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WO2025213088A1 - Compounds, compositions, and methods - Google Patents

Compounds, compositions, and methods

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Publication number
WO2025213088A1
WO2025213088A1 PCT/US2025/023251 US2025023251W WO2025213088A1 WO 2025213088 A1 WO2025213088 A1 WO 2025213088A1 US 2025023251 W US2025023251 W US 2025023251W WO 2025213088 A1 WO2025213088 A1 WO 2025213088A1
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WIPO (PCT)
Prior art keywords
alkyl
cycloalkyl
heterocyclyl
disease
heteroaryl
Prior art date
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Pending
Application number
PCT/US2025/023251
Other languages
French (fr)
Inventor
Alex L. BAGDASARIAN
Cyril Bucher
II Robert A. CRAIG
Javier De Vicente Fidalgo
Anthony A. ESTRADA
Benjamin J. HUFFMAN
Katrina W. Lexa
Maksim OSIPOV
John C. WIDEN
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Tenvie Therapeutics Inc
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Tenvie Therapeutics Inc
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Publication of WO2025213088A1 publication Critical patent/WO2025213088A1/en
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/86—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in position 4
    • C07D239/94—Nitrogen atoms
    • 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
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/04—Ortho-condensed systems
    • C07D491/044—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C07D491/048—Ortho-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring the oxygen-containing ring being five-membered

Definitions

  • the present disclosure relates generally to small molecule modulators of Mucolipin-1 (TRPML1), and their use as therapeutic agents.
  • Transient receptor potential mucolipin 1 also known as mucolipin-1, (TRPML1 or ML-1) is a member of the Transient receptor potential (TRP) ion channel super family. Electrophysiological experiments on isolated lysosomes have demonstrated that TRPML1 is a non-selective cation channel localized to late-endosomes and lysosomes (LEE) (Dong X., et al. Nature 2008, 455, 7215, 992.). Wild- type human TRPML1 has six transmembrane domains within each monomeric unit that come together to form a tetramer containing an ion pore located in the middle of this complex.
  • TRPML1 is permeable to a variety of mono- and divalent cations including Na + , K + , Ca 2+ , Fe 2+ , and Zn 2+ .
  • the release of Ca 2+ from LELs into the cytosol via TRPML1 has been linked to a variety of physiological processes including activation of Transcription Factor EB (TFEB), lysosomal trafficking, exocytosis, phagosome formation, autophagy, and lysosome biogenesis (Venkatachalam, K. et al., ‘TRPML1 -Dependent Process as Therapeutic Targets.’ TRP Channels as Therapeutic Targets, edited by Arpad Szallasi, Academic Press, 2015, pp. 469-482).
  • TFEB Transcription Factor EB
  • TRPML1 Conductance of other ions such as Fe 2+ and Zn 2+ by TRPML1 have a significant role in trace metal regulation within lysosomal stores (Du et al., Cell Reports 2021, 37, 109848). Loss-of- function (LoF) in TRPML1 leads to Fe 2+ and Zn 2+ accumulation and signaling impairment in lysosomes.
  • TRPML1 protein is encoded by the MCOLN-1 gene (Di Paola, S. et al., Cell Calcium 2018, 69, 112). LoF mutations in MCOLN-1 can cause an autosomal-recessive lysosomal storage disease (LSD) Mucolipidosis type IV (MLIV).
  • the two predominant mutations resulting in MLIV are in the third intron causing a splice variant missing exons four to five and a ⁇ 6 kb deletion spanning exons six to seven. These two mutations are present in approximately 95% of patients diagnosed with MLIV, which causes severe developmental defects including cognitive impairment, motor function disability, and retinal degeneration. Patients harboring these mutations typically do not survive past adolescence. The underlying cause of these devastating symptoms is related to LoF TRPMLl-dependenent lysosomal storage and dysfunction, which lead to neuroinflammation and neurodegeneration.
  • Phosphatidylinositol-3,5-bisphosphate (PI(3,5)P2) is an endogenous lipid enriched in LELs that agonize TRPML1 activity (Dong X., et al., Nature Communications 2010, 1, 38).
  • PI(4,5)P2 phosphatidylinositol 4,5-bisphosphate
  • TRPML1 is potentiated by H + concentration where the optimal pH coincides with that of mature lysosomes (pH ⁇ 4.5) (Dong et al., 2008).
  • LoF of FIG4 leads to reduced PI(3,5)P2 levels and impaired TRPML1 function.
  • Variants in VAC14 a scaffold protein in the PI(3,5)P2-metabolism complex, are also associated with Parkinson’s Disease (PD) and pediatric-onset neurological disease.
  • Dysfunctions in TRPML1 are proposed as the underlying mechanism for those PI(3,5)P2-deficient mutations.
  • Preclinical studies demonstrate TRPML1 activation can be beneficial in those disease models suggesting the therapeutic potential for TRPML1 agonism in CMT, AES, PD, and other neurodegenerative diseases (Dong X., et al. Nature 2008, 455, 7215, 992.). [0007] TRPML1 has been linked to LSDs.
  • NP diseases are inherited and primarily caused by accumulation of lipids including sphingomyelin and cholesterol, which lead to severe neurological symptoms. Sphingomyelin accumulation in lysosomes inhibits TRPML1 activity.
  • TRPML1 Activation of TRPML1 in NP disease cell models can reduce lipid accumulation thereby restoring lysosomal trafficking of TRPML1 (Shen et al., Nature Communications 2012, 3, 731).
  • Increasing TPRML-1 activity through overexpression of TFEB increases lysosomal exocytosis and rescues pathogenesis in mucopolysaccharidosis in cell-culture and in vivo models of the disease.
  • Many LSDs are described by lysosome disfunction including trafficking, maturation, exocytosis, lipid and biomolecule accumulation, and Ca 2+ homeostasis.
  • TRPML1 plays a major role in these processes and therefore is an attractive target for treating LSDs (Di Paola et al., 2018).
  • Lysosomal function in neurons is associated with neurodegenerative diseases (Cesan et al., Experimental Cell Research 2012, 318, 11, 1245; Ghavami et al., Progress in Neurobiology 2014, 112, 24).
  • TRPML1 activity is linked to hallmarks of Alzheimer’s Disease (AD).
  • AD Alzheimer’s Disease
  • Patients infected with Human Immunodeficiency Virus (HIV) can develop premature cognitive declines correlated with accumulation of AB and sphingomyelin in cellular compartments.
  • TRPML1 Agonism of TRPML1 in an HIV- pgl20-induced sphingomyelin and AB accumulation model using primary neurons demonstrated the clearance of these biomolecules (Bae et al., Journal of Neuroscience 2014, 34, 34, 11485). Moreover, TFEB activation, a downstream effect of TRPML1 activation reduces Tau pathology in preclinical models (Polito et al., EMBO Molecular Medicine 2014, 6, 9, 1142). These studies suggest a therapeutic potential for TRPML1 in AD.
  • DMD Duchenne muscular dystrophy
  • dystrophin which weakens the plasma membranes of muscle cells.
  • Patients with DMD are subject to severe muscle damage during contraction (Yu et al., Science Advances 2020, 6, eaaz2736).
  • TRPML1 activation rescues these phenotypes in a DMD mouse model suggesting a therapeutic approach to broadly treating this heterogenous disease.
  • TRPML1 Targeting TRPML1 with small molecule modulators has been shown to rescue hallmark phenotypes of neurodegenerative diseases, LSDs, prion and prion-like diseases, and muscular dystrophy- related diseases in cell-based and animal disease models.
  • TRPML1 function is intricately involved in many important cellular processes and could be a therapeutic target for a multitude of human diseases.
  • TRPML1 modulators to rescue impaired lysosomal function and cellular autophagy, particularly in neurodegenerative diseases.
  • a pharmaceutical composition comprising a compound as described herein, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and a pharmaceutically acceptable carrier.
  • a method for treating a disease or condition mediated, at least in part, by TRPML1 comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • Cycloalkylalkyl refers to the group “cycloalkyl-alkyl-”.
  • Halo refers to atoms occupying group VIIA of the periodic table, such as fluoro, chloro, bromo, or iodo.
  • haloalkyl examples include, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1 ,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1 ,2-dibromoethyl, and the like.
  • Haloalkoxy refers to an alkoxy group as defined above, wherein one or more (e.g., 1 to 6 or 1 to 3) hydrogen atoms are replaced by a halogen.
  • Heteroatomic groups include, but are not limited to, -NR y -, -O-, -S-, -S(O)-, -S(O) 2 -, and the like, wherein R y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein.
  • heteroaryl includes 5-10 membered ring systems, 5-7 membered ring systems, or 5-6 membered ring systems, each independently having 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, oxygen, and sulfur.
  • fused-heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[l,5-a]pyridinyl, and imidazo[l,5-a]pyridinyl, where the heteroaryl can be bound via either ring of the fused system. Any aromatic ring, having a single or multiple fused rings, containing at least one heteroatom, is considered a heteroaryl regardless of the attachment to the remainder of the molecule (i.e., through any one of the fused rings). Heteroaryl does not encompass or overlap with aryl as defined above.
  • Heterocyclyl refers to a saturated or partially 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.
  • fused ring systems such as 6,7-dihydro-5H-cyclopenta[b]pyridinyl, decahydroquinazolinyl, 1,2,3,4-tetrahydroquinazolinyl, and 5,6,7, 8-tetrahydroquinazolinyl are 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 a cycloalkyl, an aryl, or heteroaryl ring, regardless of the attachment to the remainder of the molecule.
  • heterocyclyl has 2 to 20 ring carbon atoms (i.e., C 2-20 heterocyclyl), 2 to 12 ring carbon atoms (i.e., C 2-12 heterocyclyl), 2 to 10 ring carbon atoms (i.e., C 2-10 heterocyclyl), 2 to 8 ring carbon atoms (i.e., C 2-8 heterocyclyl), 3 to 12 ring carbon atoms (i.e., C 3-12 heterocyclyl), 3 to 8 ring carbon atoms (i.e., C 3-8 heterocyclyl), or 3 to 6 ring carbon atoms (i.e., C 3-6 heterocyclyl); having 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, sulfur, or oxygen.
  • ring carbon atoms i.e., C 2-20 heterocyclyl
  • 2 to 12 ring carbon atoms i
  • heterocyclyl groups include, e.g., azetidinyl, azepinyl, benzodioxolyl, benzo[b][l,4]dioxepinyl, 1,4- benzodioxanyl, benzopyranyl, benzodioxinyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-
  • heterocyclyl also includes “spiroheterocyclyl” when there are two positions for substitution on the same carbon atom.
  • spiro-heterocyclyl rings include, e.g., bicyclic and tricyclic ring systems, such as oxabicyclo[2.2.2]octanyl, 2-oxa-7-azaspiro[3.5]nonanyl, 2-oxa-6-azaspiro[3.4]octanyl, and 6-oxa-l-azaspiro[3.3]heptanyl.
  • fused-heterocyclyl rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3- c]pyridinyl, indolinyl, and isoindolinyl, where the heterocyclyl can be bound via either ring of the fused system.
  • Heterocyclylalkyl refers to the group “heterocyclyl-alkyl-.”
