mmol) was added. The reaction was stirred for 16h at 60 °C. The reaction mixture was diluted with EtOAc and stirred while NH4Q solution was added until pH=10. The layers were separated, and the aqueous layer was extracted with EtOAc (20mL*3). The combined layers were purified by SGC (petroleum etherethyl acetate=2: l) to afford the desired product 5-bromo-2-isopropoxy- pyrimidine (1g, yield 89.1%) as a yellow oil.
[001960JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 98.35% (214 nm), Mass: found peak 219.1 (M+l) at 1.231 min.
[001961] Step 2. Synthesis of tert-butyl 4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000390_0003
[001962] Linder argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (100 mg, 0.375 mmol), 5-bromo-2-isopropoxy-pyrimidine (163 mg, 0.751 mmol), copper(I) iodide (18 mg, 0.09 mmol), 2-(dimethylamino)acetic acid (19 mg, 0.188 mmol) and potassium phosphate tribasic (240 mg, 1.13 mmol) in DMSO (3 mL) was stirred at 120 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was directly purified by flash chromatography (Biotage, 40 g silica gel column @75 mL/min, eluting with 0-4% MeOH in DCM) to afford the desired product tert-butyl 4-[l-(2- isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate (70 mg, yield 46%) as a yellow solid.
[001963JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 98.35% (214 nm), Mass: found peak 403.4 (M+l) at 1.362 min.
[001964] Step 3. Synthesis of 2-isopropoxy-5-(5-methyl-3-piperazin-l-yl-pyrazol-l- yl)pyrimidine.
Figure imgf000391_0001
[001965] To a solution of tert-butyl 4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.248 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 2-isopropoxy-5-(5-methyl-3-piperazin-l-yl-pyrazol- l-yl)pyrimidine (84 mg, yield 99%) as a yellow oil.
[001966] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 75.2% (214 nm),
Mass: found peak 303.1 (M+l) at 1.705 min.
[001967] Step 4. Synthesis of 4-[2-[4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 109).
Figure imgf000391_0002
[001968] To a solution of 2-isopropoxy-5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)pyrimidine (40 mg, 0.13 mmol), potassium carbonate (73 mg, 0.53 mmol) and KI (22 mg, 0.13 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (49 mg, 0.27 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (21.8 mg, yield 38.9 %) as a white solid.
[001969JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.24 % (214 nm) Mass: found peak 416.70(M+l) at 1.551 min.
[001970] 'H NMR (400 MHz, CDCh) 5 8.58 (s, 2H), 5.72 (s, 1H), 5.28 (dt, J = 12.4, 6.0 Hz, 1H), 3.72(t, J = 4.4 Hz, 4H), 3.25(t, J = 4.4 Hz, 4H), 2.61 (t, J = 4.4 Hz, 4H), 2.59-2.54 (m, 4H), 2.51 (t, J = 4.4 Hz, 4H), 2.27 (s, 3H), 1.42 (s, 3H), 1.41 (s, 3H) ppm.
Example S110. Synthesis of 4 4-[2-[4-[5-methyl-l-[5-(trifluoromethyl)pyrazin-2-yl]pyrazol-
3-yl]piperazin-l-yl]ethyl]morpholine (Compound 110).
[001971] Compound 110 was prepared as outlined below.
Figure imgf000392_0001
[001972] Step 1. Synthesis of tert-butyl 4-[l-(2-isopropoxypyrimidin-5-yl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000392_0002
[001973]Under argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (200 mg, 0.75 mmol), 2-bromo-5-(trifluoromethyl)pyrazine (340 mg, 1.5 mmol), copper(I) iodide (36 mg, 0.18 mmol), 2-(dimethylamino)acetic acid (38 mg, 0.4 mmol) and potassium phosphate tribasic (478 mg, 2.25 mmol) in DMSO (8 mL) was stirred at 120 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was directly purified by flash chromatography (Biotage, 40 g silica gel column @75 mL/min, eluting with 0-4% MeOH in DCM) to afford the desired product tert-butyl 4-[l-(2- isopropoxypyrimidin-5-yl)-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate (40 mg, yield 13.2%) as a yellow solid. [001974JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 70% (214 nm) Mass: found peak 357.3 (M- 55)+ at 1.537 min.
[001975] Step 2. Synthesis of 2-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)-5- (trifluoromethyl)pyrazine.
Figure imgf000393_0001
[001976] To a solution of tert-butyl 4-[5-methyl-l-[5-(trifluoromethyl)pyrazin-2-yl]pyrazol-3- yl]piperazine-l -carboxylate (40 mg, 0.096 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 2-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)-5- (trifluoromethyl)pyrazine (30 mg, yield 92%) as a yellow oil.
[001977] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (254 nm) Mass: found peak 313.2 (M+l) at 1.416 min.
[001978] Step 3. Synthesis of 4-[2-[4-[5-methyl-l-[5-(trifluoromethyl)pyrazin-2-yl]pyrazol- 3-yl]piperazin-l-yl]ethyl]morpholine (Compound 110).
Figure imgf000393_0002
[001979] To a solution of 2-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)-5- (trifluoromethyl)pyrazine (30 mg, 0.096 mmol), potassium carbonate (53 mg, 0.38 mmol) and KI (15 mg, 0.096 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (35 mg, 0.19 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NFLHCCh/water /acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[5- (trifluoromethyl)pyrazin-2-yl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (25.4 mg, yield 62.1 %) as a yellow solid.
[001980JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100.00% (214 nm) Mass: found peak 425.7 (M+l) at 1.817 min.
[001981] XH NMR (400 MHz, CDCh) 5 9.25 (s, 1H), 8.58 (s, 1H), 5.80 (s, 1H), 3.73 (t, J = 4.4 Hz, 4H), 3.34 (t, J = 4.4 Hz, 4H), 2.66 (s, 3H), 2.65 - 2.46 (m, 12H) ppm.
Example Sill. Synthesis of 4-[2-[4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 111).
[001982] Compound 111 was prepared as outlined below.
Figure imgf000394_0001
[001983] Step 1. Synthesis of 3-bromo-5-[(E)-prop-l-enyl]-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000394_0002
[001984] To a solution of 3 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole (200 mg, 0.518 mmol) in dioxane (10 mL) was added ImL water, [(E)-prop-l-enyl]boronic acid (40 mg, 0.446 mmol), disodium carbonate (110 mg, 1.04 mmol), and [1,T- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (38 mg, 0.052 mmol). The reaction mixture was stirred at room temperature for 16h. The mixture was filtered and purified by flash chromatography (PE/DCM=1/1) to afford desired product 3-bromo-5-[(E)-prop-l-enyl]-l-[4- (trifluoromethoxy) phenyl ]pyrazole (160 mg, yield 88.9%) as a colorless oil.
[001985] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 52.6% (214 nm) Mass: found peak 347.0 (M+l) at 2.311 min.
[001986] Step 2. Synthesis of 3-bromo-5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000395_0001
[001987] To a solution of 3-bromo-5-[(E)-prop-l-enyl]-l-[4-(trifluoromethoxy)phenyl]pyrazole (160 mg, 0.404 mmol) in THF (8 mL) was added PdCh (15%, 24 mg). The reaction mixture was stirred at room temperature for 0.5h under H2 atmosphere. The mixture was filtered, and the filtrate was concentrated in vacuo to afford the desired product 3-bromo-5-propyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (80 mg, yield 53.1%) as a white solid.
[001988JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 44.37 % (214 nm) Mass: found peak 349.0 (M+l) at 2.327 min.
[001989] Step 3. Synthesis of tert-butyl 4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl] piperazine- 1-carboxylate.
Figure imgf000395_0002
[001990]Under argon atmosphere, a mixture of 3-bromo-5-propyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (80 mg, 0.229 mmol), sodium tert-butoxide (66 mg, 0.687 mmol), tert-butyl piperazine- 1-carboxylate (85 mg, 0.458 mmol) and Pd PEPPSI IPENT (18.2 mg, 0.023 mmol) in anhydrous 1,4-dioxane (5 mL) was stirred at 100 °C for 16h. The reaction was cooled down to room temperature and directly purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 2-30% ethyl acetate in petroleum ether) to afford tert-butyl 4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (20 mg, yield 13.2%) as a yellow oil.
[001991JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 21% (214 nm) Mass: found peak 455.2 (M+l) at 2.401 min.
[001992] Step 4. Synthesis of l-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000396_0001
[001993] To a solution of tert-butyl 4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (20 mg, 0.044 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperazine (12 mg, yield 76%) as a yellow oil.
[001994JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100 % (214 nm) Mass: found peak 354.9 (M+l) at 1.794 min.
[001995] Step 5. Synthesis of 4-[2-[4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 111).
Figure imgf000396_0002
[001996] To a solution of l-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (12 mg, 0.034 mmol), potassium carbonate (19 mg, 0.135 mmol) and KI (6 mg, 0.0934mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (13 mg, 0.068 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (4.1 mg, yield 24.1 %) as a yellow oil.
[001997] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94.33 % (214 nm) Mass: found peak 468.2 (M+l) at 2.179 min.
[001998] XH NMR (400 MHz, CDCh) 5 7.47-7.41 (m, 2H), 7.25-7.3 l(m, 2H), 5.71 (s, 1H), 3.72 (t, J = 4.8, 4H), 3.27 (t, J = 4.8, 4H), 2.62 (t, J = 4.8, 4H), 2.60-2.54 (m, 6H), 2.51 (t, J = 4.8, 4H), 1.61 (dd, J = 4.8, 7.4 Hz, 2H), 0.94 (t, J = 4.8 , 3H) ppm.
Example S112. Synthesis of 4-[2-[4-[5-methyl-l-[6-(trifluoromethyl)indan-l-yl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 112).
[001999] Compound 112 was prepared as outlined below.
Figure imgf000397_0001
[002000] Step 1. Synthesis of 6-(trifluoromethyl)indan-l-ol.
Figure imgf000397_0002
[002001] To a solution of 6-(trifluoromethyl)indan-l-one (1.5 g, 7.5 mmol) in ethanol (15 mL) and THF (50 mL) was added NaBH4 (427 mg, 11.3 mmol). The reaction was stirred at room temperature for 2h. The reaction quenched with water, then, directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL/min, eluting with 4-20% acetone in petroleum ether) to afford the desired product 6-(trifluoromethyl)indan-l-ol (1.4 g, yield 99%) as a white solid. [002002JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 96.81 % (214 nm) Mass: found peak 185.2 (M+l) at 1.200 min.
[002003] Step 2. Synthesis of tert-butyl 4-[5-methyl-l-[6-(trifluoromethyl)indan-l- yl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000398_0001
[002004] To a solution of 6-(trifluoromethyl)indan-l-ol (1 g, 4.95 mmol) in chloroform (30 mL) was added PBo (6.69 mL, 2.47mmol). The reaction mixture was stirred at 0 °C~RT for 2h. The mixture was concentrated in vacuo. The reaction was quenched with 30 mL water, extracted with DCM (50 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product. Then tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (1.65 g, 0.68 mmol) and potassium carbonate are mixed and dissolved in DMF at 0 °C for 16h. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 0-10% MeOH in DCM) to afford the desired product tert-butyl 4-[5-methyl- 1 -[6-(trifluoromethyl)indan- 1 -yl]pyrazol-3 -yl]piperazine- 1 - carboxylate (170 mg, yield 2%) as a white solid.
[002005] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 40.6 % (214 nm) Mass: found peak 451.2 (M+l) at 2.309 min.
[002006] Step 3. Synthesis of l-[5-methyl-l-[6-(trifluoromethyl)indan-l-yl]pyrazol-3- yl] piperazine.
Figure imgf000398_0002
[002007] A solution of tert-butyl 4-[5-methyl-l-[6-(trifluoromethyl)indan-l-yl]pyrazol-3- yl]piperazine-l -carboxylate (170 mg, 0.034 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-methyl-l-[6-(trifluoromethyl)indan-l- yl]pyrazol-3-yl]piperazine (130 mg, yield 40%) as a yellow oil.
[002008JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 50 % (214 nm) Mass: found peak 351.1 (M+l) at 2.061 min.
[002009] Step 4. Synthesis of 4-[2-[4-[5-methyl-l-[6-(trifluoromethyl)indan-l-yl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 112).
Figure imgf000399_0001
[002010] To a solution of l-[5-propyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (10 mg, 0.028 mmol), potassium carbonate (15 mg, 0.114 mmol) and KI (5 mg, 0.028 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine hydrochloride (10 mg, 0.057 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NFLHCCh/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[6-(trifluoromethyl)indan-l-yl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (2.8 mg, yield 30 %) as a yellow oil.
[002011JLCMS method: Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94.33 % (214 nm) Mass: found peak 464.2 (M+l) at 2.040 min.
[002012] XH NMR (400 MHz, CDCh) 57.49-7.47 (m, 1H), 7.3-7.35 (m, 1H), 7.19 (s, 1H), 5.66 (t, J = 4.8 Hz,lH), 5.52 (s, 1H), 3.1 (t, J = 4.8 Hz, 4H), 3.28 - 3.17 (m, 1H), 3.14(t, J = 4.8 Hz, 4H), 3.02 - 2.90 (m, 1H), 2.61 - 2.53 (m, 10H), 2.49 (s,4H), 2.22 (s, 3H).
Example SI 13. Synthesis of Compound 113.
Figure imgf000399_0002
[002013] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine;hydrochloride (80 mg, 0.2211 mmol), 1- (bromomethyl)-3,3-difluoro-pyrrolidine;hydrobromide (62.1151 mg, 0.22 mmol), potassium carbonate (153 mg, 1.10 mmol) and acetonitrile (8 mL) at room temperature. The mixture was warmed to 80 °C and stirred for 18 h. The reaction mixture was filtered, and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% Formic acid) as an eluent to give the title product as a white solid (0.074 g, 66% yield). M+l (459 m/z).
[002014] 'H NMR (400 MHz, Chloroform-d) 5 8.52 (s, 1H), 8.21 (s, 1H), 7.57 - 7.38 (m, 2H), 7.40 - 7.28 (m, 2H), 6.06 (s, 1H), 3.36 - 3.23 (m, 2H), 3.02 - 2.87 (m, 2H), 2.86 - 2.59 (m, 7H), 2.56 - 2.43 (m, 2H), 2.38 - 2.20 (m, 5H), 2.00 (d, J = 36.1, 14.5, 10.8, 3.8 Hz, 4H) ppm.
Example SI 14. Synthesis of Compound 114.
Figure imgf000400_0001
[002015] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine;hydrochloride (100 mg, 0.2764 mmol), 4-(2- chloroethyl)thiomorpholine hydrochloride (59 mg, 0.27 mmol), potassium carbonate (191mg, 1.38 mmol) and acetonitrile (8 mL). The mixture was warmed to 80 °C and stirred for 18 h. The mixture was filtered, and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% Formic acid) as an eluent to give the intermediate of thiomorpholine as a white solid (0.115 mg). The intermediate was treated with 35% water (1 mL) and MeOH (1 mL) and stirred at room temperature for 18 h. The title compound was obtained as a white solid (0.035 g, 20% yield). M+l (471 m/z).
[002016] 'H NMR (400 MHz, Chloroform-d) 5 7.54 - 7.41 (m, 2H), 7.32 (d, J = 8.5 Hz, 2H), 6.08 (s, 1H), 3.48 (d, J = 11.3 Hz, 2H), 3.25 - 3.07 (m, 2H), 3.07 - 2.98 (m, 2H), 2.96 - 2.57 (m, 11H), 2.33 (s, 3H), 2.23 - 1.99 (m, 4H) ppm.
Example SI 15. Synthesis of Compound 115.
Figure imgf000400_0002
[002017] To a 100 mL round bottom flask was added 4-[l-(4-bromophenyl)-5-methyl-pyrazol-3- yl]piperidine dihydrochloride (150 mg, 0.38mmol), acetonitrile (10 mL), potassium carbonate (3 equiv., 1.1 mmol), potassium iodide (1 equiv., 0.3815 mmol) and 4-(2- chloroethyl)thiomorpholine 1,1, -dioxide (1 equiv., 0.3815 mmol). The reaction mixture was stirred at 80 °C for 18 h. The reaction mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 10-60% ACN in water (0.1% formic acid) as eluent to give the title product (0.124 g, 62% yield). M+2 (483 m/z).
[002018] 'H NMR (400 MHz, Chloroform-d) 5 8.45 (s, 1H), 7.63 - 7.53 (m, 2H), 7.37 - 7.28 (m, 2H), 6.06 (s, 1H), 3.45 - 3.13 (m, 3H), 3.13 - 2.93 (m, 8H), 2.94 - 2.80 (m, 4H), 2.75 - 2.48 (m, 2H), 2.31 (s, 3H), 2.20 - 1.88 (m, 4H) ppm.
Example SI 16. Synthesis of Compound 116.
Figure imgf000401_0001
[002019] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine;hydrochloride (70 mg, 0.1935 mmol), potassium carbonate (134mg, 0.9 mmol), potassium iodide (32mg, 0.19 mmol), 4-(2- bromoethyl)-2-methylmorpholine hydrobromide (58 mg, 0.19 mmol), and acetonitrile (6 mL). The mixture was warmed to 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.065 g, 67% yield). M+l (453 m/z).
[002020] 'H NMR (400 MHz, Chloroform-d) 5 8.55 (s, 1H), 7.52 - 7.41 (m, 2H), 7.36 - 7.28 (m, 2H), 6.07 (d, J = 0.9 Hz, 2H), 3.93 - 3.83 (m, 1H), 3.75 - 3.55 (m, 2H), 3.29 (d, J = 11.6 Hz, 2H), 2.88 - 2.74 (m, 5H), 2.74 - 2.64 (m, 2H), 2.50 (t, J = 11.3 Hz, 2H), 2.33 (d, J = 0.7 Hz, 3H), 2.27 - 2.15 (m, 1H), 2.15 - 1.94 (m, 4H), 1.94 - 1.82 (m, 1H), 1.14 (d, J = 6.4 Hz, 3H) ppm.
Example SI 17. Synthesis of Compound 117.
Figure imgf000401_0002
[002021] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine;hydrochloride (60 mg, 0.1658 mmol), potassium carbonate (114mg, 0.82 mmol), potassium iodide (27mg, 0.16 mmol), and 4-(2- chloropropyl)morpholine hydrochloride (34mg, 0.16 mmol) in acetonitrile (8 mL). The reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.045 g, 54% yield). M+l (453 m/z).
[002022] 'H NMR (400 MHz, Chloroform-d) 5 8.56 (s, 1H), 7.52 - 7.42 (m, 2H), 7.36 - 7.27
(m, 2H), 6.08 (s, 1H), 3.79 - 3.61 (m, 4H), 3.61 - 3.46 (m, 1H), 3.42 - 3.27 (m, 1H), 3.20 - 3.06 (m, 1H), 3.06 - 2.74 (m, 3H), 2.74 - 2.44 (m, 6H), 2.33 (s, 3H), 2.21 - 1.98 (m, 4H), 1.04 (d, J = 6.7 Hz, 3H) ppm.
Example SI 18. Synthesis of Compound 118.
Figure imgf000402_0001
[002023] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]piperidine hydrochloride (60 mg, 0.1658 mmol), potassium carbonate (114mg, 0.83 mmol), 4-(2-chloro-l-methylethyl)morpholine hydrochloride (33 mg, 0.15 mmol), potassium iodide (27mg, 0.16 mmol) and acetonitrile (6 mL). The mixture was warmed to 80°C and stirred for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 15-100% ACN in water (0.1% formic acid) as eluent to give the title product as a white solid (0.40 g, 48% yield). M+l (453 m/z).
[002024] 'H NMR (400 MHz, Chloroform-d) 5 8.60 (s, 1H), 8.29 (s, 1H), 7.53 - 7.43 (m, 2H), 7.37 - 7.29 (m, 2H), 6.12 - 6.03 (m, 1H), 3.71 (t, J = 4.7 Hz, 4H), 3.40 (d, J = 11.6 Hz, 1H), 3.31 - 3.18 (m, 1H), 3.12 - 2.95 (m, 1H), 2.89 - 2.75 (m, 2H), 2.69 - 2.53 (m, 5H), 2.53 - 2.41 (m, 2H), 2.33 (s, 3H), 2.13 - 1.95 (m, 4H), 1.05 (d, J = 6.6 Hz, 3H) ppm.
Example SI 19. Synthesis of Compound 119.
Figure imgf000402_0002
[002025] In a 25 mL microwave vial were placed 2-[4-[5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]-l-piperidyl]ethyl methanesulfonate (80 mg, 0.18 mmol), 3,3- difluoropiperidine hydrochloride (56 mg, 0.35 mmol), potassium carbonate (123 mg, 0.89 mmol), and acetonitrile (10 mL). The reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC chromatography using 10-100% acetonitrile in water as eluent to give the title product as a white solid (0.043 g, 51% yield). M+l (473 m/z). [002026] 'H NMR (400 MHz, Chloroform-d) 5 7.55 - 7.42 (m, 2H), 7.37 - 7.28 (m, 2H), 6.04 (s, 1H), 4.27 (t, J = 6.1 Hz, 1H), 3.78 - 3.40 (m, 1H), 3.12 - 2.97 (m, 2H), 2.79 - 2.55 (m, 6H), 2.55 - 2.39 (m, 2H), 2.32 (s, 3H), 2.26 - 2.11 (m, 2H), 2.07 - 1.93 (m, 2H), 1.93 - 1.71 (m, 5H) ppm.
Example S120. Synthesis of Compound 120.
Figure imgf000403_0001
[002027] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperidine;hydrochloride (70 mg, 0.1935 mmol), 4-(2-bromoethyl)-3,3- dimethylmorpholine hydrobromide (61 mg, 0.19 mmol), potassium carbonate (133 mg, 0.967403 mmol), and acetonitrile (6 mL). The reaction mixture was stirred at 80 °C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC chromatography using 10-100% acetonitrile in water as eluent to give the title product as a white solid (0.032 g, 32% yield). M+l (467 m/z).
[002028] 'H NMR (400 MHz, Chloroform-d) 5 8.57 (s, 1H), 7.50 - 7.41 (m, 2H), 7.31 (dq, J = 7.9, 1.0 Hz, 2H), 6.14 - 5.99 (m, 1H), 3.79 - 3.66 (m, 2H), 3.31 (s, 4H), 2.76 (ddd, J = 28.2, 19.4, 6.1 Hz, 5H), 2.66 - 2.51 (m, 4H), 2.33 (d, J = 0.7 Hz, 3H), 2.19 - 1.94 (m, 4H), 1.02 (s, 6H) ppm.
Example S121. Synthesis of Compound 121.
Figure imgf000403_0002
[002029] To a 25 mL microwave vial was added 4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine hydrochloride (100 mg, 0.28 mmol, 100 mass %), l-(2-chloroethyl)-4-isopropyl-piperazine dihydrochloride (145 mg, 0.55 mmol), potassium carbonate (191 mg, 1.38 mmol, 100 mass %), potassium iodide (45 mg, 0.27 mmol, 100 mass %), and acetonitrile (10 mL, 100 mass %) at room temperature. The reaction mixture was stirred at 80 °C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 10-100% ACN in water (0.1% formic acid) as an eluent to give the title product (formic acid salt) as a white solid (0.95 g, 65% yield). M+l (480 m/z). [002030] 'H NMR (400 MHz, Chloroform-d) 5 8.54 (s, 1H), 7.56 - 7.43 (m, 2H), 7.38 - 7.29 (m, 2H), 6.06 (s, 1H), 5.17 (s, OH), 3.33 - 3.19 (m, 2H), 3.18 - 3.05 (m, 1H), 3.03 - 2.65 (m, 13H), 2.55 - 2.42 (m, 2H), 2.32 (s, 3H), 2.16 - 1.87 (m, 4H), 1.19 (d, J = 6.6 Hz, 6H) ppm.
Example S122. Synthesis of Compound 122.
Figure imgf000404_0001
[002031] In a 25 mL microwave vial were placed l-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine;2,2,2-trifluoroacetic acid (80 mg, 0.12 mmol), potassium carbonate (78 mg, 0.58 mmol), potassium iodide (18 mg, 0.11 mmol), l-(2-bromoethyl)-4,4-difluoropiperidine hydrobromide (35mg, 0.11 mmol), and acetonitrile (8 mL) at 20 °C. The reaction mixture was stirred at 80 °C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.025 g, 44% yield). M+l (474 m/z).
[002032] 'H NMR (400 MHz, Chloroform-d) 5 7.51 - 7.40 (m, 2H), 7.31 - 7.19 (m, 2H), 5.70 (d, J = 0.8 Hz, 1H), 3.29 (t, J = 5.0 Hz, 4H), 2.78 - 2.52 (m, 12H), 2.30 (s, 3H), 2.11 - 1.92 (m, 4H) ppm.
Example S123. Synthesis of 3-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl] piperazin- 1-yl] ethyl] -8-oxa-3-azabicyclo [3.2.1] octane; hydrochloride (Compound 123).
Figure imgf000404_0002
[002033] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine hydrochloride (25 mg, 0.055 mmol), ACN (6 mL), potassium carbonate (6 equiv., 0.3277 mmol), potassium iodide (1 equiv., 0.055 mmol), then 8-oxa-3 -azabicyclo [3.2.1]octane hydrochloride (2 equiv., 0.11 mmol) was added. The reaction mixture was stirred at 65°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo off to dryness to give the title product as a HC1 salt, 3-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]-8- oxa-3-azabicyclo[3.2.1]octane;hydrochloride (20 mg, 0.0374 mmol, 68.519% yield) as a white solid. M/Z (M+H, MW 462).
[002034]Free base: 'HNMR (400 MHz, CDCI3) 5 7.21 (d, J = 1.8 Hz, 1H), 7.16 - 7.02 (m, 2H), 5.68 (d, J = 0.9 Hz, 1H), 4.27 (dq, J = 4.5, 2.3 Hz, 2H), 3.36 - 3.18 (m, 4H), 2.71 - 2.48 (m, 11H), 2.34 (dd, J = 11.2, 2.1 Hz, 2H), 2.27 (s, 3H), 1.98 - 1.72 (m, 4H) ppm.
Example S124. Synthesis of Compound 124.
Figure imgf000405_0001
[002035] In a 25 mL microwave vial were placed l-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine trihydrochloride (150 mg, 0.34 mmol), potassium carbonate (6 equiv., 2.07 mmol), postassium iodide (1 equiv., 0.34 mmol), 4-(2-chloroethyl)thiomorpholine 1,1- dioxide (1 equiv., 0.34 mmol) and ethanol (6 mL). The reaction mixture was stirred at 80 °C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-60% ACN in water (0.1% formic acid) as eluent to give the title product (0.097 g, 58% yield). M+l (488 m/z).
[002036] 'H NMR (400 MHz, Chloroform-d) 5 7.52 - 7.41 (m, 2H), 7.34 - 7.23 (m, 2H), 5.70 (d, J = 0.9 Hz, 1H), 3.34 (dd, J = 6.1, 4.0 Hz, 4H), 3.07 (s, 8H), 2.86 - 2.75 (m, 6H), 2.71 (dd, J = 7.0, 5.2 Hz, 2H), 2.31 (s, 3H) ppm.
Example S125. Synthesis of 4-[2-[4-[l-(4-chlorophenyl)-5-methyl-pyrazol-3-yl]piperazin-l- yl] ethyl] -1,4-thiazinane 1,1-dioxide (Compound 125).
Figure imgf000405_0002
[002037] To a 25 mL microwave vial was added 3-bromo-l-(4-chlorophenyl)-5-methyl-pyrazole (100 mg, 0.37 mmol), 4-(2 -piperazin- 1-ylethyl)- 1,4-thiazinane l,l-dioxide;hydrochloride (1.2 equiv., 0.44 mmol), tetrahydrofuran (10 mL, 99.5 mass %), dichloro [ 1 ,3-bis(2,6-di-3- pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.02 equiv., 0.007 mmol) and potassium tert-butoxide (6 equiv., 2.2096 mmol) at 20 °C under N2, then the reaction mixture was stirred at 40 °C for 20 h in a sealed vial. The mixture was diluted with EtOAc (20 mL), filtered, and concentrated to give a residue, which was purified by chromatography using 10- 100% ACN and water as eluent to give the desired product of 4-[2-[4-[l-(4-chlorophenyl)-5- methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (2.5 mg, 0.0057 mmol, 1.5% yield) as a white solid. M+l (438 m/z).
[002038] 'H NMR (400 MHz, Chloroform-d) 5 7.46 - 7.32 (m, 4H), 5.68 (d, J = 0.8 Hz, 1H), 3.33 (t, J = 5.1 Hz, 4H), 3.15 - 2.98 (m, 8H), 2.83 - 2.63 (m, 8H), 2.29 (d, J = 0.7 Hz, 3H) ppm.
Example S126. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]-l-imino-l,4-thiazinane l-oxide;hydrochloride (Compound 126).
Figure imgf000406_0001
[002039] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine hydrochloride (45 mg, 0.10 mmol), ACN (6 mL), potassium carbonate (6 equiv., 0.59 mmol), and potassium iodide (1 equiv., 0.10 mmol), then 1-imino-thiomorpholin-l-one HCI (2 equiv., 0.20 mmol) was added. The reaction mixture was stirred at 65°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HCI in MeOH (1 mL), then concentrated in vacuo to dryness to give the title product as a HCI salt, 4-[2-[4-[l-(2,2-difluoro- 1 ,3 -benzodioxol-5-yl)-5-methyl-pyrazol-3 -yl]piperazin- 1 -yl]ethyl]- 1 -imino- 1 ,4-thiazinane 1 - oxide;hydrochloride (47 mg, 0.08 mmol, 86% yield), as a white solid. M/Z (M+H, MW 483). [002040] 'H NMR (400 MHz, CDCh) 5 7.19 (t, J = 1.3 Hz, 1H), 7.10 (d, J = 1.2 Hz, 2H), 5.69 (s, 1H), 3.63 (t, J = 5.1 Hz, 6H), 3.29 - 2.87 (m, 15H), 2.28 (s, 3H) ppm.
Example S127. Synthesis of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3- yl]-4-[2-(4-fluoro-l-piperidyl)ethyl]piperazine;hydrochloride (Compound 127).
Figure imgf000406_0002
[002041] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine;hydrochloride (40 mg, 0.09 mmol), ACN (6 mL), potassium carbonate (6 equiv., 0.52 mmol), and potassium iodide (1 equiv., 0.09 mmol), then 4-fluoropiperidine hydrochloride (2 equiv., 0.17 mmol) was added. The reaction mixture was stirred at 60°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo off to give the title product as a HC1 salt, l-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]-4-[2-(4-fluoro-l- piperidyl)ethyl]piperazine;hydrochloride (42 mg, 0.080 mmol, 91% yield), as a white solid. M/Z (M+H, MW 452).
[002042] XH NMR (400 MHz, CDCh) 5 7.21 (d, J = 1.8 Hz, 1H), 7.15 - 7.02 (m, 2H), 5.68 (d, J = 0.9 Hz, 1H), 4.71 (ddq, J = 48.6, 6.2, 3.2 Hz, 1H), 3.37 - 3.16 (m, 4H), 2.85 - 2.50 (m, 12H), 2.27 (s, 3H), 2.16 - 1.77 (m, 4H) ppm.
Example S128. Synthesis of Compound 128.
Figure imgf000407_0001
[002043] In a 25 mL microwave vial were placed 1 -[5 -methyl- 1- [4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine;2,2,2-trifluoroacetic acid (100 mg, 0.15 mmol), potassium carbonate (103 mg, 0.75 mmol), potassium iodide (25 mg, 0.15 mmol), l-(2- bromoethyl)-3, 3 -difluoropyrrolidine hydrobromide (46 mg, 0.15 mmol), and acetonitrile (8 mL). the reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.020 g, 29% yield). M+l (460 m/z).
[002044] 'H NMR (400 MHz, Chloroform-d) 5 7.51 - 7.43 (m, 2H), 7.29 - 7.24 (m, 2H), 5.70 (s, 1H), 3.35 - 3.21 (m, 4H), 2.95 (t, J = 13.3 Hz, 2H), 2.78 (t, J = 6.9 Hz, 2H), 2.72 - 2.53 (m, 8H), 2.35 - 2.18 (m, 5H) ppm.
Example S129. Synthesis of l-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl] piperazin- 1-yl] ethyl] piperidin-4-ol; hydrochloride (Compound 129).
Figure imgf000407_0002
[002045] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine;hydrochloride (50 mg, 0.11 mmol), ACN (8 mL), potassium carbonate (6 equiv., 0.6554 mmol), and potassium iodide (1 equiv., 0.11 mmol), then 4-hydroxypiperidine (2 equiv., 0.22 mmol) was added. The reaction mixture was stirred at 60°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo to dryness to give the title product as a HC1 salt, l-[2-[4-[l-(2,2-difhioro-l,3-benzodioxol- 5-yl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]piperidin-4-ol;hydrochloride (53 mg, 0.10 mmol, 92% yield), as a white solid. M/Z (M+H, MW 450).
[002046] Formic acid salt: 'HNMR (400 MHz, CDC13) 5 8.50 (s, 1H), 7.20 (dd, J = 1.7, 0.8 Hz, 1H), 7.15 - 6.99 (m, 2H), 5.68 (d, J = 0.9 Hz, 1H), 3.93 (dd, J = 7.0, 3.7 Hz, 1H), 3.25 (t, J = 5.0 Hz, 4H), 3.13 (ddd, J = 12.1, 8.7, 3.5 Hz, 2H), 2.91 (t, J = 6.6 Hz, 4H), 2.77 (t, J = 6.6 Hz, 2H), 2.65 (t, J = 5.0 Hz, 4H), 2.27 (d, J = 0.7 Hz, 3H), 2.10 (ddd, J = 13.5, 8.3, 3.8 Hz, 2H), 1.77 (dtd, J = 13.7, 6.8, 3.5 Hz, 2H) ppm.
Example S130. Synthesis of Compound 130.
Figure imgf000408_0001
[002047] To a 100 mL round flask was added 4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]thiomorpholine (202 mg, 0.44 mmol), Methanol (4 mL) and hydrogen peroxide (16.9 mol/L) in water (2 mL, 27.0 mmol) at 20 °C, then the reaction mixture was stirred for one h. The mixture was concentrated to give a residue, which was purified by reverse HPLC using 10-100% ACN in water (0.1% formic acid) as an eluent to give the title product as a white solid (0.001 g, 0.5% yield). M+l (472 m/z).
[002048] 'H NMR (400 MHz, Chloroform-d) 5 7.49 - 7.41 (m, 2H), 7.34 - 7.27 (m, 2H), 5.69 (s, 1H), 3.40 - 3.21 (m, 4H), 3.22 - 2.99 (m, 3H), 2.95 - 2.73 (m, 6H), 2.73 - 2.55 (m, 7H), 2.30 (s, 3H) ppm.
Example S131. Synthesis of l-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl- pyrazol-3-yl]piperazin-l-yl]ethyl]-3-fluoro-piperidin-4-ol;hydrochloride (Compound 131).
Figure imgf000408_0002
[002049] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine;hydrochloride (40 mg, 0.09 mmol), ACN (6 mL), potassium carbonate (6 equiv., 0.52 mmol), and potassium iodide (1 equiv., 0.09 mmol), then 3-fluoropiperidin-4-ol hydrochloride (2 equiv., 0.1748 mmol) was added. The reaction mixture was stirred at 60°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo to dryness to give the title product as a HC1 salt, 1 -[2-[4-[ 1 - (2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]-3-fluoro- piperidin-4-ol;hydrochloride (30 mg, 0.05 mmol, 63% yield), as a white solid. M/Z (M+H, MW 468).
[002050] Formic acid salt: 'HNMR (400 MHz, CDCh) 5 8.35 (s, 1H), 7.19 (s, 1H), 7.09 (d, J = 1.3 Hz, 2H), 5.68 (s, 1H), 4.70 (ddt, J = 47.9, 6.7, 3.1 Hz, 1H), 3.91 (d, J = 16.6 Hz, 1H), 3.41 (t, J = 5.0 Hz, 4H), 3.12 - 2.42 (m, 12H), 2.27 (s, 3H), 1.87 (dtt, J = 17.7, 9.7, 4.7 Hz, 2H) ppm.
Example S132. Synthesis of Compound 132.
Figure imgf000409_0001
[002051] In a 25 mL microwave vial were placed l-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine;2,2,2-trifluoroacetic acid (80 mg, 0.12 mmol), potassium carbonate (82.7151 mg, 0.598500 mmol), potassium iodide (19.8700 mg, 0.12 mmol), 4-(2-chloro-l- methylethyl)morpholine hydrochloride (24 mg, O. l lmmol), and acetonitrile (8 mL). The reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.025 g, 46% yield). M+l (454 m/z).
[002052] 'H NMR (400 MHz, Chloroform-d) 5 7.54 - 7.41 (m, 2H), 7.30 - 7.24 (m, 2H), 5.70 (s, 1H), 3.70 (t, J = 4.7 Hz, 4H), 3.32 - 3.16 (m, 4H), 2.94 - 2.36 (m, 10H), 2.30 (s, 4H), 1.07 (dd, J = 6.6, 2.9 Hz, 3H) ppm.
Example S133. Synthesis of 4-[2-[4-[5-methyl-l-[[3-
(trifluoromethyl)phenyl] methyl] pyrazol-3-yl] piperazin-l-yl]ethyl] morpholine (Compound 133).
Figure imgf000410_0001
[002053] To a 25 mL microwave vial was added 3-bromo-5-methyl-l-[[3- (trifluoromethyl)phenyl]methyl]pyrazole (200 mg, 0.63 mmol), 4-(2-(piperazin-l- yl)ethyl)morpholine (1.2 equiv., 0.75207 mmol), tTetrahydrofuran (15 mL, 99.5 mass %), dichloro [l,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium (II) (0.02 equiv., 0.012 mmol) and potassium tert-butoxide (1.5 equiv., 0.94 mmol) at 20 °C under N2, then the reaction mixture was stirred at 50 °C for 20 h in a sealed vial. The mixture was diluted with EtOAc (60 mL) and washed with water (3 x 40 mL), brine (20 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give a residue, which was purified by reverse phase HPLC using 0-60% ACN in water as eluent to give 4-[2-[4-[5-methyl- l-[[3-(trifluoromethyl)phenyl]methyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (70 mg, 0.1600 mmol, 25.53% yield) as a solid. M+l (439 m/z).
[002054] 'H NMR (400 MHz, Chloroform-d) 5 7.50 (d, J = 7.7 Hz, 1H), 7.46 - 7.32 (m, 2H), 7.20 (d, J = 7.7 Hz, 1H), 5.55 (s, 1H), 5.17 (s, 2H), 3.76 - 3.66 (m, 4H), 3.27 - 3.13 (m, 4H), 2.69 - 2.44 (m, 12H), 2.13 (s, 3H) ppm.
Example S134. Synthesis of formic acid;4-[2-[4-[5-methyl-l-[[4- (trifluoromethyl)phenyl] methyl] pyrazol-3-yl] piperazin-l-yl]ethyl] morpholine (Compound 134).
Figure imgf000410_0002
[002055] To a 25 mL microwave vial was added 3-bromo-5-methyl-l-[[4-(trifluoromethyl) phenyl]methyl]pyrazole (248 mg, 0.77 mmol), 4-(2-(piperazin-l-yl)ethyl) morpholine (1.2 equiv., 0.93 mmol), tetrahydrofuran (15 mL, 99.5 mass%), dichlorofl, 3-bis(2,6-di-3- pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.02 equiv., 0.015 mmol) and potassium tert-butoxide (1.5 equiv., 1.1657 mmol) at 20 °C under N2, then the reaction mixture was stirred at 60 °C for 20 h in a sealed vial. The mixture was diluted with EtOAc (60 mL) and washed with water (40 mL) and brine (20 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give a residue, which was purified by reverse phase HPLC chromatography using 0-60% ACN in water as eluent to give formic acid;4-[2-[4-[5-methyl-l- [[4-(trifluoromethyl)phenyl]methyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (9 mg, 0.0186 mmol, 2% Yield). M+l (438 m/z).
[002056] 'H NMR (400 MHz, Chloroform-d) 5 9.22 (s, 1H), 8.35 (s, 1H), 7.55 (d, J = 8.1 Hz, 2H), 7.15 (d, J = 8.0 Hz, 2H), 5.54 (s, 1H), 5.17 (s, 2H), 3.86 - 3.64 (m, 4H), 3.29 (dd, J = 6.2, 3.9 Hz, 4H), 2.89 - 2.67 (m, 8H), 2.61 (t, J = 4.6 Hz, 4H), 2.12 (s, 3H) ppm.
Example S135. Synthesis of rac-(3S)-l-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5- methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]piperidin-3-ol;hydrochloride (Compound 135).
Figure imgf000411_0001
[002057] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine;hydrochloride (40 mg, 0.10 mmol), ACN (6 mL), potassium carbonate (6 equiv., 0.52 mmol), potassium iodide (1 equiv., 0.10 mmol), then (S)-3 -hydroxypiperidine hydrochloride (2 equiv., 0.17 mmol) was added. The reaction mixture was stirred at 65°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo off to dryness to give the title product as a HC1 salt, rac- (3 S)-l-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazin-l- yl]ethyl]piperidin-3-ol;hydrochloride (40 mg, 0.076 mmol, 87% yield), as a white solid. M/Z (M+H, MW 450).
[002058] Formic acid salt: 'HNMR (400 MHz, CDCh) 5 8.48 (s, 1H), 7.25 - 7.17 (m, 1H), 7.17
- 7.08 (m, 2H), 7.00 (d, J = 51.0 Hz, 2H), 5.68 (d, J = 0.9 Hz, 1H), 3.96 (p, J = 4.7 Hz, 1H), 3.39
- 3.24 (m, 4H), 3.01 - 2.52 (m, 12H), 2.27 (s, 3H), 2.13 - 1.92 (m, 1H), 1.78 - 1.53 (m, 3H) ppm.
Example S136. Synthesis of (lR,4R)-5-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5- methyl-pyrazol-3-yl]piperazin-l-yl]ethyl]-2-oxa-5-azabicyclo[2.2.1]heptane;hydrochloride (Compound 136).
Figure imgf000412_0001
[002059] In a 25 mL microwave vial were placed l-(2-chloroethyl)-4-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazine;hydrochloride (40 mg, 0.09 mmol), ACN (8 mL), potassium carbonate (6 equiv., 0.52 mmol), potassium iodide (1 equiv., 0.09 mmol), then (lS,4S)-2-oxa-5-azabicyclo[2.2.1]heptane hydrochloride (2 equiv., 0.17 mmol) was added. The reaction mixture was stirred at 65°C for 20 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 20-100% ACN in water as eluent to give the title product as a formic acid salt, which was treated with MeOH (2 mL) and 5% HC1 in MeOH (1 mL), then concentrated in vacuo to dryness to give the title product as a HC1 salt, (lR,4R)-5-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-5-methyl-pyrazol-3-yl]piperazin-l- yl]ethyl]-2-oxa-5-azabicyclo[2.2.1]heptane;hydrochloride (38 mg, 0.073 mmol, 83% yield), as a white solid. M/Z (M+H, MW 448).
[002060] Formic acid salt: 'HNMR (400 MHz, CDCh) 5 8.45 (s, 2H), 7.20 (d, J = 1.8 Hz, 1H), 7.16 - 7.03 (m, 2H), 5.68 (s, 1H), 4.47 (t, J = 2.1 Hz, 1H), 4.13 (d, J = 9.0 Hz, 1H), 4.01 (s, 1H), 3.71 (dd, J = 9.0, 1.9 Hz, 1H), 3.42 - 3.17 (m, 5H), 3.04 (ddt, J = 33.7, 12.6, 6.5 Hz, 2H), 2.85 - 2.58 (m, 7H), 2.10 (dd, J = 10.6, 2.2 Hz, 1H), 1.89 (dt, J = 10.6, 2.6 Hz, 1H) ppm.
Example S137. Synthesis of 4-[2-[4-[5-methyl-l-[[3-(trifluoromethyl)cyclobutyl] methyl] pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 137).
Figure imgf000412_0002
[002061] To a 25 mL microwave vial was added 3-bromo-5-methyl-l-[[3- (trifluoromethyl)cyclobutyl]methyl]pyrazole;5-bromo-3-methyl-l-[[3- (trifluoromethyl)cyclobutyl]methyl]pyrazole (100 mg, 0.17 mmol), 4-(2-(piperazin-l- yl)ethyl)morpholine (1.2 equiv., 0.20 mmol), tetrahydrofuran (15 mL, 99.5 mass %), dichlorofl, 3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.02 equiv., 0.0034 mmol) and potassium tert-butoxide (1.5 equiv., 0.2524 mmol) at 20 °C under N2, then the reaction mixture stirred at 50 °C for 2 days in a sealed vial. The mixture was diluted with EtOAc (60 mL) and washed with water (3 x 40 mL) and brine (20 mL), then dried over sodium sulfate, filtered, and concentrated to give a residue, which was purified by chromatography using 10% EtOH and 90% heptane as eluent and Chiralpak IB column to give the product, 4-[2-[4-[5-methyl-l-[[3-(trifluoromethyl)cyclobutyl] methyl] pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (58 mg, 0.14 mmol, 83% yield) as a solid in the second fraction. M+l (416 m/z).
[002062] 'H NMR (400 MHz, Chloroform-d) 5 5.44 (s, 1H), 3.92 (d, J = 7.1 Hz, 2H), 3.76 - 3.64 (m, 4H), 3.17 (dd, J = 6.2, 3.9 Hz, 4H), 2.94 - 2.73 (m, 2H), 2.67 - 2.39 (m, 12H), 2.33 - 2.00 (m, 7H) ppm.
Example S138. Synthesis of 4-[2-[4-[5-methyl-2-[[3-(trifluoromethyl)cyclobutyl] methyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 138).
Figure imgf000413_0001
[002063] To a 25 mL microwave vial was added 3-bromo-5-methyl-l-[[3-(trifluoromethyl) cyclobutyl]methyl]pyrazole;5-bromo-3-methyl-l-[[3-(trifluoromethyl)cyclobutyl] methyl]pyrazole (100 mg, 0.17 mmol), 4-(2-(piperazin-l-yl)ethyl) morpholine (1.2 equiv., 0.2019 mmol), tetrahydrofuran (15 mL, 99.5 mass%), dichlorofl, 3-bis(2,6-di-3- pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.02 equiv., 0.0034 mmol) and potassium tert-butoxide (1.5 equiv., 0.2524 mmol) at 20 °C under N2, then the reaction mixture was stirred at 50 °C for 2 days in a sealed vial. The mixture was diluted with EtOAc (60 mL) and washed with water (3 x 40 mL) and brine (20 mL), then dried over sodium sulfate, filtered, and concentrated to give a residue, which was purified by chromatography using 10% EtOH and 90% heptane as eluent and Chiralpak IB column to give the product, 4-[2-[4-[5-methyl-2-[[3- (trifluoromethyl)cyclobutyl] methyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (20 mg, 0.048 mmol, 28% yield) in a first fraction. M+l (416 m/z).
[002064] 'H NMR (400 MHz, Chloroform-d) 5 5.36 (s, 1H), 3.79 (d, J = 6.8 Hz, 2H), 3.73 - 3.55 (m, 4H), 3.16 - 2.99 (m, 4H), 2.83 - 2.61 (m, 2H), 2.59 - 2.33 (m, 12H), 2.23 - 2.03 (m, 5H), 1.96 - 1.80 (m, 2H) ppm.
Example S139. Synthesis of 4-[(9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a] pyrazin-7-yl]-l,4-thiazinane 1,1-dioxide (Compound 139).
[002065] Compound 139 was prepared as outlined below.
Figure imgf000414_0001
[002066] Step 1. Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
Figure imgf000414_0002
[002067]Under argon atmosphere, a mixture of (2S,5S)-5-hydroxypiperidine-2-carboxylic acid (500 mg, 3.44 mmol) and thionyl chloride (615 mg, 5.17 mmol) in MeOH (20 mL) was stirred at 65 °C for 2h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluted with 0-40% acetone in petroleum ether) to afford the desired product methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate (540 mg, yield 98.5%) as a colorless oil.
[002068JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; Mass: found peak 160 (M + H) at 0.399 min.
[002069] Step 2. Synthesis of methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate.
Figure imgf000414_0003
[002070]Under argon atmosphere, a mixture of methyl (2S,5S)-5-hydroxypiperidine-2- carboxylate(540 mg, 3.39 mmol), potassium carbonate (1.41 g, 10.2 mmol), tert-butoxycarbonyl tert-butyl carbonate (830 mg, 3.78 mmol) in THF/water (20 mL/5 mL) was stirred at room temperature for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product methyl (2S, 5S)-5-hydroxypiperidine-2-carboxylate (800 mg, yield 90.9%) as a colorless oil.
[002071JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 100% (214 nm); Mass: found peak 160 (M -100) + at 1.061 min.
[002072] Step 3. Synthesis of Ol-tert-butyl O2-methyl (2S)-5-oxopiperidine-l,2- dicarboxylate.
Figure imgf000415_0001
[002073] To a solution of Ol-tert-butyl O2-methyl (2S, 5S)-5-hydroxypiperidine-l,2- dicarboxylate (700 mg, 2.7 mmol) in DCM (30 mL) at 0 °C was added DMP (2.29 g, 5.4 mmol). The reaction mixture was stirred at this temperature for Ih. The reaction was diluted with DCM (30 mL), washed with 5% Na2S20s aqueous (20 mL) and 5% sodium bicarbonate aqueous (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-45% ethyl acetate in petroleum ether) to afford the desired product Ol-tert-butyl O2-methyl (2S)-5- oxopiperidine-l,2-dicarboxylate (500 mg, yield 70%) as a yellow oil.
[002074JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak Mass: found peak 158.3 (M- 100)+ at 1.111 min.
[002075] Step 4. Synthesis of Ol-tert-butyl O2-methyl (2S)-5-(l,l-dioxo-l,4-thiazinan-4- yl)piperidine- 1 ,2-dicarboxylate.
Figure imgf000415_0002
[002076] Under argon atmosphere, a mixture of Ol-tert-butyl O2-methyl (2S)-5-oxopiperidine- 1,2-dicarboxylate (200 mg, 0.78 mmol), acetic acid (24 mg, 0.39 mmol),l,4-thiazinane 1,1- dioxide (211 mg, 1.56 mmol) and sodium triacetoxyborohydride (590 mg, 3.12 mmol) in DCE (10 mL) was stirred at rt for 16h. The reaction mixture was filtered and diluted with water (20 mL), then extracted with ethyl acetate (50 mL*3), dried over sodium sulfate, filtered, and concentrated, then purified by flash chromatography 2-30% ethyl acetate in petroleum ether to afford Ol-tert-butyl O2-methyl (2S)-5-(l,l-dioxo-l,4-thiazinan-4-yl)piperidine-l,2- dicarboxylate (120 mg, 41% yield) as a yellow solid.
[002077] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.23% (214 nm) Mass: found peak 377.3 (M+l) at 1.602 min.
[002078] Step 5. Synthesis of methyl (2S)-5-(l, l-dioxo-l,4-thiazinan-4-yl)piperidine-2- carboxylate.
Figure imgf000416_0001
[002079] To a solution of Ol-tert-butyl O2-methyl (2S)-5-(l,l-dioxo-l,4-thiazinan-4- yl)piperidine-l,2-dicarboxylate (455 mg, 1.21 mmol) in 1,4-di oxane (10 mL) was added a solution of HCl/dioxane (4 M, 3.43 mL, 1.37 mmol). The reaction was stirred at room temperature for 2h. The reaction was concentrated in vacuo. The residue was washed with EtOAc (20 mL), dried in vacuo to afford the desired product methyl (2S)-5-(l, 1 -di oxo- 1,4- thiazinan-4-yl)piperidine-2-carboxylate (270 mg, 67.1% yield) as a white solid. The crude product was used directly in the next step.
[002080] Step 6. Synthesis of methyl l-[2-(l, 3-dioxoisoindolin-2-yl)ethyl]-4-(l,l-dioxo-l,4- thiazinan-4-yl)piperidine-2-carboxylate.
Figure imgf000416_0002
[002081] To a cooled (0 °C) solution of 2-(2-hydroxyethyl)isoindoline-l, 3-dione (166 mg, 0.87 mmol) in DCM (5 mL) was added trifluoromethanesulfonic anhydride (270 mg, 0.96 mmol) under argon atmosphere. After 10 min 2, 6-lutidine (94 mg, 0.87 mmol) and after another lOmin a solution of methyl 4-(l,l-dioxo-l,4-thiazinan-4-yl)piperidine-2-carboxylate (240 mg, 0.87 mmol) and TEA (89 mg, 0.87 mmol) in DCM (5 mL) were added. The reaction mixture was stirred at room temperature for 16h. The residue was diluted with water (10 mL), extracted with dichloromethane (20 mL* 3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica column chromatography (dichloromethane: methanol = 10 / 1) to give methyl l-[2-(l, 3-dioxoisoindolin-2-yl)ethyl]-4-(l,l-dioxo-l,4-thiazinan-4-yl)piperidine-2- carboxylate (70 mg, 8.43% yield) as a yellow oil.
[002082JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 47% (214 nm); Mass: found peak 450.2 (M + H) at 1.397 min.
[002083] Step 7. Synthesis of (9aS)-7-(l,l-dioxo-l,4-thiazinan-4-yl)-2,3,4,6,7,8,9,9a- octahydropyrido [1,2-a] pyrazin-l-one.
Figure imgf000417_0001
[002084] To a solution of methyl (2S)-l-[2-(l,3-dioxoisoindolin-2-yl)ethyl]-5-(l,l-dioxo-l,4- thiazinan-4-yl)piperidine-2-carboxylate (70 mg, 0.156 mmol) in toluene (10 mL) was added NH2NH2 water (17.5 mg, 0.34 mmol). The reaction mixture was stirred at room temperature for 16h. The reaction mixture was concentrated in vacuo. The residue was dissolved with dichloromethane (30 mL), filtered, and concentrated in vacuo to afford (9aS)-7-(l,l-dioxo-l,4- thiazinan-4-yl)-2,3,4,6,7,8,9,9a-octahydropyrido[l,2-a]pyrazin-l-one (40 mg, crude). The crude product was used directly in the next step.
[002085] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak.
[002086] Step 8. Synthesis of 4-[(9aS)-2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin- 7-yl]-l,4-thiazinane 1,1-dioxide.
Figure imgf000417_0002
[002087] At 0 °C, to a solution of 4-[(9aS)-2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin- 7-yl]-l,4-thiazinane 1,1-dioxide (40 mg, 0.14 mmol) in dry THF (20 mL) was added Li Al FL (2.3 mL, 2.3 mmol) under Argon atmosphere. The reaction was stirred at 60 °C for 3h, then were slowly added 2 drops of water, 2 drops of 15% NaOH and 6 drops of water. The product was dried over sodium sulfate, filtered, and concentrated to dryness to afford 4-[(9aS)- 2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-7-yl]-l,4-thiazinane 1,1-dioxide (38 mg, crude) as a yellow oil. The crude product was used directly in the next step.
[002088JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: no peak.
[002089] Step 9. Synthesis of 4-[(9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a] pyrazin-7-yl]-l,4-thiazinane 1,1-dioxide (Compo
Figure imgf000418_0001
[002090] Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (68 mg, 0.21 mmol), 4-(2,3,4,6,7,8,9,9a-octahydro-lH-pyrido[l,2-a]pyrazin-7- yl)-l,4-thiazinane 1,1-dioxide (57.9 mg, 0.21 mmol), tBuXPhos Pd G3 (16.8 mg, 0.021 mmol) and sodium tert-butoxide (61 mg, 0.635 mmol) in 1,4-dioxane (4 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% MeOH in DCM) to afford the crude product, which was further purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[(9aS)-2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- l,3,4,6,7,8,9,9a-octahydropyrido[l,2-a] pyrazin-7-yl]-l,4-thiazinane 1,1-dioxide (4.2 mg, yield 3.78%) as a white solid.
[002091JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89.21% (214 nm) Mass: found peak 514.2 (M+l) at 1.923 min.
[002092] 'H NMR (400 MHz, CDCh) 5 7.39 (d, J = 9.0 Hz, 2H), 7.20 (d, J = 7.3 Hz, 2H), 5.62 (s, 1H), 3.55-3.39 (m, 2H), 3.31-3.19 (m, 2H), 3.18-3.07 (m, 2H), 3.00-2.91 (m, 4H), 2.92 - 2.76 (m, 2H), 2.63 (d, J = 8.8 Hz, 2H), 2.50 (t, J = 13.1 Hz, 1H), 2.27 (dd, J = 14.6, 7.1 Hz, 2H), 2.25- 2.20 (m, 3H), 2.19 (d, J = 4.6 Hz, 1H), 2.07 (t, J = 11.5 Hz, 2H), 1.84 (d, J = 13.3 Hz, 2H) ppm.
Example S140. Synthesis of 4-[2-[4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol-3-yl] piperazin-l-yl]ethyl]morpholine (Compound 140).
[002093] Compound 140 was prepared as outlined below.
Figure imgf000419_0001
[002094] Step 1. Synthesis of tert-butyl 3-cyclobutyl-3-oxo-propanoate.
Figure imgf000419_0002
THF, 0°C-rt, 24h
[002095] To a solution of cyclobutanecarboxylic acid (2 g, 20 mmol) in THF (40 mL) at 0°C was added di(imidazol-l-yl)methanone (3.6 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24 h. In a separate flask, 2M isopropyl magnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (4.8 g, 30 mmol) in THF (20 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 3-cyclobutyl-3-oxo-propanoate (3.1 g, yield: 31.3%) as a yellow oil.
[002096] XH NMR (400 MHz, CDCh) 5 3.44-3.33 (m, 1H), 3.31 (s, 2H), 2.30-2.10 (m, 4H), 2.02-1.92 (m, 1H), 1.9-1.76 (m, 1H), 1.47 (s, 9H) ppm.
[002097] Synthesis of tert-butyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l-carboxylate.
Figure imgf000419_0003
[002098] A mixture of tert-butyl 3-cyclobutyl-3-oxo-propanoate (1.50 g, 7.56 mmol) and tertbutyl piperazine- 1 -carboxylate (1.7 g, 9.12 mmol) in toluene (100 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l -carboxylate (3.3 g, 68% yield) as a yellow solid.
[002099JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.73% (214 nm) Mass: found peak 333.2 (M + Na) at 1.666 min.
[002100] Step 3. Synthesis of tert-butyl 4-(3-cyclobutyl-3-oxo-propanethioyl)piperazine-l- carboxylate. o o s o
K 1 (0.5 eq) Lawesson's reagent
Figure imgf000420_0002
" Boc toluene, 75°c, 16h
Figure imgf000420_0001
[002101] To a solution of tert-butyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l -carboxylate (3.3 g, 10.6 mmol) in toluene (70 mL) was added Lawesson's reagent (2.06 g, 5.1 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-cyclobutyl-3-oxo- propanethioyl)piperazine-l -carboxylate (1.5 g, yield 45%) as a yellow solid.
[002102JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 96% (214 nm) Mass: found peak 327.3 (M + H) at 1.879 min.
[002103] Step 4. Synthesis of tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000420_0003
[002104] To a solution of tert-butyl 4-(3-cyclobutyl-3-oxo-propanethioyl)piperazine-l- carboxylate (1.50 g, 4.59 mmol) in toluene (50 mL) was added NELNEU water (704 mg, 13.8 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (1.0 g, yield 71%) as a white solid. [002105] LCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 307.2 (M + H) at 1.557 min.
[002106] Step 5. Synthesis of tert-butyl 4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000421_0001
[002107] To a solution of tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (300 mg, 0.98 mmol) in chloroform (80 mL) was added (2, 2-difluoro-l, 3-benzodioxol-5- yl)boronic acid (403 mg, 1.96 mmol), anhydrous copper acetate (356 mg, 1.96 mmol), pyridine (380 mg, 4.75 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% dichloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3- benzodioxol-5-yl)pyrazol-3-yl]piperazine-l-carboxylate (405 mg, yield 83.2%) as a yellow solid.
[002108JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 93% (214 nm) Mass: found peak 463.3 (M + 1) at 2.206 min.
[002109] Step 6. Synthesis of l-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000421_0002
[002110] To a solution of tert-butyl 4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol -3 -yl]piperazine-l -carboxylate (200 mg, 0.43 mmol) in dichloromethane (5 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (160 mg, crude). The crude product was used directly in the next step.
[002111JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 92.02% (214 nm) Mass: found peak 363.1 (M + 1) + at 1.591min.
[002112] Step 7. Synthesis of 4-[2-[4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol-3-yl] piperazin-l-yl]ethyl]morpholine.
Figure imgf000422_0001
[002113] To a solution of l-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)pyrazol-3- yl]piperazine (60 mg, 0.166 mmol), potassium carbonate (114 mg, 0.828 mmol) and KI (27.5 mg, 0.166 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (37.2 mg, 0.248 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)pyrazol-3-yl] piperazin-1- yl]ethyl]morpholine (29.6 mg, yield: 35.7%) as a gray solid.
[002114JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 476.2 (M + H) at 2.160 min.
[002115] 'H NMR (400 MHz, CDCh) 5 7.07 (d, J = 1.8 Hz, 1H), 6.98 (dd, J = 11.0, 5.2 Hz, 2H), 5.72 (s, 1H), 3.76 - 3.48 (m, 4H), 3.33 (s, 1H), 3.26 - 3.13 (m, 4H), 2.59 - 2.53 (m, 4H), 2.52 - 2.47 (m, 4H), 2.44 (s, 4H), 2.18 (dt, J = 11.4, 5.7 Hz, 2H), 2.12 -1.99 (m, 2H), 1.86 (dt, J = 17.8, 10.1 Hz, 2H) ppm.
Example S141. Synthesis of 4-[2-[4-[5-cyclobutyl-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 141). [002116] Compound 141 was prepared as outlined below.
Figure imgf000423_0001
[002117] Step 1. Synthesis of l-(2-chloroethyl)-4-[5-cyclobutyl-l-(2,2-difluoro-l,3- benzodioxol-5-yl)pyrazol-3-yl]piperazine.
Figure imgf000423_0002
[002118]Under argon atmosphere, a mixture of l-[5-cyclobutyl-l-(2,2-difluoro-l,3- benzodioxol-5-yl)pyrazol-3-yl]piperazine (124 mg, 0.338 mmol), 2-chloroacetaldehyde (133 mg, 0.667 mmol) and sodium cyanoborohydride(43 mg, 0.68 mmol) in methanol (5 mL) was stirred at rt for 16h. The mixture was filtered. The reaction mixture was filtered and diluted with water (5 mL), then extracted with ethyl acetate (20 mL*3), dried over sodium sulfate, filtered, and concentrated, then purified by flash chromatography 2-30% ethyl acetate in petroleum ether to afford l-(2-chloroethyl)-4-[5-cyclobutyl-l-(2,2-difluoro-l,3-benzodioxol-5-yl)pyrazol-3- yl]piperazine (104 mg, 68.1% yield) as a yellow oil.
[002119JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95% (214 nm) Mass: found peak 425.3 (M+l) at 1.530 min.
[002120] Step 2. Synthesis of 4-[2-[4-[5-cyclobutyl-l-(2,2-difluoro-l,3-benzodioxol-5- yl)pyrazol-3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 141).
Figure imgf000423_0003
[002121] A mixture of l-(2-chloroethyl)-4-[5-cyclobutyl-l-(2,2-difluoro-l,3-benzodioxol-5-yl) pyrazol-3-yl]piperazine (104 mg, 0.245 mmol), 1,4-thiazinane 1,1-dioxide (66.2 mg, 0.49 mmol) and N-ethyl-N-isopropyl-propan-2-amine (158 mg, 1.22 mmol) in NMP (5 mL) was treated with microwave reactor and stirred at 160 °C for 2h. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[5-cyclobutyl-l-(2,2-difluoro-l,3-benzodioxol-5-yl)pyrazol-3-yl]piperazin-l-yl]ethyl]- 1,4-thiazinane 1,1-dioxide (30 mg, yield: 61.9%) as a white solid.
[002122] LCMS method: Column X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: WATER(10 mM ammonium hydrogen carbonate) (A) / ACN (B) Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Temperature: 50°C; Detection: UV (214 , 4 nm) and MS (ESI, POS mode, 103 to 1000 amu) LC purity: 100 % (214 nm); Mass: found peak 524.2 (M + H) at 2.109 min.
[002123] 'H NMR (400 MHz, CDCh) 5 7.13 (d, J = 1.8 Hz, 1H), 7.07 (d, J = 8.4 Hz, 1H), 7.03 (dd, J = 8.5, 2.0 Hz, 1H), 5.79 (s, 1H), 3.45 - 3.33 (m, 1H), 3.26 (s, 4H), 3.07 (d, J = 3.2 Hz, 8H), 2.72 (t, J = 6.8 Hz, 2H), 2.57 (dd, J = 15.5, 8.4 Hz, 4H), 2.26 (dd, J = 8.3, 3.2 Hz, 2H), 2.19 - 2.05 (m, 2H), 1.93 (dd, J = 17.7, 8.7 Hz, 2H) ppm.
Example S142. Synthesis of 4-[2-[4-[5-cydobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 142).
[002124] Compound 142 was prepared as outlined below.
Figure imgf000424_0001
[002125] Step 1. Synthesis of tert-butyl 4-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000425_0001
[002126] To a solution of tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (200 mg, 0.65 mmol) in chloroform (80 mL) was added [4-(trifluoromethoxy) phenyl]boronic acid (274 mg, 1.31 mmol), anhydrous copper acetate (237 mg, 1.31 mmol), pyridine (258 mg, 3.26 mmol) and molecular sieves 4A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (140 mg, yield 42.7%) as a yellow solid.
[002127] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 96.3% (214 nm) Mass: found peak 467.1 (M + 1) + at 2.391 min.
[002128] Step 2. Synthesis of tert-butyl 4-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000425_0002
[002129] To a solution of tert-butyl 4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (120 mg, 0.257 mmol) in dichloromethane (5 mL) was added 2,2,2- trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl 4-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (104 mg, crude). The crude product was used directly in the next step.
[002130JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 363.1 (M + 1) + at 1.634 min.
[002131] Step 3. Synthesis of l-(2-chloroethyl)-4-[5-cyclobutyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000426_0001
[002132]Under argon atmosphere, a mixture of l-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine (104 mg, 0.284 mmol), 2-chloroacetaldehyde (111 mg, 0.567 mmol) and sodium cyanoborohydride (35.7 mg, 0.567 mmol) in methanol (5 mL) was stirred at RT for 16h. The mixture was filtered. The reaction mixture was filtered and diluted with water (5 mL), then extracted with ethyl acetate (20 mL*3), dried over sodium sulfate, filtered, and concentrated, then purified by flash chromatography 2-30% ethyl acetate in petroleum ether to afford l-(2- chloroethyl)-4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (104 mg, 81.2% yield) as a yellow oil.
[002133JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 95% (214 nm) Mass: found peak 429.2 (M+l) at 1.550 min.
[002134] Step 4. Synthesis of 4-[2-[4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 142).
Figure imgf000426_0002
[002135] A mixture of l-(2-chloroethyl)-4-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (120 mg, 0.28 mmol), 1,4-thiazinane 1,1- dioxide (80 mg, 0.56 mmol) and N-ethyl-N-isopropyl-propan-2-amine (181 mg, 1.4 mmol) in NMP (3 mL) was treated with microwave reactor and stirred at 160 °C for 2h. The reaction was cooled to room temperature and directly purified by prep-HPLC (NFLHCCh/water/acetonitrile) to afford the desired product 4-[2-[4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (34.6 mg, yield: 23.4%) as a yellow solid. [002136] LCMS method: Column X-BRIDGE Cl 8 (4.6x 50 mm, 3.5 gm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min; Temperature: 50 °C; Detection: UV (214 nm) and MS (ESI, POS mode, 103 to 1000 amu); LC purity: 100 % (214 nm); Mass: found peak 528.0 (M + H) at 1.888 min.
[002137] 'H NMR (400 MHz, CDCh) 5 7.41-7.35 (m, 2H), 7.25 (d, J = 6.9 Hz, 2H), 5.82 (s, 1H), 3.53-3.35 (m, 1H), 3.28 (s, 4H), 3.07 (d, J = 2.8 Hz, 8H), 2.73 (s, 2H), 2.59 (d, J = 16.7 Hz, 4H), 2.34-2.22 (m, 2H), 2.22-2.07 (m, 2H), 2.05-1.82 (m, 2H) ppm.
Example S143. Synthesis of 4-[2-[4-[5-(methoxymethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 143). [002138] Compound 143 was prepared as outlined below.
Figure imgf000427_0001
[002139] Step 1. Synthesis of tert-butyl 3-cyclobutyl-3-oxo-propanoate.
Figure imgf000427_0002
THF, O°C-rt, 24h
[002140] To a solution of cyclobutanecarboxylic acid (2 g, 20 mmol) in THF (40 mL) at 0°C was added di(imidazol-l-yl)methanone (3.6 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (4.8 g, 30 mmol) in THF (20 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 3-cyclobutyl-3-oxo-propanoate (3 g, yield: 75.7%) as a yellow oil.
[002141] ‘H NMR (400 MHz, CDCh) 5 3.34 (s, 2H), 2.53 (t, J = 7.4 Hz, 2H), 1.66-1.52 (m, 2H), 1.47 (s, 9H), 1.39-1.28 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H) ppm.
[002142] Step 2. Synthesis of tert-butyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l- carboxylate.
Figure imgf000428_0001
[002143] A mixture of tert-butyl 3-cyclobutyl-3-oxo-propanoate (1.50 g, 7.56 mmol) and tertbutyl piperazine- 1 -carboxylate (1.7 g, 9.12 mmol) in toluene (100 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l -carboxylate (2.0 g, 89% yield) as a yellow solid.
[002144JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 97% (214 nm) Mass: found peak 301 (M + Na) + at 1.473 min.
[002145] Step 3. Synthesis of tert-butyl 4-(3-oxoheptanethioyl)piperazine-l-carboxylate.
(0.5 eq) Lawesson's reagent
Figure imgf000428_0002
toluene, 75°c, 16h
Figure imgf000428_0003
[002146] To a solution of tert-butyl 4-(3-oxoheptanoyl)piperazine-l -carboxylate (1.39 g, 4.44mmol) in toluene (50 mL) was added Lawesson's reagent (900 mg, 2.22 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-oxoheptanethioyl)piperazine-l- carboxylate (878 mg, yield 60.1%) as a yellow solid.
[002147] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 87.52% (214 nm) Mass: found peak 261.0 (M -55) + at 1.645 min.
[002148] Step 4. Synthesis of tert-butyl 4-[5-(methoxymethyl)-lH-pyrazol-3-yl]piperazine- 1-carboxylate.
Figure imgf000429_0001
[002149] To a solution of tert-butyl 4-(4-methoxy-3-oxo-butanethioyl)piperazine-l -carboxylate (0.51 g, 1.61 mmol) in toluene (30 mL) was added NH2NH2 water (242 mg, 4.84 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 40 g silica gel column @70mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-[5-(methoxymethyl)-lH-pyrazol-3-yl]piperazine-l-carboxylate (340 mg, yield 71.2%) as a yellow oil.
[002150JLCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min.
Column X-BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C. LC purity: 95.31% (214 nm) Mass: found peak 297.2 (M + H) at 1.649 min.
[002151] Step 5. Synthesis of tert-butyl 4-[5-(methoxymethyl)-l-[4-(trifluoromethoxy) phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000429_0002
[002152] To a solution of tert-butyl 4-[5-(methoxymethyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (100 mg, 1.15 mmol) in chloroform (80 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (482 mg, 2.29 mmol), anhydrous copper acetate (417 mg, 2.29 mmol), pyridine (454 mg, 5.74 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, contrated in vacuo and the residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-(methoxymethyl)- 1 -[4-(trifluorom ethoxy) phenyl]pyrazol-3-yl]piperazine- 1 -carboxylate (294 mg) as a yellow solid.
[002153JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 87% (214 nm) Mass: found peak 457.0 (M + 1) + at 2.163 min.
[002154] Step 6. Synthesis of l-[5-(methoxymethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-
3-yl] piperazine.
Figure imgf000430_0001
[002155] To a solution of tert-butyl 4-[5-(methoxymethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 0.219 mmol) in dichloromethane (5 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-(methoxymethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (78 mg, crude). The crude product was used directly in the next step.
[002156] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 357.1 (M + 1) + at 1.583 min.
[002157] Step 7. Synthesis of 4-[2-[4-[5-(methoxymethyl)-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 143).
Figure imgf000430_0002
[002158] To a solution of l-[5-(methoxymethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (97 mg, 0.27 mmol), potassium carbonate (188 mg, 1.36 mmol) and KI (45 mg, 0.27 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (61.1 mg, 0.41 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- (methoxymethyl)- 1 -[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin- 1 -yl]ethyl]morpholine (82.1 mg, yield: 64.2%) as a yellow solid.
[002159JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.92 % (214 nm); Mass: found peak 469.9 (M + H) at 1.765 min.
[002160] 'H NMR (400 MHz, CDCh) 5 7.67-7.60 (m, 2H), 7.31-7.23 (m, 2H), 5.95 (s, 1H), 4.33 (s, 2H), 3.71 (dd, J = 14.7, 10.0 Hz, 4H), 3.40 (s, 3H), 3.35-3.26 (m, 4H), 2.63 (dd, J = 10.5, 5.6 Hz, 4H), 2.60-2.55 (m, 4H), 2.52 (d, J = 4.0 Hz, 4H) ppm.
Example S144. Synthesis of 4-[2-[4-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 144).
[002161] Compound 144 was prepared as outlined below.
Figure imgf000431_0001
[002162] Step 1. Synthesis of tert-butyl 3-oxoheptanoate.
Figure imgf000431_0002
THF, O C-rt, 24h
[002163] To a solution of pentanoic acid (1 g, 9.79 mmol) in THF (20 mL) at 0°C was added di (imidazol-l-yl)m ethanone (1.75 g, 10.8 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (16.2 mL, 32.3 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (2.35 g, 14.7 mmol) in THF (20 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 3-oxoheptanoate (1.74 g, yield: 88.7%) as a yellow oil.
[002164] ‘H NMR (400 MHz, CDCh) 5 3.34 (s, 2H), 2.53 (t, J = 7.4 Hz, 2H), 1.66 -1.52 (m,
2H), 1.47 (s, 9H), 1.39 -1.28 (m, 2H), 0.91 (t, J = 7.3 Hz, 3H) ppm.
[002165] Step 2. Synthesis of tert-butyl 4-(3-oxoheptanoyl)piperazine-l-carboxylate.
Figure imgf000432_0001
[002166] A mixture of tert-butyl 3-oxoheptanoate (1.74 g, 8.69 mmol) and tert-butyl piperazine- 1-carboxylate (1.78 g, 9.56 mmol) in toluene (100 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tert-butyl 4-(3- oxoheptanoyl)piperazine-l -carboxylate (2.39 g, 88.1% yield) as a yellow solid.
[002167] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 98.44% (214 nm) Mass: found peak 257.2 (M -55) + at 1.879 min.
[002168] Step 3. Synthesis of tert-butyl 4-(3-oxoheptanethioyl)piperazine-l-carboxylate.
(0.5 eq) Lawesson's reagent
Figure imgf000432_0003
toluene, 75°c, 16h
Figure imgf000432_0002
[002169] To a solution of tert-butyl 4-(3-oxoheptanoyl)piperazine-l -carboxylate (1.39 g, 4.44mmol) in toluene (50 mL) was added Lawesson's reagent (900 mg, 2.22 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography, eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-oxoheptanethioyl)piperazine-l- carboxylate (878 mg, yield 60.1%) as a yellow solid.
[002170JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 96% (214 nm) Mass: found peak 273.1 (M -55) + at 1.871 min.
[002171] Step 4. Synthesis of tert-butyl 4-(5-butyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000433_0001
,
[002172] To a solution of tert-butyl 4-(3-oxoheptanethioyl)piperazine-l-carboxylate(878 mg, 2.67 mmol) in toluene (50 mL) was added NELNEh water (410 mg, 8.02 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tertbutyl 4-(5-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (550 mg, yield 66.7%) as a white solid.
[002173JLCMS method: Mobile phase: WATER (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C. LC purity: 100% (214 nm) Mass: found peak 309.1 (M + H) at 1.741 min.
[002174] Step 5. Synthesis of tert-butyl 4-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000433_0002
[002175] To a solution of tert-butyl 4-(5-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (550 mg, 1.78 mmol) in chloroform (80 mL) was added [4-(trifluoromethoxy)phenyl] boronic acid (734 mg, 3.56 mmol), anhydrous copper acetate (644 mg, 3.56 mmol), pyridine (704 mg, 8.9 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, and concentrated in vacuo The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (380 mg, yield 42.3%) as a white solid.
[002176] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 98.61% (214 nm) Mass: found peak 469.3 (M + 1) + at 2.249 min.
[002177] Step 6. Synthesis of l-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000434_0001
[002178] To a solution of tert-butyl 4-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.21 mmol) in dichloromethane (5 mL) was added 2,2,2- trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 1 -[5-butyl- 1 -[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (100 mg, crude). The crude product was used directly in the next step.
[002179JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 369.2 (M + 1) + at 1.674 min.
[002180] Step 7. Synthesis of 4-[2-[4-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 144).
Figure imgf000434_0002
[002181] To a solution of l-[5-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (100 mg, 0.27 mmol), potassium carbonate (188 mg, 1.36 mmol) and KI (45 mg, 0.27 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (60.9 mg, 0.41 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-butyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (86.8 mg, yield: 66.4%) as a white solid.
[002182JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.56 % (214 nm); Mass: found peak 481.9 (M + H) at 1.999 min. [002183] 'H NMR (400 MHz, CDCh) 5 7.46-7.41 (m, 2H), 7.30 -7.25 (m, 2H), 5.71 (s, 1H), 3.75- 3.68 (m, 4H), 3.32 -3.20 (m, 4H), 2.64-2.60 (m, 4H), 2.56 (dd, J = 11.3, 5.4 Hz, 6H), 2.51 (d, J = 4.2 Hz, 4H), 1.60-1.51 (m, 2H), 1.34 (dd, J = 14.9, 7.4 Hz, 2H), 0.88 (t, J = 7.3 Hz, 3H) ppm.