  • substituted means any of the above groups (i.e., alkyl, alkenyl, alkynyl, alkylene, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, and/or heteroalkyl) wherein at least one (e.g., 1 to 5 or 1 to 3) hydrogen atom is replaced by a bond to a non-hydrogen atom such as, but not limited to, halo, hydroxy, cyano, nitro, alkyl, alkenyl, alkynyl, alkoxy, heteroalkyl, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, oxo, thioxo, N-oxide, azido, oxime, thiocyanate, amidino, guanadino, cycloal
  • any compound or structure given herein is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. These forms of compounds may also be referred to as “isotopically enriched analogs.” Isotopically labeled compounds have structures depicted 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, ”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 3 H and 14 C are incorporated.
  • Deuterium labelled or substituted therapeutic compounds of the disclosure may have improved DMPK (drug metabolism and pharmacokinetics) properties, relating to distribution, metabolism, and excretion (ADME). Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life, reduced dosage requirements, and/or an improvement in therapeutic index.
  • An 18 F, 3 H, or ”C labeled compound may be useful for PET or SPECT or other imaging studies.
  • the compounds of this disclosure are capable of forming acid and/or base salts by virtue of the presence of amino, and/or carboxyl groups, or groups similar thereto.
  • “Pharmaceutically acceptable” or “physiologically acceptable” refer 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” or “physiologically acceptable salts” include, for example, salts with inorganic acids, and salts with organic acids.
  • 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.
  • Salts derived from organic acids include, e.g., acetic acid, propionic acid, gluconic 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 or organic bases.
  • Suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.
  • the compounds of the disclosure, or their pharmaceutically acceptable salts 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 (S)- or, as (D)- or (L)- for amino acids.
  • the present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms.
  • Optically active (+) and (-), (R )- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and/or 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, or mixtures thereof, and includes “enantiomers,” which refers to two stereoisomers whose molecules are nonsuperimposeable mirror images of one another.
  • Prodrugs include compounds described herein wherein a hydroxy, amino, carboxyl, or sulfhydryl group in a compound described herein is bonded to any group that may be cleaved in vivo to regenerate the free hydroxy, amino, or sulfhydryl group, respectively.
  • Examples of prodrugs include, but are not limited to esters (e.g., acetate, formate, and benzoate derivatives), amides, guanidines, carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups in compounds described herein, and the like. Preparation, selection, and use of prodrugs is discussed in T. Higuchi and V.
  • a compound of Formula I or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
  • X 1 is N or CR 7 ;
  • X 2 is N or CR 6 ; and the dashed bond is a double bond; or
  • X 2 is NR 2 ; R 1 is oxo; and the dashed bond is a single bond;
  • X 3 is N or CR 5 ;
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 3 is: ; wherein X is N or CH; each R 4 is independently hydroxy, halo, cyano, -CD3, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O)C, 6 alkyl, -C(O)OC, 6 alkyl, -C(O)NR a R b , -S(O) 2 C 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy;
  • R a and R b are each independently hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, -(CH 2 ) 0-2 -C 3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R 8 is independently C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkoxy, or C 3-7 cycloalkyl; or two R 8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C 3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C 3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substitute
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , -NR 12a C(O)OR 12 , or heteroaryl optionally substituted with one to five substituents independently selected from halo, hydroxy, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, or C 3-6 cycloalkyl; provided that when X is N, then R 10 is not -NR 12a C(O)R 12 or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl,
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3;
  • a compound of Formula I or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
  • X 1 is N or CR 7 ;
  • X 2 is N or CR 6 ; and the dashed bond is a double bond; or
  • X 2 is NR 2 ; R 1 is oxo; and the dashed bond is a single bond;
  • X 3 is N or CR 5 ;
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 2 is hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl
  • R 3 is: each R 4 is independently hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O) C 1-6 alkyl, -C(O)OC 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy; R 5 , R 6 and R 7 are each independently hydrogen, hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , or C 3-7 cycloalkyl;
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3;
  • X 1 is N or CR 7 ;
  • X 2 is N or CR 6 ; and the dashed bond is a double bond; or
  • X 2 is NR 2 ; R 1 is oxo; and the dashed bond is a single bond;
  • X 3 is N or CR 5 ;
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -0-, -S-, -C(0)-, -S(0)-, -S(0) 2 -, -C(0)0-*, -0C(0)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 1 is hydrogen, halo, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , -OR 11 , -SR 11 , -C(O)R 11 , -C(O)OR 11 , -S(O)R 11 , -S(O) 2 R 11 , -C(O)N(R 11 ) 2 , -NR 11 C(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O) 2 R 11 , -S(O)N(R 11 ) 2 , -S(O) 2 N(R 11 ) 2 , -NR 11 C(0)N(R 11 ) 2 , -NR 11 S(O)N(R 11 ) 2 ,
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C
  • X 1 is N or CR 7 . In certain embodiments, X 1 is N. In certain embodiments, X 1 is CR 7 .
  • X 2 is N or CR 6 ; and the dashed bond, i.e., the bond between X 2 and C(R’), is a double bond.
  • X 2 is N; and the dashed bond is a double bond.
  • X 2 is CR 6 ; and the dashed bond is a double bond.
  • X 2 is NR 2 ; R 1 is oxo; and the dashed bond, i.e., the bond between X 2 and C(R’), is a single bond.
  • X 3 is N or CR 5 . In certain embodiments, X 3 is N. In certain embodiments, X 3 is CR 5 .
  • a compound of Formula IA or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, X 2 , X 3 , R 1 , R 3 , R 4 , and q are each independently as defined herein.
  • a compound of Formula IB or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R 1 , R 3 , R 4 , and q are each independently as defined herein.
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 1 is hydrogen, halo, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , -OR 11 , -SR 11 , -C(O)R 11 , -C(O)OR 11 , -S(O)R 11 , -S(O) 2 R 11 , -C(O)N(R 11 ) 2 , -NR 11 C(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O) 2 R 11 , -S(O)N(R 11 ) 2 , -S(O) 2 N(R 11 ) 2 , -NR 11 C(O)N(R 11 ) 2 , -NR 11 S(O)N(R 11 ) 2
  • R 3 is: each R 4 is independently hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O)C 1-6 alkyl, -C(O)OC 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy;
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z 1a ; each R 12 is independently C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3;
  • a compound of Formula IC or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein:
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • R 3 is: each R 4 is independently hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O)C 1-6 alkyl, -C(O)OC 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy;
  • R a and R b are each independently hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, -(CH 2 ) 0-2 - C 3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R 8 is independently C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkoxy, or C 3-7 cycloalkyl; or two R 8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C 3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C 3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substitute
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2 -6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z 1a ; each R 12 is independently C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 1 is hydrogen, halo, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , -OR 11 , -SR 11 , -C(O)R 11 , -C(O)OR 11 , -S(O)R 11 , -S(O) 2 R 11 , -C(O)N(R 11 ) 2 , -NR 11 C(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O) 2 R 11 , -S(O)N(R 11 ) 2 , -S(O) 2 N(R 11 ) 2 , -NR 11 C(O)N(R 11 ) 2 , -NR 11 S(O)N(R 11 ) 2
  • R 2 is hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl
  • R 3 is: each R 4 is independently hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O)C 1-6 alkyl, -C(O)OC 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy;
  • R a and R b are each independently hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, -(CH 2 ) 0-2 -C 3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R 8 is independently C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkoxy, or C 3-7 cycloalkyl; or two R 8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C 3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C 3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substitute
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z 1a ; each R 12 is independently C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3;
  • a compound of Formula IE or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R 1 , R 3 , R 4 , and q are each independently as defined herein.
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 1 is hydrogen, halo, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , -OR 11 , -SR 11 , -C(O)R 11 , -C(O)OR 11 , -S(O)R 11 , -S(O) 2 R 11 , -C(O)N(R 11 ) 2 , -NR 11 C(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O) 2 R 11 , -S(O)N(R 11 ) 2 , -S(O) 2 N(R 11 ) 2 , -XR 11 C(O )N( R 11 ) 2 .
  • R a and R b are each independently hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, -(CH 2 ) 0-2 -C 3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R 8 is independently C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkoxy, or C 3-7 cycloalkyl; or two R 8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C 3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C 3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substitute
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, -O-C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z 1a ;
  • R 10 is -C(O)OR 12 , -NR 12a C(O)R 12 , -C(O)N(R 12 ) 2 , or -NR 12a C(O)OR 12 ; each R 11 is independently hydrogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z 1a ; each R 12 is independently C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 1-6 alkyl, C 3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five
  • Ring A is C 3-10 cycloalkyl.
  • Ring A is aryl
  • Ring A is heteroaryl
  • Ring A is monocyclic C 3-7 cycloalkyl, monocyclic heterocyclyl, phenyl, or monocyclic heteroaryl.
  • Ring A is monocyclic C 3-7 cycloalkyl.
  • Ring A is monocyclic heterocyclyl.
  • Ring A is bicyclic heterocyclyl.
  • Ring A is phenyl
  • Ring A is monocyclic heteroaryl.
  • Ring A is bicyclic heteroaryl.
  • Ring A is monocyclic C 3-7 cycloalkyl, 5- or 6-membered monocyclic heterocyclyl, phenyl, or 5- or 6-membered monocyclic heteroaryl. [0089] In certain embodiments, Ring A is 5- or 6-membered monocyclic heterocyclyl.
  • Ring A is 5- or 6-membered monocyclic heteroaryl.
  • Ring A is C 3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl.
  • Ring A is 4- or 5 -membered heterocyclyl or 5- or 6-membered membered heteroaryl.
  • Ring A is cyclopropyl, cyclobutyl, azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
  • Ring A is azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
  • Ring A is phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
  • Ring A is pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
  • Ring A is pyrazolyl, triazolyl, oxadiazolyl, or pyridyl; and at least one
  • R 4 is cyano
  • q is 0, 1, or 2. In certain embodiments, q is 0. In certain embodiments, q is 1. In certain embodiments, q is 2.
  • each R 4 is independently halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, -C(O)OC 1-6 alkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkyl, or C 1-6 alkoxy.
  • each R 4 is independently selected from cyano, C 1-6 alkyl, or -C(O)OC1 6 alkyl.
  • each R 4 at least one R 4 is cyano.
  • q is 1, 2, or 3; and at least one R 4 is cyano.
  • L is a bond
  • a compound of Formula IF or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein R 1 , R 3 , and each R 4 are independently as defined herein.
  • a compound of Formula IGG or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein q, R 1 , R 3 , and each R 4 are independently as defined herein.
  • R 11 is hydrogen or C 1-6 alkyl optionally substituted with one to five independently selected halo.
  • R 1 is hydrogen, halo, cyano, C 1-6 alkyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , or -OR 11 ; wherein the C 1-6 alkyl or C3 10 cycloalkyl is independently optionally substituted with one to five Z 1a .
  • R 1 is hydrogen, halo, cyano, C 1-6 alkyl, C 3-10 cycloalkyl, -N(R 11 ) 2 , or -OR 11 ; wherein the C 1-6 alkyl or C310 cycloalkyl is independently optionally substituted with one to five Z 1a . In certain embodiments, R 1 is hydrogen.
  • R 3 is
  • n is 1. In certain embodiments, m is 2.
  • n is 1. In certain embodiments, n is 2. In certain embodiments, m is 3.
  • n + n is 2. In certain embodiments, m and n are both 1.
  • m2 is 1 or 2.
  • each R 8 is independently C 1-6 alkyl.
  • p is 1 or 2; and each R 8 is methyl.
  • p is 2; and each R 8 is methyl.