Example S145. Synthesis of 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-3-methyl-azetidin-3-yl]oxyethyl]morpholine (Compound 145).
[002184] Compound 145 was prepared as outlined below.
Figure imgf000435_0001
[002185] Step 1. Synthesis of 3, 5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000435_0002
[002186] To a solution of 3,5-dibromo-lH-pyrazole (1 g, 4.43 mmol) in chloroform (100 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (1.86 g, 8.85 mmol), anhydrous copper acetate (1.61 g, 8.85 mmol), pyridine (1.75 g,22.1 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10- 50% dichloromethane in petroleum ether) to afford the desired product 3, 5-dibromo-l-[4- (trifluoromethoxy)phenyl]pyrazole (600 mg, yield 35.1%) as a colorless oil.
[002187] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 386.9 (M + 1) + at 2.233 min.
[002188] Step 2. Synthesis of 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole.
Figure imgf000436_0001
2 3( . eq) dioxane/H2O(10:1 )
[002189] To a solution of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole (500 mg, 1.3 mmol) and 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (196 mg, 1.17 mmol) in 1,4- di oxane (5 mL) was added sodium carbonate (412 mg, 3.89 mmol) and PdC12(dppf) dichloromethane complex (106 mg, 0.13 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (360 mg, yield 64%) as a colorless oil.
[002190JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 85 % (214 nm); Mass: found peak 347.0 (M + H) at 2.103 min.
[002191] Step 3. Synthesis of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000436_0002
[002192] To a solution of 3-bromo-5-isopropenyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (360 mg, 1.04 mmol) in 5mL THF was added platinum dioxide (50 mg). The mixture was stirred under H2 at rt for Ih. The reaction mixture was filtered and concentrated under vacuo to give the crude product.
[002193JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 m 4.6*30mm; Column Temperature: 40 °C. LC purity: 82.62 % (214 nm); Mass: found peak 349.0 (M + H) at 2.241 min.
[002194] Step 4. Synthesis of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-3- methyl-azetidin-3-ol.
Figure imgf000436_0003
[002195] To a solution of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 0.43 mmol), tris(dibenzylideneacetone)dipalladium(0) (3 mg, 0.0028 mmol), cesium carbonate (46 mg, 0.14mmol) and xantphos (4 mg, 0.0056 mmol) in 1,4-dioxane (2 mL) was added tert-butyl piperazine- 1 -carboxylate (174 mg, 0.935 mmol). The reaction was stirred at 100 °C in tube for 16h. The mixture was filtered, the filtrate was washed with water (30mL) and extracted by EA (30mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (PE/EA=4/1) to give l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-3-methyl- azetidin-3-ol (90 mg, 55.7% yield) as a yellow solid.
[002196] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column:
X-Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm);
Mass: found peak 356.2 (M + H) at 1.964 min.
[002197] Step 5. Synthesis of 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-3-methyl-azetidin-3-yl]oxyethyl]morpholine (Compound 145).
Figure imgf000437_0001
[002198] To a solution of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3-methyl- azetidin-3-ol (80 mg, 0.225 mmol) in dry DMF (3 mL) was added sodium hydride (60%, 90 mg, 2.25 mmol). The reaction was stirred at 80 °C for 2h. The reaction was cooled and 4-(2- chloroethyl)morpholine hydrochloride (50.5 mg, 0.338 mmol) was added and stirred at 80 °C for 16h. The reaction was cooled to room temperature, quenched with water (5 mL), extracted with DCM (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3-methyl- azetidin-3-yl]oxyethyl]morpholine (40.3 mg, yield 38.2%) as a yellow solid.
[002199JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 469.2 (M+l) at 2.061 min.
[002200] 'H NMR (400 MHz, CDCh) 5 7.45-7.37 (m, 2H), 7.27 (d, J = 8.7 Hz, 2H), 5.55 (s, 1H), 3.90 (d, J = 7.7 Hz, 2H), 3.72 (dd, J = 11.6, 6.5 Hz, 6H), 3.54 (t, J = 6.0 Hz, 2H), 3.00-2.81 (m, 1H), 2.59 (t, J = 6.0 Hz, 2H), 2.54 -2.33 (m, 4H), 1.57 (s, 3H), 1.17 (d, J = 6.8 Hz, 6H) ppm. Example S146. Synthesis of 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]azetidin-3-yl] oxy ethyl] morpholine (Compound 146).
[002201] Compound 146 was prepared as outlined below.
Figure imgf000438_0001
[002202] Step 1. Synthesis of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000438_0002
[002203] To a solution of 3,5-dibromo-lH-pyrazole (1 g, 4.43 mmol) in chloroform (100 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (1.86 g, 8.85 mmol), anhydrous copper(II) acetate (1.61 g, 8.85 mmol), pyridine (1.75 g, 22.1 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product 3, 5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole (600 mg, yield 35.1%) as a colorless oil.
[002204JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 386.9 (M + 1) + at 2.233 min.
[002205] Step 2. Synthesis of 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole.
Figure imgf000438_0003
[002206] To a solution of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole (500 mg, 1.3 mmol) and 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (196 mg, 1.17 mmol) in 1,4- di oxane (5 mL) was added sodium carbonate (412 mg, 3.89 mmol) and PdC12(dppf) dichloromethane complex (106 mg, 0.13 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (360 mg, yield 64%) as a colorless oil.
[002207] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 85 % (214 nm); Mass: found peak 347.0 (M + H) at 2.103 min.
[002208] Step 3. Synthesis of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
J. /^ o F PtO2, THF, rt, 1h
AA J AF -
Br F
Figure imgf000439_0001
[002209] To a solution of 3-bromo-5-isopropenyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (360 mg, 1.04 mmol) in 5mL THF was added platinum dioxide (50 mg). The mixture was stirred under H2 at rt for Ih. The reaction mixture was filtered and concentrated under vacuum to give the crude product.
[002210JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 82.62 % (214 nm); Mass: found peak 349.0 (M + H) at 2.241 min.
[002211] Step 4. Synthesis of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]azetidin-3-ol.
Figure imgf000439_0002
[002212] To a solution of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (500 mg, 1.43 mmol), tris(dibenzylideneacetone)dipalladium(0) (13.1 mg, 0.0143 mmol), tBuNa (55 mg, 0.573 mmol) and xantphos (16.5 mg, 0.0286 mmol) in 1,4-dioxane (2 mL) was added azetidin-3-ol (15.7 mg, 0.215 mmol). The reaction was stirred at 100 °C in tube for 16h. The reaction was cooled to room temperature, quenched with water (5 mL), extracted with DCM (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% dichloromethane in petroleum ether) to afford the desired product l-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]azetidin-3-ol (180 mg, yield 36.8%) as a yellow solid. [002213JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 36% (214 nm)
Mass: found peak 342.1 (M+l) at 1.223 min.
[002214] Step 5. Synthesis of 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]azetidin-3-yl] oxy ethyl] morpholine (Compound 146).
Figure imgf000440_0001
[002215] To a solution of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]azetidin-3- ol (136 mg, 0.398 mmol) in dry DMF (3 mL) was added sodium hydride (60%, 159 mg, 3.98 mmol). The reaction was stirred at 80 °C for Ih. The reaction was cooled and 4-(2- chloroethyl)morpholine hydrochloride (50.5 mg, 0.338 mmol) was added and stirred at 80 °C for 16h. . The reaction was cooled to room temperature, quenched with water (5 mL), extracted with DCM (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]azetidin-3- yl]oxyethyl]morpholine (60 mg, yield 33.1%) as a yellow oil.
[002216] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 455.0 (M+l) at 1.900 min.
[002217] 'H NMR (400 MHz, CDCh) 5 7.45-7.36 (m, 2H), 7.28 (d, J = 8.7 Hz, 2H), 5.54 (s, IH), 4.42 (t, J = 5.7 Hz, IH), 4.17-4.06 (m, 2H), 3.82 (dd, J = 8.5, 5.1 Hz, 2H), 3.75-3.69 (m, 4H), 3.55 (t, J = 5.7 Hz, 2H), 2.93 (dd, J = 13.9, 7.1 Hz, IH), 2.59 (t, J = 5.7 Hz, 2H), 2.54-2.46 (m, 4H), 1.17 (d, J = 6.8 Hz, 6H) ppm.
Example S147. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l,4-diazepan-l-yl]ethyl]morpholine (Compound 147).
[002218] Compound 147 was prepared as outlined below.
Figure imgf000441_0001
[002219] Step 1. Synthesis of 3-bromo-5-isopropenyl-l-[4-
(trifluoromethoxy)phenyljpyrazole.
Figure imgf000441_0002
2 3 . eq dioxane/H2O(10:1 )
[002220] To a solution of 3,5-dibromo-l-[4-(trifluoromethoxy)phenyl]pyrazole (500 mg, 1.3 mmol) and 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (196 mg, 1.17 mmol) in 1,4- di oxane (5 mL) was added sodium carbonate (412 mg, 3.89 mmol) and PdC12(dppf) dichloromethane complex (106 mg, 0.13 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 3-bromo-5-isopropenyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (360 mg, yield 64%) as a colorless oil.
[002221JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C, LC purity: 85 % (214 nm); Mass: found peak 347.0 (M + H) at 2.103 min.
[002222] Step 2. Synthesis of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000441_0003
[002223] To a solution of 3-bromo-5-isopropenyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (360 mg, 1.04 mmol) in 5 mL THF was added platinum dioxide (50 mg). The mixture was stirred under Lb at rt for Ih. The reaction mixture was filtered and concentrated in vacuo to give the crude product.
[002224JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 82.62 % (214 nm); Mass: found peak 349.0 (M + H) at 2.241 min.
[002225] Step 3. Synthesis of tert-butyl 4-[5-isopropyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-diazepane-l-carboxylate.
Figure imgf000442_0001
[002226] To a solution of 3-bromo-5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, 0.86 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladipm;ditert-butyl-[2- (2,4,6-triisopropylphenyl)phenyl]phosphane (69 mg, 0.086 mmol), and sodium tert-butoxide (248 mg, 2.58 mmol) in 1,4-dioxane (2 mL) was added tert-butyl 1,4-diazepane-l -carboxylate (344 mg, 1.72 mmol). The reaction was stirred at 100 °C in tube for 16h. The mixture was filtered, and the filtrate was washed with water (30 mL) and extracted by EA (30 mL x 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (PE/EA=4/1) to give tert-butyl 4-[5- isopropyl- 1 -[4-(trifluorom ethoxy )phenyl]pyrazol-3 -yl ] - 1 ,4-diazepane- 1 -carboxylate (250 mg, 43.5% yield) as a yellow oil.
[002227] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X-Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 70% (214 nm); Mass: found peak 469.2 (M + H) at 2.174 min.
[002228] Step 4. Synthesis of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]- 1,4-diazepane.
Figure imgf000442_0002
[002229] To a solution of tert-butyl 4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 1,4-diazepane-l -carboxylate (250 mg, 0.53 mmol) in dichloromethane (10 mL) was added 2,2,2- trifluoroacetic acid (4 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-isopropyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]-l,4-diazepane (126 mg, crude). The crude product was used directly in the next step.
[002230JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 60% (214 nm) Mass: found peak 369.1 (M + 1) + at 1.612 min.
[002231] Step 5. Synthesis of 4-[2-[4-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l,4-diazepan-l-yl]ethyl]morpholine (Compound 147).
Figure imgf000443_0001
[002232] To a solution of l-[5-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4- diazepane (126 mg, 0.342 mmol), potassium carbonate (189 mg, 1.36 mmol) and KI (34.2 mg, 0.034 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (95 mg, 0.513 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,4-diazepan-l-yl]ethyl]morpholine (14.3 mg, yield: 9%) as a yellow oil.
[002233JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 99.56 % (214 nm); Mass: found peak 482.3 (M + H) at 1.972 min.
[002234] 'H NMR (400 MHz, CDCh) 5 7.43 (d, J = 9.0 Hz, 2H), 7.27 (d, J = 3.2 Hz, 2H), 5.57 (s, 1H), 3.77-3.66 (m, 4H), 3.63 - 3.53 (m, 2H), 3.48 (s, 1H), 3.01-2.90 (m, 2H), 2.86- 2.77 (m, 1H), 2.76-2.71 (m, 2H), 2.68 (d, J = 7.8 Hz, 4H), 2.50 (dd, J = 13.7, 6.6 Hz, 6H), 1.94 (s, 2H), 1.19 (d, J = 6.8 Hz, 6H) ppm.
Example S148. Synthesis of 4-[2-[4-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 148).
[002235] Compound 148 was prepared as outlined below.
Figure imgf000444_0001
[002236] Step 1. Synthesis of tert-butyl4-(4-methyl-3-oxopentanoyl)piperazine-l- carboxylate.
Figure imgf000444_0002
[002237] A mixture of tert-butyl piperazine- 1 -carboxylate (11.3 g, 60.7 mmol) and tertbutyl acetoacetate (10.6 g, 66.7mmol) in toluene (100 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @200mL/min, eluting with 0-60% acetone in petroleum ether for 8 CV) to afford tert-butyl 4-(3-oxobutanoyl)piperazine-l- carboxylate (18 g, 97.4% yield) as a yellow oil.
[002238] LCMS method: Mobile Phase: A: water (A 10 mM NH4HCO3); B: MeCN; Gradient: from 5 to 95% of B in 1.4min at 1.8ml/min; Column: HALO C18 3.5um 4.6*30mm; Column Temperature: 50 °C; LC purity: 97.06% (214 nm) Mass: 243.2 (M-55)+ at 1.700 min.
[002239] Step 2. Synthesis of tert-butyl4-(4-methyl-3-thioyl)piperazine-l-carboxylate. Lawesson's reagent
(0.5 eq) _
Figure imgf000444_0004
toluene, 75°c, 16h
Figure imgf000444_0003
[002240] To a solution of tert-butyl 4-(3-oxobutanoyl)piperazine-l-carboxylate (19 g, 63.75 mmol) in toluene (500 mL) was added Lawesson's reagent (3.0 g, 7.3 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tert-butyl 4-(3 -oxobutanethioyl) piperazine- 1 -carboxylate (10 g, yield 46.5%) as a yellow solid. [002241] LCMS method: Mobile Phase: A: water (A 10 mM NH4HCO3) B: MeCN; Gradient: from 5 to 95% of B in 1.4 min at 1.8 mL/min; Column: HALO C18 3.5um 4.6*30mm; Column Temperature: 50 °C; LC purity: 64% (214 nm) Mass: 315.3 (M+l)+ at 1.867 min.
[002242] Step 3. Synthesis of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000445_0001
70 c, 16h
[002243] To a solution of tert-butyl 4-(4-methyl-3-oxo-pentanethioyl)piperazine-l- carboxylate (5.7 g, 18.1 mmol) in toluene (110 mL) was added NH2NH2 H2O (2.72 g, 54.39 mmol) and the reaction mixture was stirred at 70°C overnight. The mixture was concentrated in vacuo and purified by flash chromatography (Biotage, 80 g silica gel column @80mL/min, eluting with 0-6% MeOH in DCM) to afford the desired product tert-butyl 4-(5-isopropyl-lH- pyrazol-3-yl)piperazine-l -carboxylate (4.7 g, yield 83%) as a yellow solid.
[002244] LCMS method: Mobile Phase: A: water (0.01% TFA) B: MeCN (0.01% TFA); Gradient: from 5% B increase to 95% B within 1.3 min, 95% B for 1.2 min; Flow Rate: 2.2 mL/min; Column Temperature: 40°C; LC purity: 97.40% (214 nm) Mass: 295.3 (M+l)+ at 1.577 min.
[002245] Step 4. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000445_0002
[002246] A solution of tert-butyl 4-(5-isopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (150 mg, 0.51 mmol), [4-(difhioromethoxy)phenyl]boronic acid (128 mg, 1.02 mmol), pyridine (57.8 mg, 1.02 mmol), copper(II)acetate (128 mg, 1.02 mmol) in chloroform (3 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered, the filter cake washed with di chloromethane (10 mL), the solution concentrated, and the mixture was purified by flash chromatography (Biotage, 100 g silica gel column @100mL/min, eluting with 0%-30% ethyl acetate in petroleum ether for 30 min) to afford tertbutyl 4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (100 mg, 45%) as a white oil. [002247] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (254 nm); Mass: No peak at 2.162 min.
[002248] Step 5. Synthesis of l-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3- yl] piperazine.
Figure imgf000446_0001
[002249] To a solution of tert-butyl 4-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.229 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 1 -[ 1 -[4- (difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (70 mg, yield 91%) as a yellow oil.
[002250JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 91% (214 nm) Mass: No peak, at 1.271 min.
[002251] Step 6. Synthesis of 4-[2-[4-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 148).
Figure imgf000446_0002
[002252] To a solution of l-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3-yl]piperazine (70 mg, 0.208 mmol), potassium carbonate (115 mg, 0.83 mmol) and KI (34 mg, 0.21 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (58 mg, 0.31 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH-tHCCWwater /acetonitrile) to afford the desired product 4-[2-[4-[l-[4-(difluoromethoxy)phenyl]-5-isopropyl-pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (35.8 mg, yield 38%) as a yellow solid.
[002253JLCMS method: Mobile Phase: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm) Mass: found peak 450.3 (M+l)+ at 1.785 min.
[002254] 'H NMR (400 MHz, CDCh) 5 7.40 (d, J = 8.8 Hz, 2H), 7.18 (d, J = 8.8 Hz, 2H), 6.52 (t, J = 73.6 Hz, 1H), 5.69 (s, 1H), 3.75 - 3.67 (m, 4H), 3.26 (d, J = 5.0 Hz, 4H), 2.98 - 2.87 (m, 1H), 2.57 (t, J = 23.1 Hz, 12H), 1.17 (d, J = 6.8 Hz, 6H) ppm.
Example S149. Synthesis of 4-[2-[4-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6- yl)pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 149).
[002255] Compound 149 was prepared as outlined below.
Figure imgf000447_0001
[002256] Step 1. Synthesis of l,4-dibromo-2-(2-bromo-l,l,2,2-tetrafluoro-ethoxy)benzene.
Figure imgf000447_0002
[002257] To a solution of 2-bromo-5-iodo-phenol (2.0 g, 6.69 mmol) in acetonitrile (20 mL) was added potassium carbonate (1.11 g, 8.03 mmol) and bromomethylbenzene (1.37 g, 8 mmol). The mixture was warmed to 80°C and stirred at 80°C overnight. The mixture was diluted with dichloromethane (50 mL), washed with water (30 mL), and dried over sodium sulfate. The organic phase was concentrated in vacuo and purified by flash chromatography (eluting with 2% ethyl acetate in petroleum ether for 5 CV) to give the desired product l,4-dibromo-2-(2-bromo- l,l,2,2-tetrafluoro-ethoxy)benzene (2.6 g, yield: 99.9%) as a white solid.
[002258JLCMS Method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30mm; Column Temperature: 40 °C; purity: 100% (254 nm); no mass detected.
[002259] Step 2. Synthesis of tert-butyl 4-[l-(3-benzyloxy-4-bromo-phenyl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000448_0001
[002260]Under argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (1.2 g, 0.188 mmol), 2-benzyloxy-l-bromo-4-iodo-benzene (1.75 g, 4.51mmol), copper(I) iodide (85.8 mg, 4.5 mmol), 2-(dimethylamino)acetic acid (92.9 mg, 0.9 mmol) and potassium carbonate (1.87 g, 13.5 mmol) in dry DMSO (30 mL) was stirred at 120 °C for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (30 mL) and brine (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 4-[l-(3-benzyloxy-4-bromo-phenyl)-5-methyl-pyrazol-3-yl]piperazine-l -carboxylate (200 mg, yield 8.42%) as a yellow solid.
[002261JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 67.20% (214nm); Mass: found peak 527.2 (M + H) at 1.548 min.
[002262] Step 3. Synthesis of 2-bromo-5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)phenol.
Figure imgf000448_0002
[002263] A mixture of tert-butyl 4-[l-(3-benzyloxy-4-bromo-phenyl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (300 mg, 0.569 mmol) in HC1 (5 mL) was stirred at room temperature for 3h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product 2-bromo-5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)phenol (125 mg, yield 65.2%) as a yellow solid.
[002264JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 73% (214nm); Mass: found peak 337.1 (M + H) at 1.368 min.
[002265] Step 4. Synthesis of tert-butyl 4-[l-(4-bromo-3-hydroxy-phenyl)-5-methyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000449_0001
[002266]Under argon atmosphere, a mixture of 2-bromo-5-(5-methyl-3-piperazin-l-yl-pyrazol- l-yl)phenol (125 mg, 0.371 mmol) and tert-butoxycarbonyl tert-butyl carbonate (890 mg, 0.4 mmol) in THF (10 mL) was stirred at room temperature for 16h. The reaction was cooled to room temperature, diluted with EtOAc (50 mL), washed with water (10 mL) and brine (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @75 mL/min, eluting with 0-40% acetone in petroleum ether) to afford the desired product tert-butyl 4-[l-(4-bromo-3-hydroxy- phenyl)-5-methyl-pyrazol-3-yl]piperazine-l -carboxylate (120 mg, yield 74%) as a colorless oil. [002267] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 67.20% (214nm); Mass: found peak 437.2 (M + H) at 1.333 min.
[002268] Step 5. Synthesis of tert-butyl 4-[l-[4-bromo-3-(2-bromo-l,l,2,2-tetrafluoro- ethoxy)phenyl]-5-methyl-pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000449_0002
[002269] To a solution of tert-butyl 4-[l-(4-bromo-3-hydroxy-phenyl)-5-methyl-pyrazol-3- yl]piperazine-l -carboxylate (120 mg, 0.28 mmol) in DMSO (10 mL) was added cesium carbonate (134 mg, 0.41 mmol) and l,2-dibromo-l,l,2,2-tetrafhroro-ethane (145 mg, 0.56 mmol). The mixture was warmed to 50°C and stirred at 50°C overnight. The mixture was diluted with dichloromethane (50 mL), washed with water (30 mL), and dried over sodium sulfate. The organic phase was filtered and concentrated in vacuo and purified by flash chromatography (eluting with 2% ethyl acetate in petroleum ether for 5 CV) to give the desired product tert-butyl 4-[l-[4-bromo-3-(2-bromo-l,l,2,2-tetrafluoro-ethoxy)phenyl]-5-methyl- pyrazol-3-yl]piperazine-l -carboxylate (140 mg, yield: 24.8%) as a colorless oil.
[002270JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 25% (254nm); Mass: found peak 615.1(M+1) at 1.639 min.
[002271] Step 6. Synthesis of tert-butyl 4-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6- yl)pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000450_0001
[002272] To a solution of l,4-dibromo-2-(2-bromo-l,l,2,2-tetrafluoro-ethoxy)benzene (100 mg, 0.232 mmol) in DMSO (5 mL) was added copper (73.8 mg, 1.16 mmol) and 2-(2- pyridyl)pyridine (73 mg, 1.15 mmol). The mixture was warmed to 150°C and stirred at 150°C for 1.5h. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product tert-butyl 4-[5-methyl-l-(2, 2,3,3- tetrafluorobenzofuran-6-yl)pyrazol-3-yl]piperazine-l -carboxylate (20 mg, yield: 18.9%) as a yellow solid.
[002273JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 96.64%(214nm); Mass: found peak 457.3 (M+l) at 1.616 min.
[002274] Step 7. Synthesis of l-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6-yl)pyrazol-3- yl] piperazine.
Figure imgf000450_0002
[002275] To a solution of tert-butyl 4-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6-yl)pyrazol- 3 -yl]piperazine-l -carboxylate (20 mg, 0.038 mmol) in dichloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-methyl-l-(2, 2,3,3- tetrafluorobenzofuran-6-yl)pyrazol-3-yl]piperazine (13 mg, 95.7%) as a yellow solid.
[002276] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: from 10 to 95% of B in 1.5 min at 1.8 mL/min; Column: X-BRIDGE C18 3.5 pm 4.6*50 mm; Column Temperature: 50 °C; LC purity: 91% (254 nm); Mass: found peak 357.1 (M + 1)+ at 1.085 min.
[002277] Step 8. Synthesis of 4-[2-[4-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6- yl)pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 149).
Figure imgf000451_0001
[002278] To a solution of l-[5-methyl-l-(2,2,3,3-tetrafluorobenzofuran-6-yl)pyrazol-3- yl]piperazine (13 mg, 0.036 mmol), potassium carbonate (25.2 mg, 0.18 mmol) and KI (6 mg, 0.036 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (6.55 mg, 0.043 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- methyl-l-(2,2,3,3-tetrafluorobenzofuran-6-yl)pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (4.4 mg, yield: 25.7%) as a yellow oil.
[002279JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 469.9 (M + H) at 1.807 min.
[002280] XH NMR (400 MHz, CDCh) 5 7.51 (d, J = 8.3 Hz, 1H), 7.27 (d, J = 8.3 Hz, 1H), 7.16 (s, 1H), 5.68 (s, 1H), 3.69-3.61 (m, 4H), 3.20 (d, J = 4.3 Hz, 4H), 2.55 (s, 4H), 2.51 (s, 4H), 2.44 (s, 4H), 2.33 (s, 3H) ppm.
Example S150. Synthesis of 4-[2-[4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 150).
[002281] Compound 150 was prepared as outlined below.
Figure imgf000452_0001
[002282] Step 1. Synthesis of tert-butyl 3-cyclobutyl-3-oxo-propanoate.
Figure imgf000452_0002
[002283] To a solution of cyclopropanecarboxylic acid (2 g, 20 mmol) in THF (40 mL) at 0°C was added di(imidazol-l-yl)methanone (3.6 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (4.8 g, 30 mmol) in THF (20 mL) at 0°C. the reaction mixture was stirred at room temperature for 2h. Then this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous Sodium bicarbonate, dried over Sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 3-cyclobutyl-3-oxo-propanoate (3 g, yield: 75.7%) as a yellow oil.
[002284] 'H NMR (400 MHz, CDCh) 5 3.32 (p, J = 8.5 Hz, 1H), 3.23 (s, 2H), 2.25-2.01 (m, 4H), 1.99-1.84 (m, 1H), 1.81-1.69 (m, 1H), 1.40 (s, J = 2.3 Hz, 9H) ppm.
[002285] Step 2. Synthesis of tert-butyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l- carboxylate.
Figure imgf000452_0003
[002286] A mixture of tert-butyl 3-cyclobutyl-3-oxo-propanoate (1.50 g, 7.56 mmol) and tertbutyl piperazine- 1 -carboxylate (1.7 g, 9.12 mmol) in toluene (75 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l -carboxylate (2.0 g, 79.2% yield) as a yellow solid.
[002287] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 79% (214 nm) Mass: found peak 333.2 (M + Na) + at 1.850 min.
[002288] Step 3. Synthesis of tert-butyl 4-(3-cyclobutyl-3-oxo-propanethioyl)piperazine-l- carboxylate. o
O°W — ' — v' (0.5 eq)
Lawesson's reagent toluene, 75°c, 16h
Figure imgf000453_0001
Boc/
[002289] To a solution of tert-butyl 4-(3-cyclobutyl-3-oxo-propanoyl)piperazine-l -carboxylate (1 g, 3.09 mmol) in toluene (40 mL) was added Lawesson's reagent (0.625 g, 1.55 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-cyclobutyl-3-oxo- propanethioyl)piperazine-l -carboxylate (0.53 g, yield 28.3%) as a yellow solid.
[002290JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 54% (214 nm) Mass: found peak 327.3 (M + H) at 1.295 min.
[002291] Step 4. Synthesis of tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000453_0002
[002292] To a solution of tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl)piperazine-l- carboxylate (0.53 g, 2.08 mmol) in toluene (50 mL) was added NFENFU water (249 mg, 4.87 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (480 mg, yield 96.5%) as a white solid. [002293JLCMS method: Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X- BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 307.4 (M + H) at 1.651 min.
[002294] Step 5. Synthesis of tert-butyl 4-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000454_0001
[002295] To a solution of tert-butyl 4-(5-cyclobutyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (580 mg, 1.89 mmol) in chloroform (100 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (796 mg, 3.79 mmol), anhydrous copper acetate (688 mg, 3.79 mmol), pyridine (749 mg, 9.46 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-cyclobutyl-l- [4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (460 mg, yield 84.3%) as a yellow solid.
[002296] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 56% (214 nm) Mass: found peak 466.8 (M + 1) + at 1.411 min.
[002297] Step 6. Synthesis of l-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl] piperazine.
Figure imgf000454_0002
[002298] To a solution of tert-butyl 4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.186 mmol) in dichloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-cyclobutyl-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine (75 mg, yield 90%) as a yellow solid. [002299JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 82% (214 nm) Mass: found peak 367.3 (M + 1)+ at 1.074 min.
[002300] Step 7. Synthesis of 4-[2-[4-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperazin-l-yl]ethyl]morpholine (Compound 150).
Figure imgf000455_0001
[002301] To a solution of l-[5-cyclobutyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (75 mg, 0.205 mmol), potassium carbonate (141 mg, 1.02 mmol) and KI (34 mg, 0.205 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine (61.3 mg, 0.178 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- cyclobutyl- 1 -[4-(trifluoromethoxy)phenyl]pyrazol-3 -yl]piperazin- 1 -yl]ethyl]morpholine (35 mg, yield: 35.7%) as a white solid.
[002302JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 480.1 (M + H) at 2.173 min.
[002303] 'H NMR (500 MHz, CDCh) 5 7.42-7.35 (m, 2H), 7.25 (d, J =10.6 Hz, 2H), 5.82 (s, 1H), 3.75-3.68 (m, 4H), 3.43 (p, J =8.5 Hz, 1H), 3.32-3.24 (m, 4H), 2.66-2.60 (m, 4H), 2.60- 2.54 (m, 4H), 2.51 (s, 4H), 2.31-2.23 (m, 2H), 2.19-2.08 (m, 2H), 1.99-1.84 (m, 2H) ppm.
Example S151. Synthesis of 4-[2-[4-[5-(cyclopropylmethyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 151). [002304] Compound 151 was prepared as outlined below.
Figure imgf000456_0001
[002305] Step 1. Synthesis of tert-butyl tert-butyl 4-cyclopropyl-3-oxo-butanoate.
Figure imgf000456_0002
[002306] To a solution of 2-cyclopropylacetic acid (2 g, 20 mmol) in THF (200 mL) at 0°C was added di(imidazol-l-yl)methanone (3.56 g, 22 mmol) and the reaction mixture was stirred at room temperature for 24h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-tert-butoxy-3-oxo-propanoic acid (4.8 g, 30 mmol) in THF (40 mL) at 0°C, and the reaction mixture was stirred at room temperature for 2h. Then this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to room temperature and stirred 16h. The mixture was quenched by addition of 10 % aqueous citric acid (200 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl tert-butyl 4-cyclopropyl-3-oxo-butanoate (3.1 g, yield: 78.3%) as a yellow oil. [002307] 'H NMR (400 MHz, CDCh) 5 3.26 (s, 2H), 2.27 (d, J = 7.0 Hz, 2H), 1.32 (s, 9H), 0.92-0.77 (m, 1H), 0.51-0.37 (m, 2H), 0.07-0.06 (m, 2H) ppm.
[002308] Step 2. Synthesis of tert-butyl 4-(4-cyclopropyl-3-oxo-butanoyl)piperazine-l- carboxylate.
Figure imgf000456_0003
[002309] A mixture of tert-butyl tert-butyl 4-cyclopropyl-3-oxo-butanoate (1.5 g, 7.6 mmol) and tert-butyl piperazine- 1 -carboxylate (1.55 g, 8.32 mmol) in toluene (75 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tert-butyl 4-(4-cyclopropyl-3-oxo-butanoyl)piperazine-l -carboxylate (1.4 g, 55.4% yield) as a yellow oil.
[002310JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 91.7% (254 nm) Mass: found peak 311.1 (M + H) at 2.136 min.
[002311] Step 3. Synthesis of tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl)piperazine-l- carboxylate.
Figure imgf000457_0003
,
Boc
[002312] To a solution of tert-butyl 4-(3-cyclopropyl-3-oxo-propanoyl)piperazine-l -carboxylate (0.75 g, 2.43 mmol) in toluene (40 mL) was added Lawesson's reagent (0.491 g, 1.21mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-(3-cyclopropyl-3-oxo- propanethioyl)piperazine-l -carboxylate (0.65 g, yield 46.2%) as a yellow oil.
[002313JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 54% (214 nm) Mass: found peak 327.3 (M + H) at 1.653 min.
[002314] Step 4. Synthesis of tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
NH2NH2 H2O (3.0 eq)
Boc^
Figure imgf000457_0002
luene, 70°c, 16h
Figure imgf000457_0001
[002315] To a solution of tert-butyl 4-(3-cyclopropyl-3-oxo-propanethioyl)piperazine-l- carboxylate (0.65 g, 2.08 mmol) in toluene (50 mL) was added NFLNFU water (0.3 mL, 6.24 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (406 mg, yield 48.3%) as a yellow oil.
[002316] LCMS method: Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min. Column X-
BRIDGE C18 (4.6x 50 mm, 3.5pm). Temperature: 50°C; LC purity: 100% (214 nm) Mass: found peak 307.3 (M + H) at 1.789 min.
[002317] Step 5. Synthesis of tert-butyl 4-[5-cyclopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000458_0002
[002318] To a solution of tert-butyl 4-(5-cyclopropyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (100 mg, 0.342 mmol) in chloroform (15 mL) were added [4-(trifluoromethoxy) phenyl]boronic acid (144 mg, 0.684 mmol), anhydrous copper acetate (124 mg, 0.684 mmol), pyridine (0.14 mL, 1.73 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-cyclopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3 -yl]piperazine-l -carboxylate (150 mg, yield 84.3%) as a yellow solid.
[002319JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 89% (214 nm) Mass: found peak 467.3 (M + 1) + at 1.555min.
[002320] Step 6. Synthesis of l-[5-(cyclopropylmethyl)-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000458_0001
[002321] To a solution of tert-butyltert-butyl 4-[5-(cyclopropylmethyl)-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 0.186 mmol) in dichloromethane (3 mL) was added 2,2,2-trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-(cyclopropylmethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (80 mg, crude). The crude product was used directly in the next step.
[002322JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 82% (214 nm) Mass: found peak 367.3 (M + 1)+ at 1.075 min.
[002323] Step 7. Synthesis of 4-[2-[4-[5-(cyclopropylmethyl)-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound 151).
Figure imgf000459_0001
[002324] To a solution of l-[5-(cyclopropylmethyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperazine (80 mg, 0.218 mmol), potassium carbonate (151 mg, 1.09 mmol) and KI (36.2 mg, 0.218 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (39.2 mg, 0.262 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5- (cyclopropylmethyl)- 1 -[4-(trifluoromethoxy)phenyl]pyrazol-3 -yl]piperazin- 1 - yl]ethyl]morpholine (29.8 mg, yield: 28.5%) as a white solid.
[002325] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 479.9 (M + H) at 1.891 min.
[002326] 'H NMR (400 MHz, DMSO-d6) 5 7.44 (d, J = 9.0 Hz, 2H), 7.32 (d, J = 8.5 Hz, 2H),5.88 (s, IH), 3.48-3.36 (m, 4H), 3.23 (s, 2H), 3.00 (s, 4H), 2.44 (d, J = 6.8 Hz, 2H), 2.34 (d, J = 21.6 Hz, 6H), 2.26 (s, 4H), 0.85-0.73 (m, IH), 0.34 (dt, J = 5.5, 5.0 Hz, 2H), 0.01 (q, J = 4.9 Hz, 2H) ppm.
Example S152. Synthesis of Compound 152.
Figure imgf000460_0001
[002327] In a 25 mL microwave vial were placed l-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperazine; 2,2,2-trifluoroacetic acid (80 mg, 0.12 mmol), potassium carbonate (82 mg, 0.60 mmol), potassium iodide (20 mg, 0.12 mmol), 4-(2-chloropropyl)morpholine hydrochloride (24 mg, 0.11 mmol), and acetonitrile (8 mL). The reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.032 g, 59% yield). M+l (454 m/z).
[002328] 'H NMR (400 MHz, Chloroform-d) 5 7.51 - 7.43 (m, 2H), 7.30 - 7.23 (m, 2H), 5.70 (s, 1H), 3.86 - 3.70 (m, 4H), 3.45 - 3.29 (m, 4H), 3.20 - 3.03 (m, 1H), 3.03 - 2.53 (m, 9H), 2.51 - 2.34 (m, 1H), 2.30 (s, 3H), 1.16 (dd, J = 21.5, 6.6 Hz, 3H) ppm.
Example S153. Synthesis of 4-[2-[[l-[4-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-4-piperidyl]oxy]ethyl]morpholine (Compound 153).
[002329] Compound 153 was prepared as outlined below.
Figure imgf000460_0002
MeCN, 80 C, 3h
[002331] To a solution of 4-methyl-lH-pyrazole (1.0 g, 12.2 mmol) in ACN (20 mL) was added N-iodosuccinimide (2.74 g, 12.2 mmol). The mixture was stirred at 80°C for 3 h. The residue was purified by SGC to afford the desired product 3-iodo-4-methyl-lH-pyrazole (1.0g, 39.5%) as a white solid.
[002332JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95% B within 1.3 min, 95% B for 1.7 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 86% (214 nm) Mass: 208.9 (M+l) at 1.448 min.
[002333] Step 2. Synthesis of 3-iodo-4-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000461_0001
[002334] To a solution of 4-methyl-lH-pyrazole (300 mg, 1.44 mmol) in chloroform (10 mL) was added [3 -(trifluoromethoxy) phenyl] boronic acid (600 mg, 2.88 mmol), anhydrous copper acetate (524 mg, 2.88 mmol), pyridine (0.46 mL, 5.77 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24 h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @ 80mL/min, eluting with 10-50% dichloromethane in petroleum ether) to afford the desired product 3-iodo-4- methyl-l-[4-(trifluorom ethoxy) phenyl] pyrazole (200 mg, yield 37.7%) as a colorless oil.
[002335] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 96% (214 nm) Mass: found peak 368.7 (M+l) at 1.988 min.
[002336] Step 3. Synthesis of 4-[2-[[l-[4-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3- yl]-4-piperidyl]oxy]ethyl]morpholine (Compound 153).
Figure imgf000461_0002
[002337] To a solution of 3-iodo-4-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole (50 mg, 0.13 mmol) in dry 1,4-dioxane (3 mL) was added 4-[2-(4-piperidyloxy) ethyl] morpholine (58 mg, 0.27 mmol), Pd-PEPPSLipent (10.7 mg, 0.01 mmol), sodium tert-butoxide (39 mg, 0.4 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product 4-[2-[[l-[4-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4- piperidyl] oxy] ethyl] morpholine (19.8 mg, yield 32.1%) as a colorless oil.
[002338JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 364.0 (M+l) at 2.134 min. LC purity: 100% (214 nm), Mass: 455.5 (M+l) at 1.099 min.
[002339] 'H NMR (400 MHz, MeOD-d4) 5 7.84 (d, J = 0.8 Hz, 1H), 7.72-7.64 (m, 2H), 7.29 (d, J = 8.5 Hz, 2H), 3.75-3.65 (m, 6H), 3.49 (dt, J = 13.2, 4.4 Hz, 3H), 3.31 (dt, J = 3.2, 1.6 Hz, 1H), 2.94 (ddd, J = 12.4, 9.6, 2.8 Hz, 2H), 2.61 (t, J = 5.6 Hz, 2H), 2.59-2.49 (m, 4H), 2.08 (d, J = 0.5 Hz, 3H), 2.05-1.95 (m, 2H), 1.78-1.63 (m, 2H).
Example S154. Synthesis of 4-[2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 154).
[002340] Compound 154 was prepared as outlined below.
Figure imgf000462_0001
[002341] Step 1. Synthesis of 3-iodo-4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazole.
Figure imgf000462_0002
[002342] To a solution of 3-iodo-4-methyl-lH-pyrazole (700 mg, 3.37 mmol) in chloroform (30 mL) was added [4-(trifhroromethyl)phenyl]boronic acid (1.28 g, 6.73 mmol), anhydrous copper (II) acetate (1.62 g, 6.73 mmol), pyridine (1.08 mL, 13.5 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 16 h. The mixture was filtered, concentrated in vacuo and residue was purified by flash chromatography (Biotage, 120 g silica gel column @100mL/min, eluting with 0-30% ethyl acetate in petroleum ether) to afford the desired product 3-iodo-4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazole (0.97 g, yield 82%) as a yellow solid.
[002343JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 343.4 (M+l) at 1.395 min.
[002344JLCMS method: Mobile phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B within 1.3 min, 95% B for 1.2min; Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e, 50*3 mm, Column Temperature: 40 °C; LC purity: 72.82% (214 nm), Mass: found peak 353.0 (M+l) at 1.948 min.
[002345] Step 2. Synthesis of tert-butyl 4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000463_0001
dioxane, reflux, o/n
[002346] To a solution of 3-iodo-4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazole (200 mg, 0.559 mmol), sodium tert-butoxide (161 mg, 1.68 mmol), tert-butyl piperazine-l-carboxylate (161 mg, 1.68 mmol) and tert-butyl piperazine-l-carboxylate (208 mg, 1.12 mmol) in acetonitrile (10 mL) was added Pd PEPPSI IPENT (88 mg, 0.112 mmol). The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was directly purified by flash chromatography (Biotage, 120 g silica gel column @200mL/min, eluting with 10-50% ethyl acetate in petroleum ether for 8 CV) to afford the desired product tert-butyl 4-[4-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (160 mg, yield 66.0%) as a white solid.
[002347] LCMS method: Mobile Phase: A: water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm) Mass: found peak 355.3 (M+l) at 1.533 min.
[002348] Step 3. Synthesis of tert-butyl 4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000463_0002
[002349] To a solution of tert-butyl 4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (40 mg, 0.097 mmol) in di chloromethane (5 mL) was added TFA (2 mL, 26.9 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with di chloromethane (15 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product tert-butyl 4- [4- methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l -carboxylate (35 mg, yield 92%) as a yellow solid.
[002350JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 85.4% (254 nm) Mass: found peak 311.1 (M+l) at 1.496 min.
[002351] Step 4. Synthesis of 4-[2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 154).
Figure imgf000464_0001
[002352] To a solution of l-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (35 mg, 0.113 mmol), potassium carbonate (62.4 mg, 0.451 mmol) and KI (18.7 mg, 0.113 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (42 mg, 0.226 mmol). The reaction was stirred at 90 °C for 16 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (25.5 mg, yield 51.7%) as a white solid.
[002353JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95% B within 1.5 min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5 pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 96.75% (214 nm) Mass: found peak 424.1 (M+l) at 2.053 min.
[002354] 'H NMR (500 MHz, CDCh) 5 7.68-7.59 (m, 5H), 3.73 (t, J = 4.0 Hz, 4H), 3.28 (t, J = 4.0 Hz, 4H), 2.67 (t, J = 4.0 Hz, 4H), 2.63-2.55 (m, 4H), 2.52 (t, J = 4.0 Hz, 4H), 2.11 (s, 3H) ppm.
Example S155. Synthesis of 4-[2-[4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol- l-yl]-l-piperidyl]ethyl]morpholine (Compound 155). [002355] Compound 155 was prepared as outlined below.
Figure imgf000465_0001
[002356] Step 1. Synthesis of 3-ethoxy-lH-pyrazole.
Figure imgf000465_0002
[002357] To a solution of diethyl 2-(ethoxymethylene)propanedioate (1 g, 4.62 mmol) in EtOH (20 mL) at room temperature was added hydrazine (0.148 g, 4.62 mmol) and the reaction mixture was stirred at 100 °C for 16 hrs. The mixture did not need further purification and was directly used in the next step after vacuum drying. A solution of ethyl 3-hydroxy-lH-pyrazole-5- carboxylate (0.7 g, 4.48 mmol) in HC1 (5 mL, 6 M) was stirred at 100 °C for 16 hrs. The mixture was quenched by sodium bicarbonate to pH=7 and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford 3-ethoxy-lH- pyrazole (0.3 g, yield: 79.6%) as a yellow oil.
[002358JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 85 (M - Et)+ at 0.38 min.
[002359] Step 2. Synthesis of tert-butyl 4-(3-ethoxy-lH-pyrazol-l-yl)piperidine-l- carboxylate.
Figure imgf000466_0001
[002360] To a solution of 3-ethoxy-lH-pyrazole (0.3 g, 3.57 mmol) in DMF (5 mL) at room temperature was added tert-butyl 4-methylsulfonyloxypiperidine-l -carboxylate (0.997 g, 3.57 mmol) and cesium carbonate (3.49 g,107 mmol) and the reaction mixture was stirred at 100 °C for 16 hrs. The mixture was quenched by addition of water (20 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-(3-ethoxy-lH-pyrazol-l-yl)piperidine-l-carboxylate (0.1 g, yield: 10.5%) as a yellow solid.
[002361JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 296 (M +Na)+ at 1.82 min.
[002362] Step 3. Synthesis of tert-butyl 4-(3-hydroxypyrazol-l-yl)piperidine-l-carboxylate.
Figure imgf000466_0002
[002363] l-piperidin-4-yl-l,2-dihydro-pyrazol-3-one (1.2 mg, 4.06 mmol) and 48% HBr (60 mL) was heated in a sealed flask at 140°C for 4 h. The solution was dispersed in water and treated with a saturated sodium carbonate solution and the aqueous layer was extracted with EtOAc (100 mL). The organic layer was dried over sodium sulfate and concentrated in vacuo. A solution of the residue and di-tert-butyl dicarbonate (1.77 g, 8.13 mmol) in DCM (50 mL) was stirred at room temperature overnight. The reaction mixture was purified by chromatography on silica (petroleum ether/EtOAc = 100: 1 - 1 : 1) to give tert-butyl 4-(3-hydroxypyrazol-l- yl)piperidine-l -carboxylate (0.86 g, yield 79.2%) as a colorless oil.
[002364JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 268 (M + H) at 1.62 min.
[002365] Step 4. Synthesis of tert-butyl 4-[3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine-l-carboxylate.
Figure imgf000467_0001
,
Figure imgf000467_0002
150 oC,1 hr,
[002366] To a solution of tert-butyl 4-(3-hydroxypyrazol-l-yl)piperidine-l -carboxylate (0.86 g, 3.22 mmol) in DMF (20 mL) at room temperature was added l-iodo-4- (trifluoromethoxy)benzene (0.927 g, 3.22 mmol), N,N-dimethylglycine (0.171 g,1.49 mmol), copper(I) iodide (0.0945g, 0.496 mmol), and potassium carbonate (2.06 g,14.9 mmol) and the reaction mixture was stirred at 150 °C for 1 h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine-l -carboxylate (0.64 g, yield: 46.5%) as a yellow solid.
[002367] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 428 (M + H) at 1.93 min.
[002368] Step 5. Synthesis of tert-butyl 4-[4-bromo-3-[4-
(tr ifluoromethoxy)phenoxy] pyrazol- 1-yl] piperidine- 1-carboxylate.
Figure imgf000467_0003
[002369] To a solution of tert-butyl 4-[3-[4-(trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine- 1-carboxylate (0.32 g, 0.749 mmol) in DMF (5 mL) at room temperature was added 1- bromopyrrolidine-2, 5-dione (0.133 g, 0.749 mmol) and the reaction mixture was stirred at room temperature for 4 hrs. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-[4-bromo-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine-l- carboxylate (0.36 g, yield: 95.0 %) as a yellow solid. [002370JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 433 (M -t-Bu)+ at 2.40 min.
[002371] Step 6. Synthesis of tert-butyl 4-[4-isopropenyl-3-[4-
(tr ifluoromethoxy)phenoxy] pyrazol- 1-yl] piperidine- 1-carboxylate.
Figure imgf000468_0001
[002372] To a solution of tert-butyl 4-[4-bromo-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine- 1-carboxylate (0.415 g, 0.821 mmol) in dioxane/water (10/1, 10 mL) at room temperature was added 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (0.166 g, 0.985 mmol), bis(tri-tert-butylphosphine)palladium(0) (0.245 g, 0.289 mmol), potassium phosphate tribasic (0.523g, 2.46 mmol) and the reaction mixture was stirred at 100 °C for 2 h. The mixture was quenched by water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1: 4) to afford tert-butyl 4-[4-isopropenyl-3-[4- (trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine- 1-carboxylate (0.2 g, yield: 54.3%) as a white solid.
[002373JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 412 (M - t-Bu)+ at 2.35 min.
[002374] Step 7. Synthesis of tert-butyl 4-[4-isopropyl-3-[4-
(tr ifluoromethoxy)phenoxy] pyrazol- 1-yl] piperidine- 1-carboxylate.
Figure imgf000468_0002
[002375] To a solution of tert-butyl 4-[4-isopropenyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol- l-yl]piperidine- 1-carboxylate (208 mg, 0.446 mmol) in 5 mL THF was added platinum dioxide (20 mg). The mixture was stirred under H2 at room temperature for 1 h. The reaction mixture was filtered and concentrated in vacuo to afford the crude product.
[002376] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B:
Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 414 (M -t-Bu)+ at 2.40 min.
[002377] Step 8. Synthesis of 4-[4-isopropyl-3-[4-(trifluoromethoxy) phenoxy]pyrazol-l- yl] piperidine.
Figure imgf000469_0001
[002378] To a solution of tert-butyl 4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine-l -carboxylate (200 mg, 0.426 mmol) in dichloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 4-[4-isopropyl-3-[4- (trifluoromethoxy) phenoxy]pyrazol-l-yl]piperidine (150 mg, crude). The crude product was used directly in the next step.
[002379JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 351 (M + H) at 2.13 min.
[002380] Step 9. Synthesis of 4-[2-[4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol- l-yl]-l-piperidy
Figure imgf000469_0002
[002381] To a solution of 4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl]piperidine (140 mg, 0.40 mmol), potassium carbonate (166 mg, 1.20 mmol) and KI (6 mg, 0.04 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)morpholine (60.8 mg, 0.40 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[4- isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l-yl]-l-piperidyl]ethyl]morpholine (130 mg, yield: 66.3%) as a white solid.
[002382JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99.56 % (214 nm); Mass: found peak 512 (M + H) at 1.47 min.
[002383] 'H NMR (400 MHz, DMSO) 5 7.60 (s, 1H), 7.34 (m, 2H), 7.13 - 7.02 (m, 2H), 4.04 - 3.86 (m, 1H), 3.51 (m, 4H), 2.94 (m, 2H), 2.62 (m, 1H), 2.40 (m, 8H), 2.05 (m, 2H), 1.86 (m, 4H), 1.10 (m, 6H).
Example S156. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 156).
[002384] Compound 156 was prepared as outlined below.
Figure imgf000470_0001
[002385] Step 1. Synthesis of 3-bromo-l-[4-(difluoromethyl)phenyl]pyrazole.
Figure imgf000470_0002
[002386] To a solution of 3-bromo-4-iodo-lH-pyrazole (550 mg, 3.74 mmol) in chloroform (10 mL) was added[4-(difluoromethyl)phenyl]boronic acid (708 mg, 4.12 mmol), anhydrous copper acetate (1.36 g, 7.48 mmol), pyridine (592 mg, 7.48 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered and concentrated in vacuo. The filtrate was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product 3-bromo-l-[4-(difluoromethyl)phenyl]pyrazole (0.63 g, yield 61.6%) as a yellow solid.
[002387] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 275 (M + H) at 1.83 min.
[002388] Step 2. Synthesis of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000471_0001
,
[002389] To a solution of 3-bromo-l-[4-(difluoromethyl)phenyl]pyrazole (0.63 g, 2.31 mmol) in dioxane (30 mL) at room temperature was added tert-butyl piperazine- 1 -carboxylate (0.592 g, 3.18 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladipm; dicyclohexyl-[2-(2,4,6- triisopropylphenyl)phenyl]phosphane (0.245 g, 0.289 mmol), and sodium tert-butoxide (0.833g ,8.67 mmol) and the reaction mixture was stirred at 100 °C for 8 h. The mixture was quenched by water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[l-[4-(difluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (0.83 g, yield: 95.1%) as a white solid.
[002390JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 322 (M-tBu)+ at 1.91 min.
[002391] Step 3. Synthesis of tert-butyl 4-[4-bromo-l-[4-(difluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000472_0001
[002392] To a solution of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (0.83 g, 2.19 mmol) in DMF (5 mL) at room temperature was added 1- bromopyrrolidine-2, 5-dione (0.39 g, 2.19 mmol) and the reaction mixture was stirred at room temperature for 4 h. The mixture was quenched by water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-[4-bromo-l-[4-(difluoromethyl)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (0.78 g, yield: 77.8 %) as a yellow solid.
[002393JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); LCMS: LC retention time 2.22 min. MS (ESI) m/z 403 [M-t-Bu]+.
[002394] Step 4. Synthesis of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-4-isopropenyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000472_0002
[002395] To a solution of tert-butyl 4-[4-bromo-l-[4-(difluoromethyl)phenyl]pyrazol-3- yl]piperazine-l -carboxylate (0.77 g, 1.69 mmol) in dioxane/water (10/1, 10 mL) at room temperature was added 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (0.34 g, 2.02 mmol), bis(tri-tert-butylphosphine)palladium(0) (0.086 g, 0.169 mmol), and potassium phosphate tribasic (1.07 g, 5.06 mmol) and the reaction mixture was stirred at 100 °C for 2 h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[l-[4- (difluoromethyl)phenyl]-4-isopropenyl-pyrazol-3-yl]piperazine-l -carboxylate (0.6 g, yield: 84.8%) as a white solid.
[002396] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 419 (M + H) at 2.29 min.
[002397] Step 5. Synthesis of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-4-isopropyl- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000473_0001
[002398] To a solution of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-4-isopropenyl-pyrazol-3- yl]piperazine-l -carboxylate (600 mg, 1.43 mmol) in 5mL THF was added platinum dioxide (60 mg). The mixture was stirred under H2 at room temp for Ih, filtered, and concentrated in vacuo to give the crude product.
[002399JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 420 (M + H) at 2.18 min.
[002400] Step 6. Synthesis of 4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl] piperidine.
Figure imgf000473_0002
[002401] To a solution of tert-butyl 4-[l-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-3- yl]piperazine-l -carboxylate (200 mg, 0.476 mmol) in di chloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 4-[4-isopropyl-3-[4- (trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine (150 mg, crude). The crude product was used directly in the next step. [002402] Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile;
Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 321 (M + H) at 1.49 min.
[002403] Step 7. Synthesis of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (Compound 156).
Figure imgf000474_0001
[002404] To a solution of l-[l-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-3-yl]piperazine (150 mg, 0.468 mmol), potassium carbonate (166 mg, 1.20 mmol) and KI (6 mg, 0.04 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine (60.8 mg, 0.40 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4- (difluoromethyl)phenyl]-4-isopropyl-pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (60 mg, yield: 29.6%) as a white solid.
[002405] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99.56 % (214 nm); Mass: found peak 434 (M + H) at 2.02 min.
[002406] 'H NMR (400 MHz, CDCh) 5 7.66 (d, J = 8.4 Hz, 2H), 7.60 (s, 1H), 7.51 (d, J = 8.4 Hz, 2H), 6.64 (t, J = 56.6 Hz, 1H), 3.82 - 3.60 (m, 4H), 3.35 - 3.17 (m, 4H), 2.94 - 2.76 (m, 1H), 2.68 - 2.61 (m, 4H), 2.61 - 2.55 (m, 4H), 2.55 - 2.46 (m, 4H), 1.27 (d, J = 6.8 Hz, 6H).
Example S157. Synthesis of 4-[2-[4-[3-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-l- yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 157).
[002407] Compound 157 was prepared as outlined below.
Figure imgf000475_0001
[002408] Step 1. Synthesis of tert-butyl 4-(3-bromopyrazol-l-yl)piperidine-l-carboxylate.
Figure imgf000475_0002
[002409] A mixture of 3-bromo-lH-pyrazole (1.5 g, 6.8 mmol), cesium carbonate (4.43 g,13.6 mmol) and tert-butyl 4-methylsulfonyloxypiperidine-l -carboxylate (2.09 g, 7.48 mmol) in DMF (40 mL) was heated at 80°C for 16h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (ethyl acetate/petroleum ether = 1 : 4) to afford tert-butyl 4-(3 -brom opyrazol-l-yl)piperi dine- 1 -carboxylate (0.71 g, yield: 31.6%) as an oil.
[002410JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); LCMS: LC retention time 1.85 min. MS (ESI) m/z 331.1 (M+H).
[002411] Step 2. Synthesis of tert-butyl 4-[3-[4-(difluoromethyl)phenyl]pyrazol-l- yl]piperidine-l-carboxylate.
Figure imgf000476_0001
[002412] To a solution of tert-butyl 4-(3 -brom opyrazol-l-yl)piperi dine- 1 -carboxylate (0.71 g, 2.15 mmol) in dioxane/water (10/1, 10 mL) at room temperature was added [4- (difluoromethyl)phenyl]boronic acid (0.370 g, 2.15 mmol), bis(tri-tert- butylphosphine)palladium(O) (0.11 g, 0.215mmol), and potassium phosphate tribasic (1.37 g, 6.45mmol) and the reaction mixture was stirred at 100 °C for 2 h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[3-[4-(difluoromethyl)phenyl]pyrazol-l- yl]piperidine-l -carboxylate (0.73 g, yield: 90 %) as a white solid.
[002413JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 322 (M - t-Bu)+ at 1.90 min.
[002414] Step 3. Synthesis of tert-butyl 4-[4-bromo-3-[4-(difluoromethyl)phenyl]pyrazol-l- yl]piperidine-l-carboxylate.
Figure imgf000476_0002
[002415] To a solution of tert-butyl 4-[3-[4-(difluoromethyl)phenyl]pyrazol-l-yl]piperidine-l- carboxylate (0.73 g, 1.93 mmol) in DMF (5 mL) at room temperature was added 1- bromopyrrolidine-2, 5-dione (0.344 g, 1.93 mmol) and the reaction mixture was stirred at room temperature for 4 h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (acetone / petroleum ether = 1 : 4) to afford tert-butyl 4-[4-bromo-3-[4-(difluoromethyl)phenyl]pyrazol-l-yl]piperidine-l-carboxylate (0.77 g, yield: 87.2 %) as a yellow solid. [002416] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
LCMS: LC retention time 2.22 min. MS (ESI) m/z 457 (M+H).
[002417] Step 4. Synthesis of tert-butyl 4-[3-[4-(difluoromethyl)phenyl]-4-isopropenyl- pyrazol-l-yl]piperidine-l-carboxylate.
Figure imgf000477_0001
[002418] To a solution of tert-butyl 4-[4-bromo-3-[4-(difluoromethyl)phenyl]pyrazol-l- yl]piperidine-l -carboxylate (0.77 g, 1.69 mmol) in dioxane/water (10/1, 10 mL) at room temperature was added 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (0.34 g, 2.02 mmol), bis(tri-tert-butylphosphine)palladium(0) (0.086 g, 0.169 mmol), and potassium phosphate tribasic (1.07 g, 5.06 mmol) and the reaction mixture was stirred at 100 °C for 2 h. The mixture was quenched by water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1: 4) to afford tert-butyl 4-[3-[4-(difluoromethyl)phenyl]-4- isopropenyl-pyrazol-l-yl]piperidine-l-carboxylate (0.65 g, yield: 92.3%) as a white solid. [002419JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 418 (M + H) at 2.15 min.
[002420] Step 5. Synthesis of tert-butyl 4-(3-(4-(difluoromethyl)phenyl)-4-isopropyl-lH- pyrazol-l-yl)piperidine-l-carboxylate.
Figure imgf000477_0002
[002421] To a solution of tert-butyl 4-[3-[4-(difluoromethyl)phenyl]-4-isopropenyl-pyrazol-l- yl]piperidine-l -carboxylate (650 mg, 1.56 mmol) in 5mL THF was added platinum dioxide (60 mg). The mixture was stirred under EE at rt for Ih, filtered, and concentrated in vacuo to give the crude product. [002422JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 420 (M + H) at 2.18 min.
[002423] Step 6. Synthesis of 4-[4-isopropyl-3-[4-(trifluoromethoxy)phenoxy]pyrazol-l- yl] piperidine.
Figure imgf000478_0001
[002424] To a solution of tert-butyl 4-(3-(4-(difluoromethyl)phenyl)-4-isopropyl-lH-pyrazol-l- yl)piperidine-l -carboxylate (200 mg, 0.476 mmol) in dichloromethane (3 mL) was added 2,2,2- trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 4-[4-isopropyl-3-[4- (trifluoromethoxy)phenoxy]pyrazol-l-yl]piperidine (150 mg, crude). The crude product was used directly in the next step.
[002425] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 320 (M + H) at 1.49 min.
[002426] Step 7. Synthesis of 4-[2-[4-[3-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-l- yl]-l-piperidyl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 157).
Figure imgf000478_0002
[002427] To a solution of 4-[3-[4-(difluoromethyl)phenyl]-4-isopropyl-pyrazol-l-yl]piperidine (128 mg, 0.40 mmol), potassium carbonate (166 mg, 1.20 mmol) and KI (6 mg, 0.04 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl) thiomorpholine 1,1 -di oxide (103 mg, 0.44 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[3-[4- (difluoromethyl)phenyl]-4-isopropyl-pyrazol- 1 -yl]- 1 -piperidyl]ethyl]- 1 ,4-thiazinane 1 , 1 -dioxide (50 mg, yield: 24.5%) as a white solid. [002428JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99.56 % (214 nm); Mass: found peak 512 (M + H) at 1.47 min.
[002429] 'H NMR (400 MHz, CDCh) 5 7.64 (d, J = 6.8 Hz, 2H), 7.45 (s, 1H), 7.34 (d, J = 6.8 Hz, 2H), 6.73 - 6.71 (m, 1H), 4.17 (s, 1H), 3.58 (s, 2H), 3.29 - 2.74 (m, 14H), 2.65 (d, J = 6.4 Hz, 1H), 2.20 (s, 4H), 1.14 - 1.11 (m, 6H).
Example S158. Synthesis of 4-[2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 158).
[002430] Compound 158 was prepared as outlined below.
Figure imgf000479_0001
[002431] Step 1. Synthesis of l-(2-chloroethyl)-4-[4-methyl-l-[4- (trifluoromethyl)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000479_0002
[002432] Under argon atmosphere, a mixture of l-[4-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (70 mg, 0.225 mmol), acetic acid (6.7 mg, 0.113 mmol), 2-chloroacetaldehyde (88 mg, 0.451 mmol) and sodium cyanoborohydride (28 mg, 0.452 mmol) in methanol (3 mL) was stirred at rt for 16h. Then the mixture was filtered and concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with sodium bicarbonate to pH = 8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-(2-chloroethyl)-4-[4-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (30 mg, yield 35.7%) as a yellow oil.
[002433JLCMS Method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 56.35% (254 nm); Mass: found peak 372.9 (M+H) at 2.021 min.
[002434] Step 2. Synthesis of 4-[2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound 158).
Figure imgf000480_0001
[002435] A solution of l-(2-chloroethyl)-4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (30 mg, 0.081 mmol), 1,4-thiazinane 1,1-dioxide (21 mg, 0.161 mmol) and DIPEA (52 mg, 0.41 mmol) in NMP (2 mL) was stirred at 160 °C for 2h in MW. The reaction was cooled to room temperature and directly purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[4-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (11.2 mg, yield: 29.8%) as a yellow oil. [002436] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 472.1 (M+l) at 2.034 min.
[002437] XH NMR (400 MHz, CDCh) 5 7.68-7.60 (m, 5H), 3.28 (t, J = 4.4 Hz, 4H), 3.12-3.04 (m, 8H), 2.77-2.71 (m, 2H), 2.70-2.55 (m, 6H), 2.11 (s, 3H) ppm.
Example S-al. Synthesis of 4-[2-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound al).
[002438] Compound al was prepared as outlined below.
Figure imgf000480_0002
[002439] Step 1. Synthesis of tert-butyl 6-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]-2,6-diazaspiro [3.3] heptane-2-carboxylate.
Boc
Figure imgf000480_0003
[002440]Under argon atmosphere, a mixture of 3-bromo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (200 mg, 0.62 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane- 2-carboxylate (185 mg, 0.934 mmol), tBuXPhos Pd G3 (50 mg, 0.062 mmol) and sodium tert- butoxide (180 mg, 1.87 mmol) in 1,4-dioxane (16 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0-60% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 6-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-2,6-diazaspiro[3.3]heptane- 2-carboxylate (240 mg, yield 88%) as a yellow solid. LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 439.3 (M+l) at 1.432 min.
[002441] Step 2. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-2,6- diazaspiro [3.3] heptane.
Figure imgf000481_0001
[002442] To a solution of tert-butyl 6-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 2,6-diazaspiro[3.3]heptane-2-carboxylate (240 mg, 0.547 mmol) in dichloromethane (20 mL) was added TFA (4 mL, 53.8 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[5- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-2,6-diazaspiro[3.3]heptane (190 mg, yield 98.5%) as a yellow oil.
[002443JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.34% (214 nm) Mass: found peak 339.1 (M+l) at 1.844 min.
[002444] Step 3. Synthesis of 4-[2-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound al).
Figure imgf000482_0001
[002445] To a solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-2,6- diazaspiro[3.3]heptane (180 mg, 0.53 mmol), potassium carbonate (294 mg, 2.13 mmol) and KI (88 mg, 0.53 mmol) in 95% ethanol (20 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (148 mg, 0.8 mmol). The reaction was stirred at 90 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[2-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (119.6 mg, yield 48.4%) as a yellow oil.
[002446] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.27% (214 nm) Mass: found peak 452.2 (M+l) at 1.934 min.
[002447] 'H NMR (400 MHz, CD3OD) 5 7.57-7.52 (m, 2H), 7.42 (d, J = 8.4 Hz, 2H), 5.66 (s, 1H), 3.97 (s, 4H), 3.70 (t, J = 4.4 Hz, 4H), 3.48 (s, 4H), 2.65 (t, J = 7.2 Hz, 2H), 2.49 (t, J = 4.4
Hz, 4H), 2.38 (t, J = 7.2 Hz, 2H), 2.28 (s, 3H) ppm.
Example S-a2. Synthesis of 7-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-9-(2- morpholinoethyl)-3-oxa-7,9-diazabicyclo[3.3.1]nonane (Compound a2).
[002448] Compound a2 was prepared as outlined below.
Figure imgf000482_0002
[002449]Under argon atmosphere, a mixture of 3-iodo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (120 mg, 0.326 mmol), 9-(2-morpholinoethyl)-3-oxa-7,9- diazabicyclo[3.3.1]nonane trihydrochloride (172 mg, 0.489 mmol), PEPPSI-IPent (13 mg, 0.0163 mmol) and sodium tert-butoxide (188 mg, 1.96 mmol) in 1,4-dioxane (10 mL) was stirred at 100 °C for 16h. The reaction mixture was cooled to room temp, concentrated in vacuo and the residue was chromatographed on a silica gel column @75mL/min, eluting with 0-15% methanol in dichloromethane) to afford crude product, which was further purified by prep- HPLC (NH4HCO3/water/acetonitrile) to afford the desired product 7-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-9-(2-morpholinoethyl)-3-oxa-7,9- diazabicyclo[3.3.1]nonane (6.5 mg, yield 4.14%) as a yellow solid.
[002450JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 482.2 (M+l) at 1.952 min.
[002451] 'H NMR (400 MHz, CD3OD) 5 7.60-7.55 (m, 2H), 7.42 (d, J = 8.4 Hz, 2H), 5.84 (s, 1H), 4.03 (d, J = 11.2 Hz, 2H), 3.88 (d, J = 11.2 Hz, 2H), 3.71 (t, J = 4.4 Hz, 4H), 3.51 (d, J = 12 Hz, 2H), 3.45-3.37 (m, 2H), 3.00 (t, J = 6.8 Hz, 2H), 2.92-2.88 (m, 2H), 2.61-2.52 (m, 6H), 2.31 (s, 3H) ppm.
Example S-a4. Synthesis of 4-[2-(5-methyl-l-phenyl-pyrazol-3-yl)oxyethyl]morpholine
(Compound a4).
[002452] Compound a4 was prepared as outlined below.
Figure imgf000483_0001
[002453] Step 1. Synthesis of 5-methyl-l-phenyl-pyrazol-3-ol.
Figure imgf000483_0002
[002454] A mixture of N'-phenylacetohydrazide (1.5 g, 10.0 mmol), ethyl 3-oxobutanoate (1.3 g, 10.0 mmol) and trichlorophosphine (1.37 g, 10.0 mmol) was stirred at 50 °C for 2 h. After this time, ice was added, and the solution was stirred for 1 h. The suspension was poured into ethyl acetate and the precipitate filtered. The organic layer of the filtrate was washed with water (30 mL) and brine (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was suspended in EtOH, filtered, and washed with cool EtOH to afford the desired product 5-methyl-l-phenyl-pyrazol-3-ol (415 mg, yield 23.3%) as white solid. [002455] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.94% (214 nm), Mass: found peak 175.1 (M+l) at 1.659 min.
[002456] 'H NMR (400 MHz, CDCh) 5 7.48-7.44 (m, 2H), 7.39-7.34 (m, 3H), 5.59 (s, 1H), 2.25 (s, 3H) ppm
[002457] Step 2. Synthesis of 4- [2-(5-methyl-l-phenyl-pyrazol-3-yl)oxy ethyl] morpholine (Compound a4).
Figure imgf000484_0001
[002458] To a solution of 5-methyl-l-phenyl-lH-pyrazol-3-ol (348 mg, 2.0 mmol, 98%) in DMF (8 mL) was added sodium hydride (240 mg, 6.0 mmol, 60% in mineral oil) at 0 °C, the mixture was stirred at room temperature for 30 min. 4-(2-chloroethyl)morpholine hydrochloride (484 mg, 2.6 mmol) was added and the mixture was stirred at 60 °C for 5h. After being cooled to room temperature, the reaction mixture was filtered, and the filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-(5- methyl-l-phenyl-pyrazol-3-yl)oxyethyl]morpholine (371 mg, yield 64.6%) as colorless oil. [002459JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 288.1 (M+l) at 1.866 min.
[002460] 'H NMR (500 MHz, DMSO-d6) 5 7.48-7.47 (m, 4H), 7.36-7.33 (m, 1H), 5.79 (s, 1H), 4.20 (t, J = 5.5 Hz, 2H), 3.57 (t, J = 5.0 Hz, 4H), 2.66 (t, J = 5.5 Hz, 2H), 2.45-2.43 (m, 4H), 2.28 (s, 3H) ppm.
Example S-a5. Synthesis of [5-methyl-3-(2-morpholinoethoxy)pyrazol-l-yl]-phenyl- methanone (Compound a5).
[002461] Compound a5 was prepared as outlined below.
Figure imgf000485_0001
[002463] To a solution of 5-methyl-l,2-dihydro-3H-pyrazol-3-one (0.981 g, 10 mmol) and triethylamine (2.02 g, 20 mmol) in DCM (20 mL) was added benzoyl chloride (1.41 g, 10 mmol) dropwise at 0 °C, the mixture was stirred at room temperature overnight. The mixture was poured into water (50 mL), extracted with DCM (30 mL x 3). The combined organic layers were washed with brine (30 mL x 2), water (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-50% ethyl acetate in petroleum ether for 8 CV) to afford the desired product 2-benzoyl-3 -methyl- lH-pyrazol-5-one (404 mg, yield 19.2%) as a white solid.
[002464JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96.3% (254 nm), Mass: found peak 203.1 (M+l) at 1.528 min.
[002465] Step 2. Synthesis of [5-methyl-3-(2-morpholinoethoxy)pyrazol- 1-yl] -phenylmethanone (Compound a5).
Figure imgf000485_0002
[002466] To a solution of 2-benzoyl-3-methyl-lH-pyrazol-5-one (295 mg, 1.46 mmol) in dry DMF (10 mL) was added sodium hydride (87.5 mg, 2.19 mmol, 60% in mineral oil) at 0 °C, the mixture was stirred at room temperature for 30min. 4-(2-chloroethyl)morpholine;hydrochloride (298 mg, 1.6 mmol) was added and the mixture was stirred at 60 °C for 4h. After cooled to room temperature, the mixture was filtered, and the filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product [5-methyl-3-(2- morpholinoethoxy)pyrazol-l-yl]-phenyl-methanone (65.7 mg, yield 14.3%) as a colorless oil. [002467] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE
C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 316.2 (M+l) at 1.389 min.
[002468] 'H NMR (400 MHz, DMSO-d6) 5 7.89 (td, J = 6.8, 1.6 Hz, 2H), 7.62 (t, J = 7.2 Hz, 1H), 7.52 (t, J = 7.6 Hz, 1H), 6.11 (s, 1H), 4.17 (t, J = 5.6 Hz, 2H), 3.53 (t, J = 4.4 Hz, 4H), 2.62 (t, J = 5.6 Hz, 2H), 2.56 (s, 3H), 2.40-2.38 (m, 4H) ppm
Example S-a6. Synthesis of [5-methyl-3-(2-morpholinoethoxy)pyrazol-l-yl]-phenyl- methanone (Compound a6).
[002469] Compound a6 was prepared as outlined below.
Figure imgf000486_0001
[002470] Step 1. Synthesis of 2-benzyl-3-methyl-lH-pyrazol-5-one.
Figure imgf000486_0002
[002471] To a solution of 5-methyl-l,2-dihydropyrazol-3-one (1.96 g, 20 mmol) and NaOH (0.8 g, 20 mmol) in water (20 mL) was added chloromethylbenzene (2.62 g, 20 mmol) dropwise at room temperature, the mixture was heated to 70 °C and stirred for 3h. After cooled to room temperature, the mixture was filtered, and the solid was collected. The solid was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 2- benzyl-3-methyl-lH-pyrazol-5-one (120 mg, yield 3.11%) as a white solid.
[002472JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm Column Temperature: 50 °C; LC purity: 97.7% (214 nm), Mass: found peak 189.1 (M+l) at 1.694 min.
[002473] 'H NMR (500 MHz, DMSO-d6) 5 9.44 (br, 1H), 7.32 (t, J = 7.5 Hz, 2H), 7.25 (t, J = 7.5 Hz, 1H), 7.09 (d, J = 7.5 Hz, 2H), 5.32 (s, 1H), 5.02 (s, 2H), 2.12 (s, 3H) ppm.
[002474] Step 2. Synthesis of 4- [2-(l-benzyl-5-methyl-pyrazol-3yl)oxy ethyl] morpholine
(Compound a6).
Figure imgf000487_0001
[002475] To a solution of l-benzyl-5-methyl-l,2-dihydro-3H-pyrazol-3-one (110 mg, 0.57 mmol) in DMF (7 mL) was added sodium hydride (68.5 mg, 1.71 mmol, 60% in mineral oil) at 0 °C, the mixture was stirred at room temperature for 30min. 4-(2- chloroethyl)morpholine;hydrochloride (138 mg, 0.74 mmol) was added and the mixture was stirred at 60 °C for 5h. After cooled to room temperature, filtered, and the filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-(l-benzyl-5-methyl-pyrazol-3yl)oxyethyl]morpholine (115 mg, yield 66.5%) as a white solid.
[002476] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 302.1 (M+l) at 1.850 min.
[002477] 'H NMR (500 MHz, DMSO-d6) 5 7.32 (t, J = 7.0 Hz, 2H), 7.26 (t, J = 7.0 Hz, 1H), 7.08 (d, J =7.0 Hz, 2H), 5.53 (s, 1H), 5.10 (s, 2H), 4.09 (t, J = 6.0 Hz, 2H), 3.55 (t, J = 4.5 Hz, 4H), 2.60 (t, J = 6.0 Hz, 2H), 2.42-2.40 (m, 4H), 2.14 (s, 3H) ppm.
Example S-a7. Synthesis of 5-[5-methyl-3-[4-(2-morpholinoethyl)piperazin-l-yl]pyrazol-l- yl]indan-l-one (Compound a7).
[002478] Compound a7 was prepared as outlined below.
Figure imgf000487_0002
[002479] Step 1. Synthesis of tert-butyl 4-[5-methyl-l-(l-oxoindan-5-yl)pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000488_0001
[002480]Under argon atmosphere, a mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3- yl)piperazine-l -carboxylate (100 mg, 0.37 mmol), 5 -bromoindan- 1 -one (158 mg, 0.75 mmol), copper(I) iodide (18 mg, 0.09 mmol), 2-(dimethylamino)acetic acid (20 mg, 0.18 mmol) and potassium phosphate tribasic (240 mg, 1.13 mmol) in DMF (3 mL) was stirred at 120 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was directly purified by flash chromatography (Biotage, 40 g silica gel column @75 mL/min, eluting with 0- 4% MeOH in DCM) to afford the desired product tert-butyl 4-[5-methyl-l-(l-oxoindan-5- yl)pyrazol-3-yl]piperazine-l -carboxylate (30 mg, yield 20.2%) as a yellow oil.
[002481JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm) Mass: found peak 397.4 (M+l) at 1.321 min.
[002482] Step 2. Synthesis of 5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)indan-l-one.
Figure imgf000488_0002
[002483] To a solution of tert-butyl 4-[5-methyl-l-(l-oxoindan-5-yl)pyrazol-3-yl]piperazine-l- carboxylate (30 mg, 0.11 mmol) in dichloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the mixture was concentrated, and neutralized with a solution of potassium carbonate to pH=10. The resulting mixture was extracted with DCM and dried over sodium sulfate and filtered. The filtrate was concentrated to afford the desired product 5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)indan-l-one (22 mg, yield 98.1 %) as a yellow oil.
[002484JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 98 % (214 nm) Mass: found peak 297.3 (M+l) at 0.940 min. [002485] Step 3. Synthesis of 5-[5-methyl-3-[4-(2-morpholinoethyl)piperazin-l-yl]pyrazol- l-yl]indan-l-one (Compound a7).