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five substituents independently selected from hydroxy, halo, cyano, C 1-6 alkyl, -O-C 1-6 alkyl, C 3-7 cycloalkyl, or -S(O) 2 C 1-6 alkyl; wherein each C 1-6 alkyl, -O-C 1-6 alkyl, C 3-7 cycloalkyl, or -S(O) 2 C 1-6 alkyl is independently optionally substituted with one to five independently selected halo.
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z 1a .
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z 1a ; and each Z 1a is independently halo, cyano, C 1-6 alkyl, or -OR 13 .
  • R 9 is C 1-6 alkyl, C 1-6 alkoxy, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 alkoxy, or C 3-7 cycloalkyl is independently optionally substituted with one to five halo, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, or C 3-7 cycloalkyl.
  • R 9 is C 1-6 alkyl.
  • R 9 is C 1-6 alkyl optionally substituted with one to five halo, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, or C 3-7 cycloalkyl. In certain embodiments, R 9 is C 1-6 alkyl.
  • a compound of Formula II or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein each X 2 , X 3 , L, Ring A, R 1 , R 4 , R 8 , R 9 , m, n, p, and q are independently as defined herein.
  • X 3 is N or CR 5 ;
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • X 2 is N; and the dashed bond is a double bond.
  • X 2 is CR 6 ; and the dashed bond is a double bond.
  • R 6 is hydrogen, hydroxy, halo, or C 3-7 cycloalkyl.
  • X 3 is N.
  • X 3 is CR 7 .
  • R 7 is hydrogen
  • X 1 is N; and X 3 is CH.
  • n and n are each 1.
  • a compound of Formula IID or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein p, R 1 , R 9 , each R 4 , and each R 8 are independently as defined herein.
  • a compound of Formula IIE or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein R 9 and each R 4 are independently as defined herein; and ring B is R 1 is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is independently optionally substituted with one to five Z 1a .
  • X 2 is N or CR 6 ;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 6 is hydrogen, hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , or C 3-7 cycloalkyl;
  • R a and R b are each independently hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, -(CH 2 ) 0-2 -C 3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R 8 is independently C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl, C 1-6 haloalkyl, or C 3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C 1-6 alkoxy, or C 3-7 cycloalkyl; or two R 8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C 3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C 3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substitute
  • X 2 is N or CR 6 ;
  • L is a bond, C 1-2 alkylene, C 2 alkenylene, C 2 alkynylene, -NR 11 -, -O-, -S-, -C(O)-, -S(O)-, -S(O) 2 -, -C(O)O-*, -OC(O)-*, -C(O)NR 11 -*, -NR 11 C(O)-*, -NR 11 S(O)-*, -NR 11 S(O) 2 -*, -S(O)NR 11 -*, -S(O) 2 NR 11 -*, -NR 11 C(O)NR 11 -, -NR 11 S(O)NR 11 -, -NR 11 S(O) 2 NR 11 -, -OC(O)NR 11 -*, or -NR 11 C(O)O-*; wherein the * bond is attached to Ring A;
  • Ring A is C 3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
  • R 1 is hydrogen, halo, cyano, nitro, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , -OR 11 , -SR 11 , -C(O)R 11 , -C(O)OR 11 , -S(O)R 11 , -S(O) 2 R 11 , -C(O)N(R 11 ) 2 , -NR 11 C(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O)R 11 , -NR 11 S(O) 2 R 11 , -S(O)N(R 11 ) 2 , -S(O) 2 N(R 11 ) 2 , -XR 11 C(O )N( R 11 ) 2 .
  • each R 4 is independently hydroxy, halo, cyano, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylthio, -NR a R b , -C(O)C 1-6 alkyl, -C(O)OC 1-6 alkyl, C 3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo
  • R 12a is hydrogen, C 1-6 alkyl, or C 3-7 cycloalkyl; wherein the C 1-6 alkyl or C 3-7 cycloalkyl is independently optionally substituted with one to five Z 1a ; or two R 12 , R 12 and R 12a , or R 12a and an R 8 , together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z 1b ; m3 is 0 or 1 ; n3 is 0, 1, or 2; wherein m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z 1a is independently halo, cyano, -NO 2 , C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 13
  • L is a bond
  • Ring A is C 3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl
  • at least one R 4 is cyano
  • each R 8 is independently C 1-6 alkyl.
  • L is a bond
  • Ring A is C 3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl
  • R 1 is hydrogen, halo, cyano, C 1-6 alkyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , or -OR 11 ; wherein the C 1-6 alkyl or C 3-10 cycloalkyl is independently optionally substituted with one to five Z 1a ; at least one R 4 is cyano; each R 8 is independently C 1-6 alkyl; and R 9 is C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl; wherein the C 1-6 alkyl, C 1-6 alkoxy, C 3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z 1a
  • L is a bond
  • Ring A is C 3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl
  • R 1 is hydrogen, halo, cyano, C 1-6 alkyl, C 3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R 11 ) 2 , or -OR 11 ; wherein the C 1-6 alkyl or C 3-10 cycloalkyl is independently optionally substituted with one to five Z 1a ; at least one R 4 is cyano; each R 8 is independently C 1-6 alkyl; and R 9 is C 1-6 alkyl, C 3-7 cycloalkyl, or heterocyclyl; wherein the C 1-6 alkyl, C 3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z 1a .
  • a compound selected from Table 1 or a pharmaceutically acceptable salt, isotopically enriched analog, prodrug, stereoisomer, or a mixture of stereoisomers thereof:
  • a compound selected from Table 2 or a pharmaceutically acceptable salt thereof is provided.
  • TRP Transient receptor potential channels are integral membrane proteins consisting of a superfamily of cation channels that allow permeability of both monovalent and divalent cations. TRP channels are subdivided into six subfamilies: TRPC, TRPV, TRPM, TRPP, TRPML, and TRPA, and are expressed in almost every cell and tissue. TRPs play an instrumental role in the regulation of various physiological processes. TRP channels are extensively represented in brain tissues and are present in both prokaryotes and eukaryotes, exhibiting responses to several mechanisms, including physical, chemical, and thermal stimuli.
  • TRP channels are involved in the perturbation of Ca 2+ homeostasis in intracellular calcium stores, both in neuronal and non-neuronal cells, and its discrepancy leads to several neuronal disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), and Amyotrophic lateral sclerosis (ALS). TRPs participate in neurite outgrowth, receptor signaling, and excitotoxic cell death in the central nervous system.
  • AD Alzheimer’s disease
  • PD Parkinson’s disease
  • HD Huntington’s disease
  • ALS Amyotrophic lateral sclerosis
  • prevention means any treatment of a disease or condition that causes the clinical symptoms of the disease or condition not to develop.
  • Compounds may, in certain embodiments, be administered to a subject (including a human) who is at risk or has a family history of the disease or condition.
  • terapéuticaally effective amount or “effective amount” of a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof means an amount sufficient to effect treatment when administered to a subject, to provide a therapeutic benefit such as amelioration of symptoms or slowing of disease progression.
  • a therapeutically effective amount may be an amount sufficient to decrease a symptom of a disease or condition of as described herein.
  • the therapeutically effective amount may vary depending on the subject, and disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the manner of administering, which can readily be determined by one of ordinary skill in the art.
  • the methods described herein may be applied to cell populations in vivo or ex vivo.
  • “In vivo” means within a living individual, as within an animal or human. In this context, the methods described herein may be used therapeutically in an individual.
  • “Ex vivo” means outside of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples including fluid or tissue samples obtained from individuals. Such samples may be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein may be used for a variety of purposes, including therapeutic and experimental purposes.
  • modulation of TRPML1 comprises activation of TRPML1.
  • a method of treating a disease or disorder that can be treated by activation of TRPML1 comprising administering to a subject in need thereof a compound described herein (e.g., a compound of Formula I, or subformula thereof) or pharmaceutically acceptable salts, solvates, hydrates, tautomers, stereoisomers, isotopically labeled derivatives thereof, or a composition described herein.
  • a compound described herein e.g., a compound of Formula I, or subformula thereof
  • the compounds provided herein e.g., a compound of Formula I, or subformula thereof, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, activates TRPML1.
  • a method of activating TRPML1 activity comprising contacting a cell with an effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • the activating can be in vitro or in vivo.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof for use in activating TRPML1 activity (e.g., in vitro or in vivo).
  • a method for treating a disease or disorder mediated, at least in part, by TRPML1 comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof to a subject in need thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof for use in treating a disease or disorder mediated, at least in part, by TRPML1 in a subject in need thereof.
  • a method of treating a disease or disorder which can be treated by modulation of lysosomes comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof to a subject in need thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof
  • TRPML1 The transient receptor potential channel mucolipin 1 (TRPML1) is a cation channel that can transport Ca 2+ , Fe 2+ and Zn 2+ .
  • the primary function of TRPML1 is to induce Ca 2+ release from endolysosomal compartments, a vital process for endolysosomal function.
  • Dysfunction in TRPML1- relevant endolysosomal pathways have been widely observed in preclinical and clinical samples for late- onset neurodegenerative diseases.
  • TRPML1 The functions regulated by TRPML1 include lysosomal biogenesis dependent on TFEB family, maturation of late endosomes to lysosomes; endolysosomal trafficking; nutrient sensing and adaptation; positioning, exocytosis, fission, clearance, and reformation of lysosomes; autophagy, phagocytosis and clearance of aggregated proteins and pathogens.
  • Loss-of- function mutations in the human TRPML1 gene MCOLN1 cause mucolipidosis type IV (MLIV), a rare recessive lysosomal storage disorder (LSD).
  • MLIV is characterized by neurodegeneration, psychomotor impairment, ophthalmologic and gut defects.
  • AD Alzheimer’ s disease
  • the underlying pathology involves the accumulation of extracellular amyloid aggregates called senile plaques and intracellular neurofibrillary tangles, leading to the selective loss of synapses and neurons in the hippocampal and cerebral cortical regions. While the amyloid and tau hypotheses have been the traditional theories explaining AD, the dysregulation of calcium homeostasis has recently gained attention as a critical factor in AD pathogenesis.
  • Calcium is an essential intracellular messenger that binds to multiple proteins, receptors, and ion channels to regulate various physiological functions. During neurodegeneration, neurons become inefficient in regulating calcium levels. Enhanced pathological lesions induce neurotoxicity and cytokines, leading to dysregulation of calcium homeostasis and leaving neurons prone to excitotoxicity and apoptosis.
  • a method for treating Alzheimer’s disease comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. Parkinson’s disease
  • Parkinson’ s disease is the second most predominant neurodegenerative disorder related to aging after AD. It is a movement disorder, depicted by the disintegration of dopaminergic neurons (DNs) in the substantia nigra (SN), that worsens over time. Eewy bodies are the cytoplasmic inclusions formed by a-synuclein protein and are considered the main pathological hallmark of PD.
  • DNs dopaminergic neurons
  • SN substantia nigra
  • Eewy bodies are the cytoplasmic inclusions formed by a-synuclein protein and are considered the main pathological hallmark of PD.
  • Several factors in PD can lead to the loss of DNs in SN, such as oxidative stress, mitochondrial dysfunction, protein aggregation, and alteration in calcium homeostasis. Studies reported that TRP channels could facilitate some of the mechanisms that advance the progression of the disease.
  • Huntington’s disease is an autosomal neurodegenerative disorder triggered by polyglutamine expansion in Huntingtin protein and represented by cognitive impairment, medium spiny neurons (MSN) loss in the striatum, and convulsive movements.