Figure imgf000489_0001
[002486] To a solution of 5-(5-methyl-3-piperazin-l-yl-pyrazol-l-yl)indan-l-one (22 mg, 0.074 mmol), potassium carbonate (51 mg, 0.37 mmol) and KI (12.3 mg, 0.074 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl) morpholine (16.7 mg, 0.11 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 5-[5-methyl-3-[4-(2- morpholinoethyl)piperazin-l-yl]pyrazol-l-yl]indan-l-one (4.3 mg, yield 14.0%) as a yellow oil. [002487] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 410.1 (M+l) at 1.665 min.
[002488] 'H NMR (400 MHz, CDCh) 5 7.79 (d, J = 8.3 Hz, 1H), 7.61 (s, 1H), 7.47 (d, J = 8.2 Hz, 1H), 5.76 (s, 1H), 3.78 - 3.64 (m, 4H), 3.29 (d, J = 4.5 Hz, 4H), 3.20 - 3.13 (m, 2H), 2.76 - 2.70 (m, 2H), 2.67 - 2.46 (m, 12H), 2.40 (s, 3H).
Example S-a8. Synthesis of 4-[2-[4-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl) -l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound a8). [002489] Compound a8 was prepared as outlined below.
Figure imgf000490_0001
[002490] Step 1. Synthesis of tert-butyl 3-(3-methyl-l-bicyclo[l.l.l]pentanyl)-3-oxo- propanoate.
Figure imgf000490_0002
[002491] To a solution of 3 -methylbicyclofl.1.1 ]pentane-l -carboxylic acid (500 mg, 3.96 mmol) in THF (25 mL) at 0°C was added CDI (707 mg, 4.36 mmol) and the reaction mixture was stirred at RT for 3h. In a separate flask, 2M isopropylmagnesium chloride in THF (33 mL, 66 mmol) was added dropwise to a solution of 3-(tert-butoxy)-3-oxopropanoic acid (952 mg, 5.95 mmol) in THF (25 mL) at 0°C, and the reaction mixture was stirred at RT for 3h. Then, this solution was added dropwise to the acyl imidazole solution at 0°C and the resulting mixture was allowed to warm up to RT and stirred overnight. The mixture was quenched by addition of 10 % aqueous citric acid (50 mL) and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by flash column chromatography on silica gel (cyclohexane/EtOAc 1/0 to 7/3) to afford tert-butyl 3-(3- methyl-l-bicyclo[l. l.l]pentanyl)-3-oxo-propanoate (660 mg, yield 63%) as a yellow oil.
[002492] 'H NMR (500 MHz, CDCh) 5 3.36 (s, 2H), 1.92 (s, 4H), 1.82 (s, 2H), 1.46 (s, 9H), 1.18 (s, 3H) ppm. [002493] Step 2. Synthesis of tert-butyl 4-[3-(3-methyl-l-bicyclo[l.l.l]pentanyl)-3-oxo- propanoyl]piperazine-l-carboxylate.
Figure imgf000491_0001
[002494] A mixture of tert-butyl 3-(3-methyl-l-bicyclo[l.l. l]pentanyl)-3-oxo-propanoate (660 mg, 2.94 mmol) in toluene (15 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-20% acetone in petroleum ether to afford tert-butyl 4-[3-(3-methyl-l- bicyclo[l.l. l]pentanyl)-3-oxo-propanoyl]piperazine-l -carboxylate (800 mg, 73% yield) as a yellow oil.
[002495] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90.07% (214 nm) Mass: found peak 281.1 (M-55)+ at 1.97 min.
[002496] Step 3. Synthesis of tert-butyl 4-[3-(3-methyl-l-bicyclo[l.l.l]pentanyl)-3-oxo- propanethioyl]piperazine-l-carboxylate.
Figure imgf000491_0003
Figure imgf000491_0002
[002497] To a solution of tert-butyl 4-[3-(3-methyl-l-bicyclo[l.l. l]pentanyl)-3-oxo- propanoyl]piperazine-l -carboxylate (800 mg, 9.28 mmol) in toluene (30 mL) was added Lawesson's reagent (481 mg, 1.2 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-[3 -(3 -methyl- 1 -bicyclo[ 1.1.1 ]pentanyl)-3-oxo-propanethioyl]piperazine- 1 -carboxylate (140 mg, yield 12.7%) as a yellow oil.
[002498JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 75.93% (214 nm) Mass: found peak 297.1 (M-55)+ at 2.116 min.
[002499] Step 4. Synthesis of tert-butyl 4-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-lH- pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000492_0001
[002500] To a solution of tert-butyl 4-[3-(3-methyl-l-bicyclo[l.l. l]pentanyl)-3-oxo- propanethioyl]piperazine-l -carboxylate (140 mg, 0.397 mmol) in toluene (10 mL) was added NH2NH2 water (61 mg, 1.2 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tertbutyl 4-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-lH-pyrazol-3-yl]piperazine-l-carboxylate (100 mg, yield 69.3%) as a white solid.
[002501JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 91.44% (254 nm) Mass: found peak 333.2 (M+l) at 2.103 min.
[002502] Step 5. Synthesis of tert-butyl 4-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000492_0002
,
[002503] To a solution of tert-butyl 4-[5-(3-methyl-l-bicyclo[l.l. l]pentanyl)-lH-pyrazol-3- yl]piperazine-l -carboxylate (100 mg, 0.3 mmol) in chloroform (10 mL) was added [4- (trifluoromethoxy)phenyl]boronic acid (126 mg, 0.6 mmol), anhydrous copper acetate (109 mg, 0.6 mmol), pyridine (120 mg, 1.5 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 30°C for 16h. The mixture was filtered, concentrated in vacuo and the residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10- 50% dichloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-(3- methyl-l-bicyclo[l .1. l]pentanyl)-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (90 mg, yield 60%) as a yellow solid.
[002504JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (254 nm) Mass: found peak 493.1 (M+l) at 2.493 min. [002505] Step 6. Synthesis of l-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-l-[4- (tr ifluoromethoxy)phenyl] pyrazol-3-yl] piperazine.
Figure imgf000493_0001
[002506] To a solution of tert-butyl 4-[5-(3-methyl-l-bicyclo[l.l. l]pentanyl)-l-[4- (trifluoromethoxy)phenyl] (mg, 0.183 mmol) in di chloromethane (3 mL) was added TFA (0.5 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (60 mg, yield 77.3%) as a yellow oil.
[002507] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 92.39% (214 nm) Mass: found peak 393.3 (M+l) at 1.132 min.
[002508] Step 7. Synthesis of 4-[2-[4-[5-(3-methyl-l-bicyclo[l.l.l]pentanyl)-l-[4- (trifluoromethoxy)phenyl] pyrazol-3-yl] piperazin- 1-yl] ethyl] morpholine (Compound a8).
Figure imgf000493_0002
[002509] To a solution of l-[5-(3-methyl-l-bicyclo[l. l.l]pentanyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine (60 mg, 0.153 mmol), potassium carbonate (106 mg, 0.764 mmol) and KI (26 mg, 0.153 mmol) in 95% ethanol (3 mL) was added 4-(2- chloroethyl)morpholine hydrochloride (43 mg, 0.23 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-[5-(3-methyl-l-bicyclo[l.l. l]pentanyl)-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (66.7 mg, yield 86.1%) as a white solid.
[002510JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5% increase to 95%B within 1.4 min; 95% B for 1.6 min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 45 °C. LC purity: 99.80% (214 nm) Mass: found peak 506.2 (M+l) at 2.265 min.
[002511] 'H NMR (400 MHz, MeOD-d4-d) 5 7.50-7.38 (m, 4H), 5.77 (s, 1H), 3.69 (t, J = 4.4 Hz, 4H), 3.20 (t, J = 4.4 Hz, 4H), 2.62 (t, J = 4.4 Hz, 4H)„2.60-2.56 (m, 4H), 2.52 (t, J = 4.4 Hz, 4H), 1.76 (s, 6H), 1.10 (s, 3H) ppm.
Example S-a9. Synthesis of product 4-[2-[5-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3-dioxan-2-yl]ethyl]morpholine (Compound a9- Trans and Compound a9-Cis).
[002512] Compounds a9 (a9-Trans and a9-Cis) were prepared as outlined below.
Figure imgf000494_0001
[002513] Step 1. Synthesis of (2,2-dimethyl-4H-l,3-dioxin-5-yl)trifluoromethanesulfonate.
TEA (1 eq)
Figure imgf000494_0002
DCM,-23oC~0oC,3h
[002514] To a solution of 2,2-dimethyl-l,3-dioxan-5-one (2.74 g, 21.1 mmol) in dichloromethane (200 mL) was added N,N-dimethylpyridin-4-amine (2.57 g, 21.1 mmol) and N,N-diethylethanamine (6.39 g, 63.3 mmol) at 0 °C, and the resulting reaction mixture was stirred for 3 h at RT. Then trifluoromethyl sulfonyl trifluoromethanesulfonate (8.91 g, 31.6 mmol) was added, and the solution stirred at room temperature for 16h. After completion of the reaction, di chloromethane (200 mL) was added to the solution, the solution washed with HC1 solution three times, and the solution concentrated to dryness to afford (2,2-dimethyl-4H-l,3- di oxin-5-yl)trifluorom ethanesulfonate (3 g, crude) as a yellow oil. The crude product was used in the next step directly.
[002515] XH NMR (400 MHz, CDCh): 5 6.80 (t, J = 1.2 Hz, 1H), 4.36 (d, J = 1.6 Hz, 2H), 156
(d, J = 32.4 Hz, 6H).
[002516] Step 2. Synthesis of 4-(3, 3-dimethoxypropyl) morpholine.
Figure imgf000495_0001
[002517] A mixture of 3 -bromo- 1, m 1 -dimethoxy-propane (2 g, 10.9 mmol), morpholine (1.9 g, 21.9 mmol) and potassium carbonate (4.53 g, 32.8 mmol) in N,N-dimethylformamide (30 mL) was stirred at 50 °C for 16h. The reaction was cooled to room temperature, diluted in EtOAc (200 mL), washed with water (30 mL) and brine (30 mL X 4), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 0-30% acetone in petroleum ether) to afford the desired product 4-(3,3-dimethoxypropyl)morpholine (1.95 g, yield: 94.3%) as a yellow oil. [002518JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 190.1 (M+l) + at 1.445 min.
[002519] 'H NMR (400 MHz, CDCh): 5 4.46 (t, J = 5.6 Hz, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.33 (s, 6H), 2.44 (t, J = 4.0 Hz, 4H), 2.42-2.37 (m, 2H), 1.84-1.79 (m, 2H) ppm.
[002520] Step 3. Synthesis of 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000495_0002
[002521] To a solution of 3-bromo-5-methyl-lH-pyrazole (5 g, 31.1 mmol) in chloroform (300 mL) was added [4-(trifluoromethoxy) phenyl]boronic acid (12.8 g, 62.1 mmol), anhydrous copper (II) acetate and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 16h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 330g silica gel column @100mL/min, eluting with 0-38% di chloromethane in petroleum ether) to afford the desired product 3-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (3.8 g, yield: 38.1%) as a colorless oil.
[002522JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 323.0 (M+l) +, at 1.422 min.
[002523] 'H NMR (500 MHz, CDCh): 5 7.49-7.45 (m, 2H), 7.32 (d, J = 8.5 Hz, 2H), 6.24 (d, J = 1.0 Hz, 1H), 2.34 (s, 3H) ppm.
[002524] Step 4. Synthesis of tributyl-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl] stannane.
Figure imgf000496_0002
, ,
[002525] To a solution of 3-bromo-l-(4-methoxyphenyl)-5-methyl- pyrazole (2 g, 7.49 mmol), tetrakis(triphenylphosphane)palladium (865 mg, 0.75 mmol) in 200 mL toluene was added tributyl (tributyl stannyl)stannane (18.9 mL, 37.4 mmol). The reaction mixture was stirred at 110 °C for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: di chloromethane = 4 / 1) to afford the desired product tributyl-[5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]stannane (0.81g, yield: 20%) as a colorless oil.
[002526] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 533.3 (M+l) + at 4.22 min.
[002527] Step 5. Synthesis of 3-(2,2-dimethyl-4H-l,3-dioxin-5-yl)-5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000496_0001
[002528] To a solution of tributyl-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]stannane (1.2 g, 2.26 mmol), tris(dibenzylideneacetone)dipalladium (207 mg, 0.226 mmol), and triphenylarsane (69 mg, 0.226 mmol) in 40 mL NMP was added (2, 2-dimethyl-4H-l, 3- dioxin-5-yl)trifluoromethanesulfonate (592 mg, 2.26 mmol). The reaction mixture was stirred at room temperature for 48h. The crude was directly purified by flash chromatography (Biotage, 80 g silica gel column @100 mL / min, eluting with 20% di chloromethane in petroleum ether) to afford the desired product 3-(2,2-dimethyl-4H-l,3-dioxin-5-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (370 mg, 0.9 mmol) as a yellow oil.
[002529JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 87 % (214 nm); Mass: found peak 355.2 (M + H) at 1.47 min.
[002530] Step 6. Synthesis of 3-(2, 2-dimethyl-l, 3-dioxan-5-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyljpyrazole.
Figure imgf000497_0001
[002531] Under H2 atmosphere, a mixture of 3-(2, 2-dimethyl-4H-l,3-dioxin-5-yl)-5-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazole (200 mg, 0.564 mmol), Pd2(dba)s (103 mg, 0.113 mmol) and Pd/C (10%, 40 mg) in methanol (20 mL) was stirred at room temperature for 8h. The reaction was filtered to give 3-(2, 2-dimethyl-l, 3-dioxan-5-yl)-5-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazole (200 mg, crude). The crude product was used directly in the next step without further purification.
[002532JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 22.97% (214 nm) Mass: found peak 357.3 (M+l) at 1.375 min.
[002533] Step 7. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl] propane-1, 3-diol.
Figure imgf000497_0002
[002534] A mixture of 3-(2, 2-dimethyl-l, 3-dioxan-5-yl)-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (crude, 200 mg) and HC1 aqueous (IN, 2.5 mL, 2.5 mmol) in methanol (20 mL) was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with DCM (20 mL), neutralized with sodium bicarbonate aqueous, and separated. The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0-10% methanol in di chloromethane) to afford the desired product 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]propane-l,3-diol (130 mg, yield: 72.9% (2 steps)) as a yellow oil.
[002535] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 317.0 (M + 1) + at 1.796 min.
[002536] XH NMR (400 MHz, CDCh): 5 7.50-7.45 (m, 2H), 7.32 (d, J = 8.0 Hz, 2H), 6.15 (s, 1H), 4.09-3.99 (m, 4H), 3.12-3.05 (m, 1H), 2.35 (s, 3H) ppm.
[002537] Step 8. Synthesis of 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l,3-dioxan-2-yl]ethyl]morpholine (a9-Trans) and 4-[2-[5-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3-dioxan-2-yl]ethyl]morpholine (a9-Cis).
Figure imgf000498_0001
[002538] To a mixture of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]propane-l, 3- diol (60 mg, 0.19 mmol) and 4-(3,3-dimethoxypropyl)morpholine (72 mg, 0.379 mmol) in 2 mL toluene was added 4-methylbenzenesulfonic acid (16 mg, 0.0949 mmol). The mixture was stirred at 120 °C for 16h. The reaction was cooled to room temperature and directly purified by prep-HPLC (ammonium hydrogen carbonate / water / acetonitrile) to afford the desired product 4-[2-[5-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3-dioxan-2- yl]ethyl]morpholine (a9-Trans, 27.8 mg, yield: 33.2%) as a yellow oil and the by-product 4-[2- [5-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-l,3-dioxan-2-yl]ethyl]morpholine (a9-Cis, 17.1 mg, yield: 20.4%) as a yellow oil.
[002539] a9-Trans. LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 442.1 (M + 1) + at 2.05 min. 1 H NMR (400 MHz, CD3OD): 5 7.58 (dd, J = 8.8, 2.4 Hz, 2H), 7.46 (d, J = 8.8 Hz, 2H), 6.20 (s, 1H), 4.72 (t, J = 4.8 Hz, 1H), 4.23 (dd, J = 11.6, 4.4 Hz, 2H), 3.86 (t, J = 12.0 Hz, 2H), 3.72 (t, J = 4.4 Hz, 4H), 3.24-3.29 (m, 1H), 2.51-2.55 (m, 6H), 2.32 (s, 3H), 1.84-1.89 (m, 2H) ppm.
[002540] a9-Cis. LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 442.1 (M + 1) + at 2.00 min. 1 H NMR (400 MHz, CD3OD): 5 7.59-7.63 (m, 2H), 7.46-7.48 (m, 2H), 6.52 (s, 1H), 4.80 (t, J = 5.2 Hz, 1H), 4.16-4.23 (m, 4H),
3.72 (t, J = 4.4 Hz, 4H), 2.76 (s, 1H), 2.50-2.56 (m, 6H), 2.36 (s, 3H), 1.85-1.90 (m, 2H) ppm.
Example S-alO. Synthesis of l-[2-(3-methylsulfonylazetidin-l-yl)ethyl]-4-[5-methyl-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl] piperazine (Compound alO).
[002541] Compound alO was prepared as outlined below.
Figure imgf000499_0001
[002542] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), potassium carbonate (92 mg, 0.671 mmol) and KI (22 mg, 0.134 mmol) in 95% ethanol (2 mL) was added 3-methylsulfonylazetidine; hydrochloride (46 mg, 0.268 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 1- [2-(3-methylsulfonylazetidin-l-yl)ethyl]-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (54.1 mg, yield 81.4%) as a yellow oil.
[002543JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 472.1 (M+l) at 1.696 min.
[002544] XH NMR (400 MHz, CDCh) 5 5.73 (s, 1H), 3.92 (s, 1H), 3.68 (t, J = 8.2 Hz, 2H), 3.55 (dd, J = 8.4, 6.8 Hz, 2H), 3.26 (dd, J = 11.3, 6.3 Hz, 4H), 2.89 (s, 3H), 2.68 (t, J = 6.6 Hz, 2H), 2.63 - 2.55 (m, 4H), 2.41 (t, J = 6.5 Hz, 2H), 2.36 (d, J = 2.0 Hz, 3H) ppm.
Example S-all. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-4-[2- [3-(trifluoromethoxy)azetidin-l-yl]ethyl] piperazine (Compound all).
[002545] Compound all was prepared as outlined below.
Figure imgf000500_0001
[002546] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), potassium carbonate (92 mg, 0.671 mmol) and KI (22 mg, 0.134 mmol) in 95% ethanol (2 mL) was added 3 -(trifluoromethoxy) azetidine;hydrochloride (48 mg, 0.268 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-4-[2-[3-(trifluoromethoxy)azetidin-l- yl]ethyl] piperazine (18.7 mg, yield 29.2%) as a yellow oil.
[002547] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 478.1 (M+l) at 2.101 min.
[002548] 'H NMR (400 MHz, CDCh) 5 7.67 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 8.3 Hz, 2H), 5.73 (s, 1H), 4.82 - 4.70 (m, 1H), 3.75 (t, J = 7.3 Hz, 2H), 3.34 - 3.21 (m, 4H), 3.13 (t, J = 6.9 Hz, 2H), 2.63 (dt, J = 9.0, 5.5 Hz, 6H), 2.41 (t, J = 6.6 Hz, 2H), 2.36 (s, 3H) ppm.
Example S-al2. Synthesis of l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-4-[2- [3-(trifluoromethylsulfonyl)azetidin-l-yl]ethyl] piperazine (Compound a!2).
[002549] Compound a!2 was prepared as outlined below.
Figure imgf000500_0002
[002550] To a solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (50 mg, 0.134 mmol), potassium carbonate (92 mg, 0.671 mmol) and KI (22 mg, 0.134 mmol) in 95% ethanol (2 mL) was added 3 21ambda6-thia-6-azaspiro[3.3]heptane 2,2- dioxide (39 mg, 0.268 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-[5-methyl-l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]-4-[2-[3- (trifluoromethylsulfonyl)azetidin-l-yl]ethyl] piperazine (22.3 mg, yield 31.6%) as a white solid. [002551JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 526.2 (M+l) at 1.156 min.
[002552] 'H NMR (400 MHz, CDCh) 5 7.70 (d, J = 8.5 Hz, 2H), 7.56 (d, J = 8.4 Hz, 2H), 5.74 (s, 1H), 4.26 - 4.20 (m, 1H), 3.87 (t, J = 8.2 Hz, 2H), 3.73 (s, 2H), 3.62 (s, 4H), 3.36 (s, 4H), 3.05 (s, 4H), 2.37 (s, 3H) ppm.
Example S-al3. Synthesis of 4-[2-[4-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3- yl)piperazin-l-yl]ethyl]morpholine (Compound a!3).
[002553] Compound a!3 was prepared as outlined below.
Figure imgf000501_0001
[002554] Step 1. Synthesis of tert-butyl 4-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3- yl)piperazine-l-carboxylate.
Figure imgf000501_0002
[002555] A mixture of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l -carboxylate (280 mg, 1.05 mmol), tetrahydropyran-4-yl methanesulfonate (379 mg, 2.1 mmol) and cesium carbonate (1.03 g, 3.15 mmol) in N,N-dimethylformamide (10 mL) was stirred at 90 °C for 48h. The reaction mixture was cooled to room temperature and diluted with EtOAc (150 mL), washed with water (15 mL), brine (15 mL*4), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100mL/min, eluting with 10-50% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3-yl)piperazine-l- carboxylate (140 mg, yield 36.3%) as a yellow solid. [002556] LCMS method: Mobile Phase: Water (0.01%TFA) (A) / Acetonitrile (0.01%TFA) (B); Gradient: 5 % - 95 % B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 95.44% (214 nm) Mass: found peak 351.3 (M+l) at 1.238 min.
[002557] XH NMR (400 MHz, DMSO-d6) 5 5.50 (s, 1H), 4.20-4.11 (m, 1H), 3.91 (dd, J = 11.2, 4.0 Hz, 2H), 3.47-3.35 (m, 6H), 2.98 (t, J = 4.8 Hz, 4H), 2.18 (s, 3H), 2.04-1.91 (m, 2H), 1.66 (dd, J = 12.4, 2.4 Hz, 2H), 1.40 (s, 9H) ppm.
[002558] Step 2. Synthesis of l-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3-yl)piperazine.
Figure imgf000502_0001
[002559] To a solution of tert-butyl 4-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3-yl)piperazine- 1-carboxylate (140 mg, 0.399 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 2.69 mmol). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-(5-methyl-l-tetrahydropyran- 4-yl-pyraz°l-3-yl)piperazine (95 mg, yield 85.8%) as a yellow solid.
[002560JLCMS method: Mobile Phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B); Gradient: from 10% to 95% of B ini.5 min at 1.8 mL/min; Column: X- BRIDGE C18 (4.6x 50 mm, 3.5pm); Column Temperature: 50 °C; LC purity: 90.34% (214 nm), Mass: found peak 251.1 (M+l)+ at 1.262 min.
[002561] Step 3. Synthesis of 4-[2-[4-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3- yl)piperazin-l-yl]ethyl]morpholine (Compound al3).
Figure imgf000502_0002
[002562] To a solution of l-(5-methyl-l-tetrahydropyran-4-yl-pyrazol-3-yl)piperazine (100 mg, 0.379 mmol), potassium carbonate (210 mg, 1.52 mmol) and KI (63 mg, 0.379 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (106 mg, 0.569 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-(5- methyl-l-tetrahydropyran-4-yl-pyrazol-3-yl)piperazin-l-yl]ethyl]morpholine (89.3 mg, yield 64.5%) as a white solid.
[002563JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within Temperature: 50 °C; LC purity: 99.66% (214 nm) Mass: found peak 364.2 (M+l) at in 1.5 min; Flow Rate: 1.5 mL/min; Column: X- Bridge C18, 50*4.6mm, 3.5pm; Column temperature: 50 °C, 1.612 min.
[002564] XH NMR (400 MHz, DMSO-d6) 5 5.45 (s, 1H), 4.19-4.09 (m, 1H), 3.91 (dd, J = 11.2, 4.0 Hz, 2H), 3.55 (t, J = 4.4 Hz, 4H), 3.42 (td, J = 11.6, 1.6 Hz, 2H), 2.99 (t, J = 4.4 Hz, 4H), 2.46 (t, J = 4.8 Hz, 4H), 2.43-2.41 (m, 4H), 2.38 (t, J = 4.4 Hz, 4H), 2.17 (s, 3H), 1.97 (qd, J = 12.4, 4.4 Hz, 2H), 1.65 (dd, J = 12.4, 2.4 Hz, 2H) ppm.
Example S-al4. Synthesis of 4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]piperazin-2-one (Compound a!4).
[002565] Compound a!4 was prepared as outlined below.
Figure imgf000503_0001
[002566] A solution of l-(2-chloroethyl)-4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3- yl]piperazine (80 mg, 0.206 mmol), piperazin-2-one (30 mg, 0.309 mmol) and DIPEA (133 mg, 1.03 mmol) in NMP (2 mL) was stirred at 160 °C for 2h in MW, then concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3- yl]piperazin-l-yl]ethyl]piperazin-2-one (36.1 mg, yield 38%) as a yellow solid.
[002567] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 100.0% (214 nm); Mass: found peak 453.2(M+H) at 1.87 min.
[002568] 'H NMR (400 MHz, MeOD-d4) 5 7.55 (t, J = 8.0 Hz, 2H), 7.39 (d, J = 8.0 Hz, 2H), 5.84 (s, 1H), 3.36-3.31 (m, 2H), 3.23 (t, J = 8.0 Hz, 4H), 3.17 (s, 2H), 2.75 (t, J = 8.0 Hz, 2H), 2.70-2.57 (m, 8H), 2.29 (s, 3H) ppm. Example S-al5. Synthesis of l-[2-[4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3- yl)pyrazol-3-yl]-l-piperidyl] ethyl]piperidin-4-ol (Compound a!5).
[002569] Compound a!5 was prepared as outlined below.
Figure imgf000504_0001
[002571] To a solution of 3,5-dibromo-lH-pyrazole (1 g, 4.46 mmol) and (4- chlorophenyl)boronic acid (628 mg, 4.0 mmol) in 1,4-dioxane (10 mL) was added sodium carbonate (1.4 g, 13.38 mmol) and PdC12(dppf) dichloromethane complex (326 mg, 0.446 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product 4-bromo-2-(4-chlorophenyl)-lH-pyrrole (557 mg, yield 49%) as a white solid.
[002572JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 82% (214 nm) Mass: found peak 257.1(M +H) + at 1.524 min.
[002573] Step 2. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]-3,6-dihydro- 2H-pyridine-l-carboxylate.
Figure imgf000505_0001
[002574] To a solution of 4-bromo-2-(4-chlorophenyl)-lH-pyrrole (450 mg, 01.76 mmol) and tert-butyl 4-(4, 4,5, 5-tetramethyl- 1,3, 2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyri dine- 1- carboxylate (816 mg, 2.64 mmol) in 1,4-di oxane (5 mL) was added sodium carbonate (560 mg, 5.28 mmol) and PdC12(dppf) dichloromethane complex (129 mg, 0.176mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]-3,6-dihydro-2H-pyridine-l- carboxylate (630 mg, yield 99.8%) as a colorless oil.
[002575] LCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 82% (214 nm) Mass: found peak 304.0 (M -55) + at 1.659 min.
[002576] Step 3. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]piperidine-l- carboxylate.
Figure imgf000505_0002
[002577]Under H2 atmosphere, a mixture of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]- 3, 6-dihydro-2H-pyridine-l -carboxylate (600 mg, 1.67 mmol) and PtCh (100 mg) in MeOH (30 mL) was stirred at room temperature for Ih. The reaction was filtered and concentrated in vacuo. The residue was used directly in the next step without further purification.
[002578JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 84% (214 nm) Mass: found peak 306.0 (M -55) + at 2.094 min.
[002579] Step 4. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-chloropyridazin-3- yl)pyrazol-3-yl] piperidine-l-carboxylate.
Figure imgf000505_0003
[002580] A mixture of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]piperidine-l- carboxylate (300 mg, 0.48 mmol), 3,6-dichloropyridazine (247 mg, 0.96 mmol) and sodium hydride (50 mg, 1.24 mmol) in THF (10 mL) was stirred at 60 °C for 16h.The filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0%-10% EtOAc in PE) to afford the desired product tertbutyl 4-[5-(4-chlorophenyl)-l-(6-chloropyridazin-3-yl)pyrazol-3-yl] piperidine- 1 -carboxylate (328 mg, yield 58.4%) as a yellow solid.
[002581JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: found peak 374.0(M - 100)+ at 2.410 min.
[002582] Step 5. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3-yl) pyrazol-3-yl] piperidine-l-carboxylate.
Figure imgf000506_0001
[002583] To a solution of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]piperidine-l- carboxylate (328 mg, 0.69 mmol) and 2,4,6-trimethyl-l,3,5,2,4,6-trioxatriborinane (156 mg, 0.62 mmol) in 1,4-dioxane (5 mL) was added cesium carbonate (886 mg, 2.07 mmol) and PdC12(dppf) dichloromethane complex (51 mg, 0.069 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered and concentrated in vacuo. The filtrate was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluted with 30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3-yl) pyrazol-3-yl] piperidine-l- carboxylate (72 mg, yield 14%) as a colorless oil.
[002584JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 93% (214 nm); Mass: found peak 354.3 (M -100)+ at 2.422 min.
[002585] Step 6. Synthesis of 3-[5-(4-chlorophenyl)-3-(4-piperidyl)pyrazol-l-yl]-6-methyl- pyridazine.
Figure imgf000506_0002
[002586] To a solution of tert-butyl 4-(5-(4-chlorophenyl)-l-(6-methylpyridazin-3-yl)-lH- pyrazol-3-yl)piperidine-l -carboxylate (94 mg, 0.216 mmol) in dichloromethane (5 mL) was added 2,2,2-trifluoroacetic acid (ImL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 3-[5-(4-chlorophenyl)-3-(4- piperidyl)pyrazol-l-yl]-6-methyl-pyridazine (75 mg, crude). The crude product was used directly in the next step.
[002587] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 84% (214 nm) Mass: found peak 354.0 (M + 1) + at 1.887 min.
[002588] Step 7. Synthesis of 3-[3-[l-(2-chloroethyl)-4-piperidyl]-5-(4- chlorophenyl)pyrazol-l-yl]-6-methyl-pyridazine.
Figure imgf000507_0001
[002589]Under argon atmosphere, a mixture of 13-[5-(4-chlorophenyl)-3-(4-piperidyl)pyrazol- l-yl]-6-methyl-pyridazine (72 mg, 0.2 mmol), 2-chloroacetaldehyde (80 mg, 0.4 mmol)and sodium cyanoborohydride (25.6 mg, 0.4 mmol) in methanol (6 mL) was stirred at rt for 16h. The mixture was quenched by sodium bicarbonate, then concentrated in vacuo to remove most of the solvent. The mixture was diluted with DCM (5 mL) and 3 mL water was added, extracted with DCM (5 x 3 mL), the organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @75 mL/min, eluting with 0-10% methanol in dichloromethane) to afford the desired product 3 -[3 -[ 1 - (2-chloroethyl)-4-piperidyl]-5-(4-chlorophenyl)pyrazol-l-yl]-6-methyl-pyridazine (80 mg, yield: 94.4%) as a yellow oil.
[002590JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73% (214 nm); Mass: found peak 416.1(M+1) at 1.708 min.
[002591] Step 8. Synthesis of l-[2-[4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3- yl)pyrazol-3-yl]-l-piperidyl] ethyl]piperidin-4-ol (Compound al5).
Figure imgf000508_0001
[002592] To a solution of 3-[3-[l-(2-chloroethyl)-4-piperidyl]-5-(4-chlorophenyl)pyrazol-l-yl]- 6-methyl-pyridazine (72 mg, 0.2 mmol), potassium carbonate (120 mg, 0.8 mmol) and KI (28.7 mg, 0.2 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine (38.8 mg, 2.59 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product l-[2-[4-[5-(4- chlorophenyl)-l-(6-methylpyridazin-3-yl)pyrazol-3-yl]-l-piperidyl] ethyl]piperidin-4-ol (11.1 mg, yield: 13.3%) as a white solid.
[002593JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 481.4 (M + H) at 1.736 min.
[002594] 'H NMR (400 MHz, CDCI3) 5 8.16 (d, J = 9.0 Hz, 1H), 7.80 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 9.0 Hz, 1H), 7.40 (d, J = 8.4 Hz, 2H), 6.62 (s, 1H), 3.83 (s, 1H), 3.74 (t, J = 8.2 Hz, 1H), 3.08 (d, J = 11.1 Hz, 2H), 2.92 - 2.80 (m, 2H), 2.76 (s, 3H), 2.26 (t, J = 11.3 Hz, 4H), 2.14 (d, J = 12.5 Hz, 2H), 1.90-1.95 (m, 2H), 1.88-1.75 (m, 4H), 1.56-1.68 (m, J = 18.4, 9.2 Hz, 4H) ppm.
Example S-al6. Synthesis of 4-[2-[4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3- yl)pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound a!6).
[002595] Compound a!6 was prepared as outlined below.
Figure imgf000508_0002
[002596] Step 1. Synthesis of tert-butyl 4-[3-(4-chlorophenyl)-3-oxo-propanoyl]piperazine-
1-carboxylate.
Figure imgf000508_0003
[002597] A mixture of ethyl 3-(4-chlorophenyl)-3-oxo-propanoate (1.0 g, 4.4 mmol) and tertbutyl piperazine- 1-carboxylate (894 mg, 4.8 mmol) in toluene (100 mL) was heated at 100°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 0-30% acetone in petroleum ether to afford tertbutyl 4-[3-(4-chlorophenyl)-3-oxo-propanoyl]piperazine-l-carboxylate (878 mg, 54.2% yield) as a yellow solid.
[002598JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 98.44% (214 nm) Mass: found peak 257.2 (M -55)+ at 1.944 min.
[002599] Step 2. Synthesis of tert-butyl 4-[3-(4-chlorophenyl)-3-oxo- propanethioyl]piperazine-l-carboxylate.
Figure imgf000509_0001
[002600] To a solution of tert-butyl 4-(3-oxoheptanoyl)piperazine-l -carboxylate (1.9 g, 4.44mmol) in toluene (50 mL) was added Lawesson's reagent (890mg, 2.2 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography eluting with 10-65% ethyl acetate in petroleum ether to afford tert-butyl 4-[3-(4-chlorophenyl)-3-oxo- propanethioyl]piperazine-l -carboxylate (855 mg, yield 43.1%) as a yellow solid.
[002601JLCMS method: Mobile Phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min;
Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 87.52% (214 nm) Mass: found peak 383.2 (M + H) at 2.024 min.
[002602] Step 3. Synthesis of tert-butyl 4-(5-butyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Figure imgf000509_0002
[002603] To a solution of tert-butyl 4-[3-(4-chlorophenyl)-3-oxo-propanethioyl]piperazine-l- carboxylate (636 mg, 1.66 mmol) in toluene (50 mL) was added NH2NH2 water (250 mg, 4.99 mmol) and the mixture was stirred at 75°C for 16h. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 10 CV) to afford tert-butyl 4-(5-butyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (550 mg, yield 66.7%) as a yellow solid. [002604JLCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B), Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min.
Column X-B RIDGE Cl 8 (4.6x 50 mm, 3.5pm). Temperature: 50°C. LC purity: 96% (214 nm) Mass: found peak 363.2 (M + H) at 1.927 min.
[002605] Step 4. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-chloropyridazin-3- yl)pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000510_0001
[002606] A mixture of tert-butyl 4-[5-(4-chlorophenyl)-lH-pyrazol-3-yl]piperazine-l- carboxylate (224 mg, 0.6 mmol), 3,6-dichloropyridazine (184 mg, 1.2 mmol) and sodium hydride (50 mg, 1.2 mmol) in THF (10 mL) was stirred at 60 °C for 16h. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @75mL/min, eluting with 0%-10% EtOAc in PE) to afford the desired product tertbutyl 4-[5-(4-chlorophenyl)-l-(6-chloropyridazin-3-yl)pyrazol-3-yl]piperazine-l-carboxylate (332 mg, yield 79.2%) as a yellow solid.
[002607] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 100% (214nm); Mass: found peak 475.2(M +H)+ at 2.762 min.
[002608] Step 5. Synthesis of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3- yl)pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000510_0002
[002609] To a solution of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-chloropyridazin-3-yl)pyrazol-3- yl]piperazine-l -carboxylate (431 mg, 0.9 mmol) and 2,4,6-trimethyl-l,3,5,2,4,6- tri oxatrib orinane (341 mg, 2.72 mmol) in 1,4-di oxane (5 mL) was added cesium carbonate (886 mg, 2.72 mmol) and PdC12(dppf) dichloromethane complex (74 mg, 0.09 mmol) under argon atmosphere. The reaction mixture was stirred at 80 °C for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @40mL/min, eluting with 30% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3-yl)pyrazol-3-yl]piperazine-l- carboxylate (94 mg, yield 18.2%) as a colorless oil.
[002610JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C. LC purity: 93% (214 nm); Mass: found peak 455.3 (M +H)+ at 2.458 min.
[002611] Step 6. Synthesis of 3-[5-(4-chlorophenyl)-3-piperazin-l-yl-pyrazol-l-yl]-6-methyl- pyridazine.
Figure imgf000511_0001
[002612] To a solution of tert-butyl 4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3-yl)pyrazol-3- yl]piperazine-l -carboxylate (94 mg, 0.216 mmol) in di chloromethane (5 mL) was added 2,2,2- trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 3-[5-(4-chlorophenyl)-3- piperazin-l-yl-pyrazol-l-yl]-6-methyl-pyridazine (60 mg, crude). The crude product was used directly in the next step.
[002613JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 70% (214 nm) Mass: found peak 355.0 (M + 1)+ at 1.748 min.
[002614] Step 7. Synthesis of 4-[2-[4-[5-(4-chlorophenyl)-l-(6-methylpyridazin-3- yl)pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (Compound al6).
Figure imgf000511_0002
[002615] To a solution of 3-[5-(4-chlorophenyl)-3-piperazin-l-yl-pyrazol-l-yl]-6-methyl- pyridazine (60 mg, 0.17 mmol), potassium carbonate (70.1 mg, 0.5 mmol) and KI (28.1 mg, 0.17 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine (37.9 mg, 0.25 mmol). The reaction was stirred at 95 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[5-(4- chlorophenyl)-l-(6-methylpyridazin-3-yl)pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (18.4 mg, yield: 23.3%) as a white solid.
[002616] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98.39 % (214 nm);
Mass: found peak 448.3 (M + H) at 1.822 min.
[002617] 'H NMR (400 MHz, CDCh) 5 8.07 (d, J = 9.0 Hz, 1H), 7.80 (d, J = 8.5 Hz, 2H), 7.46
(d, J = 9.0 Hz, 1H), 7.39 (d, J = 8.5 Hz, 2H), 6.27 (s, 1H), 3.67-3.79 (m, 4H), 3.15-3.40 (m, 4H), 2.98-2.33 (m, 15H) ppm.
Example S-al7. Synthesis of l-[2-[4-(difluoromethyl)phenyl]-5-[4-(2-morpholinoethyl) piperazin-l-yl]pyrazol-3-yl]ethanol (Compound al7).
[002618] Compound al7 was prepared as outlined below.
Figure imgf000512_0001
[002619] To a solution of 4-[2-[4-[l-[4-(difluoromethyl)phenyl]-5-(l -methoxy ethyl)pyrazol-3- yl]piperazin-l-yl]ethyl]morpholine (100 mg, 0.222 mmol) in DCM (10 mL) at -78°C was added tribromoborane (167 mg, 0.667 mmol) and the reaction mixture was stirred at -78°C for 16h. The residue was purified by prep-HPLC (NH4HCO4 /water/acetonitrile) to afford the desired product l-[2-[4-(difluoromethyl)phenyl]-5-[4-(2-morpholinoethyl) piperazin- l-yl]pyrazol-3- yl]ethanol (10.1 mg, yield 10.4%) as a white solid.
[002620JLCMS method: Mobile phase: Water (10 mM ammonium hydrogen carbonate) (A) / Acetonitrile (B); Elution program: Gradient from 10 to 95% of B inl.5min at 1.8mL/min;
Temperature: 50°C; LC purity: 100% (214 nm); Mass: found peak 436.4 (M-55)+ at 1.54 min. [002621] 'H NMR (400 MHz, CDCh) 5 7.71 (d, J = 8.4 Hz, 2H), 7.58 (d, J = 8.2 Hz, 2H), 6.68 (t, J = 56.4 Hz, 1H), 5.97 (s, 1H), 4.85 (d, J = 6.4 Hz, 1H), 3.74 - 3.71 (m, 4H), 3.30 (d, J = 4.4 Hz, 4H), 2.67 - 2.44 (m, 15H).
Example S-al8. Synthesis of 2-[5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]propan-2-ol (Compound al8).
[002622] Compound al8 was prepared as outlined below.
Figure imgf000513_0001
[002623] Step 1. Synthesis of 3,5-dibromo-l-[4-(trifluoromethyl)phenyl] pyrazole.
Figure imgf000513_0002
[002624] A solution of 3,5-dibromo-lH-pyrazole (1.5 g, 6.64 mmol), [4- (trifluoromethyl)phenyl]boronic acid (2.5 g, 13.3 mmol), pyridine (26.2 g, 33.2 mmol), and copper(II)acetate (36.0 g, 19.9 mmol) in chloroform (50 mL) was stirred at RT for 16h under the atmosphere of O2, and LCMS showed the reaction was complete. The mixture was filtered, the filter cake washed with dichloromethane (100 mL), the solution was concentrated, and the mixture was purified by flash chromatography (Biotage, 330 g silica gel column @100mL/min, eluting with 0%-40% ethyl acetate in petroleum ether for 30 min) to afford 3, 5 -dibromo- 1- [4- (trifluoromethyl)phenyl] pyrazole (1.2 g, 13.0%) as an oil.
[002625] LCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 87.0% (214 nm) Mass: found peak 370.9 (M+l) at 2.630 min.
[002626] Step 2. Synthesis of l-[5-bromo-2-[4-(trifluoromethyl)phenyl]pyrazol-3- yljethenone.
Figure imgf000513_0003
[002627]Under argon atmosphere, to a solution of 3,5-dibromo-l-[4-(trifluoromethyl)phenyl] pyrazole (100 mg, 0.27 mmol) in 1,4-di oxane (3 mL) was added tributyl(l-ethoxyvinyl) stannane (88 mg, 0.24 mmol) and triphenylphosphine (31.2 mg, 0.027 mmol). The reaction mixture was stirred at 110 °C for 16 h. The reaction was purified by SGC (PE : DCM = 5: 1) to afford the desired product l-[5-bromo-2-[4-(trifluoromethyl) phenyl]pyrazol-3-yl]ethanone (69 mg, 54.3%) as a white solid.
[002628JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 69.29% (214 nm) Mass: found peak 459.9 (M+l) at 1.846 min.
[002629] Step 3. Synthesis of 2-[5-bromo-2-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]propan-2-ol.
Figure imgf000514_0001
[002630] Under argon atmosphere, to a solution of l-[5-bromo-2-[4-(trifluoromethyl) phenyl] pyrazol-3-yl]ethanone (320 mg, 0.99 mmol) in THF (15 mL) was added bromo(methyl)magnesium (4 mL, 4.0 mmol) at 0 °C. The reaction mixture was stirred at rt for 16 h. The reaction mixture was quenched with ice water (3 mL), extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @36 mL/min, eluting with 0- 20% ethyl acetate in petroleum ether) to afford the desired product 2-[5-bromo-2-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]propan-2-ol (220 mg, yield 63.6%) as a yellow solid. [002631JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 351.0 (M+l) at 2.029 min.
[002632] Step 4. Synthesis of tert-butyl 4-[5-(l-hydroxy-l-methyl-ethyl)-l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000514_0002
[002633] Under argon atmosphere, a mixture of 2-[5-bromo-2-[4-(trifluoromethyl) phenyl]pyrazol-3-yl]propan-2-ol (120 mg, 0.5 mmol), tert-butyl piperazine- 1 -carboxylate (144 mg, 0.77 mmol), tBuXPhos Pd G3 (41 mg, 0.05 mmol) and sodium tert-butoxide (148 mg, 1.54 mmol) in 1,4-di oxane (8 mL) was stirred 100 °C for 16h, concentrated and purified by flash chromatography using 2-30% ethyl acetate in petroleum ether to afford tert-butyl 4-[5-(l - hydroxy- 1 -methyl-ethyl)- 1 -[4-(trifluoromethyl)phenyl]pyrazol-3 -yl]piperazine- 1 -carboxylate (100 mg, 41% yield) as a yellow solid.
[002634JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: X- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 96.25% (214 nm) Mass: found peak 455.1 (M+l) at 2.136 min.
[002635] Step 5. Synthesis of 2-[5-piperazin-l-yl-2-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]propan-2-ol.
Figure imgf000515_0001
[002636] To a solution of tert-butyl 4-[5-(l-hydroxy-l-methyl-ethyl)-l-[4-(trifluoromethyl) phenyl]pyrazol-3-yl]piperazine-l -carboxylate (100 mg, 0.22 mmol) in dichloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (30 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford 2-[5-piperazin-l-yl-2-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]propan-2-ol (60 mg, yield 56.4%) as a yellow oil.
[002637] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 73.23% (214 nm) Mass: found peak 355.1 (M+l) at 1.725 min.
[002638] Step 6. Synthesis of 2-[5-[4-(2-morpholinoethyl)piperazin-l-yl]-2-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]propan-2-ol (Compound a!8).
Figure imgf000515_0002
[002639] To a solution of 2-[5-piperazin-l-yl-2-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]propan- 2-ol (30 mg, 0.08 mmol), potassium carbonate (59 mg, 0.4 mmol) and KI (14 mg, 0.08 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (24 mg, 0.13 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 2-[5-[4-(2- morpholinoethyl)piperazin-l-yl]-2-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]propan-2-ol (21.3 mg, yield 52.7%) as a yellow solid.
[002640JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97.94% (214 nm) Mass: found peak 468.2 (M+l) at 1.858 min.
[002641] 'H NMR (400 MHz, MeOD-d4) 5 7.83-7.69 (m, 4H), 5.98 (s, 1H), 3.70 (t, J = 4.4 Hz, 4H), 3.23 (t, J = 4.4 Hz, 4H), 2.72-2.44 (m, 12H), 1.42 (s, 6H) ppm.
Example S-al9. Synthesis of 4-[2-[4-(5-methyl-lH-pyrazol-3-yl)piperazin-l- yl] ethyl] morpholine (Compound al9).
[002642] Compound al9 was prepared as outlined below,
Figure imgf000516_0001
[002643] Step 1. Synthesis of tert-butyl 4-(3-oxobutanoyl)piperazine-l-carboxylate.
Figure imgf000516_0002
[002644] A mixture of tert-butyl piperazine- 1 -carboxylate (20.0 g, 10.7 mmol) and tert-butyl 3- oxobutanoate (18.7 g, 118 mmol) in toluene (400 mL) was heated at 100°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-30% acetone in petroleum ether for 8 CV) to afford tert-butyl 4-(3-oxobutanoyl)piperazine-l- carboxylate (29 g, 99% yield) as a yellow solid. [002645] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm;
Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 231.1 (M-55)+ at 1.278 min.
[002646] Step 2. Synthesis of tert-butyl 4-(3-oxobutanethioyl)piperazine-l-carboxylate.
Lawesson's reagent
(0.5 eq) toluene, 75°c, 16h
Figure imgf000517_0001
Figure imgf000517_0002
[002647] To a solution of tert-butyl 4-(3-oxobutanoyl)piperazine-l -carboxylate (29 g, 107 mmol) in toluene (400 mL) was added Lawesson's reagent (21.7 g, 53.6 mmol) and the mixture was heated at 75°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 10-65% ethyl acetate in petroleum ether for 8 CV) to afford tertbutyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (22 g, yield 71.6%) as a brown oil.
[002648JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 88% (214 nm), Mass: found peak 231.0 (M-55)+ at 1.867 min.
[002649] Step 3. Synthesis of tert-butyl 4-(5-methyl-lH-pyrazol-3-yl)piperazine-l- carboxylate.
Boc
Figure imgf000517_0003
[002650] To a solution of tert-butyl 4-(3-oxobutanethioyl)piperazine-l -carboxylate (22 g, 76.8 mmol) in toluene (400 mL) was added hydrazine monohydrate (11.4 mL, 230 mmol) and the mixture was stirred at 70°C overnight. The mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 330 g silica gel column @200mL/min, eluting with 0-6% MeOH in DCM for 8 CV) to afford tert-butyl 4-(5- methyl-lH-pyrazol-3-yl)piperazine-l-carboxylate (13 g, 63.5% yield) as a yellow solid. [002651JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94% (214 nm), Mass: found peak 267.3 (M+l) at 1.554 min. [002652] Step 4. Synthesis of l-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine.
Figure imgf000518_0001
[002653] To a solution of tert-butyl 4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (5 g, 12.6 mmol) in dichloromethane (60 mL) was added TFA (20 mL). The reaction mixture was stirred at room temperature for 2h. Then the reaction mixture was concentrated to remove solvent and potassium carbonate was added to give pH=10. It was extracted with DCM and dried with sodium sulfate. The reaction mixture was concentrated to afford the desired product l-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (3.7 g, yield 99 %) as a yellow oil.
[002654JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; Mass: found peak 166.2 (M+l) at 0.18 min.
[002655] Step 5. Synthesis of 4-[2-[4-(5-methyl-lH-pyrazol-3-yl)piperazin-l- yl] ethyl] morpholine (a!9).
Figure imgf000518_0002
[002656] To a solution of l-(5-methyl-lH-pyrazol-3-yl)piperazine (3.12 g, 18.8 mmol) and N- ethyl-N-isopropyl-propan-2-amine (11.7 g, 0.2 mmol) in NMP (50 mL) was added 4-(2- chloroethyl)morpholine (2.81 g, 18.8 mmol). The reaction was stirred at 100 °C under microwave for 2h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford 4-[2-[4-(5-methyl-lH-pyrazol-3-yl)piperazin-l- yl]ethyl]morpholine (1.5 g, yield 28.3%) as a yellow solid.
[002657] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 100% (214 nm) Mass: found peak 280.2 (M+l) at 1.461 min. [002658] 'H NMR (400 MHz, CDCh) 5 5.50 (s, 1H), 3.71 (t, J = 4.4 Hz, 4H), 3.20 (t, J = 4.8 Hz, 4H), 2.61 (t, J = 4.8 Hz, 4H, 4H), 2.59 - 2.54 (m, 4H), 2.50 (t, J = 4.4 Hz, 4H), 2.24 (s, 3H) ppm.
Example S-a20. Synthesis of Compound a20.
Figure imgf000519_0001
[002659] To a 100 mL round flask was added tert-butyl (3R,4R)-3-fluoro-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (40 mg, 0.09 mmol), methylene chloride (1 mL) and 4.0 M HC1 in dioxane (2 mL). The reaction mixture was stirred at room temperature for 2 h, then concentrated to dryness to give a solid, which was treated with acetonitrile (6 mL), 4-(2-chloroethyl)morpholine hydrochloride (16.96 mg, 0.09023 mmol), potassium carbonate (62mg, 0.45 mmol) and potassium iodide (14 mg, 0.09 mmol) at 20 °C. The reaction mixture was stirred at 80 °C in a sealed vial for 18 h. The reaction mixture was filtered and concentrated to give a residue, which was which was purified by reverse phase HPLC using 20-100 ACN in water (0.1% formic acid) to give the title product (0.015 g, 36% yield) M/Z (M+H, MW 457).
[002660] 'H NMR (400 MHz, Chloroform-d) 5 7.53 - 7.43 (m, 2H), 7.38 - 7.28 (m, 2H), 6.14 (s, 1H), 4.86 - 4.58 (m, 1H), 4.17 - 3.84 (m, 1H), 3.79 - 3.64 (m, 4H), 3.41 - 3.26 (m, 1H), 2.98 - 2.74 (m, 2H), 2.65 - 2.45 (m, 7H), 2.34 (s, 3H), 2.28 - 2.00 (m, 3H), 1.93 - 1.75 (m, 1H) ppm. Example S-a21. Synthesis of Compound a21.
Figure imgf000519_0002
[002661] In a 25 mL microwave vial were placed (3R,4R)-4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidin-3-ol;2,2,2-trifluoroacetic acid (70 mg, 0.1537 mmol), potassium carbonate (106 mg, 0.77 mmol), potassium iodide (25 mg, 0.15 mmol), 4-(2- chloroethyl) morpholine hydrochloride (28 mg, 0.15 mmol), and acetonitrile (8 mL). The reaction mixture was stirred at 80°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse phase HPLC using 10-100% ACN in water (0.1% formic acid) as eluent to give the title product (0.046g, 66% yield). M+l (455 m/z).
[002662] 'H NMR (400 MHz, Chloroform-d) 5 7.52 - 7.44 (m, 2H), 7.35 - 7.30 (m, 2H), 6.19 - 6.01 (m, 1H), 4.28 - 4.12 (m, 1H), 4.10 - 3.87 (m, 1H), 3.84 - 3.68 (m, 4H), 3.63 - 3.45 (m, 2H), 3.11 (ddt, J = 36.4, 13.1, 6.1 Hz, 2H), 2.96 - 2.78 (m, 4H), 2.70 (t, J = 4.7 Hz, 4H), 2.34 (s, 4H), 2.25 - 2.07 (m, 1H) ppm.
Example S-a22. Synthesis of Compound a22.
Figure imgf000520_0001
[002663] In a 25 mL microwave vial were placed 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperidine;hydrochloride (90 mg, 0.2488 mmol), 2-(chloromethyl) pyrazine hydrochloride (56 mg, 0.33 mmol), potassium carbonate (152 mg, 1.10 mmol), and acetonitrile (8 mL, 100 mass %). The reaction mixture was stirred at 20°C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC chromatography using 10-100% acetonitrile in water as eluent to give the title product as a white solid (0.079 g, 76% yield). M+l (418 m/z).
[002664] 'H NMR (400 MHz, Chloroform-d) 5 8.72 (d, J = 1.4 Hz, 1H), 8.59 - 8.52 (m, 1H), 8.47 (d, J = 2.5 Hz, 1H), 7.51 - 7.42 (m, 2H), 7.36 - 7.28 (m, 2H), 6.05 (s, 1H), 3.75 (s, 2H), 3.01 (dt, J = 11.9, 3.4 Hz, 2H), 2.80 - 2.65 (m, 1H), 2.36 - 2.22 (m, 5H), 2.03 - 1.93 (m, 2H), 1.93 - 1.77 (m, 2H) ppm.
Example S-a23. Synthesis of Compound a23.
Figure imgf000520_0002
[002665] To a 25 mL microwave vial was added 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]piperidine hydrochloride (100 mg, 0.28 mmol, 100 mass %), 2- (chloromethyl)pyridine (54 mg, 0.41 mmol), potassium carbonate (190 mg, 1.38 mmol, 100 mass %), and acetonitrile (10 mL, 100 mass %) at room temperature. The reaction mixture was stirred at 20 °C for 18 h. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC using 10-100% ACN in water (0.1% formic acid) as an eluent to give the title product (formic acid salt) as a white solid (0.075 g, 65% yield). M+l (417 m/z).
[002666] 'H NMR (400 MHz, Chloroform-d) 5 8.66 - 8.47 (m, 1H), 7.75 - 7.62 (m, 1H), 7.56 - 7.40 (m, 3H), 7.38 - 7.26 (m, 2H), 7.24 - 7.08 (m, 1H), 6.05 (s, 1H), 3.72 (s, 2H), 3.07 - 2.97 (m, 2H), 2.97 - 2.80 (m, 1H), 2.80 - 2.65 (m, 1H), 2.32 (s, 3H), 2.30 - 2.18 (m, 2H), 2.02 - 1.77 (m, 4H) ppm. Example S-a24. Synthesis of [4-[3-[4-[2-(l,l-dioxo-l,4-thiazinan-4-yl)ethyl]piperazin-l-yl]- 5-methyl-pyrazol-l-yl]phenyl]-pentafluoro-k6-sulfane;hydrochloride (Compound a24).
Figure imgf000521_0001
[002667] To a 25 mL microwave vial was added [4-(3-bromo-5-methyl-pyrazol-l-yl)phenyl]- pentafluoro-L’-sulfane (100 mg, 0.27 mmol), 4-(2 -piperazin- l-ylethyl)-l,4-thiazinane 1,1- dioxide;hydrochloride (1.5 equiv., 0.41 mmol), tetrahydrofuran (10 mL, 99.5 mass%), and potassium tert-butoxide (6 equiv., 1.65 mmol) at 20 °C under N2. Next, dichloro[l,3-bis(2,6-di- 3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (0.02 equiv., 0.005 mmol) was added and the reaction mixture was stirred at 50 °C for 20 h in a sealed vial. The mixture was filtered and concentrated to give a residue, which was purified by reverse HPLC chromatography using 10-100% ACN in water as eluent to give the product [4-[3-[4-[2-(l , 1 - di oxo- 1 ,4-thiazinan-4-yl)ethyl]piperazin- 1 -yl]-5-methyl-pyrazol- 1 -yl]phenyl]-pentafluoro-X6- sulfane;hydrochloride (25 mg, 0.041 mmol, 15% yield). M+l (530 m/z).
[002668] 'H NMR (400 MHz, Methanol-d4) 5 8.00 (d, J = 8.5 Hz, 2H), 7.76 (d, J = 8.7 Hz, 2H), 6.13 (s, 1H), 3.94 (d, J = 6.4 Hz, 4H), 3.89 - 3.45 (m, 16H), 2.40 (s, 3H) ppm.
Example S-bl. Synthesis of 4-[2-[[l-[l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4- piperidyl] oxy] ethyl] morpholine (Compound bl).
[002669] Compound bl was prepared as outlined below.
Figure imgf000521_0002
[002670] Step 1. Synthesis of 3-iodo-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000521_0003
[002671] To a solution of 3-iodo-lH-pyrazole (1.0 g, 5.16 mmol) in chloroform (40 mL) was added [3 -(trifluoromethoxy) phenyl] boronic acid (2.17 g, 10.3 mmol), anhydrous copper acetate (1.87 g, 10.3 mmol), pyridine (1.66 mL, 20.6 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10- 50% dichloromethane in petroleum ether) to afford the desired product 3-iodo-l-[4- (trifluoromethoxy) phenyl] pyrazole (1.2 g, yield 65%) as a yellow solid.
[002672JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 89% (254 nm), Mass: found peak 354.8 (M+l) at 2.090 min.
[002673] Step 2. Synthesis of 4-[2-[[l-[l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4- piperidyl] oxy] ethyl] morpholine (Compound bl).
Figure imgf000522_0001
[002674] To a solution of 3-iodo-l-[4-(trifluoromethoxy) phenyl] pyrazole (100 mg, 0.28 mmol) in dry 1,4-dioxane (3 mL) was added 4-[2-(4-piperidyloxy) ethyl] morpholine (90 mg, 0.42 mmol), Pd-PEPPSI-ipent (6 mg, 0.008 mmol), and sodium tert-butoxide (81 mg, 0.8 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 16 h. The mixture was filtered. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product 4-[2-[[l-[l-[4-(trifluoromethoxy) phenyl] pyrazol-3-yl]-4-piperidyl] oxy] ethyl] morpholine (16.9 mg, yield 13.6%) as a colorless oil.
[002675] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 92% (214 nm), Mass: found peak 194.0 (M+l) at 2.103 min.
[002676] 'H NMR (400 MHz, MeOD-d4) 5 8.05 (d, J = 2.8 Hz, 1H), 7.76 (d, J = 9.2 Hz, 2H), 7.34 (d, J = 8.4 Hz, 2H), 6.08 (d, J = 2.4 Hz, 1H), 3.74 - 3.63 (m, 8H), 3.54 (ddd, J = 12.4, 8.4, 3.6 Hz, 1H), 3.09 - 2.99 (m, 2H), 2.61 (dd, J = 16.8, 11.2 Hz, 6H), 2.01 (d, J = 8.8 Hz, 2H), 1.73 - 1.62 (m, 2H) ppm. Example S-b2. Synthesis of 4-[2-[4-[3-isopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-4- yl] piperazin-l-yl] ethyl] morpholine (Compound b2).
[002677] Compound b2 was prepared as outlined below.
Figure imgf000523_0001
[002678] Step 1. Synthesis of 4-bromo-3-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000523_0002
pyridine(4.0 eq) CHCI3, 30°C, o/n
[002679]Under argon atmosphere, a mixture of 4-bromo-3-isopropyl-lH-pyrazole (400 mg, 2.12 mmol), [4-(trifluoromethoxy) phenyl] boronic acid (871 mg, 4.23 mmol), copper acetate (769 mg, 4.23 mmol) and pyridine (669 mg, 8.46 mmol) in chloroform (10 mL) was stirred at RT for 16h. The reaction mixture was then concentrated and purified by SGC (PE:DCM = 2: 1) to afford the desired product 4-bromo-3-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (800 mg, 61.9%) as a colorless oil.
[002680JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2mL/min; Column: HALO Cl 8, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 349.1 (M+l) at 1.633 min.
[002681] Step 2. Synthesis of tert-butyl 4-[3-isopropyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine-l-carboxylate.
Pd-PEPPSI4pent(0 1 eq) Boc
Figure imgf000523_0003
[002682]Under argon atmosphere, a mixture of 3-iodo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (400 mg, 1.13 mmol), tert-butyl 2,2-dimethylpiperazine-l- carboxylate(420 mg, 2.25 mmol), [l,3-Bis(2,6-Diisopropylphenyl)imidazol-2-ylidene](3- chloropyridyl)palladium(II) dichloride (27 mg, 0.038 mmol) and sodium tert-butoxide (38.5 mg, 0.4 mmol) in anhydrous THF (2 mL) in a sealed tube was stirred at 60 °C for 16 h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate =1 : 1) to afford the product tert-butyl 4-[3-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazine-l -carboxylate (30 mg, 5.55%) as a yellow oil.
[002683JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 88% (214 nm),
Mass: found peak 455.3 (M+l) at 2.502 min.
[002684] Step 3. Synthesis of l-[3-isopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-4-yl] piperazine.
Figure imgf000524_0001
[002685] To a solution of tert-butyl 4-[3-isopropyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazine-l -carboxylate (40 mg, 0.088 mmol) in di chloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2 h. Then the reaction mixture was concentrated to remove solvent, and potassium carbonate was added to pH=10. The product was extracted with DCM and dried over sodium sulfate. The organic layer was concentrated to afford the desired product l-[3-isopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-4-yl] piperazine (30 mg, yield 96.2 %) as a yellow oil.
[002686] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 80% (214 nm), Mass: found peak 355.3 (M+l) at 1.105 mm.
[002687] Step 4. Synthesis of 4-[2-[4-[3-isopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol- 4-yl] piperazin-l-yl] ethyl] morpholine (Compound b2).
Figure imgf000525_0001
, ,
[002688] To a solution of 5-piperazin-l-yl-2-[4-(trifluoromethoxy)phenyl]pyrazole-3- carbonitrile (30 mg, 0.084 mmol), potassium carbonate (58 mg, 0.42 mmol) and KI (14 mg, 0.084 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (23 mg, 0.127 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water /acetonitrile) to afford 4-[2-[4-[3-isopropyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-4-yl] piperazin- 1-yl] ethyl] morpholine (6.6 mg, yield 19.6%) as a white solid.
[002689JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 94% (214 nm), Mass: found peak 468.2 (M+l) at 2.259 min.
[002690] 'H NMR (400 MHz, MeOD-d4) 5 7.96 (s, 1H), 7.85-7.74 (m, 2H), 7.36 (d, J = 8.8 Hz, 2H), 3.79-3.67 (m, 4H), 3.09 (dt, J = 13.6, 6.8 Hz, 1H), 2.98 (s, 4H), 2.77-2.50 (m, 12H), 1.35 (d, J = 6.8 Hz, 6H) ppm.
Example S-b3. Synthesis of 4-[2-[4-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound b3).
[002691] Compound b3 was prepared as outlined below.
Boc
Figure imgf000525_0002
[002692] Step 1. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl] pyrazol-4- yl] piperazine-l-carboxylate. Boc
Figure imgf000526_0001
tBuONa(3.0 eq) THF, 60°c, o/n
[002693] Under argon atmosphere, a mixture of 4-bromo-l-[4-(trifluorom ethoxy )phenyl] pyrazole (300 mg, 0.97 mmol), tert-butyl 2,2-dimethylpiperazine-l -carboxylate (364 mg, 1.95 mmol), [l,3-Bis(2,6-Diisopropylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) dichloride (27 mg, 0.038 mmol) and sodium tert-butoxide (282 mg, 2.93 mmol) in anhydrous dioxane (2 mL) in a sealed tube was stirred at 90 °C for 16h. Then the reaction mixture was concentrated and purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the product tertbutyl 4-[l-[4-(trifluoromethoxy)phenyl] pyrazol-4-yl]piperazine-l -carboxylate (80 mg, 20%) as a colorless oil.
[002694JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2mL/min; Column: HALO Cl 8, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (254 nm), Mass: found peak 357.2 (M-55) at 1.462 min.
[002695] Step 2. Synthesis of tert-butyl 4-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol- 4-y I] piperazine- 1-carboxylate.
Figure imgf000526_0002
[002696] Under argon atmosphere, a mixture of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl] pyrazol-4-yl]piperazine- 1-carboxylate (70 mg, 0.16 mmol), N-Chloro-succinimide (44 mg, 0.33 mmol) in anhydrous DMF (5 mL) was stirred at rt for 16h. Water (10 mL) was added and extracted with ethyl acetate (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over sodium sulfate, and concentrated in vacuo. The product was purified by SGC (petroleum etherethyl acetate =2: 1) to afford the desired product tert-butyl 4-[5-chl oro-1 - [4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine-l-carboxylate (50 mg, 67%).
[002697] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2mL/min; Column: HALO Cl 8, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 97% (254 nm), Mass: found peak 391.0 (M-55)+ at 1.533 min. [002698] Step 3. Synthesis of l-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperazine.
Figure imgf000527_0001
[002699] To a solution of tert-butyl 4-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazine-l -carboxylate (50 mg, 0.088 mmol) in di chloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the reaction mixture was concentrated to remove solvent, and potassium carbonate was added to pH=10. The product was extracted with DCM and dried over sodium sulfate. The solution was concentrated to afford the desired product l-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (35 mg, yield 90.2 %) as a yellow oil.
[002700JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 75% (214 nm), Mass: found peak 347.1 (M+l) at 1.556 min.
[002701] Step 4. Synthesis of 4-[2-[4-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound b3).
Figure imgf000527_0002
[002702] To a solution of l-[5-chloro-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (30 mg, 0.084 mmol), potassium carbonate (58 mg, 0.42 mmol) and KI (14 mg, 0.084 mmol) in 95% ethanol (5 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (23 mg, 0.127 mmol). The reaction was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate /water/acetonitrile) to afford 4-[2-[4-[5-chloro-l- [4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazin-l-yl]ethyl]morpholine (12.1 mg, yield 22.5%) as a yellow solid.
[002703JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 460.3 (M+l) at 1.041 min.
[002704] 'H NMR (400 MHz, MeOD-d4) 5 7.69 (s, 2H), 7.67-7.65 (m, 1H), 7.48 (d, J = 8.4 Hz, 2H), 3.76-3.69 (m, 4H), 3.16-3.10 (m, 4H), 2.73 (s, 4H), 2.67-2.60 (m, 4H), 2.58 (d, J = 10.8 Hz, 4H) ppm.
Example S-b4. Synthesis of 4-[2-[2-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b4).
[002705] Compound b4 was prepared as outlined below.
Figure imgf000528_0001
[002706] Step 1. Synthesis of 4-bromo-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000528_0002
. q pyridine(4.0 eq) CHCI3, 30°C, o/n
[002707] To a solution of 4-bromo-lH-pyrazole (1 g, 6.8 mmol) in chloroform (20 mL) was added [3-(trifluoromethoxy)phenyl]boronic acid (2.86 g, 13.6 mmol), anhydrous copper acetate (2.47 g, 13.6 mmol), pyridine (2.19 mL, 27.2 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 40g silica gel column @80mL/min, eluting with 10- 50% dichloromethane in petroleum ether) to afford the desired product 4-bromo-l-[4- (trifluoromethoxy)phenyl]pyrazole (1.3 g, yield 62%) as a white solid.
[002708JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95% B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 91% (214 nm), Mass: found peak 307.0 (M+l) at 2.214 min. [002709] Step 2. Synthesis of tert-butyl 6-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro [3.3] heptane-2-carboxylate.
Figure imgf000529_0001
dioxane, reflux, o/n
[002710] Under argon atmosphere, a mixture of 4-bromo-l-[4-(trifluoromethoxy)- phenyl]pyrazole (200 mg, 0.64 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (127 mg, 0.64 mmol), [l,3-Bis(2,6-Diisopropylphenyl)imidazol-2-ylidene](3- chloropyridyl)palladium(II) dichloride (43.7 mg, 0.064 mmol) and sodium tert-butoxide (185 mg, 1.92 mmol) in anhydrous THF (10 mL) in a sealed tube was stirred at 60 °C for 16h. Then concentrated and purified by SGC (petroleum ether: ethyl acetate = 1 : 1) to afford the desired product tert-butyl 6-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6-diazaspiro [3.3]heptane-2- carboxylate (180 mg, 66.2%).
[002711JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95% B within 1.3min, 95% B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 88% (214 nm) Mass: found peak 425.1 (M+l) at 1.964 min.
[002712] Step 3. Synthesis of 2-[l-[4-(trifluoromethoxy)phenyl] pyrazol-4-yl]-2,6- diazaspiro [3.3] heptane.
Boc
Figure imgf000529_0002
[002713] A solution of tert-butyl 6-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptane-2-carboxylate (185 mg, 0.28 mmol) in DCM (10 mL) was added TFA (3 mL). The mixture was stirred for 2 h at ambient temperature. The reaction was concentrated in vacuo and water (10 mL) was added. Then potassium carbonate was added until pH=10. It was extracted with DCM (2x20mL) and dried over sodium sulfate. The combined organic layers were concentrated to afford the desired product 2-[l-[4-(trifluoromethoxy)phenyl] pyrazol-4-yl]- 2,6-diazaspiro[3.3]heptane (140 mg, 99%) as a yellow solid.
[002714JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95% B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 90% (214 nm), Mass: found peak 325.0 (M+l) at 1.664 min.
[002715] Step 4. Synthesis of 4-[2-[2-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b4).
Figure imgf000530_0001
[002716] A solution of 2-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptane (140 mg, 0.43 mmol), 4-(2-chloroethyl)morpholine hydrochloride (120 mg, 0.65 mmol), potassium carbonate (179 mg, 1.3 mmol) and KI (71 mg, 0.43 mmol) in EtOH (10 mL) was stirred at 90 °C overnight. Then concentrated and purified by prep-HPLC (ammonium hydrogen carbonate) to afford the desired product 4-[2-[2-[l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (32.4 mg, 17.2%) as a white solid.
[002717] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 438.3 (M+l) at 1.000 min.
[002718] 'H NMR (400 MHz, MeOD-d4) 5 7.79 (d, J = 9.6 Hz, 3H), 7.38 (d, J = 10.2 Hz, 3H), 5.51 (s, 1H), 3.89 (s, 3H), 3.73-3.66 (m, 4H), 3.49 (s, 4H), 2.66 (d, J = 7.6 Hz, 2H), 2.49 (s, 4H), 2.41-2.36 (m, 2H) ppm.
Example S-b5. Synthesis of 4-[2-[3-[4-fluoro-5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] -3,8-diazabicyclo [3.2.1] octan-8-yl] ethyl] morpholine (Compound b5).
[002719] Compound b5 was prepared as outlined below.
Figure imgf000531_0001
[002720] Step 1. Synthesis of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Br
Figure imgf000531_0002
[002721] To a solution of 3-bromo-5-methyl-lH-pyrazole (1.55 g, 9.62 mmol) in chloroform (100 mL) was added [4-(trifluoromethyl)phenyl]boronic acid (3.96 g, 19.2 mmol), anhydrous copper acetate (3.49 g, 19.2 mmol), pyridine (3.87 mL, 48.1 mmol) and molecular sieves 4 A. The reaction mixture was stirred at 40 °C for 48h. The mixture was filtered. The filtrate was purified by flash chromatography (Biotage, 120 g silica gel column @100 mL/min, eluting with 0-38% dichloromethane in petroleum ether) to afford the desired product 3-bromo-5-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazole (0.73 g, yield 23.6%) as a colorless oil.
[002722JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2mL/min; Column: HALO C18 2.7 pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm) Mass: found peak 323.0 (M+l)+ at 1.422 min.
[002723] 'H NMR (400 MHz, CDCh) 5 7.50-7.45 (m, 2H), 7.32 (d, J = 8.4 Hz, 2H), 6.24 (s, 1H), 2.34 (s, 3H) ppm.
[002724] Step 2. Synthesis of 3-bromo-4-fluoro-5-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000532_0001
[002725] To a solution of 3-bromo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (500 mg, 1.56 mmol) in acetonitrile (20 mL) was added N-Fluoro-N'-chloromethyltriethylenediamine (827 mg, 2.34 mmol). The mixture was stirred at room temperature for 64h. The reaction was filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @100 mL/min, eluting with 0-10% ethyl acetate in petroleum ether) to afford the crude product, which was further purified by prep- HPLC (ammonium hydrogen carbonate /water/acetonitrile) to afford the desired product 3- bromo-4-fluoro-5-methyl-l-[4-(trifluoromethoxy) phenyl ]pyrazole (160 mg, yield 30.1%) as a yellow oil.
[002726] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO C18 2.7 pm 4.6*30 mm; Column Temperature: 40 °C; LC purity: 99.37% (214 nm) Mass: found peak 341.1 (M+l)+ at 1.478 min.
[002727] 1 H NMR (400 MHz, CDCI3) 5 7.49-7.44 (m, 2H), 7.36-7.31 (m, 2H), 2.34 (d, J = 1.6 Hz, 3H) ppm.
[002728] Step 3. Synthesis of tert-butyl 3-[4-fluoro-5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] -3,8-diazabicyclo [3.2.1] octane-8-carboxylate.
Figure imgf000532_0002
[002729] A mixture of 3-bromo-4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (140 mg, 0.413 mmol), tert-butyl 3,8-diazabicyclo[3.2.1]octane-8-carboxylate (131 mg, 0.619 mmol), PEPPSI-IPent (10 mg, 0.012 mmol) and sodium tert-butoxide (119 mg, 1.24 mmol) in 1,4- di oxane (10 mL) was stirred at 100 °C for 16h under argon atmosphere. The reaction mixture was directly purified by flash chromatography (Biotage, 40 g silica gel column @75 mL/min, eluting with 0-20% ethyl acetate in petroleum ether) to afford the desired product tert-butyl 3-[4- fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,8-diazabicyclo[3.2.1]octane-8- carboxylate (110 mg, yield 50.4%) as a yellow oil.
[002730JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO Cl 8 2.7 pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 88.94% (214 nm) Mass: found peak 471.1 (M+l) at 2.439 min.
[002731] Step 4. Synthesis of 3-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol- 3-yl]-3,8-diazabicyclo [3.2.1] octane.
Figure imgf000533_0001
[002732] To a solution of tert-butyl 3-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (110 mg, 0.234 mmol) in dichloromethane (10 mL) was added TFA (2 mL, 26.9 mmol). The reaction was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH 9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 3-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl] pyrazol-3-yl]- 3,8-diazabicyclo[3.2.1]octane (86 mg, yield 93%) as a yellow oil.
[002733JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2 min at 2.2 mL/min; Column: HALO Cl 8 2.7 pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 93.59% (214 nm), Mass: found peak 371.3 (M+l) at 1.099 min.
[002734] Step 5. Synthesis of 4-[2-[3-[4-fluoro-5-methyl-l-[4-
(tr ifluoromethoxy)phenyl] pyrazol-3-yl] -3,8-diazabicyclo [3.2.1] octan-8-yl] ethyl] morpholine (Compound b5).
Figure imgf000534_0001
[002735] To a solution of 3-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]- 3,8-diazabicyclo[3.2.1]octane (86 mg, 0.232 mmol), potassium carbonate (128 mg, 0.929 mmol) and KI (38.5 mg, 0.232 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (64.8 mg, 0.348 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[3-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-3,8- diazabicyclo[3.2.1]octan-8-yl]ethyl] morpholine (41.9 mg, yield 36.9%) as a yellow solid.
[002736] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95% B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 99.74% (214 nm) Mass: found peak 484.2 (M+l) at 2.202 min.
[002737] 'H NMR (400 MHz, CD3OD) 5 7.60-7.54 (m, 2H), 7.40 (d, J = 8.4 Hz, 2H), 3.71 (t, J = 4.4 Hz, 4H), 3.48-3.37 (m, 4H), 3.11 (d, J = 10.8 Hz, 2H), 2.67-2.57 (m, 4H), 2.55 (t, J = 4.4 Hz, 4H), 2.29 (d, J = 1.6 Hz, 3H), 2.10-2.02 (m, 2H), 1.92-1.84 (m, 2H) ppm.
Example S-b6. Synthesis of 7-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-9-(2-morpholinoethyl)-3-oxa-7,9-diazabicyclo [3.3.1]nonane (Compound b6).
[002738] Compound b6 was prepared as outlined below.
Figure imgf000534_0002
[002739] A mixture of 3-bromo-4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (100 mg, 0.295 mmol), 9-(2-morpholinoethyl)-3-oxa-7,9-diazabicyclo[3.3.1] nonane (107 mg, 0.442 mmol), PEPPSI-IPent (7 mg, 0.0088 mmol) and sodium tert-butoxide (85 mg, 0.885 mmol) in 1,4-di oxane (10 mL) was stirred at 100 °C for 16h under argon atmosphere. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 7-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]-9-(2- morpholinoethyl)-3-oxa-7,9-diazabicyclo [3.3.1]nonane (25.9 mg, yield 17.2%) as a yellow solid.
[002740JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95% B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 98.07% (214 nm) Mass: found peak 500.2 (M+l) at 2.026 min.