  • Various channels have been reported to alter the K + homeostasis, such as the Kir4.1 channel expressing striatal astrocytes in mutated HTT protein that disintegrates the extracellular K + homeostasis hence provokes hyperexcitability in neurons, i.e., HD motor symptoms in striatal neurons.
  • the normal Kir4.1 channel is a prevalent astrocytic K + channel that plays a prominent role in balancing the cells resting membrane potential and buffering K + ions in the brain.
  • HD mHTT protein modifies the high voltage stimulated Ca 2+ channels. Besides dysfunction in the Ca 2+ channel, other ion channels have also exhibited reduced expression in several HD mouse models. Thus, modification in these ion channels disintegrates the ion homeostasis in cortical pyramidal neurons, due to which synaptic integration, neurotransmitter release, and genetic expression gets affected, which plays a central role in cortical dysfunction in HD.
  • a method for treating Huntington’s disease comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • ALS Amyotrophic lateral sclerosis
  • AVCCs voltage-gated calcium channels
  • a method for treating amyotrophic lateral sclerosis comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • the lysosomal storage disease or disorder is selected from the group consisting of Niemann-Pick disease, Gaucher’s disease, neuronopathic Gaucher’s disease, neuronal ceroid lipofuscinosis, sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, gangliosidoses, Gaucher Disease, Lysosomal acid lipase deficiency, sulfatidoses, mucopolysaccharidoses, mucolipidoses, lipidoses, and oligosaccharidoses.
  • the lysosomal storage disorder is selected from the group consisting of sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, Fabry disease, Schindler disease, beta-galactosidase disorder, GM1 gangliosidosis, GM2 gangliosidosis AB variant, GM2 gangliosidosis activator deficiency, Sandhoff disease, Tay- Sachs disease, Gaucher disease, lysosomal acid lipase deficiency, Niemann-Pick disease, metachromatic leukodystrophy, Saposin B deficiency, multiple sulfatase deficiency, Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome, Hunter syndrome, Sanfilippo syndrome, Morquio syndrome, Maroteaux-Lamy syndrome, Sly syndrome, hyaluronidase deficiency, sialidosis, I-cell disease, pseudo-Hurler polydystrophy
  • a method for treating a glycogen storage disease or disorder comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a compound as disclosed herein e.g., a compound of Formula I, or subformula thereof
  • a pharmaceutically acceptable salt isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • the glycogen storage disease or disorder is selected from the group consisting of Pompe disease and Danon disease.
  • a method of treating a ciliopathy comprising administering to a subject in need thereof a therapeutically effective amount of a compound capable of modulating TRPML, or a therapeutically effective amount of a pharmaceutical composition comprising the compound and a pharmaceutically acceptable excipient.
  • the ciliopathy is selected from the group consisting of polycystic kidney disease, pancreatic cysts in polycystic kidney disease, Bardet-Biedl syndrome, nephronophthisis, Joubert Syndrome, Meckel-Gruber Syndrome, oral-facial- digital syndrome, Senior Loken Syndrome, Birt-Hogg- Dube syndrome, Leber’s congenital amaurosis, Alstrom syndrome, Jeune asphyxiating thoracic dystrophy, Ellis van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia.
  • ambroxol and afegostat amantadine
  • agents capable of treating Alzheimer’s e.g., acetylcholinesterase inhibitors such as tacrine, rivastigmine, galantamine, donepezil, and NMD A receptor antagonists such as memantine).
  • kits that include a compound of the disclosure, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and suitable packaging.
  • a kit further includes instructions for use.
  • a kit includes a compound of the disclosure, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and a label and/or instructions for use of the compounds in the treatment of the indications, including the diseases or conditions, described herein.
  • articles of manufacture that include a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof in a suitable container.
  • the container may be a vial, jar, ampoule, preloaded syringe, or intravenous bag.
  • compositions that contain one or more of the compounds described herein, or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, or prodrug thereof, and one or more pharmaceutically acceptable vehicles selected from carriers, adjuvants, and excipients.
  • suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers, and adjuvants.
  • Such compositions are prepared in a manner well known in the pharmaceutical art.
  • the pharmaceutical compositions may be administered in either single or multiple doses.
  • the pharmaceutical composition may be administered by various methods including, for example, rectal, buccal, intranasal, and transdermal routes.
  • the pharmaceutical composition may be administered by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.
  • One mode for administration is parenteral, for example, by injection.
  • the forms in which the pharmaceutical compositions described herein may be incorporated for administration by injection include, for example, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles.
  • Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets.
  • the active ingredient is usually diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper, or other container.
  • a carrier that can be in the form of a capsule, sachet, paper, or other container.
  • the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient.
  • compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.
  • excipients include, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose.
  • the formulations can additionally include lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxy- benzoates; sweetening agents; and flavoring agents.
  • the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • a pharmaceutical excipient for preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
  • the active ingredient may be dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules.
  • the tablets or pills of the compounds described herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach.
  • the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former.
  • the two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release.
  • enteric layers or coatings such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
  • compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable, aqueous, or organic solvents, or mixtures thereof, and powders.
  • the liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein.
  • the compositions are administered by the oral or nasal respiratory route for local or systemic effect.
  • compositions in pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, in one embodiment, orally or nasally, from devices that deliver the formulation in an appropriate manner.
  • the amount of the compound in a pharmaceutical composition or formulation can vary within the full range employed by those skilled in the art. Typically, the formulation will contain, on a weight percent (wt %) basis, from about 0.01-99.99 wt % of a compound of this disclosure based on the total formulation, with the balance being one or more suitable pharmaceutical excipients. In one embodiment, the compound is present at a level of about 1-80 wt %. Representative pharmaceutical formulations are described below.
  • a suppository of total weight 2.5 g is prepared by mixing the compound of this disclosure with Witepsol® H-15 (triglycerides of saturated vegetable fatty acid; Riches-Nelson, Inc., New York), and has the following composition:
  • a dosage of from about 0.0001 to about 100 mg per kg of body weight per day, from about 0.001 to about 50 mg of compound per kg of body weight, or from about 0.01 to about 10 mg of compound per kg of body weight may be appropriate.
  • body weight is particularly useful when adjusting dosages between subjects of widely disparate size, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.
  • the compounds may be prepared using the methods disclosed herein and routine modifications thereof, which will be apparent given the disclosure herein and methods well known in the art. Conventional and well-known synthetic methods may be used in addition to the teachings herein. The synthesis of typical compounds described herein may be accomplished as described in the following examples. If available, reagents and starting materials may be purchased commercially, e.g., from Sigma Aldrich or other chemical suppliers.
  • protecting groups for alcohols include silyl ethers (including trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso- propylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ethers), which can be removed by acid or fluoride ion, such as NaF, TBAF (tetra-n-butylammonium fluoride), HF-Py, or HF-NEt3.
  • TMS trimethylsilyl
  • TDMS tert-butyldimethylsilyl
  • TOM tri-iso- propylsilyloxymethyl
  • TIPS triisopropylsilyl
  • Other protecting groups for alcohols include acetyl, removed by acid or base, benzoyl, removed by acid or base, benzyl, removed by hydrogenation, methoxyethoxymethyl ether, removed by acid, dimethoxytrityl, removed by acid, methoxymethyl ether, removed by acid, tetrahydropyranyl or tetrahydrofuranyl, removed by acid, and trityl, removed by acid.
  • protecting groups for amines include carbobenzyloxy, removed by hydrogenolysis p-methoxybenzyl carbonyl, removed by hydrogenolysis, tert-butyloxycarbonyl, removed by concentrated strong acid (such as HC1 or CF3COOH), or by heating to greater than about 80 °C, 9-fluorenylmethyloxycarbonyl, removed by base, such as piperidine, acetyl, removed by treatment with a base, benzoyl, removed by treatment with a base, benzyl, removed by hydrogenolysis, carbamate group, removed by acid and mild heating, p-methoxybenzyl, removed by hydrogenolysis, 3,4-dimethoxybenzyl, removed by hydrogenolysis, p-methoxyphenyl, removed by ammonium cerium(IV) nitrate, tosyl, removed by concentrated acid (such as HBr or H2SO4) and strong reducing agents (sodium in liquid ammonia or sodium naphthalenide
  • the compounds of this disclosure may contain one or more chiral centers. Accordingly, if desired, such compounds can be prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or diastereomers or as stereoisomer-enriched mixtures. All such stereoisomers
  • stereoisomers are included within the scope of this disclosure, unless otherwise indicated.
  • Pure stereoisomers may be prepared using, for example, optically active starting materials or stereoselective reagents well-known in the art.
  • racemic mixtures of such compounds can be separated using, for example, chiral column chromatography, chiral resolving agents, and the like.
  • the starting materials for the following reactions are generally known compounds or can be prepared by known procedures or obvious modifications thereof.
  • many of the starting materials are available from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA), Bachem (Torrance, California, USA), Emka-Chemce or Sigma (St. Louis, Missouri, USA).
  • compounds of Formula I can be prepared by contacting compound 1-1 with an amine 1-2 under suitable coupling reaction conditions, followed by optional functionalization or deprotection when required. Upon reaction completion, compounds of Formula I can be recovered and purified by conventional techniques such as neutralization, extraction, precipitation, chromatography, filtration and the like.
  • Scheme II shows an exemplary methods for further derivatization of R 3 , for Example.
  • each R 8 , R 9 , p, m, n, ml, m2, nl, and n2 are independently as defined herein, and LG is a leaving group (e.g., halo, alkoxy, etc.).
  • a process for providing a compound of Formula I comprising contacting a compound of Formula 1-1 : with a compound of Formula 1-2: under conditions sufficient to provide the compound of Formula I; wherein Ring A, L, X 1 , X 2 , X 3 , the dashed line, R 1 , R 3 , R 4 , R 8 , and q are each independently as defined herein.
  • (S)-2-cyano-3,3-difluoro-2- methylpropanoic acid (Intermediate 4a): To a solution of Intermediate 3a (5 g, 12.90 mmol) in MeCN (60 mL) and H 2 O (20 mL) was added CsF (3.92 g, 25.81 mmol) at 20 °C and stirred for 1 h. NaOH (1.03 g, 25.81 mmol) in H 2 O (8 mL) was added to the reaction and stirred at 20 °C for 1 h. Seven batches were combined and the combined reaction mixture was poured into water (I L). The reaction mixture was extracted with DCM (600 mL x 4).
  • (2R,5S)-tert -butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate To a solution of (2R,5S)-tert-butyl 4-(5-bromo-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (350 mg, 0.77 mmol) and cyclopropylboronic acid (66 mg, 0.77 mmol) in dioxane (3 mL) was added Pd(dppf)Ch (56 mg, 0.07 mmol) and K3PO4 (489 mg, 2.30 mmol), Ag2O (89 mg, 0.38 mmol) under N 2 .
  • Pd(dppf)Ch 56 mg, 0.07 mmol
  • K3PO4 489 mg, 2.30 mmol
  • Ag2O 89 mg, 0.38 mmol
  • (2R ,5S)-tert -butyl 4-(5-cyclopropyl-8-phenylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (220 mg, 0.53 mmol) and phenylboronic acid (320 mg, 2.64 mmol) in dioxane (5 mL) and H 2 O (0.2 mL) was added Pd(dppf)Ch (77 mg, 0.11 mmol) and CS 2 CO 3 (516 mg, 1.58 mmol).