[002741] 'H NMR (400 MHz, CD3OD) 5 7.60-7.54 (m, 2H), 7.40 (d, J = 8.4 Hz, 2H), 4.07 (d, J = 11.2 Hz, 2H), 3.89 (d, J = 11.2 Hz, 2H), 3.71 (t, J = 4.4 Hz, 4H), 3.65 (d, J = 12.4 Hz, 2H), 3.49 (d, J = 12.4 Hz, 2H), 3.03 (t, J = 7.2 Hz, 2H), 2.86 (s, 2H), 2.65-2.50 (m, 6H), 2.30 (d, J = 2.0 Hz, 3H) ppm.
Example S-b7. Synthesis of 4-[2-[4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l- yl] ethyl] morpholine (Compound b7).
[002742] Compound b7 was prepared as outlined below.
Figure imgf000535_0001
[002743] Step 1. Synthesis of 3-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000535_0002
pyridine(4.0 eq)
DCM, rt, 16h
[002744] To a solution of 3-iodo-lH-pyrazole (1.0 g, 5.16 mmol) in chloroform (40 mL) were added [3-(trifluoromethyl)phenyl]boronic acid (2.0 g, 10.3 mmol), anhydrous copper acetate (1.87 g, 10.3 mmol), pyridine (1.66 mL, 20.6 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24h. The mixture was filtered, concentrated in vacuo and the residue was purified by flash chromatography (Biotage, 40g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product 3-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole (1.2 g, yield 65%) as a yellow solid. [002745] LCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2 mL/min; Column: HALO C18, 2.7 pm, 4.6*30 mm; Column Temperature: 40 °C; LC purity: 70% (214 nm) Mass: found peak 339.0(M+l) at 1.427 min.
[002746] Step 2. Synthesis of tert-butyl 4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine-l-carboxylate.
Figure imgf000536_0001
[002747] To a solution of 3-iodo-l-[4-(trifluoromethyl)phenyl]pyrazole (200 mg, 0.3 mmol) in dry 1,4-dioxane (8 mL) were added tert-butyl piperazine-l-carboxylate (220 mg, 1.2 mmol), tBuXPhos Pd G3 (50 mg, 0.06 mmol), and sodium tert-butoxide (171 mg, 1.77 mmol) in a microwave tube. The reaction mixture was stirred at 100 °C for 24h. The mixture was filtered. The filtrate was purified by SGC (petroleum ether: ethyl acetate=l : 1) to afford the desired product tert-butyl 4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l-carboxylate (200 mg, yield 85.3%) as a colorless oil.
[002748JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 87% (214 nm) Mass: found peak 341.1 (M-55)+ at 1.507 min.
[002749] Step 3. Synthesis of l-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine.
Figure imgf000536_0002
[002750] To a solution of tert-butyl 4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine-l- carboxylate (70 mg, 0.17 mmol) in dichloromethane (6 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. Then the mixture was concentrated to remove solvent, treated with potassium carbonate aqueous solution to neutralized to pH=10. The resulting mixture was extracted with DCM (3* 10 mL) and the combined DCM layers were dried over sodium sulfate, filtered and concentrated to afford the desired product l-[l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (40 mg, yield 76.5 %) as a yellow oil. [002751JLCMS method: Mobile Phase: A: Water (0.01% TFA) B: Acetonitrile (0.01% TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2 mL/min; Column: HALO Cl 8, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 95% (254 nm) Mass: found peak 297.2 (M+l) at 0.983 min.
[002752] Step 4. Synthesis of 4-[2-[4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin- l-yl]ethyl] morpholine (Compound b7).
Figure imgf000537_0001
[002753] To a suspension of l-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (40 mg, 0.13 mmol), potassium carbonate (93 mg, 0.67 mmol) and KI (22.4 mg, 0.13 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine (37 mg, 0.2 mmol) in one portion. The reaction mixture was stirred at 95 °C for 16h. The reaction mixture was cooled to room temperature, filtered, the filtrate was concentrated in vacuo. The residue was purified by prep- HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4- [l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l-yl]ethyl] morpholine (18.4 mg, yield 33%) as a white solid.
[002754JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95% B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 96% (214 nm), Mass: found peak 410.1 (M+l) at 2.009 min.
[002755] 'H NMR (400 MHz, Methanol-d4) 5 8.14 (d, J = 2.8 Hz, 1H), 7.84 (d, J = 8.8 Hz, 2H), 7.69 (d, J = 8.8 Hz, 2H), 6.11 (d, J = 2.8 Hz, 1H), 3.69 (t, J = 4.8 Hz, 4H), 3.38-3.31 (m, 4H), 2.72 - 2.46 (m, 12H) ppm.
Example S-b8. Synthesis of 4-[2-[4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin-l- yl] ethyl] -1,4-thiazinane 1,1-dioxide (Compound b8).
[002756] Compound b8 was prepared as outlined below.
Figure imgf000538_0001
[002757] Step 1. Synthesis of l-(2-chloroethyl)-4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl] piperazine.
Figure imgf000538_0002
[002758] A mixture of l-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (100 mg, 0.34 mmol), 2-chloroacetaldehyde (40% aqueous, 100 mg, 0.5 mmol), sodium cyanoborohydride (42 mg, 0.67 mmol) and acetic acid (10 mg, 0.17 mmol) in methanol (8 mL) was stirred at room temperature for 16h. The reaction mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25g silica gel column @70mL/min, eluting with 0-10% methanol in dichloromethane) to afford the desired product l-(2-chloroethyl)-4-[l-[4- (trifluoromethyl)phenyl]pyrazol-3-yl]piperazine (40 mg, yield 6.6%) as a yellow oil. [002759JLCMS method: Mobile Phase: A: water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 17% (214 nm), Mass: found peak 359.1 (M+l) at 1.515 min.
[002760] Step 2. Synthesis of 4-[2-[4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3-yl]piperazin- l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound b8).
Figure imgf000538_0003
[002761] A mixture of l-(2-chloroethyl)-4-[l-[4-(trifluoromethyl)phenyl]pyrazol-3- yl]piperazine (40 mg, 0.03 mmol), 1,4-thiazinane 1,1-dioxide (9 mg, 0.06 mmol) and DIPEA (21 mg, 0.16 mmol) in NMP (3 mL) was heated to 160 °C in Biotage microwave reactor and stirred for 2h. The reaction mixture was cooled to room temperature and directly purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2- [4- [ 1 -[4-(trifl uoromethyl )phenyl ]pyrazol-3 -yl]piperazin- 1 -y 1 ] ethyl ] - 1 ,4-thiazinane 1,1- dioxide (7.1 mg, yield 46%) as a yellow solid.
[002762JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 458.3 (M+l) at 1.021 min.
[002763] 'H NMR (400 MHz, CDCh) 5 7.82 (d, J = 2.8 Hz, 1H), 7.70 (d, J = 8.8 Hz, 2H), 7.64 (d, J = 8.8 Hz, 2H), 5.97 (d, J = 2.8 Hz, 1H), 3.33 (t, J = 4.8 Hz 4H), 3.07 (s, 8H), 2.73 (t, J = 6.8 Hz, 2H), 2.62 (t, J = 4.8 Hz, 4H), 2.56(t, J = 6.4 Hz, 2H) ppm.
Example S-b9. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-l- piperidyljethyljmorpholine (Compound b9).
[002764] Compound b9 was prepared as outlined below.
Figure imgf000539_0001
[002765] Step 1. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6- dihydro-2H-pyridine-l-carboxylate.
Figure imgf000539_0002
Pd(dppf)CI2 (0.1eq) dioxane/H20, 80°C, 16h
[002766] Under argon atmosphere, a mixture of 4-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, 0.847 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydro- 2H-pyridine-l -carboxylate (393 mg, 1.27 mmol), [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (62 mg, 0.0847 mmol) and potassium carbonate (234 mg, 1.69 mmol) was stirred in 1,4-dioxane / water (15mL / 6 mL) at 80 °C for 2h. The mixture was filtered, and the filtrate was washed with water (10 mL) and extracted by EtOAc (20 mL X 3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum etherethyl acetate=4/l) to give tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6- dihydro-2H-pyridine-l -carboxylate (339 mg, 97.7% yield) as a white solid.
[002767] LCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column:
Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 94% (214 nm); Mass: found peak 410.2 (M + H) at 1.50 min.
[002768] Step 2. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperidine-l-carboxylate.
Figure imgf000540_0001
[002769] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6-dihydro- 2H-pyridine-l -carboxylate (339 mg, 0.828 mmol) in MeOH (10 mL) was added Pd/C (10%, 88.1 mg) under N2. The mixture was degassed with H2 and stirred under H2 balloon atmosphere at RT for 0.5h. The mixture was filtered to remove Pd/C. The filtrate was concentrated in vacuo to give tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine-l -carboxylate (0.309g, crude) as a colorless oil. The crude product was used directly in the next step without purification.
[002770JLCMS method: Mobile Phase : A:water (0.01%TFA) B ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 97% (214 nm); Mass: found peak 412.3 (M + H) at 1.49 min.
[002771] Step 3. Synthesis of 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine.
Figure imgf000540_0002
[002772] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine-l- carboxylate (309 mg, 0.75 mmol) was added HCI (3mol/L, 1.25 mL). The mixture was stirred at rt for Ih. The mixture was concentrated to dryness to give 4-[l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine (230 mg, crude) as a white solid. The crude product was used directly in the next step without purification.
[002773JLCMS method: Mobile Phase : A: water(0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99% (214 nm); Mass: found peak 312.3 (M + H) at 0.99 min. [002774] Step 4. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-l- piperidyl]ethyl]morpholine (Compound b9).
Figure imgf000541_0001
[002775] To a solution of 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine (258 mg, 0.829 mmol), potassium carbonate (573 mg, 4.14 mmol) and KI (138 mg, 0.829 mmol) in 95% ethanol / water (10 mL /0.5 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (231 mg, 1.24 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (192.3mg, yield 54.7%) as a yellow solid.
[002776] LCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.5min, 95%B for 1.5min, back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column : Sunfire C18, 4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 424.9 (M + H) at 1.85 min. [002777] XH NMR (400MHz, CDC13): 5 7.67-7.71 (m, 3H), 7.61 (s, 1H), 7.32 (s, 1H), 7.29 (s, 1H), 3.74 (t, J =4.8Hz, 4H), 3.05 (d, J =11.6Hz, 2H), 2.55-2.61 (m, 5H), 2.52 (d, J =4.4Hz, 4H), 2.14 (d, J =10.8 Hz, 2H), 1.97 (d, J =12.4Hz, 2H), 1.71-1.78 (m, 2H) ppm.
Example S-blO. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound blO).
[002778] Compound blO was prepared as outlined below.
Figure imgf000541_0002
[002779] Step 1. Synthesis of 4-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole. F3CO B(OH)2
Figure imgf000542_0001
Cu(OAc)2 (2.0eq) dry DCM, RT, 16h
[002780] To a solution of 4-iodo-lH-pyrazole (0.97 g, 5.0 mmol) in di chloromethane (50 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (2.06 g, 10 mmol), anhydrous copper acetate (1.82 g, 10 mmol), pyridine (1.98 mg, 25 mmol) and molecular sieves 4 A. The reaction mixture was stirred at rt for 16h. The mixture was filtered and concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: ethyl acetate=20/l) to afford the desired product 4-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole (1.58 g, yield 89.1%) as a white solid.
[002781JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99 % (214 nm); Mass: found peak 355.0 (M + H) at 1.43 min.
[002782] 'H NMR (400MHz, CDC13): 5 7.96 (d, J =0.4Hz, 1H), 7.75 (s, 1H), 7.70 (dd, J =6.8Hz, 2Hz, 2H), 7.34 (dd, J =8.8Hz, 0.8Hz, 2H) ppm.
[002783] Step 2. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperazine-l-carboxylate.
Figure imgf000542_0002
L-(-)-Proline (0.2eq)
Cui (0.1 eq)
DMSO, 100°C
[002784] Under argon atmosphere, a mixture of 4-iodo-l-[4-(trifluoromethoxy)phenyl]pyrazole (250 mg, 0.706 mmol), tert-butyl piperazine-l-carboxylate (197 mg, 1.06 mmol), L(-)-Proline (16.3 mg, 0.141 mmol), potassium carbonate (195 mg, 1.41 mmol) and cuprous iodide (13.4 mg, 0.0706 mmol) was stirred in DMSO (2 mL) at 100 °C for 3h. The mixture was filtered, and the filtrate was washed with water (20 mL) and extracted by EtOAc (20 mLx3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum ether: ethyl acetate=4/l) to give tertbutyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine-l -carboxylate (128 mg, 44% yield) as a yellow solid.
[002785] LCMS method: Mobile Phase : A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99%
(214 nm); Mass: found peak 413.3 (M + H) at 1.43 min.
[002786] 'H NMR (400 MHz, Chloroform): 5 7.65 (dd, J =6.8Hz, 2.0Hz, 2H), 7.47 (s, 1H), 7.41
(s, 1H), 7.29 (s, 2H), 3.59 (t, J =5.2Hz, 4H), 2.96 (t, J =5.2Hz, 4H), 1.48 (s, 9H) ppm.
[002787] Step 3. Synthesis of l-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine.
Figure imgf000543_0001
[002788] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine-l- carboxylate (128 mg, 0.31 mmol) was added HC1 (3mol/L, 0.517 mL). The mixture was stirred at rt for Ih. The mixture was concentrated to dryness to give 1 -[ 1 -[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (95 mg, crude) as a white solid. The crude product was used directly in the next step without purification.
[002789JLCMS method: Mobile Phase : A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min,95%B for 1.2min. Flow Rate :2.2 mL/min; Column:
Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99% (214 nm); Mass: found peak 313.3 (M + H) at 0.98 min.
[002790] Step 4. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound blO).
Figure imgf000543_0002
,
[002791] To a solution of l-[l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (97 mg, 0.311 mmol), potassium carbonate (215 mg, 1.55 mmol) and KI (51.6 mg, 0.311 mmol) in 95% ethanol / water (10 mL /0.5 mL) was added 4-(2-chloroethyl)morpholine (69.7 mg, 0.466 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazin-l-yl]ethyl]morpholine (93.4 mg, yield 70.7%) as a white solid.
[002792JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%Bincrease to 95%B within 1.5min, 95%B for 1.5min,back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column : Sunfire C18 ,4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 425.9 (M + H) at 1.68 min. [002793] 'H NMR (400 MHz, CDCI3): 5 7.67 (dd, J =6.8Hz, 2.0Hz, 2H), 7.48 (d, J =0.4Hz, 1H), 7.41 (d, J =0.8Hz, 1H), 7.30-7.31 (m, 2H), 3.74 (t, J =4.4Hz, 4H), 3.06 (t, J =5.2Hz, 4H), 2.69 (t, J =4.8Hz, 4H), 2.53-2.63 (m, 8H) ppm.
Example S-bll. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (Compound bll).
[002794] Compound bl 1 was prepared as outlined below.
Figure imgf000544_0001
[002795] Step 1. Synthesis of give 4-iodo-3-(trifluoromethyl)-lH-pyrazole.
Figure imgf000544_0002
[002796] To a solution of 3-(trifluoromethyl)-lH-pyrazole (200 mg, 1.47 mmol) in 2 mL 50% H2SO4 was added NIS (397 mg, 1.76mmol). The reaction was stirred at 0 °C for 10 min and rt for 3h. The mixture was quenched by adding water, then extracted by EA (10 mLX3). The combined organic layer was washed by water (10 mL*3) and sodium carbonate (10 mLX3). The organic layer was then dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum ether: ethyl acetate=10/l) to give 4-iodo-3-(trifluoromethyl)-lH-pyrazole (0.348 g, 90.4% yield) as a white solid.
[002797] LCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3 min, 95%B for 1.2 min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 91 % (214 nm); Mass: found peak 263.0 (M + H) at 1.10 min.
[002798] 'H NMR (400 MHz, CDCI3): 5 12.08 (t, J =2.8Hz, 1H), 1.77 (s, 1H) ppm.
[002799] Step 2. Synthesis of 4-iodo-l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazole. F3CO B(OH)2
Figure imgf000545_0001
y ne ( eq) DCM, RT, 48h
[002800] To a solution of 4-iodo-3-(trifluoromethyl)-lH-pyrazole (0.2 g, 0.763 mmol) in dichloromethane (10 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (0.314 g, 1.53 mmol), anhydrous copper acetate (0.277 g, 1.53 mmol), pyridine (302 mg, 3.82 mmol) and molecular sieves 4 A. The reaction mixture was stirred at rt for 16h. The mixture was filtered. The filtrate was purified by silica column chromatography (petroleum ether: ethyl acetate=10/l) to afford the desired product 4-iodo-l-[4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazole (299 mg, yield 92.8%) as a colorless oil.
[002801JLCMS method: Mobile Phase : A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99% (214 nm); Mass: found peak 423.0 (M + H) at 1.50 min.
[002802] 'H NMR (400 MHz, CDC13): 5 8.03 (s, 1H), 7.73 (dd, J =6.8Hz, 2.0Hz, 2H), 7.37 (d, J =8.4Hz, 2H) ppm.
[002803] Step 3. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000545_0002
[002804]Under argon atmosphere, a mixture of 4-iodo-l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazole (299 mg, 0.708 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l -carboxylate (329 mg, 1.06 mmol), [1,1' bis(diphenylphosphino)ferrocene]dichloropalladium(II) (51.8 mg, 0.0708 mmol) and potassium carbonate (196 mg, 1.42 mmol) was stirred in 1,4-dioxane / water (5mL / 2.5 mL) at 80 °C for 2h. The mixture was filtered, the filtrate was washed with water (10 mL) and extracted by EtOAC (20 mLX3). The combined organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica column chromatography (petroleum etherethyl acetate=10/l) to give tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]-3,6-dihydro-2H-pyridine-l-carboxylate (335 mg, 99% yield) as a colorless oil.
[002805] LCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA)
Gradient:5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate :2.2 mL/min;
Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40C; LC purity: 87% (214 nm); Mass: found peak 478.4 (M + H) at 1.59 min.
[002806] Step 4. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-3-
(trifluoromethyl)pyrazol-4-yl]piperidine-l-carboxylate.
Boc
Figure imgf000546_0001
[002807] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]-3,6-dihydro-2H-pyridine-l-carboxylate (342 mg, 0.716 mmol) in MeOH (5 mL) were added Pd/C (10%, 76.2 mg) under N2. The mixture was degassed with H2 and stirred under H2 balloon atmosphere at RT for 0.5h. The mixture was filtered to remove Pd/C. The filtrate was concentrated in vacuo to give tert-butyl 4-[l-[4- (trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazol-4-yl]piperidine-l-carboxylate (0.350 g, crude) as a colorless oil. The crude product was used directly in the next step without purification.
[002808JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; ColumnTemperature: 40 °C; LC purity: 96% (214 nm); Mass: found peak 480.3 (M + H) at 1.60 min.
[002809] Step 5. Synthesis of 4-[l-[4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazol- 4-yl]piperidine.
Figure imgf000546_0002
[002810] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]piperidine-l -carboxylate (343 mg, 0.715 mmol) was added HC1 (3 mol/L, 1.19 mL). The mixture was stirred at rt for Ih. The mixture was concentrated to dryness to give 4-[l-[4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazol-4-yl]piperidine (280 mg, crude) as a white solid. The crude product was used directly in the next step without purification. [002811JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA) Gradient:5%B increase to 95%B within 1.3min, 95%B for 1.2min. Flow Rate: 2.2 mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 99% (214 nm); Mass: found peak 380.2 (M + H) at 1.11 min.
[002812] Step 6. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-3- (trifluoromethyl)pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (Compound bll).
Figure imgf000547_0001
[002813] To a solution of 4-[l-[4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazol-4- yl]piperidine (268 mg, 0.707 mmol), potassium carbonate (488 mg, 3.53 mmol) and KI (117 mg, 0.707 mmol) in 95% ethanol / water (10 mL /0.5 mL) was added 4-(2-chloroethyl)morpholine (159 mg, 1.06 mmol). The reaction was stirred at 90 °C for 16h. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica column chromatography (DCM:MeOH=10/l) to afford the desired product 4-[2-[4-[ 1 - [4-(trifluoromethoxy)phenyl]-3-(trifluoromethyl)pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (238 mg, yield 68.3%) as a white solid.
[002814JLCMS Method: Mobile Phase: A: water (10 mMammonium hydrogen carbonate) B: Acetonitrile; Gradient: 5%B increase to 95%B within 1.5min, 95%B for 1.5min, back to 5%B within O.Olmin. Flow Rate: 1.8mL/min; Column: Sunfire C18, 4.6*50mm, 3.5pm; Oven Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 492.8 (M + H) at 2.08 min. [002815] 'H NMR (400 MHz, CDCh): 5 8.42-8.47 (m, 1H), 7.97 (d, J =8.8Hz, 2H), 7.48 (d, J =8.4Hz, 2H), 3.76 (t, J =4.4Hz, 4H), 3.67 (d, J =11.6Hz, 2H), 3.27 (s, 2H), 3.04-3.15 (m, 3H), 2.81 (t, J =6.0Hz, 2H), 2.63 (s, 4H), 2.21 (d, J =13.6Hz, 2H), 1.98-2.11 (m, 2H) ppm.
Example S-bl2. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound bl2).
[002816] Compound bl2 was prepared as outlined below.
Figure imgf000548_0001
[002817] Step 1. Synthesis of 4-bromo-3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000548_0002
chloroform, 30°c, 16h
Figure imgf000548_0003
[002818] To a solution of 4-bromo-5-methyl-lH-pyrazole (2 g, 12.4 mmol) in chloroform (150 mL) was added [4-(trifhioromethyl)phenyl]boronic acid (5.12 g, 24.8 mmol), anhydrous copper acetate (4.51 g, 24.8 mmol), pyridine (5 mL, 62.1 mmol) and molecular sieves 4A. The reaction mixture was stirred at 30 °C for 48h. The mixture was filtered, concentrated in vacuo, and the residue was purified by flash chromatography (PE/DCM=1/1) to afford the desired product 4- bromo-3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (2.49 g, yield 62.4%) as a colorless oil. [002819JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100% (214 nm); Mass: found peak 321.1(M+H) at 1.510 min.
[002820] 'H NMR (400 MHz, MeOD-d4) 5 8.76 (s, 1H), 7.91 (d, J = 8.0 Hz, 2H), 7.51 (d, J = 4.0 Hz, 3H), 2.25 (s, 3H) ppm.
[002821] Step 2. Synthesis of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 4-y I] piperazine- 1-carboxylate.
Figure imgf000548_0004
[002822]Under argon atmosphere, a mixture of 4-bromo-3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (200 mg, 0.623 mmol), tert-butyl piperazine- 1 -carboxylate (174 mg, 0.934 mmol), tBuXPhos Pd G3 (49 mg, 0.062 mmol) and sodium tert-butoxide (180 mg, 1.87 mmol) in 1,4-di oxane (15 mL) was stirred at 100 °C for 16h. The residue was purified by flash chromatography (PE/EA= 2/1) to give tert-butyl 4-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine-l -carboxylate (26 mg, yield 9%) as a colorless oil.
[002823JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 427.3(M+H) at 1.516 min.
[002824] Step 3. Synthesis of l-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperazine.
Figure imgf000549_0001
[002825] A solution of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazine-l -carboxylate (26 mg, 0.060 mmol) and TFA (69 mg, 0.610 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (19 mg, yield 93.6%) as a yellow oil.
[002826] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 2.0mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 50 °C; LC purity: 98.4% (214 nm); Mass: found peak 327.0(M+H) at 1.54 min.
[002827] Step 4. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound b!2).
Figure imgf000549_0002
[002828] A solution of l-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (19 mg, 0.058 mmol), l,4-oxazepane;hydrochloride (16 mg, 0.087 mmol), potassium carbonate (24 mg, 0.175 mmol) and KI (9 mg, 0.058 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h. Then the mixture was concentrated in vacuo. The residue was purified by prep-HPLC (TFA/water/acetonitrile) to afford the desired product 4-[2-[4-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazin-l-yl]ethyl]morpholine (16.1 mg, yield 62.9%) as a yellow oil.
[002829JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA); Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 440.3 (M+H) at 1.03 min.
[002830] 'H NMR (400 MHz, MeOD-d4) 5 7.93 (s, 1H), 7.77 (t, J = 8.0 Hz, 2H), 7.37 (d, J = 8.0 Hz, 2H), 3.73 (t, J = 8.0 Hz, 4H), 3.01 (t, J = 4.0 Hz„ 4H), 2.75 (s, 4H), 2.65 (td, J = 8.0, 4.0 Hz, 4H), 2.57 (t, J = 4.0 Hz, 4H), 2.31 (s, 3H) ppm.
Example S-bl3. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound b!3).
[002831] Compound bl3 was prepared as outlined below.
Figure imgf000550_0001
[002832] Step 1. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl] morpholine.
Figure imgf000550_0002
dioxane 100°C 16h
[002833] Under argon atmosphere, a mixture of 4-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (400 mg, 1.25 mmol), 4-(2-piperazin-l-ylethyl)morpholine (372 mg, 1.87 mmol), tBuXPhos Pd G3 (99 mg, 0.125 mmol) and sodium tert-butoxide (359 mg, 3.74 mmol) in 1,4-di oxane (10 mL) was stirred at 100 °C for 16h. The residue was purified by flash chromatography (PE/EA= 2/1) to give 4-[2-[4-[5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazin-l-yl]ethyl] morpholine (150 mg, 7% yield) as a colorless oil.
[002834JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 29.9% (214 nm); Mass: found peak 427.3(M+H) at 1.453 min.
[002835] Step 2. Synthesis of l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperazine.
Figure imgf000551_0001
[002836] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazine-l -carboxylate (150 mg, 0.352 mmol) and TFA (401 mg, 3.52 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product l-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (100 mg, yield 93.6%) as a yellow oil.
[002837] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 28.8% (214 nm); Mass: found peak 327.2(M+H) at 1.011 min.
[002838] Step 3. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperazin-l-yl]ethyl]morpholine (Compound b!3).
Figure imgf000551_0002
[002839] A solution of l-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperazine (19 mg, 0.058 mmol), l,4-oxazepane;hydrochloride (16 mg, 0.087 mmol), potassium carbonate (24 mg, 0.175 mmol) and KI (9 mg, 0.058 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h, then the mixture was concentrated in vacuo. The residue was purified by prep-HPLC (TFA/water/acetonitrile) to afford the desired product 4-[2-[4-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperazin-l-yl]ethyl]morpholine (16.1 mg, yield 62.9%) as a yellow oil.
[002840JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 100.0% (214 nm); Mass: found peak 440.3 (M+H) at 1.009 min.
[002841] 'H NMR (400 MHz, MeOD-d4) 5 7.60 (d, J = 8.0 Hz, 2H), 7.58 (s, 1H), 7.47 (d, J = 8.0 Hz, 2H), 3.73 (t, J = 8.0 Hz, 4H), 3.01 (t, J = 12.0 Hz, 4H), 2.73 (d, J = 20.0 Hz, 4H), 2.67- 2.59 (m, 4H), 2.56 (t, J = 8.0 Hz, 4H), 2.29 (s, 3H) ppm.
Example S-bl4. Synthesis of 4-[2-[2-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b!4).
[002842] Compound b!4 was prepared as outlined below.
Figure imgf000552_0001
[002843] Step 1. Synthesis of tert-butyl 6-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
4-yl]-2,6-diazaspiro [3.3] heptane-2-carboxylate.
Boo
Figure imgf000552_0002
,
[002844]Under argon atmosphere, a mixture of 4-bromo-3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (200 mg, 0.623 mmol), tert-butyl 2,6-diazaspiro[3.3]heptane- 2-carboxylate (185 mg, 0.934 mmol), tBuXPhos Pd G3 (49 mg, 0.062 mmol) and sodium tert- butoxide (180 mg, 0.365 mmol) in 1,4-dioxane (15 mL) was stirred 100 °C for 16h. The reaction mixture was directly purified by flash chromatography to afford the desired product tert-butyl 6- [3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6-diazaspiro[3.3]heptane-2- carboxylate (180 mg, yield 58%) as a yellow oil.
[002845] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA;
Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 89.8% (214 nm); Mass: found peak 439.3 (M+H) at 1.407 min.
[002846] Step 2. Synthesis of 2-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro [3.3] heptanes.
Boc
Figure imgf000553_0001
[002847] A solution oftert-butyl 6-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptane-2-carboxylate (180 mg, 0.411 mmol) and TFA (468 mg, 4.11 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]- 2,6-diazaspiro[3.3]heptanes (120 mg, yield 66.5%) as a yellow oil.
[002848JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 77.0% (214 nm); Mass: found peak 339.3(M+H) at 1.03 min.
[002849] Step 3. Synthesis of 4-[2-[2-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b!4).
Figure imgf000553_0002
[002850] A solution of 2-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptanes (120 mg, 0.355 mmol), 1,4-oxazepane-hydrochloride (99 mg, 0.532 mmol), potassium carbonate (147 mg, 1.06 mmol), and KI (58 mg, 0.355 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h, then concentrated in vacuo. The residue was purified by prep- HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[2- [3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6-diazaspiro[3.3]heptan-6- yl]ethyl]morpholine (91.8 mg, yield 52.2%) as a colorless oil.
[002851JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.5 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 91.5% (214 nm); Mass: found peak 452.2(M+H) at 1.938 min.
[002852] 'H NMR (400 MHz, MeOD-d4) 5 7.73 (dd, J = 8.0, 4.0 Hz, 3H), 7.36 (d, J = 8.0 Hz, 2H), 3.90 (s, 4H), 3.71 (t, J = 8.0 Hz, 4H), 3.49 (s, 4H), 2.73 (d, J = 16.0 Hz, 2H), 2.49 (s, 4H), 2.39 (t, J = 12.0 Hz, 2H), 2.24 (s, 3H) ppm.
Example S-bl5. Synthesis of 4-[2-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b!5).
[002853] Compound b!5 was prepared as outlined below.
Figure imgf000554_0001
[002854] Step 1. Synthesis of tert-butyl 6-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
4-yl]-2,6-diazaspiro [3.3] heptane-2-carboxylate.
Figure imgf000554_0002
Figure imgf000554_0003
[002855] Under argon atmosphere, a mixture of 4-bromo-5-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (200 mg, 0.623 mmol), tert-butyl 2,6-diazaspiro [3.3]heptane-2-carboxylate (185 mg, 0.934 mmol), tBuXPhos Pd G3 (49mg, 0.062 mmol) and sodium tert-butoxide (180 mg, 0.365 mmol) in 1,4-dioxane (15 mL) was stirred 100 °C for 16h. The reaction mixture was purified by flash chromatography (PE/EA=2/1) to afford the desired product tert-butyl 6-[5- methyl- l-[4-(tri fluoromethoxy )phenyl]pyrazol-4-yl]-2,6-diazaspiro[3.3]heptane-2-carboxylate (140 mg, yield 46%) as a yellow oil.
[002856] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA;
Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 90.6% (214 nm); Mass: found peak 439.3(M+H) at 1.30 min.
[002857] Step 2. Synthesis of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro [3.3] heptanes.
Boc
Figure imgf000555_0001
[002858] A solution of tert-butyl 6-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptane-2-carboxylate (140 mg, 0.319 mmol) and TFA (364 mg, 3.19 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]- 2,6-diazaspiro[3.3]heptanes (90 mg, yield 55%) as a yellow oil.
[002859JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5% B increase to 95% B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 66.5% (214 nm); Mass: found peak 339.2 (M+H) at 0.988 min.
[002860] Step 3. Synthesis of 4-[2-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-2,6-diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (Compound b!5).
Figure imgf000555_0002
[002861] A solution of 2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptanes (90 mg, 0.266 mmol), 1,4-oxazepane hydrochloride (74 mg, 0.399 mmol), potassium carbonate (110 mg, 0.798 mmol) and KI (44 mg, 0.266 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h, then the mixture was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[2-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-2,6- diazaspiro[3.3]heptan-6-yl]ethyl]morpholine (22 mg, yield 17.8%) as a white solid. [002862JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B:
Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min;
Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 97.7% (214 nm); Mass: found peak 452.1(M+H) at 1.804 min.
[002863] 'H NMR (400 MHz, MeOD-d4) 5 7.55 (d, J = 8.0 Hz, 2H), 7.46 (d, J = 8.0 Hz, 2H), 7.39 (s, 1H), 3.93 (s, 4H), 3.71 (d, J = 8.0 Hz, 4H), 3.50 (s, 4H), 2.67 (d, J = 8.0 Hz, 2H), 2.46 (s, 4H), 2.40 (t, J = 8.0 Hz, 2H), 2.24 (s, 3H) ppm.
Example S-bl6. Synthesis of 4-[2-[(lR,5S)-3-[3-methyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,8-diazabicyclo[3.2.1]octan-8-yl]ethyl]morpholine (Compound b!6).
[002864] Compound b!6 was prepared as outlined below.
Figure imgf000556_0001
[002865] Step 1. Synthesis of tert-butyl (lR,5S)-3-[3-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazol-4-yl]-3,8-diazabicyclo [3.2. l]octane-8-carboxylate.
Figure imgf000556_0002
[002866] Under argon atmosphere, a mixture of 4-bromo-3-methyl-l-[4-(trifluoromethoxy) phenyl]pyrazole (150 mg, 0.467 mmol), tert-butyl(lR,5S)-3,8-diazabicyclo[3.2.1]octane-8- carboxylate(149 mg, 0.701 mmol), PEPPSI-IPent (37 mg, 0.046 mmol) and sodium tert- butoxide (135 mg, 1.40 mmol) in 1,4-dioxane (15 mL) was stirred 100 °C for 16h. The residue was purified by flash chromatography (PE/EA= 2/1) to give tert-butyl (lR,5S)-3-[3-methyl-l-[4- (trifluoromethoxy) phenyl]pyrazol-4-yl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (50 mg, yield 17%) as a colorless oil.
[002867] LCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA;
Gradient: 5%B increase to 95%B within l.Omin; Flow Rate:2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 72.2% (214 nm); Mass: found peak 453.3 (M+H) at 1.579 min.
[002868] Step 2. Synthesis of (lR,5S)-3-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-3,8-diazabicyclo[3.2.1]octane.
Figure imgf000557_0001
[002869] A solution of tert-butyl (lR,5S)-3-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (55 mg, 0.122 mmol) and TFA (139 mg, 1.22 mmol) in DCM (10 mL) was stirred at rt for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (20 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product (lR,5S)-3-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]-3,8-diazabicyclo[3.2.1]octane (40 mg, yield 68.2%) as a yellow oil.
[002870JLCMS Method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01%TFA; Gradient: 5%B increase to 95%B within 1.0 min; Flow Rate: 2.2 mL/min; Column: Sunfire, 4.6*30 mm, 2.7 pm; Column Temperature: 40 °C; LC purity: 73.0% (214 nm); Mass: found peak 353.3 (M+H) at 1.081 min.
[002871] Step 3. Synthesis of 4-[2-[(lR,5S)-3-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]-3,8-diazabicyclo[3.2.1]octan-8-yl]ethyl]morpholine (Compound bl6).
Figure imgf000557_0002
[002872] A solution of (lR,5S)-3-[3-methyl-l-[4-(trifhioromethoxy)phenyl]pyrazol-4-yl]-3,8- diazabicyclo[3.2.1]octane (40 mg, 0.114 mmol), l,4-oxazepane;hydrochloride (31 mg, 0.170 mmol), potassium carbonate (47 mg, 0.341 mmol) and KI (18 mg, 0.114 mmol) in EtOH (10 mL) was stirred at 90 °C for 16h, then the mixture was concentrated in vacuo. The filtrate was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[(lR,5S)-3-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,8- diazabicyclo[3.2.1]octan-8-yl]ethyl]morpholine (18.8 mg, yield 34.9%) as a colorless oil. [002873JLCMS Method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10%B increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; Column: Sunfire, 4.6*50 mm, 3.5 pm; Column Temperature: 50 °C; LC purity: 98.3% (214 nm); Mass: found peak 466.2(M+H) at 2.153 min.
[002874] 'H NMR (400 MHz, MeOD-d4) 5 7.82 (s, 1H), 7.74 (dd , J = 8.0, 4.0 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 3.72 (t, J = 8.0 Hz, 4H), 3.39 (s, 2H), 3.03 (dd, J = 12.0, 4.0 Hz, 2H), 2.91 (d, J = 12.0 Hz, 2H), 2.62 (dt, J = 8.0, 4.0 Hz, 4H), 2.56 (s, 4H), 2.30 (s, 3H), 2.04-2.01 (m, 4H) ppm.
Example S-bl7. Synthesis of 4-[2-[4-[4-fluoro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound b!7). [002875] Compound b!7 was prepared as outlined below.
Figure imgf000558_0001
[002876] Step 1. Synthesis of tert-butyl 4-[4-fluoro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate.
Figure imgf000558_0002
[002877] To a solution of l-(4-pyridyl)butane-l,3-dione (200 mg, 0.3 mmol) in acetonitrile (5 mL) was added N-fluoro-N'-chloromethyltriethylenediamine (109 mg, 0.3 mmol). The mixture was stirred for 16h at RT, concentrated in vacuo and purified by SGC (petroleum ether: ethyl acetate=3: l) to afford the desired product tert-butyl 4-[4-fluoro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l-carboxylate(60mg, 28.8%) as a yellow oil. [002878JLCMS method: Mobile Phase : A: water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2min; Flow Rate :2.2mL/min; Column : Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 388.1 (M-55)+ at 1.543 min.
[002879] Step 2. Synthesis of 4-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 3-yl]piperidine.
Figure imgf000559_0001
[002880] To a solution of tert-butyl 4-[4-fluoro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine-l -carboxylate (160 mg, 0.33 mmol) in DCM (10 mL) was added TFA (2 mL), which was stirred for 2 h at ambient temperature. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, and extracted with dichloromethane (20 mL*3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated to afford the desired product 4-[4-fhioro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]piperidine (40 mg, 86.1%) as an orange oil.
[002881JLCMS method: Mobile Phase : A: water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5%B increase to 95%B within 1.3min, 95%B for 1.2min; Flow Rate: 2.2mL/min; Column: Chromolith Fast gradient RP-18e 50mm*3mm; Column Temperature: 40 °C; LC purity: 100% (214 nm), Mass: found peak 203.2 (M+l) at 0.744 min.
[002882] Step 3. Synthesis of 4-[2-[4-[4-fluoro-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-3-yl]-l-piperidyl]ethyl]morpholine (Compound b!7).
Figure imgf000559_0002
[002883] To a solution of 4-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]piperidine (40 mg, 0.12 mmol), KI (20 mg, 0.12 mmol), and potassium carbonate(48 mg, 0.35 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (32 mg, 0.17 mmol). The reaction was stirred at 90 °C overnight. The reaction was allowed to cool to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[4-fluoro-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3- yl]-l-piperidyl]ethyl]morpholine (24.1 mg, yield 45.3 %) as a yellow oil.
[002884] LCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 5% increase to 95%B within 1.3min, 95%B for 1.7min; Flow Rate: 1.8 mL/min; Column: X-Bridge C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 456.9 (M+l) at 1.900 min.
[002885] 'H NMR (400 MHz, DMSO-d6) 5 7.66 (d, J = 9.2 Hz, 2H), 7.53 (d, J = 8.8 Hz, 2H), 3.57 (s, 4H), 3.09-2.82 (m, 3H), 2.42 (s, 8H), 2.30 (s, 3H), 2.08-1.74 (m, 6H) ppm.