  • (2R ,5.S)-tert -butyl 4-(6-bromo-8-(4-cyanopyridin-2-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of (2R,5S)-tert-butyl 4-(6-bromo-8-iodoquinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.15 mmol) and 2-(trimethylstannyl)isonicotinonitrile (94 mg, 0.18 mmol) in DMF (1 mL) was added Pd(PPh 3 ) 4 (34 mg, 0.029 mmol) at 20 °C under N 2 .
  • (2R,5S)-tert -butyl 2,5-dimethyl-4-(8-phenylquinazolin-4-yl)piperazine-l-carboxylate To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.19 mmol) and phenylboronic acid (116 mg, 0.95 mmol) in dioxane (1 mL) and H 2 O (0.2 mL) was added Pd(dppf)Ch (28 mg, 0.038 mmol) and CS 2 CO 3 (186 mg, 0.57 mmol) at 20 °C under N 2 .
  • the mixture was stirred at 50 °C for 2 h.
  • the reaction mixture was diluted with H 2 O (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • 3-amino-2-chloroisonicotinic acid To a solution of methyl 3-amino-2-chloroisonicotinate (2 g, 10.72 mmol) in MeOH (10 mL) and H 2 O (10 mL) was added NaOH (1.16 g, 28.94 mmol). The mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove MeOH. The aqueous solution was adjusted pH to 3 ⁇ 4 by 3N HC1. The precipitate was filtered and dried under reduced pressure to give 3-amino-2-chloroisonicotinic acid (1.7 g, 92%).
  • 8-chloropyrido[3,4-d]pyrimidin-4(3H)-one To a solution of 3-amino-2-chloroisonicotinic acid (500 mg, 2.90 mmol) in n-BuOH (10 mL) was added formamidine acetate (1.51 g, 14.49 mmol) at 20 °C. The mixture was stirred at 140 °C for 40 h. The reaction mixture was filtered and the solid was collect o give 8-chloropyrido[3,4- d]pyrimidin-4(3H)-one (600 mg, 57%).
  • (2R,5S)-tert-butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)pyrido[3,4-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of (2R,5S)-tert-butyl 4-(8-chloropyrido[3,4- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (70 mg, 0.19 mmol) and 1H-pyrazole-4- carbonitrile (51.73 mg, 0.55 mmol) in DMF (1.5 mL) was added K 2 CO 3 (64 mg, 0.46 mmol), (15,25)- N 1 ,N 2 -dimcthylcyclohcxanc- l ,2-diaminc (8 mg, 0.06 mmol), and Cui (4 mg, 0.02 mmol) at 20
  • the mixture was stirred to 110 °C for 16 h.
  • the reaction mixture was diluted with H 2 O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • the mixture was stirred at 100 °C for 16 h.
  • the reaction mixture was diluted with water (5 mL), extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure.
  • 8-bromo-4-chloropyrido[4,3-d]pyrimidine To a solution of 8-bromopyrido[4,3-d]pyrimidin- 4(3H)-one (1.6 g, 7.08 mmol) in SOCh (14 mL) was added DMF (517 mg, 7.08 mmol) at 20 °C under N 2 . The mixture was stirred at 85 °C for 16 h. The reaction mixture was concentrated under reduced pressure.
  • trans-(2R,5S)-tert-butyl 4-(8-chloro-5-(2-(ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (300 mg, 0.66 mmol) and trans-2- (ethoxycarbonyl)cyclopropyl)boraneylidene)- ⁇ 3-fluoraneyl)potassium(III) fluoride (174 mg, 0.79 mmol) in H 2 O (0.6 mL) and toluene (2.4 mL) was added K3PO4 (279 mg, 1.32 mmol) and [2-(2- aminophenyl)phenyl]-chloro-palladium
  • (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H -l,2,4-triazol-3-yl)-5-cyclopropylquinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of (2R,5S)-tert-butyl 4-(5-cyclopropyl-8- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 0.30 mmol) in 1,4-dioxane (5 mL) and H 2 O (0.5 mL) was added 5-bromo-2-methyl-l,2,4-triazole-3- carbonitrile (110 mg, 0.59 mmol), K 2 CO 3 (122 mg, 0.89 mmol) and Pd(dppf
  • the mixture was stirred at 20 °C for 3 h.
  • the reaction mixture was quenched by water (3 mL) at 20 °C, adjusted pH to 7 ⁇ 8 using sat. NaHCO 3 . extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure.
  • the mixture was stirred at 20 °C for 16 hr.
  • the reaction mixture concentrated under reduced pressure to remove THF.
  • Water (5 mL) was added to the remaining mixture and extracted with MTBE (5 ml).
  • the aqueous phase was adjusted to pH 4-5 by IN HC1 and extracted with EtOAc (3 x 10 mL).
  • (2R,5S)-tert butyl 4-(8-chloro-5-methoxyquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-hydroxyquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (200 mg, 0.51 mmol) in THF (4 mL) was added K 2 CO 3 (140 mg, 1.02 mmol) and Mel (108 mg, 0.763 mmol). The mixture was stirred at 40 °C for 4 h. The reaction mixture was concentrated under reduced pressure.
  • (2R,5S)-tert -butyl 4-(8-bromo-5-(dimethylamino)pyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of (2R,5S)-tert-butyl 4-(5-amino-8-bromopyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (100 mg, 0.23 mmol) in DMF (2 mL) was added NaH (22 mg, 0.57 mmol) at 0 °C under N 2 .
  • the suspension was degassed and purged with N 2 for 3 times. The mixture was stirred at 100 °C for 2 h. The reaction mixture was poured into water (5mL) and extracted with EtOAc (3 x 3 mL). The combined organic phase was concentrated under reduced pressure.
  • the suspension was degassed and purged with N 2 3 times.
  • the mixture was stirred at 80 °C for 2 h under N 2 .
  • the reaction mixture was poured into water (2 mL) and extracted with EtOAc (3 x 1 mL). The combined organic phase was concentrated under reduced pressure.
  • Example 53A & 53B 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lS)- 2,2-difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4-[2- cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lR)-2,2- difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile [0368] 3-((2R,5S)-4-(5-bromo-8-iodo-quinazolin-4-yl)-2,5-dimethyl-piperazin
  • (2.S,5R )-tert -butyl 4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazine- 1-carboxylate To a solution of (2S,5R)-tert-butyl 4-(2-cyano-3-hydroxy-2-(hydroxymethyl)propanoyl)- 2,5-dimethylpiperazine-l -carboxylate (1.5 g, 4.39 mmol) in DCM (20 mL) was added dropwise DAST (2.12 g, 13.18 mmol) at -40 °C under N 2 . The mixture was stirred at 20 °C for 12 h.
  • 5-bromo-6-fluoro-8-iodoquinazolin-4-ol To a solution of 2-amino-6-bromo-5-fluoro-3-iodo- benzoic acid (700 mg, 1.94 mmol) in n-BuOH (10 mL) was added formamidine acetate (1.21 g, 11.67 mmol) at 20 °C. The mixture was stirred at 140 °C for 1 h in sealed tube. The residue was poured into H 2 O (20 mL). The suspension was filtered and the filter cake was dried under reduced pressure to give 5- bromo-6-fluoro-8-iodoquinazolin-4-ol (570 mg, 79%).
  • 5-bromo-4-chloro-6-fluoro-8-iodoquinazoline To a solution of 5-bromo-6-fluoro-8- iodoquinazolin-4-ol (450 mg, 1.22 mmol) in toluene (9 mL) was added POCl 3 (748 mg, 4.88 mmol) and DIEA (315 mg, 2.44 mmol) at 20 °C. The mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give 5-bromo-4-chloro-6-fluoro-8-iodoquinazoline (670 mg, crude), which was used directly in the next step.
  • 5-bromo-8-iodoquinazolin-4-ol To a solution of 2-amino-6-bromo-3-iodo-benzamide (17.9 g, 52.50 mmol) in diethoxymethoxyethane (110 mL) was added p-TsOH (904 mg, 5.25 mmol) at 20 °C. The mixture was stirred at 120 °C for 1 h. The residue was poured into H 2 O (60 mL). The suspension was filtered and the filter cake was dried under reduced pressure to give 5-bromo-8-iodoquinazolin-4-ol (14.5 g, 78% yield).
  • 5-bromo-4-chloro-8-iodoquinazoline To a solution of 5-bromo-8-iodoquinazolin-4-ol (5 g, 14.25 mmol) in toluene (50 mL) was added POCl 3 (8.74 g, 56.99 mmol) and DIEA (3.68 g, 28.50 mmol) at 20 °C. The mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give crude 5-bromo-4-chloro-8-iodoquinazoline (5.26 g), which was used into next step directly.
  • tert-butyl (2R,5S)-4-(5-bromo-8-(4-(ethoxycarbonyl)-5-methoxy-1H -pyrazol-l- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2A , ,5S’)-4-(5- bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 0.92 mmol) and ethyl 3- methoxy-1H-pyrazole-4-carboxylate (311 mg, 1.83 mmol) in DMF (6 mL) was added K 2 CO 3 (253 mg, 1.83 mmol) and Cui (69 mg, 0.37 mmol) A ⁇ l ,N 2 -dimcthylcyclohcxanc-l ,2-
  • tert-butyl (2R,5S)-4-(8-(4-(ethoxycarbonyl)-5-methoxy-1H -pyrazol-l-yl)-5-(5-fluoropyridin- 3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2R.5S)-4-(5- bromo-8-(4-(ethoxycarbonyl)-5-methoxy- 1H-pyrazol-l -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (860 mg, 1.46 mmol) and (5-fluoro-3-pyridyl)boronic acid (411 mg, 2.92 mmol) in 1,4- dioxane (10 mL) was added Pd(dppf)Cl 2 -CH 2 Cl 2 (119 mg, 0.15
  • tert-butyl (2R,5S)-4-(8-(4-cyano-5-methoxy-1H -pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2R,5S)-4-(8-(4- carbamoyl -5-methoxy-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (270 mg, 0.46 mmol) in THF (5 mL) was added methoxycarbonyl- (triethylammonio)sulfonyl-azanide (557 mg, 2.34 mmol).
  • Example 153 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- pyrrolidin-l-yl-quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
  • tert-butyl (2R,5S)-4-(8-chloro-5-(pyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate To a mixture of tert-butyl (2R,5S)-4-(5-amino-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (210 mg, 0.54 mmol) in DMF (3 mL) was added NaH (47 mg, 1.18 mmol, 60% purity) at 0 °C under N 2 .
  • tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(pyrrolidin-l-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-(pyrrolidin- l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (200 mg, 0.45mmol) and l-methyl-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (209 mg, 0.90 mmol) in 1,4- dioxane (5 mL) and H 2 O (1 mL) was added K3PO4 (190 mg, 0.90 mmol
  • tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(3-fluorophenyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 0.95 mmol) and (3-fluorophenyl)boronic acid (399 mg, 2.85 mmol) in 1,4-dioxane (10 mL) was added Pd(dppf)Cl 2 .
  • tert-butyl (2R,5S)-4-(5-(4-bromobutanamido)-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of tert-butyl (2R,5S)-4-(5-amino-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (840 mg, 2.14 mmol) and 4-bromobutanoyl chloride (596 mg, 3.22 mmol) in THF (10 mL) was added K 2 CO 3 (740 mg, 5.36 mmol).
  • tert-butyl (2R,5,S)-4-(8-chloro-5-(2-oxopyrrolidin-l-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of tert-butyl (2R .5.S')-4-(5-(4-bromobutanamido)-8- chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (860 mg, 1.59 mmol) in DMF (10 mL) was added NaH (287 mg, 7.17 mmol) at 0°C . The mixture was stirred at 20°C for 1 h.
  • the reaction mixture was poured into ice saturated NH4CI solution (20 mL).
  • the reaction mixture was poured into H 2 O (10 mL).
  • the aqueous phase was extracted with EtOAc (3 x 20 mL).
  • the combined organic layers were washed with brine (20 mL).
  • the combined organic phase was dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure.
  • tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(2-oxopyrrolidin-l- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2A',5S’)-4-(8- chloro-5-(2-oxopyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (180 mg, 0.4 mmol) and 2-methyl-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole-3-carbonitrile (136 mg, 0.59 mmol) in dioxane (2 mL) H 2 O (0.4 mL) was added Pd 2 (dba) 3 (36 mg,
  • tert -butyl (2R,5S)-4-(8-(3-bromo-4-methoxy-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of (4-((25,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)-5-cyclopropylquinazolin-8-yl)boronic acid (500 mg, 1.17 mmol) and 3-bromo-4- methoxy-1H-pyrazole (415 mg, 2.35 mmol) in pyridine (10 mL) was added Cu(OAc) 2 (426 mg, 2.35 mmol) under O 2 .
  • tert-butyl (2R,5S)-4-(8-(3-cyano-4-methoxy-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4-yl)- 2,5-dimethylpiperazine-l-carboxylate To a solution of tert-butyl (2R,5S)-4-(8-(3-bromo-4-methoxy- 1H-pyrazol- 1 -yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (320 mg, 0.57 mmol) in DMF (3 mL) was added Pd(dppf)Ch (42 mg, 0.06 mmol), Zn(CN) 2 (67 mg, 0.57 mmol) and Zn (38 mg, 0.57 mmol).
  • Example 200 4-chloro-2-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]thiazole-5-carbonitrile
  • Example 205 & 206 5-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-3-carbonitrile & 3-[4-[(2S,5R)-4-[2- cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-5-carbonitrile
  • tert -butyl (2R,5S)-4-(8-chloro-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (8 g, 17.55 mmol) and (5-fluoro-3-pyridyl)boronic acid (3.71 g, 26.33 mmol) in dioxane (200 mL) and H 2 O (30 mL) was added Ag2O (1.22 g, 5.27 mmol), Pd(dppf)Cl 2 -CH 2 Cl 2 (1.43 g, 1.76 mmol) and K3PO4 (11.18 g, 52.66 mmol) under N 2
  • Example 210 3-[5-(3-chloro-2-pyridyl)-4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl- piperazin-l-yl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
  • Example 214 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[2- (trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
  • Example 215 4-chloro-l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]pyrazole-3-carbonitrile
  • tert-butyl (2R ,5.S)-4-(8-(4-chloro-3-cyano-1H-pyrazol-l-yl)-5-(5-fhioropvridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate To a solution of (4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)boronic acid (300 mg, 0.63 mmol) and 4-chloro-1H-pyrazole-3-carbonitrile (159 mg, 1.25 mmol) in pyridine (5 mL) was added Cu(OAc) 2 (226 mg, 1.25 mmol).
  • Example 219A & 219B 3-chloro-l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-[(lR,2R)-2-methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile & 3-chloro-l-[4- [(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lS,2S)-2- methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile
  • Solution 3 trimethyl borate (3.32 g, 31.98 mmol) in THF (70 mL).
  • the solution 1 was pumped by Pump 1 (SI, Pl, 6.848 mL/min) to flow reactor 1 (FLR1, PFA, Dynamic mixer, 1.5 mL, 20 °C ; FLR1, PFA, Coils reactor, 3.175(1/8") mm, 14.96 mL, 20 °C).
  • the solution 2 was pumped by Pump 2 (S2, P2, 1.382 mL/min) to flow reactor 1 (FLR1, PFA, Dynamic mixer, 1.5 mL, 20 °C ; FLR1, PFA, Coils reactor 3 175(1/8") mm 14 96 mL 20 °C)
  • the residence time of flow reactor 1 was 2 min.
  • the mixture was collected with a bottle (Solution 3).
  • the Pump 1 and Pump 2 was started at the same time.
  • the Pump 3 was started after.
  • the reaction mixture was collected after running FLR1, 2 min. Stir for 2 min, quenched by aqueous NH4CI (150 mL).
  • the aqueous phase was extracted with EtOAc (3 x 100 mL).
  • reaction mixture was stirred at 80 °C under O2 (15 psi) for 12 h.
  • the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure.

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Abstract

The present disclosure relates generally to small molecule modulators of Mucolipin-1, and their use as therapeutic agents.

Description

COMPOUNDS, COMPOSITIONS, AND METHODS
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit under 35 U.S.C. §119(e) to U.S. Provisional Application Numbers 63/575,484, filed April 5, 2024, and 63/705,992, filed October 10, 2024, each of which is incorporated by reference in its entirety.
FIELD
[0002] The present disclosure relates generally to small molecule modulators of Mucolipin-1 (TRPML1), and their use as therapeutic agents.
BACKGROUND
[0003] Transient receptor potential mucolipin 1, also known as mucolipin-1, (TRPML1 or ML-1) is a member of the Transient receptor potential (TRP) ion channel super family. Electrophysiological experiments on isolated lysosomes have demonstrated that TRPML1 is a non-selective cation channel localized to late-endosomes and lysosomes (LEE) (Dong X., et al. Nature 2008, 455, 7215, 992.). Wild- type human TRPML1 has six transmembrane domains within each monomeric unit that come together to form a tetramer containing an ion pore located in the middle of this complex. TRPML1 is permeable to a variety of mono- and divalent cations including Na+, K+, Ca2+, Fe2+, and Zn2+. The release of Ca2+ from LELs into the cytosol via TRPML1 has been linked to a variety of physiological processes including activation of Transcription Factor EB (TFEB), lysosomal trafficking, exocytosis, phagosome formation, autophagy, and lysosome biogenesis (Venkatachalam, K. et al., ‘TRPML1 -Dependent Process as Therapeutic Targets.’ TRP Channels as Therapeutic Targets, edited by Arpad Szallasi, Academic Press, 2015, pp. 469-482). Conductance of other ions such as Fe2+ and Zn2+ by TRPML1 have a significant role in trace metal regulation within lysosomal stores (Du et al., Cell Reports 2021, 37, 109848). Loss-of- function (LoF) in TRPML1 leads to Fe2+ and Zn2+ accumulation and signaling impairment in lysosomes. [0004] TRPML1 protein is encoded by the MCOLN-1 gene (Di Paola, S. et al., Cell Calcium 2018, 69, 112). LoF mutations in MCOLN-1 can cause an autosomal-recessive lysosomal storage disease (LSD) Mucolipidosis type IV (MLIV). The two predominant mutations resulting in MLIV are in the third intron causing a splice variant missing exons four to five and a ~6 kb deletion spanning exons six to seven. These two mutations are present in approximately 95% of patients diagnosed with MLIV, which causes severe developmental defects including cognitive impairment, motor function disability, and retinal degeneration. Patients harboring these mutations typically do not survive past adolescence. The underlying cause of these devastating symptoms is related to LoF TRPMLl-dependenent lysosomal storage and dysfunction, which lead to neuroinflammation and neurodegeneration.
[0005] Endogenous lipids have been discovered to modulate the activity of TRPML1. Phosphatidylinositol-3,5-bisphosphate (PI(3,5)P2) is an endogenous lipid enriched in LELs that agonize TRPML1 activity (Dong X., et al., Nature Communications 2010, 1, 38). Whereas phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is predominantly found in cellular plasma membranes and inhibits TRPML1 activity on the plasma membrane during its trafficking to LELs. TRPML1 is potentiated by H+ concentration where the optimal pH coincides with that of mature lysosomes (pH~4.5) (Dong et al., 2008).
[0006] Mutations in the enzyme complex that mediates the metabolism of PI(3,5)P2 cause neurodegenerative diseases. The PIKfyve kinase is required for PI(3,5)P2 synthesis and ablation of its activity causes defects in LEE trafficking and TRPML1 activity (Zolov et al., PNAS 2012, 109, 43, 17472). LoF mutations in FIG4, a phosphoinositide phosphatase, are associated with Charcot-Marie- Tooth Disease (CMT4J). The FIG4 gene has been implicated as a genetic factor in 1-2% of patients with amyotrophic lateral sclerosis (AES) (Chow et al., American Journal of Human Genetics 2009, 84, 85). LoF of FIG4 leads to reduced PI(3,5)P2 levels and impaired TRPML1 function. Variants in VAC14, a scaffold protein in the PI(3,5)P2-metabolism complex, are also associated with Parkinson’s Disease (PD) and pediatric-onset neurological disease. Dysfunctions in TRPML1 are proposed as the underlying mechanism for those PI(3,5)P2-deficient mutations. Preclinical studies demonstrate TRPML1 activation can be beneficial in those disease models suggesting the therapeutic potential for TRPML1 agonism in CMT, AES, PD, and other neurodegenerative diseases (Dong X., et al. Nature 2008, 455, 7215, 992.). [0007] TRPML1 has been linked to LSDs. Niemann-Pick (NP) diseases are inherited and primarily caused by accumulation of lipids including sphingomyelin and cholesterol, which lead to severe neurological symptoms. Sphingomyelin accumulation in lysosomes inhibits TRPML1 activity.
Activation of TRPML1 in NP disease cell models can reduce lipid accumulation thereby restoring lysosomal trafficking of TRPML1 (Shen et al., Nature Communications 2012, 3, 731). Increasing TPRML-1 activity through overexpression of TFEB increases lysosomal exocytosis and rescues pathogenesis in mucopolysaccharidosis in cell-culture and in vivo models of the disease. Many LSDs are described by lysosome disfunction including trafficking, maturation, exocytosis, lipid and biomolecule accumulation, and Ca2+ homeostasis. TRPML1 plays a major role in these processes and therefore is an attractive target for treating LSDs (Di Paola et al., 2018).
[0008] Lysosomal function in neurons is associated with neurodegenerative diseases (Cesan et al., Experimental Cell Research 2012, 318, 11, 1245; Ghavami et al., Progress in Neurobiology 2014, 112, 24). Specifically, TRPML1 activity is linked to hallmarks of Alzheimer’s Disease (AD). Patients infected with Human Immunodeficiency Virus (HIV) can develop premature cognitive declines correlated with accumulation of AB and sphingomyelin in cellular compartments. Agonism of TRPML1 in an HIV- pgl20-induced sphingomyelin and AB accumulation model using primary neurons demonstrated the clearance of these biomolecules (Bae et al., Journal of Neuroscience 2014, 34, 34, 11485). Moreover, TFEB activation, a downstream effect of TRPML1 activation reduces Tau pathology in preclinical models (Polito et al., EMBO Molecular Medicine 2014, 6, 9, 1142). These studies suggest a therapeutic potential for TRPML1 in AD.
[0009] Duchenne muscular dystrophy (DMD) is caused by mutations in dystrophin, which weakens the plasma membranes of muscle cells. Patients with DMD are subject to severe muscle damage during contraction (Yu et al., Science Advances 2020, 6, eaaz2736). TRPML1 activation rescues these phenotypes in a DMD mouse model suggesting a therapeutic approach to broadly treating this heterogenous disease.