Example S-bl8. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-l-piperidyl]ethyl]morpholine (Compound b!8).
[002886] Compound b!8 was prepared as outlined below.
Figure imgf000560_0001
[002887] Step 1. Synthesis of 4-iodo-3-methyl-l-[4-(trifluoromethoxy) phenyl] pyrazole.
Figure imgf000560_0002
,
[002888] To a solution of 3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (200 mg, 0.82 mmol) in acetonitrile (10 mL) was added N-iodosuccinimide(743 mg, 3.3 mmol). The reaction mixture was heated to 60°C. After 3 h, the reaction mixture was diluted with ethyl acetate, washed with IN aqueous sodium hydroxide and brine, then dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (ethyl acetate/hexanes gradient) to afford the desired product 4-iodo-3-methyl-l-[4- (trifluoromethoxy) phenyl] pyrazole (150 mg, 49.3%) as a yellow solid. [002889JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 97% (214 nm), Mass: found peak 368.9 (M+l) at 2.302 min.
[002890] Step 2. Synthesis of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 4-yl]-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000561_0001
Pd(dppf)Cl2(0.2 eq) dioxane/H2O
80°c, o/n
[002891] A solution of 4-iodo-3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (120 mg, 0.32 mmol), tert-butyl 4-(4, 4,5, 5-tetramethyl- 1,3, 2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyri dine- 1- carboxylate (202 mg, 0.65 mmol), 1,1’ bis(diphenyl-phosphino)ferrocene- palladium(II)dichloride dichloromethane complex (80 mg, 0.09 mmol), potassium carbonate(135 mg, 0.9 mmol) in dioxane/water (10/2 mL) was stirred overnight at 80 °C. The solution was concentrated and purified by SGC (petroleum ether: ethyl acetate=3: l) to afford the desired product tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6-dihydro- 2H-pyridine-l -carboxylate (80 mg, 86.9%) as yellow oil.
[002892JLCMS method: Mobile Phase: A: water (0.01%TFA) B: ACN (0.01%TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0mL/min; Column: Sunfire C18, 4.6*50mm, 3.5pm; Column Temperature: 45 °C; LC purity:92% (214 nm) Mass: found peak 424.1 (M+l) at 2.164 min.
[002893] Step 3. Synthesis of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 4-yl]piperidine-l-carboxylate.
Figure imgf000561_0002
[002894] A solution of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6- dihydro-2H-pyridine-l-carboxylate(120 mg, 0.28 mmol) and Pd/C(40 mg) in methanol (20 mL) was stirred for 30 min at RT. The mixture was filtered and concentrated in vacuo to afford the desired product tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine-l- carboxylate (80 mg, 66.4%) as yellow oil. [002895] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 370.0 (M+l) at 2.349 min.
[002896] Step 4. Synthesis of 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperidine.
Figure imgf000562_0001
[002897] To a solution of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperidine-l-carboxylate(160 mg, 0.33 mmol) in dichloromethane (10 mL) was added TFA (2.78 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[3-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine (60 mg, 98.1%) as a yellow oil. [002898JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 84% (214 nm), Mass: found peak 326.2 (M+l) at 1.047 min.
[002899] Step 5. Synthesis of 4-[2-[4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-l-piperidyl]ethyl]morpholine (Compound b!8).
Figure imgf000562_0002
[002900] To a solution of 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine (60 mg, 0.18 mmol), KI (30.6 mg, 0.12 mmol), and potassium carbonate (76.5 mg, 0.55 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (51 mg, 0.27 mmol). The reaction was stirred at 90 °C overnight, allowed to cool to room temperature, then filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4-[2-[4-[3- methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (29.1 mg, yield 36 %) as yellow solid. [002901JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 93% (214 nm), Mass: found peak 439.1 (M+l) at 2.094 min.
[002902] 'H NMR (400 MHz, DMSO-d6) 5 8.25 (s, 1H), 7.86 (d, J = 9.2 Hz, 2H), 7.44 (d, J = 8.8 Hz, 2H), 3.55 (t, J = 4.8 Hz, 4H), 2.95 (d, J = 11.2 Hz, 2H), 2.45-2.37 (m, 8H), 2.21 (s, 3H), 2.02 (t, J = 11.6 Hz, 2H), 1.78 (d, J = 12.4 Hz, 2H), 1.62-1.50 (m, 2H) ppm.
Example S-bl9. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-l-piperidyl]ethyl]morpholine (Compound b!9).
[002903] Compound b!9 was prepared as outlined below.
Figure imgf000563_0001
[002904] Step 1. Synthesis of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000563_0002
pyridine(5.0 eq) DCM, rt, o/n
[002905] To a solution of 3-methyl-lH-pyrazole (1.0 g, 12.2 mmol) in dichloromethane (20 mL) was added [4-(trifluoromethoxy)phenyl]boronic acid (5.02 g, 24.4 mmol), anhydrous copper acetate (8.85 g, 48.7 mmol), pyridine (3.85 g, 48.7 mmol), and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 40 h. The mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 80g silica gel column @75mL/min, eluting with 0-20% EA in petroleum ether) to afford the product 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (300 mg, yield 8.6%)and 3-methyl-l- [4-(trifluoromethoxy)phenyl]pyrazole (1.8 g, yield 61.0%) as a yellow solid.
[002906] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1CV X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 243.1 (M+H)+ at 2.131 min. [002907] 'H NMR (400 MHz, DMSO-d6) 5 8.41 (d, J = 1.6 Hz, 1H), 7.98-7.84 (m, 2H), 7.47 (d, J = 8.8 Hz, 2H), 6.36 (s, 1H), 2.28 (s, 3H) ppm.
[002908JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column:
X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 98% (214 nm), Mass: found peak 243.1 (M+H)+ at 2.058 min.
[002909] 'H NMR (400 MHz, DMSO-d6) 5 7.71 - 7.64 (m, 2H), 7.59 (d, J = 1.6 Hz, 1H), 7.55-
7.49 (m, 2H), 6.33-6.25 (m, 1H), 2.36 (s, 3H) ppm.
[002910] Step 2. Synthesis of 4-iodo-5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole.
Figure imgf000564_0001
60 C, o/n
[002911] To a solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (150 mg, 0.62 mmol) in acetonitrile (10 mL) was added N-iodosuccinimide (279 mg, 1.24 mmol). The reaction mixture was heated to 60°C. After 3 h, the reaction mixture was diluted with ethyl acetate, washed with IN aqueous sodium hydroxide and brine, then dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel (ethyl acetate/hexanes gradient) to afford the desired product 4-iodo-5-methyl-l-[4- (trifluoromethoxy)phenyl]pyrazole (150 mg, 65.8%) as a yellow oil.
[002912] LCMS method: Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Temperature: 50 °C; LC purity: 100% (214 nm), Mass: found peak 368.9 (M+l) at 2.201 min.
[002913] Step 3. Synthesis of tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol- 4-yl]-3,6-dihydro-2H-pyridine-l-carboxylate.
Figure imgf000564_0002
[002914] A solution of 5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazole (150 mg, 0.62 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-l- carboxylate(383 mg, 1.24 mmol), l,r-bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex (152 mg, 0.18 mmol), and potassium carbonate (257 mg, 1.86 mmol) in dioxane/water (10/2 mL) was stirred for overnight at 80 °C. The mixture was concentrated in vacuo and the residue was purified by SGC (petroleum ether: ethyl acetate=3: l) to afford the desired product tert-butyl 4-[3-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6-dihydro- 2H-pyridine-l -carboxylate (130 mg, 49.6%) as a yellow oil.
[002915] LCMS method: Column: X-BRIDGE Cl 8 (4.6x 50 mm, 3.5pm); Mobile phase: water (10 mM ammonium hydrogen carbonate) (A) / ACN (B); Elution program: Gradient from 10 to 95% of B in 1.5min at 1.8mL/min; Temperature: 50°C; LC purity: 90% (214 nm), Mass: found peak 424.0 (M+l) at 2.277 min.
[002916] Step 4. Synthesis of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-
4-yl]piperidine-l-carboxylate.
Figure imgf000565_0001
[002917] A solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-3,6- dihydro-2H-pyridine-l-carboxylate(70 mg, 0.15mmol) and Pd/C(40 mg) in methanol (20 mL) was stirred for 30min at RT. Then the mixture was filtered and concentrated to afford the desired product tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine-l- carboxylate (130 mg, 99%) as a yellow oil.
[002918JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 82% (214 nm), Mass: found peak 370.0 (M+l) at 2.267 min.
[002919] Step 5. Synthesis of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl] piperidine.
Figure imgf000565_0002
[002920] To a solution of tert-butyl 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperidine-l-carboxylate(130 mg, 0.30 mmol) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (50 mL), neutralized with potassium carbonate to pH=9, extracted with dichloromethane (20 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford the desired product 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]piperidine (80 mg, 80.5%) as a yellow oil.
[002921JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: from 5 to 95% of B in 1.2min at 2.2mL/min; Column: HALO C18, 2.7pm, 4.6*30mm; Column Temperature: 40 °C; LC purity: 82% (214 nm), Mass: found peak 326.2 (M-17)+ at 1.022 min.
[002922] Step 6. Synthesis of 4-[2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4- yl]-l-piperidyl]ethyl]morpholine (Compound b!9).
Figure imgf000566_0001
[002923] To a solution of 4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]piperidine (80 mg, 0.24 mmol), KI (40.8 mg, 0.24 mmol), and potassium carbonate (102 mg, 0.74 mmol) in 95% ethanol (10 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (68 mg, 0.37 mmol). The reaction was stirred at 90 °C overnight. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[5-methyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-4-yl]-l-piperidyl]ethyl]morpholine (18.9 mg, yield 17.5 %) as a yellow solid.
[002924JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate), B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 9% (214 nm), Mass: found peak 194.0 (M+l) at 2.103 min.
[002925] 'H NMR (400 MHz, DMSO-d6) 5 7.63 (d, J = 8.8 Hz, 2H), 7.55 (s, 1H) 7.50 (d, J = 8.8 Hz, 2H), 3.55 (t, J = 4.8 Hz, 4H), 2.95 (d, J = 11.2 Hz, 2H), 2.45-2.37 (m, 8H), 2.25 (s, 3H), 2.02 (t, J = 11.6 Hz, 2H), 1.78 (d, J = 10.4 Hz, 2H), 1.62-1.50 (m, 2H) ppm.
Example S-b20. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl- pyrazol-4-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound b20).
[002926] Compound b20 was prepared as outlined below.
Figure imgf000567_0001
[002927] Step 1. Synthesis of tert-butyl 4-[5-sec-butyl-l-[4-
(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine-l-carboxylate.
Figure imgf000567_0002
[002928] To a solution of 3-bromo-4-iodo-lH-pyrazole (800 mg, 2.93 mmol) in chloroform (10 mL) was added (2,2-difluoro-l,3-benzodioxol-5-yl)boronic acid (1.18 g, 5.86 mmol), anhydrous copper acetate (1.07 g, 5.86 mmol), pyridine (928 mg, 11.7 mmol) and molecular sieves 4 A. The reaction mixture was stirred at room temperature for 24 hrs. The mixture was filtered and concentrated in vacuo. The residue was purified by flash chromatography (Biotage, 25 g silica gel column @70mL/min, eluting with 10-50% di chloromethane in petroleum ether) to afford the desired product tert-butyl 4-[5-sec-butyl-l-[4-(trifluoromethoxy)phenyl]pyrazol-3-yl]piperazine- 1-carboxylate (1.24 g, yield 98.6%) as a yellow solid.
[002929JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 429 (M + H) at 2.06 min.
[002930] Step 2. Synthesis of tert-butyl 4-[3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5- yl)pyrazol-4-yl]piperazine-l-carboxylate.
Figure imgf000567_0003
[002931] To a solution of 3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-4-iodo-pyrazole (1.24 g, 2.89 mmol) in THF (30 mL) at room temperature was added tert-butyl piperazine-1- carboxylate (0.592 g, 3.18 mmol), [2-(2-aminophenyl)phenyl]-methylsulfonyloxy-palladium, dicyclohexyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphane (0.245 g, 0.289 mmol), and sodium tert-butoxide (0.833 g,8.67 mmol) and the reaction mixture was stirred at 60 °C for 8 h. The mixture was quenched by water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[3-bromo-l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)pyrazol-4-yl]piperazine-l-carboxylate (0.4 g, yield: 28.4%) as a white solid. [002932JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 433 (M - tBu)+ at 2.31 min
[002933] Step 3. Synthesis of tert-butyl 4-[l-(2, 2-difluoro-l, 3-benzodioxol-5-yl)-3- isopropenyl-pyrazol-4-yl]piperazine-l-carboxylate.
Figure imgf000568_0001
[002934] To a solution of tert-butyl 4- [3 -bromo- 1 -(2, 2-difluoro-l, 3-benzodioxol-5-yl)pyrazol-4- yl]piperazine-l -carboxylate (0.4 g, 0.821 mmol) in dioxane/water (10/1, 10 mL) at room temperature was added 2-isopropenyl-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (0.166 g, 0.985 mmol), bis(tri-tert-butylphosphine)palladium(0) (0.042 g, 0.0821 mmol), and potassium phosphate tribasic (0.523 g, 2.46 mmol) and the reaction mixture was stirred at 100 °C for 2 h. The mixture was quenched with water and the aqueous layer was extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaCl, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (EtOAc / petroleum ether = 1 : 4) to afford tert-butyl 4-[l-(2,2- difluoro-l,3-benzodioxol-5-yl)-3-isopropenyl-pyrazol-4-yl]piperazine-l-carboxylate (0.2 g, yield: 54.3%) as a white solid.
[002935] LCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm);
Mass: found peak 449 (M + H) at 2.53 min.
[002936] Step 4. Synthesis of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl- pyrazol-4-yl]piperazine-l-carboxylate.
Figure imgf000569_0001
[002937] To a solution of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropenyl- pyrazol-4-yl]piperazine-l -carboxylate (200 mg, 0.446 mmol) in 5mL THF was added platinum dioxide (20 mg). The mixture was stirred under H2 at room temperature for Ih. The reaction mixture was filtered and concentrated under vacuo to give the crude product.
[002938JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 451 (M + H) at 2.51 min.
[002939] Step 5. Synthesis of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl-pyrazol-4- yl] piperazine.
Figure imgf000569_0002
[002940] To a solution of tert-butyl 4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl- pyrazol-4-yl]piperazine-l -carboxylate (180 mg, 0.4 mmol) in dichloromethane (3 mL) was added 2,2,2-trifluoroacetic acid (1 mL). The reaction mixture was stirred at room temperature for Ih. The reaction mixture was concentrated in vacuo. The residue was diluted with water (5 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (5 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford l-[l-(2,2-difluoro-l,3- benzodioxol-5-yl)-3-isopropyl-pyrazol-4-yl]piperazine (100 mg, crude). The crude product was used directly in the next step.
[002941JLCMS method: Mobile Phase: A: water (10 mM ammonium hydrogen carbonate) B: Acetonitrile; Gradient: 10% increase to 95%B within 1.5 min; Flow Rate: 1.5 mL/min; Column: X-Bridge C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C; LC purity: 100 % (214 nm); Mass: found peak 351 (M + H) at 2.13 min.
[002942] Step 6. Synthesis of 4-[2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl- pyrazol-4-yl]piperazin-l-yl]ethyl]-l,4-thiazinane 1,1-dioxide (Compound b20).
Figure imgf000570_0001
[002943] To a solution of l-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl-pyrazol-4- yl]piperazine (140 mg, 0.40 mmol), potassium carbonate (166 mg, 1.20 mmol) and KI (6 mg, 0.04 mmol) in 95% ethanol (5mL) was added 4-(2-chloroethyl)thiomorpholine 1,1 -di oxide (103 mg, 0.44 mmol). The reaction was stirred at 95 °C for 16 hrs. The reaction was cooled to room temperature and filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (ammonium hydrogen carbonate/water/acetonitrile) to afford the desired product 4- [2-[4-[l-(2,2-difluoro-l,3-benzodioxol-5-yl)-3-isopropyl-pyrazol-4-yl]piperazin-l-yl]ethyl]-l,4- thiazinane 1,1-dioxide (50 mg, yield: 24.5%) as a white solid.
[002944JLCMS method: Mobile Phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA); Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.7 min; Flow Rate: 2.0 mL/min; Column: SUNFIRE C18, 50*4.6mm, 3.5pm; Column Temperature: 50 °C. LC purity: 99.56 % (214 nm); Mass: found peak 512 (M + H) at 1.47 min.
[002945] 'H NMR (400 MHz, DMSO) 5 8.12 (s, 1H), 7.80 (s, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.47 (d, J = 8.4 Hz, 1H), 3.03 (m, 4H), 2.96 (s, 6H), 2.83 (s, 4H), 2.62 (m, 3H), 2.53 (m, 4H), 1.26 (m, 6H).
Example S-b21. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta[c]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound b21).
[002946] Compound b21 was prepared as outlined below.
Figure imgf000570_0002
[002947] Step 1. Synthesis of 3-iodo-l,4,5,6-tetrahydrocyclopenta[c]pyrazole.
Figure imgf000570_0003
[002948] To a solution of l,4,5,6-tetrahydrocyclopenta[c]pyrazole (500 mg, 4.6 mmol) in DMF (10 mL) was added 1 -iodopyrrolidine-2, 5-dione (1.25 g, 5.5 mmol). The reaction was stirred at rt for 16h. The reaction mixture was diluted with EtOAc (50 mL), and washed with water (15 mL) and saturated brine (30 mL * 3). Then the organic layer was purified by SGC (PE: EA = 1 : 1) to afford 3-iodo-l,4,5,6-tetrahydrocyclopenta[c]pyrazole (600 mg, yield 55.4%) as a white solid.
[002949JLCMS method: Mobile phase: A: Water (0.01%TFA), B: Acetonitrile (0.01% TFA) Gradient: 5% increase to 95%B within 1.3 min, 95%B for 1.2min; Flow Rate: 2.2 mL/min Column: Chromolith Fast gradient RP-18e, 50*3 mm, Column Temperature: 40 °C; LC purity: 72.8% (214 nm) Mass: found peak 234.8 (M+l) at 0.901 min.
[002950] Step 2. Synthesis of 3-iodo-l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta [c] pyrazole.
Figure imgf000571_0001
[002951] To a solution of 3-iodo-l,4,5,6-tetrahydrocyclopenta[c]pyrazole (500 mg, 2.14 mmol) in DCM (20 mL) was added [4-(trifluoromethyl)phenyl]boronic acid (880 mg, 4.27 mmol), copper(II) acetate (776 mg, 4.27 mmol), pyridine (676 mg, 8.55 mmol) and molecular sieves 4A. The reaction mixture was stirred at room temperature for 48h. The mixture was filtered, and purified by flash chromatography (PE/DCM=1/1) to afford desired product 3-iodo-l-[4- (trifluoromethoxy)phenyl]-5,6-dihydro-4H-cyclopenta[c]pyrazole (300 mg, yield 35.6%) as a colorless oil.
[002952JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 30% (214 nm) Mass: found peak 395.0 (M+l) at 1.550 min.
[002953] Step 3. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta [c] pyrazol-3-yl] piperazine- 1-carboxylate.
[002954]Under argon atmosphere, a mixture of 3-iodo-l-[4-(trifluoromethoxy)phenyl]-5,6- dihydro-4H-cyclopenta[c]pyrazole (100 mg, 0.460 mmol), sodium tert-butoxide (74 mg, 0.761 mmol), tert-butyl piperazine- 1-carboxylate (95 mg, 0.507 mmol) and Pd PEPPSI IPENT (21.5 mg, 0.025 mmol) in anhydrous 1,4-dioxane (5 mL) was stirred at 100 °C for 16h. The reaction was cooled down to room temperature, directly purified by flash chromatography (Biotage, 80 g silica gel column @100mL/min, eluting with 2-30% ethyl acetate in petroleum ether) to afford tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H-cyclopenta[c]pyrazol-3- yl]piperazine-l -carboxylate (60 mg, yield 52.3%) as a yellow solid.
[002955] LCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA);
Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm 4.6*30mm; Column Temperature: 40 °C; LC purity: 98% (214 nm) Mass: found peak 453.3 (M+l) at 1.613 min.
[002956] Step 4. Synthesis of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta [c] pyrazol-3-yl] piperazine- 1-carboxylate.
Figure imgf000572_0001
[002957] To a solution of tert-butyl 4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta[c]pyrazol-3-yl]piperazine- 1-carboxylate (60 mg, 0.133 mmol) in dichloromethane (3 mL) was added TFA (1 mL). The reaction mixture was stirred at room temperature for 2h. The reaction mixture was concentrated in vacuo. The residue was diluted with water (10 mL), neutralized with potassium carbonate to pH=8, extracted with dichloromethane (10 mL*3), dried over sodium sulfate, filtered, and concentrated in vacuo to afford tert-butyl 4-[l-[4- (trifluoromethoxy)phenyl]-5,6-dihydro-4H-cyclopenta[c]pyrazol-3-yl]piperazine-l-carboxylate (50 mg, yield 86%) as a yellow oil.
[002958JLCMS method: Mobile Phase: A: Water (0.01%TFA) B: Acetonitrile (0.01%TFA); Gradient: 5% increase to 95%B in 1.0 min; Flow Rate: 2.2 mL/min; Column: HALO C18 2.7pm [002959] 4.6*30mm; Column Temperature: 40 °C; LC purity: 97% (214 nm) Mass: found peak 353.2 (M+l) at 1.084 min.
[002960] Step 5. Synthesis of 4-[2-[4-[l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta[c]pyrazol-3-yl]piperazin-l-yl]ethyl]morpholine (Compound b21).
Figure imgf000572_0002
[002961] To a solution of 3-piperazin-l-yl-l-[4-(trifluoromethoxy)phenyl]-5,6-dihydro-4H- cyclopenta[c]pyrazole (50 mg, 0.142 mmol), potassium carbonate (78 mg, 0.57 mmol) and KI (24 mg, 0.14 mmol) in 95% ethanol (3 mL) was added 4-(2-chloroethyl)morpholine hydrochloride (52 mg, 0.28 mmol). The reaction was stirred at 95 °C for 16h. The reaction was filtered. The filtrate was concentrated in vacuo. The residue was purified by prep-HPLC (NH4HCO4/water/acetonitrile) to afford the desired product 4-[2-[4-[l-[4- (trifluoromethoxy)phenyl]-5,6-dihydro-4H-cyclopenta[c]pyrazol-3-yl]piperazin-l- yl]ethyl]morpholine (45 mg, yield 68.1 %) as a yellow solid.
[002962JLCMS method: Mobile Phase: A: Water (10 mM ammonium hydrogen carbonate) B: ACN; Gradient: 10% increase to 95%B within 1.5min; Flow Rate: 1.8 mL/min; ColummX- Bridge: C18, 3.5pm, 4.6*50mm; Column Temperature: 50 °C; LC purity: 100% (214 nm) Mass: found peak 466.1 (M+l) at 2.192 min.
[002963] 'H NMR (400 MHz, CDCh) 5 7.53 (d, J = 9.0 Hz, 2H), 7.20 (d, J = 8.8 Hz, 2H), 3.72(t, J = 4.8 Hz, 4H), 3.32 (t, J = 4.8 Hz, 4H), 2.93 (t, J = 7.2 Hz, 2H), 2.70 (t, J = 6.8 Hz, 2H), 2.65 - 2.54 (m, 10H), 2.51 (s, 4H) ppm.
Biological Examples
Example Bl. Human EBP Biochemical Assay.
[002964] The EBP biochemical assay was designed to measure the production of lathosterol-d7 product from zymostenol-d7 substrate in the presence of EBP enzyme. A decrease in lathosterol-d7 production indicated compound inhibition. This assay allowed for the discovery of small molecule inhibitors of EBP.
[002965] For the enzyme source, human EBP microsomes were isolated from Sf9 insect cells expressing human EBP. EBP microsomes were diluted to the final assay concentration of 20 pg/ml in assay buffer (50 mM Tris-HCl, 2 mM MgCh, 1 mM EDTA, 2 mM betamercaptoethanol, 5% glycerol, 0.1% Tween-80, pH 7.5). Zymostenol-d7 substrate (Avanti Polar Lipids) was diluted to 15 pM in assay buffer. Test compounds and controls were dispensed to Corning 3575 plates (20 nL/well) using the Beckman Coulter Echo 655 (0.1% DMSO final) in duplicate plates.
[002966] 10 pL/well of 20 ug/mL EBP microsomes were added to each plate containing test compounds with a multichannel pipette. The plates were sealed with foil seal. The plates were spun for 30 seconds at 1000 rpm. The compounds and microsomes were incubated in the plates for 15 minutes at 37°C. 10 pL/well of 15 pM zymostenol-d7 substrate was added to the plates with a multichannel pipette. The plates were sealed with foil seal and spun down for 30 seconds at 1000 rpm. The plates were incubated at 37°C for 2 hours.
[002967] To extract samples, 70 pL/well of extraction solvent (50% acetonitrile, 50% methanol, 5 mM ammonium formate, 0.2% formic acid) were added with a Multidrop. The plates were spun down for 5 minutes at 2300 rpm at room temperature. 70 pL of supernatant were transferred to 384-well Waters plates using Apricot iPipette with 125 uL tips. The Waters plates were heat sealed (3.2-3.5 seconds, 160-163°C on PlateLoc) and spun at 1500 rpm for 5 minutes at room temperature.
[002968] The plates were analyzed on Thermo Vanquish/Sciex 6500+ LC/MS for zymostenol-d7 substrate and lathosterol-d7 product using a 1 pL injection (autosampler at 10°C, column temperature 60°C, 90% isocratic, flow rate 0.8 mL/min, column Acquity CSH C18 2.1 x 100 mm, 1.7 um; Mobile phase A: water + 0.2% formic acid, Mobile phase B: acetonitrile + 0.2% formic acid).
[002969] Data was integrated using Analyst software and uploaded to IDBS Abase for validation. The ICso values for tested compounds against human EBP (hEBP) are summarized in Table 4.
Table 4.
Figure imgf000574_0001
Figure imgf000575_0001
Figure imgf000576_0001
Figure imgf000577_0001
ND = not determined
Example B2. Mouse OPC Studies.
Isolation, Culture, and Treatment
[002970] C57B1/6 mouse brains were collected on P7-8 and stored in Hibernate A (Thermofisher, A124750) medium until tissue digestion. The digestion was conducted using a Neural Tissue Dissociation Kit (Miltenyi, 130-092-628) and gentleMACs Dissociator (Miltenyi) in accordance with the manufacturer’s protocol. Anti-04 microbeads (Miltenyi, 130-094-543) and LS columns (Miltenyi, 130-042-401) were used to positively select for oligodendrocyte precursor cells (OPCs) per the manufacturer’s protocol. The isolated cells were plated 10,000 cells per well on black 96-well plates (Perkin Elmer, 6055302) and coated with 100 mg/mL Poly-D-lysine (Sigma, P6407) for at least 1 hour. Cells were cultured in DMEM/F12 (Gibco, 11330-057) with 1% N-2 (Invitrogen, 17502048), 100 U/mL Penicillin / 100 mg/mL Streptomycin (Gibco, 15140122), 0.01% bovine serum albumin (Sigma, A9576), 5 mg/mL N- Acetyl-L-cysteine (Sigma, A8199), 10 ng/mL d-Biotin (Sigma, B4639), 2% B-27 (Invitrogen, 17504044), 1 mM pyruvate (Gibco, 11360070), 5 mg/mL insulin (Sigma, 19278), 4.2 mg/mL forskolin (Sigma, F6886) and 10 ng/mL CNTF (Peprotech, 450-50). To promote proliferation, 20 ng/mL PDGF-AA (Invitrogen, PHG0035) and 1 ng/mL NT-3 (Peprotech, 450-03) were added to the medium. After two days, the proliferation medium was removed and replaced with differentiation medium (proliferation medium minus PDGF-AA and NT-3), and the test compounds were administered. Two days later, the media was replenished. Cells were fixed in 4% PFA (Electron Microscopy Sciences, 15714S) three days later for a total of 5 days of treatment. Immunostaining and Imaging
[002971]Fixed cells were incubated with 10% normal goat serum (Vector Labs, S-1000) in PBS (Gibco, 10010023) with 0.2% Triton X-100 (Sigma, T9284; PBST) for 30 min to block nonspecific binding. The primary antibodies used were rat anti-myelin basic protein (BioRad, MCA409, 1 :400 dilution) and rabbit anti-Olig2 (Millipore, AB9610 1 :400). Cells were incubated with the primary antibodies in 5% normal goat serum in PBST overnight at 4°C. The secondary antibodies used were AlexaFluor488 and AlexaFluor555 (1 : 1000, Invitrogen, Al 1006 and A21428), and cells were incubated with secondary antibodies for 1 hr at room temperature. The cells were imaged on the Opera Phenix (Perkin Elmer). 21 images per well were taken with 4 x 1 pm thick Z Stack. Images were analyzed using Harmony software 4.9 (Perkin Elmer).
Data
[002972] Mouse OPC data for select compounds is summarized in Table 5.
Table 5.
Figure imgf000578_0001
[002973] While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein may be employed in practicing the disclosure. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby. The disclosures of all patent and scientific literature cited herein are expressly incorporated herein in their entirety by reference. To the extent that any incorporated material is inconsistent with the express content of this disclosure, the express content controls.

Claims

1. A compound of Formula (I):
Figure imgf000579_0001
or a pharmaceutically acceptable salt thereof, wherein:
X1 is C and X2 is N, or X1 is N and X2 is C;
= is a single bond or a double bond, provided that a double bond and a single bond;
R1 is Ci-C6 alkyl, C3-C6 cycloalkyl, -CN, Ci-C6 haloalkyl, -(Ci-C6 alkylene)-O-(Ci-C6 alkyl),
-(Ci-Ce alkylene)-O-(Ci-Ce haloalkyl), or -(Ci-Ce alkylene)(C3-Ce cycloalkyl);
L1 is a bond, O, or -CH2-;
Ring B is C3-C6 cycloalkyl, 6-membered heteroaryl containing 1 or 2 nitrogen atoms, or Ce-Cio aryl; each R2 is independently Ci-Ce alkyl, Ci-Ce haloalkyl, halo, -O(Ci-Ce alkyl), or -O(Ci-C6 haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and Ci-Ce haloalkyl;
L2 is a bond or O;
Ring A is
Figure imgf000579_0002
, 9- to 11-membered spiro heterocyclylene, or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms;
Y1 is N or CH; Y2 is N or CH; x is 0, 1, or 2; y is 0 or 1; m is 0-5; each R3 is independently Ci-Ce alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C6 cycloalkyl;
L3 is a bond, -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, -CH(Ra)CH(Ra)N(Ra)-, 5- to 6-membered heterocyclylene, or -O-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms; each Ra is independently H or Ci-Ce alkyl;
W is O, CH2, SO2, S(O)=NH, SO, or N(H);
Z is N or CH; r is 0, 1, or 2; s is 0 or 1; each R4 is independently halo, -OH, Ci-Ce alkyl, Ci-Ce haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group; and n is 0-5.
2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000580_0001
3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000581_0001
4. The compound of any one of claims 1-3, or a pharmaceutically acceptable salt thereof, wherein:
R1 is Ci-C6 alkyl, C3-C5 cycloalkyl, -CN, C1-C3 haloalkyl, -(C1-C3 alkylene)-O-(Ci-C3 alkyl), -(C1-C3 alkylene)-O-(Ci-C3 haloalkyl), or -(C1-C3 alkylene)(C3-Ce cycloalkyl).
5. The compound of claim 4, or a pharmaceutically acceptable salt thereof, wherein:
R1 is -CN, -CH3, -CH2CH3, -CH2CH2CH3, -CH(CH3)2, -C(CH3)3, -CH2CH2CH2CH3, -CH2CH(CH3)2, -CH(CH3)CH2CH3, -CH2CH2CH(CH3)2, -CH2OCH3, -CH2CH2OCH3, -CH(CH3)OCH3, -CH(CH3)CH2OCH3, -CH2CHF2, -CF3, -CHF2,
Figure imgf000581_0002
6. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein:
L1 is a bond.
7. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein:
L1 is O.
8. The compound of any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein:
L1 is -CH2-.
9. The compound of any one of claims 1-8, or a pharmaceutically acceptable salt thereof, wherein: Ring B is C4-C6 cycloalkyl, pyridinyl, pyrazinyl, pyrimidinyl, or phenyl.
10. The compound of any one of claims 1-9, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000582_0001
11. The compound of any one of claims 1-10, or a pharmaceutically acceptable salt thereof, wherein: each R2 is independently C1-C4 alkyl, C1-C3 haloalkyl, halo, -O(Ci-C3 alkyl), or
-O(Ci-C3 haloalkyl), or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from halo and C1-C3 haloalkyl.
12. The compound of claim 11, or a pharmaceutically acceptable salt thereof, wherein: each R2 is independently -CF3, -CF2CH3, -CH2CHF2, -CHF2, F, Cl, Br, -OCF3, -OCHF2, -OCH3, or -C(CH3)3, or two R2 groups on adjacent carbon atoms are taken together to form a fused phenyl or a fused 5-membered heterocyclyl containing 1 or 2 oxygen atoms, each of which is optionally substituted by 1-5 groups selected from F or -CF3.
13. The compound of any one of claims 1-12, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000583_0001
14. The compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, wherein:
L2 is a bond.
15. The compound of any one of claims 1-13, or a pharmaceutically acceptable salt thereof, wherein:
L2 is O.
16. The compound of any one of claims 1-15, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000584_0001
Y1 is N or CH;
Y2 is N or CH; x is 0, 1, or 2; and y is 0 or 1.
17. The compound of any one of claims 1-16, or a pharmaceutically acceptable salt thereof, wherein:
Ring A is 9- to 11 -membered spiro heterocyclylene or 8- to 10-membered bicyclic fused heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms.
18. The compound of any one of claims 1-17, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000584_0002
Figure imgf000585_0001
19. The compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 0.
20. The compound of any one of claims 1-18, or a pharmaceutically acceptable salt thereof, wherein: m is 1-3.
21. The compound of any one of claims 1-18 and 20, or a pharmaceutically acceptable salt thereof, wherein: each R3 is independently C1-C3 alkyl, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro C3-C5 cycloalkyl.
22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, wherein: each R3 is independently -CH3, or two R3 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R3 groups on the same carbon atom are taken together to form a spiro cyclopropyl.
23. The compound of any one of claims 1-22, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000586_0001
24. The compound of any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein:
L3 is a bond.
25. The compound of any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein:
L3 is -CH(Ra)-, -CH(Ra)CH(Ra)-, -OCH(Ra)CH(Ra)-, or -CH(Ra)CH(Ra)N(Ra)-; and each Ra is independently H or C1-C3 alkyl.
26. The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein:
L3 is -CH2, -CH2CH2-, -CH(CH3)CH2-, -CH2CH(CH3)-, -OCH2CH2-, or -CH2CH2N(CH2CH3)-.
27. The compound of any one of claims 1-23, or a pharmaceutically acceptable salt thereof, wherein:
L3 is 5- to 6-membered heterocyclylene or -0-(4-membered heterocyclylene), wherein the heterocyclylene contains 1-2 nitrogen atoms.
28. The compound of claim 27, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000587_0001
29. The compound of any one of claims 1-28, or a pharmaceutically acceptable salt thereof, wherein:
W is O, CH2, or N(H).
30. The compound of any one of claims 1-28, or a pharmaceutically acceptable salt thereof, wherein:
W is SO2, S(O)=NH, or SO.
31. The compound of any one of claims 1-30, or a pharmaceutically acceptable salt thereof, wherein:
Z is N.
32. The compound of any one of claims 1-30, or a pharmaceutically acceptable salt thereof, wherein:
Z is CH.
33. The compound of any one of claims 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 1.
34. The compound of any one of claims 1-32, or a pharmaceutically acceptable salt thereof, wherein: r and s are each 0.
35. The compound of any one of claims 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 1; and s is 0.
36. The compound of any one of claims 1-32, or a pharmaceutically acceptable salt thereof, wherein: r is 2; and s is 1.
37. The compound of any one of claims 1-36, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000588_0001
38. The compound of any one of claims 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 0.
39. The compound of any one of claims 1-37, or a pharmaceutically acceptable salt thereof, wherein: n is 1-3.
40. The compound of any one of claims 1-37 and 39, or a pharmaceutically acceptable salt thereof, wherein: each R4 is independently halo, -OH, C1-C3 alkyl, or C1-C3 haloalkyl, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
41. The compound of claim 40, or a pharmaceutically acceptable salt thereof, wherein: each R4 is independently F, -OH, -CH3, -CH(CH3)2, or -CF3, or two R4 groups are taken together to form a bridging -CH2- or -CH2CH2- group, or two R4 groups on adjacent atoms are taken together to form a fused 5-membered heterocyclyl containing 1 oxygen atom, or two R4 groups on the same carbon atom are taken together to form a spiro 4-membered heterocyclyl containing 1 oxygen atom or SO2 group.
42. The compound of any one of claims 1-41, or a pharmaceutically acceptable salt thereof, wherein:
Figure imgf000589_0001
Figure imgf000590_0001
43. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-a), (I-b), or (I-c):
Figure imgf000590_0002
44. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-d) or (I-e):
Figure imgf000590_0003
45. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (I-f), (I-g), (I-h), (I-i), (I-j), (I-k), or (1-1):
Figure imgf000591_0001
heterocyclylene, wherein the heterocyclylene contains 1-2 nitrogen atoms.
46. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Ila) or (lib):
Figure imgf000592_0001
47. The compound of claim 46, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (lie):
Figure imgf000592_0002
48. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (Illa) or (Illb):
Figure imgf000592_0003
wherein
Figure imgf000592_0004
is 9- to 11 -membered spiro heterocyclylene containing 1-2 nitrogen atoms.
49. The compound of any one of claims 1-42, or a pharmaceutically acceptable salt thereof, wherein the compound is of Formula (IVa) or (IVb):
Figure imgf000593_0001
nitrogen atoms.
50. A compound selected from the compounds of Table 1 or a pharmaceutically acceptable salt thereof.
51. A compound selected from the compounds of Table 2 or a pharmaceutically acceptable salt thereof.
52. A compound selected from the compounds of Table 3 or a pharmaceutically acceptable salt thereof.
53. A pharmaceutical composition comprising the compound of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
54. A method of inhibiting Emopamil Binding Protein (EBP) comprising contacting EBP with an effective amount of the compound of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 53.
55. A method of remyelinating a neuronal axon comprising contacting the neuronal axon with an effective amount of the compound of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 53.
56. A method of treating a demyelinating disease in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 53.
57. The method of claim 56, wherein the demyelinating disease is multiple sclerosis (MS), neuromyelitis optica spectrum disorder (NMOSD), acute optic neuritis, transverse myelitis, chronic inflammatory demyelinating polyneuropathy (CIDP), or Guillain-Barre syndrome.
58. A method of treating multiple sclerosis (MS) in a subject in need thereof, comprising administering to the subject an effective amount of the compound of any one of claims 1-52, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 53.
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