[0010] Targeting TRPML1 with small molecule modulators has been shown to rescue hallmark phenotypes of neurodegenerative diseases, LSDs, prion and prion-like diseases, and muscular dystrophy- related diseases in cell-based and animal disease models. TRPML1 function is intricately involved in many important cellular processes and could be a therapeutic target for a multitude of human diseases. Several tool molecules exist that modulate TRPML1 but are unsuitable as human drugs due to their reduced potency, physicochemical properties, and in vivo pharmacokinetic properties. Many of the diseases described above that are linked to TRPML1 activity do not have effective, curative treatments. Therefore, there is a significant need for the development of clinically relevant small molecule modulators of TRPML1.
[0011] Therefore, it is desired to develop TRPML1 modulators to rescue impaired lysosomal function and cellular autophagy, particularly in neurodegenerative diseases.
DESCRIPTION
[0012] Provided herein are compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, that are useful in treating and/or preventing diseases mediated, at least in part, by TRPML1.
[0013] In certain embodiments, provided are compounds that activate TRPML1.
[0014] In another embodiment, provided is a pharmaceutical composition comprising a compound as described herein, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and a pharmaceutically acceptable carrier.
[0015] In another embodiment, provided is a method for treating a disease or condition mediated, at least in part, by TRPML1, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound disclosed herein, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
[0016] The disclosure also provides compositions, including pharmaceutical compositions, kits that include the compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, methods of using (or administering) and making the compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and intermediates thereof.
[0017] The disclosure further provides compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or compositions thereof for use in a method of treating a disease, disorder, or condition that is mediated, at least in part, by TRPML1. [0018] Moreover, the disclosure provides uses of the compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or compositions thereof in the manufacture of a medicament for the treatment of a disease, disorder, or condition that is mediated, at least in part, by TRPML1.
[0019] The description herein sets forth exemplary embodiments of the present technology. It should be recognized, however, that such description is not intended as a limitation on the scope of the present disclosure but is instead provided as a description of exemplary embodiments.
1. Definitions
[0020] A dash (“-”) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -C(O)NH2 is attached through the carbon atom. A dash at the front or end of a chemical group is a matter of convenience; chemical groups may be depicted with or without one or more dashes without losing their ordinary meaning. A wavy line or a dashed line drawn through a line in a structure indicates a specified point of attachment of a group. Unless chemically or structurally required, no directionality or stereochemistry is indicated or implied by the order in which a chemical group is written or named.
[0021] The prefix “Cu-v” indicates that the following group has from u to v carbon atoms. For example, “C1-6 alkyl” indicates that the alkyl group has from 1 to 6 carbon atoms.
[0022] Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. In certain embodiments, the term “about” includes the indicated amount + 10%. In other embodiments, the term “about” includes the indicated amount + 5%. In certain other embodiments, the term “about” includes the indicated amount ± 1%. Also, to the term
“about X” includes description of “X”. Also, the singular forms “a” and “the” include plural references unless the context clearly dictates otherwise. Thus, e.g., reference to “the compound” includes a plurality of such compounds and reference to “the assay” includes reference to one or more assays and equivalents thereof known to those skilled in the art.
[0023] “Alkyl” refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (i.e., C1-20 alkyl), 1 to 12 carbon atoms (i.e., C1-12 alkyl), 1 to 8 carbon atoms (i.e., C1-8 alkyl), 1 to 6 carbon atoms (i.e., C1-6 alkyl) or 1 to 4 carbon atoms (i.e., C1-4 alkyl). Examples of alkyl groups include, e.g., 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., -(CH2)3CH3), sec-butyl (i.e., -CH(CH3)CH2CH3), isobutyl (i.e., -CH2CH(CH3)2), and tert-butyl (i.e., -C(CH3)3); and “propyl” includes n-propyl (i.e., -(CH2)2CH3) and isopropyl (i.e., -CH(CH3)2).
[0024] Certain commonly used alternative chemical names may be used. For example, a divalent group such as a divalent “alkyl” group, a divalent “aryl” group, a divalent heteroaryl group, etc., may also be referred to as an “alkylene” group or an “alkylenyl” group (for example, methylenyl, ethylenyl, and propylenyl), an “arylene” group or an “arylenyl” group (for example, phenylenyl or napthylenyl for arylenyl, or quinolinyl for heteroarylene), respectively. Also, unless indicated explicitly otherwise, where combinations of groups are referred to herein as one moiety, e.g., cycloalkylalkyl, arylalkyl, heterocyclylalkyl, heteroarylalkyl, the groups are defined by their components and the last-mentioned group contains the atom by which the moiety is attached to the rest of the molecule.
[0025] “Alkenyl” refers to an alkyl group containing at least one (e.g., 1-3, or 1) carbon-carbon double bond and having from 2 to 20 carbon atoms (i.e., C2-20 alkenyl), 2 to 12 carbon atoms (i.e., C2-12 alkenyl), 2 to 8 carbon atoms (i.e., C2-8 alkenyl), 2 to 6 carbon atoms (i.e., C2-6 alkenyl), or 2 to 4 carbon atoms (i.e., C2-4 alkenyl). Examples of alkenyl groups include, e.g., ethenyl, propenyl, butadienyl (including 1,2- butadienyl and 1,3-butadienyl).
[0026] “Alkynyl” refers to an alkyl group containing at least one (e.g., 1-3, or 1) carbon-carbon triple bond and having from 2 to 20 carbon atoms (i.e., C2-20 alkynyl), 2 to 12 carbon atoms (i.e., C2-12 alkynyl), 2 to 8 carbon atoms (i.e., C2-8 alkynyl), 2 to 6 carbon atoms (i.e., C2-6 alkynyl), or 2 to 4 carbon atoms (i.e., C2-4 alkynyl). The term “alkynyl” also includes those groups having one triple bond and one double bond.
[0027] “Alkoxy” refers to the group “alkyl-O-”. Examples of alkoxy groups include, e.g., methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy, and 1 ,2-dimethylbutoxy.
[0028] “Amino” refers to the group -NRyRz wherein Ry and Rz are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein.
[0029] “Amidino” refers to -C(NRy)(NRz2), wherein Ry and Rz are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein.
[0030] “Aryl” refers to an aromatic carbocyclic group having a single ring (e.g., monocyclic) or multiple rings (e.g., bicyclic or tricyclic) including fused systems. As used herein, aryl has 6 to 20 ring carbon atoms (i.e., C6 -20 aryl), 6 to 12 carbon ring atoms (i.e., C6-12 aryl), or 6 to 10 carbon ring atoms (i.e., C6-10 aryl). Examples of aryl groups include, e.g., phenyl, naphthyl, fluorenyl, and anthryl. Aryl, however, does not encompass or overlap in any way with heteroaryl defined below. If one or more aryl groups are fused with a heteroaryl, the resulting ring system is heteroaryl regardless of point of attachment. If one or more aryl groups are fused with a heterocyclyl, the resulting ring system is heterocyclyl regardless of point of attachment. If one or more aryl groups are fused with a cycloalkyl, the resulting ring system is cycloalkyl regardless of point of attachment.
[0031] “Arylalkyl” or “Aralkyl” refers to the group “aryl-alkyl-”.
[0032] “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) and carbocyclic fused ring systems having at least one sp3 carbon atom (i.e., at least one non-aromatic ring). As used herein, cycloalkyl has from 3 to 20 ring carbon atoms (i.e., C3-20 cycloalkyl), 3 to 14 ring carbon atoms (i.e., C3-14 cycloalkyl), 3 to 12 ring carbon atoms (i.e., C3-12 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C3-10 cycloalkyl), 3 to 8 ring carbon atoms (i.e., C3-8 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C3-6 cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic groups include, for example, bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, 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. Still further, cycloalkyl also includes “spirocycloalkyl” when there are two positions for substitution on the same carbon atom, for example spiro[2.5]octanyl, spiro[4.5]decanyl, or spiro[5.5]undecanyl.
[0033] “Cycloalkylalkyl” refers to the group “cycloalkyl-alkyl-”.
[0034] “Halo” refers to atoms occupying group VIIA of the periodic table, such as fluoro, chloro, bromo, or iodo.
[0035] “Haloalkyl” refers to an unbranched or branched alkyl group as defined above, wherein one or more (e.g., 1 to 6 or 1 to 3) hydrogen atoms are replaced by a halogen. For example, where a residue is substituted with more than one halogen, it may be referred to by using a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyl substituted with two (“di”) or three (“tri”) halo groups, which may be, but are not necessarily, the same halogen. Examples of haloalkyl include, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1 ,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1 ,2-dibromoethyl, and the like.
[0036] “Haloalkoxy” refers to an alkoxy group as defined above, wherein one or more (e.g., 1 to 6 or 1 to 3) hydrogen atoms are replaced by a halogen.
[0037] “Heteroalkyl” refers to an alkyl group in which one or more of the carbon atoms (and any associated hydrogen atoms), excluding any terminal carbon atom(s), are each independently replaced with the same or different heteroatomic group, provided the point of attachment to the remainder of the molecule is through a carbon atom. The term “heteroalkyl” includes unbranched or branched saturated chain having carbon and heteroatoms. By way of example, 1, 2 or 3 carbon atoms may be independently replaced with the same or different heteroatomic group. Heteroatomic groups include, but are not limited to, -NRy-, -O-, -S-, -S(O)-, -S(O)2-, and the like, wherein Ry is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein. Examples of heteroalkyl groups include, e.g., ethers (e.g., -CH2OCH3, -CH(CH3)OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3, etc.), thioethers (e.g., -CH2SCH3, -CH(CH3)SCH3, -CH2CH2SCH3,-CH2CH2SCH2CH2SCH3, etc.), sulfones (e.g., -CH2S(O)2CH3, -CH(CH3)S(O)2CH3, -CH2CH2S(O)2CH3, -CH2CH2S(O)2CH2CH2OCH3, etc.), and amines (e.g., -CH2NRyCH3, -CH(CH3)NRyCH3, -CH2CH2NRyCH3, -CH2CH2NRyCH2CH2NRyCH3, etc., where Ry is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein). As used herein, heteroalkyl includes 2 to 10 carbon atoms, 2 to 8 carbon atoms, or 2 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.
[0038] “Heteroaryl” refers to an aromatic group 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 20 ring carbon atoms (i.e., Ci-20 heteroaryl), 3 to 12 ring carbon atoms (i.e., C3-12 heteroaryl), or 3 to 8 carbon ring atoms (i.e., C3-8 heteroaryl), and 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, oxygen, and sulfur. In certain instances, heteroaryl includes 5-10 membered ring systems, 5-7 membered ring systems, or 5-6 membered ring systems, each independently having 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include, e.g., acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, phenazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, and triazinyl. Examples of the fused-heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[l,5-a]pyridinyl, and imidazo[l,5-a]pyridinyl, where the heteroaryl can be bound via either ring of the fused system. Any aromatic ring, having a single or multiple fused rings, containing at least one heteroatom, is considered a heteroaryl regardless of the attachment to the remainder of the molecule (i.e., through any one of the fused rings). Heteroaryl does not encompass or overlap with aryl as defined above.
[0039] “Heteroarylalkyl” refers to the group “heteroaryl-alkyl-”.
[0040] “Heterocyclyl” refers to a saturated or partially 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 (e.g., 1 to 3) oxo (=0) (such as, e.g., -C(O), -S(O)-, -S(O)2-, etc.) or N-oxide (-0 ) moieties. Any non-aromatic ring or fused ring system containing at least one heteroatom and one non-aromatic ring is considered a heterocyclyl, regardless of the attachment to the remainder of the molecule. For example, fused ring systems such as 6,7-dihydro-5H-cyclopenta[b]pyridinyl, decahydroquinazolinyl, 1,2,3,4-tetrahydroquinazolinyl, and 5,6,7, 8-tetrahydroquinazolinyl are 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 a cycloalkyl, an aryl, or heteroaryl ring, regardless of the attachment to the remainder of the molecule. As used herein, heterocyclyl has 2 to 20 ring carbon atoms (i.e., C2-20 heterocyclyl), 2 to 12 ring carbon atoms (i.e., C2-12 heterocyclyl), 2 to 10 ring carbon atoms (i.e., C2-10 heterocyclyl), 2 to 8 ring carbon atoms (i.e., C2-8 heterocyclyl), 3 to 12 ring carbon atoms (i.e., C3-12 heterocyclyl), 3 to 8 ring carbon atoms (i.e., C3-8 heterocyclyl), or 3 to 6 ring carbon atoms (i.e., C3-6 heterocyclyl); having 1 to 5 ring heteroatoms, 1 to 4 ring heteroatoms, 1 to 3 ring heteroatoms, 1 to 2 ring heteroatoms, or 1 ring heteroatom independently selected from nitrogen, sulfur, or oxygen. Examples of heterocyclyl groups include, e.g., azetidinyl, azepinyl, benzodioxolyl, benzo[b][l,4]dioxepinyl, 1,4- benzodioxanyl, benzopyranyl, benzodioxinyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, oxiranyl, oxetanyl, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tetrahydropyranyl, trithianyl, tetrahydroquinolinyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. The term “heterocyclyl” also includes “spiroheterocyclyl” when there are two positions for substitution on the same carbon atom. Examples of the spiro-heterocyclyl rings include, e.g., bicyclic and tricyclic ring systems, such as oxabicyclo[2.2.2]octanyl, 2-oxa-7-azaspiro[3.5]nonanyl, 2-oxa-6-azaspiro[3.4]octanyl, and 6-oxa-l-azaspiro[3.3]heptanyl. Examples of the fused-heterocyclyl rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3- c]pyridinyl, indolinyl, and isoindolinyl, where the heterocyclyl can be bound via either ring of the fused system.
[0041] “Heterocyclylalkyl” refers to the group “heterocyclyl-alkyl-.”
[0042] “Oxime” refers to the group -CRy(=NOH) wherein Ry is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl, or heteroaryl; each of which may be optionally substituted, as defined herein.
[0043] 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. Also, the term “optionally substituted” refers to any one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms on the designated atom or group may or may not be replaced by a moiety other than hydrogen.
[0044] The term “substituted” used herein means any of the above groups (i.e., alkyl, alkenyl, alkynyl, alkylene, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, and/or heteroalkyl) wherein at least one (e.g., 1 to 5 or 1 to 3) hydrogen atom is replaced by a bond to a non-hydrogen atom such as, but not limited to, halo, hydroxy, cyano, nitro, alkyl, alkenyl, alkynyl, alkoxy, heteroalkyl, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, oxo, thioxo, N-oxide, azido, oxime, thiocyanate, amidino, guanadino, cycloalkylalkyl, arylalkyl, heterocyclylalkyl, heteroarylalkyl, -NRgRh, -NRgC(O)Rh, -NRsC(O)NRsRh, -NRgC(O)ORh, -NRgS(O)i 2Rh, -C(O)Rg, -C(O)ORg, -OC(O)ORg, -OC(O)RS, -C(O)NRsRh, -C(NRg)Rh, -OC(O)NRgRh, -C(O)NRgC(O)Rh or -N(C(O)Rg)C(O)Rh, -SRg, -S(O)RS, -S(O)2Rg, -OS(O)1-2Rg, -S(O)1-2ORg, -NRgS(O)1-2NRgRh, =NSO2Rg, =NNRgRh, -NRgNRgRh, =NORS, -S(O)1-2NRgRh, -NRgS(O)1-2Rh, -CH2S(O)1-2Rg, -CH2S(O)1-2NRgRh, -SF5, -SCF3, or -Si(Ry)3; wherein each alkyl, alkenyl, alkynyl, alkoxy, heteroalkyl, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocyclylalkyl, and heteroarylalkyl is independently optionally substituted with one to three oxo, halo, cyano, hydroxy, alkoxy, or alkyl, which alkyl is optionally substituted with oxo, halo, cyano, hydroxy, or alkoxy; each Ry is independently hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, aryl, or heteroaryl; and each Rg and Rh are independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, or heteroarylalkyl; or Rg and Rh on the same atom or any two Rg and Rh on different atoms are taken together with the atom or atoms to which they are attached to form a heterocyclyl ring optionally substituted with oxo, halo, cyano, hydroxy, alkoxy, or alkyl, which alkyl is optionally substituted with oxo, halo, cyano, hydroxy, or alkoxy. [0045] Polymers or similar indefinite structures arrived at by defining substituents with further substituents appended ad infinitum (e.g., a substituted aryl having a substituted alkyl which is itself substituted with a substituted aryl group, which is further substituted by a substituted heteroalkyl group, etc.) are not intended for inclusion herein. Unless otherwise noted, the maximum number of serial substitutions in compounds described herein is three. For example, serial substitutions of substituted aryl groups with two other substituted aryl groups are limited to ((substituted aryl)substituted aryl) substituted aryl. Similarly, the above definitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluorines or heteroaryl groups having two adjacent oxygen ring atoms). Such impermissible substitution patterns are well known to the skilled artisan. When used to modify a chemical group, the term “substituted” may describe other chemical groups defined herein.
[0046] In certain embodiments, as used herein, the phrase “one or more” refers to one to five. In certain embodiments, as used herein, the phrase “one or more” refers to one to three.
[0047] Any compound or structure given herein, is also intended to represent unlabeled forms as well as isotopically labeled forms of the compounds. These forms of compounds may also be referred to as “isotopically enriched analogs.” Isotopically labeled compounds have structures depicted 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, ”C, 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 3H and 14C are incorporated. 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.
[0048] The term “isotopically enriched analogs” includes “deuterated analogs” of compounds described herein in which one or more hydrogens is/are replaced by deuterium, such as a hydrogen on a carbon atom. Such compounds exhibit increased resistance to metabolism and are thus 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.
[0049] Deuterium labelled or substituted therapeutic compounds of the disclosure may have improved DMPK (drug metabolism and pharmacokinetics) properties, relating to distribution, metabolism, and excretion (ADME). Substitution with heavier isotopes such as deuterium may afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life, reduced dosage requirements, and/or an improvement in therapeutic index. An 18F, 3H, or ”C labeled compound may be useful for PET or SPECT or other imaging studies. Isotopically labeled compounds of this disclosure and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the schemes or in the examples and preparations described below by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. It is understood that deuterium in this context is regarded as a substituent in a compound described herein.
[0050] The concentration of such a heavier isotope, specifically deuterium, may be defined by an isotopic enrichment factor. In the compounds of this disclosure any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Unless otherwise stated, when a position is designated specifically as “H” or “hydrogen,” the position is understood to have hydrogen at its natural abundance isotopic composition. Accordingly, in the compounds of this disclosure any atom specifically designated as a deuterium (D) is meant to represent deuterium.
[0051] In many cases, the compounds of this disclosure are capable of forming acid and/or base salts by virtue of the presence of amino, and/or carboxyl groups, or groups similar thereto.
[0052] Provided are also a pharmaceutically acceptable salt, isotopically enriched analog, deuterated analog, stereoisomer, mixture of stereoisomers, and prodrugs of the compounds described herein. “Pharmaceutically acceptable” or “physiologically acceptable” refer 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.
[0053] 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” or “physiologically acceptable salts” include, for example, salts with inorganic acids, and salts with organic acids. 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 or organic acids. Salts derived from inorganic acids include, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts derived from organic acids include, e.g., acetic acid, propionic acid, gluconic 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 or organic bases. Salts derived from inorganic bases include, by way of example only, sodium, potassium, lithium, aluminum, 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 (i.e., NH2(alkyl)), dialkyl amines (i.e., HN(alkyl)2), trialkyl amines (i.e., N(alkyl)3), substituted alkyl amines (i.e., NH2(substituted alkyl)), di(substituted alkyl) amines (i.e., HN(substituted alkyl)2), tri(substituted alkyl) amines (i.e., N(substituted alkyl)3), alkenyl amines (i.e., NH2( alkenyl)), dialkenyl amines (i.e., HN(alkenyl)2), trialkenyl amines (i.e., N(alkenyl)3), substituted alkenyl amines (i.e., NH2(substituted alkenyl)), di( substituted alkenyl) amines (i.e., HN(substituted alkenyl)2), tri(substituted alkenyl) amines (i.e., N(substituted alkenyl)3, mono-, di- or tri- cycloalkyl amines (i.e., NH2(cycloalkyl), HN(cycloalkyl)2, N(cycloalkyl)3), mono-, di- or tri- arylamines (i.e., NH2(aryl), HN(aryl)2, N(aryl)3), or mixed amines, etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethyl amine, diethyl amine, tri(iso-propyl) amine, tri(n-propyl) amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.
[0054] Some of the compounds exist as tautomers. Tautomers are in equilibrium with one another. For example, amide containing compounds may exist in equilibrium with imidic acid tautomers. Regardless of which tautomer is shown and regardless of the nature of the equilibrium among tautomers, the compounds are understood by one of ordinary skill in the art to comprise both amide and imidic acid tautomers. Thus, the amide containing compounds are understood to include their imidic acid tautomers. Likewise, the imidic acid containing compounds are understood to include their amide tautomers. [0055] The compounds of the disclosure, or their pharmaceutically acceptable salts 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 (S)- or, as (D)- or (L)- for amino acids. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R )- and (S)-, or (D)- and (L)- isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and/or 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). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.
[0056] 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, or mixtures thereof, and includes “enantiomers,” which refers to two stereoisomers whose molecules are nonsuperimposeable mirror images of one another.
[0057] “Diastereomers” are stereoisomers that have at least two asymmetric atoms, but which are not mirror-images of each other.
[0058] Relative centers of the compounds as depicted herein are indicated graphically using the “thick bond” style (bold or parallel lines) and absolute stereochemistry is depicted using wedge bonds (bold or parallel lines).
[0059] “Prodrugs” means any compound which releases an active parent drug according to a structure described herein in vivo when such prodrug is administered to a mammalian subject. Prodrugs of a compound described herein are prepared by modifying functional groups present in the compound described herein in such a way that the modifications may be cleaved in vivo to release the parent compound. Prodrugs may be prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to the parent compounds. Prodrugs include compounds described herein wherein a hydroxy, amino, carboxyl, or sulfhydryl group in a compound described herein is bonded to any group that may be cleaved in vivo to regenerate the free hydroxy, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to esters (e.g., acetate, formate, and benzoate derivatives), amides, guanidines, carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups in compounds described herein, and the like. Preparation, selection, and use of prodrugs is discussed in T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Vol. 14 of the A.C.S. Symposium Series; “Design of Prodrugs,” ed. H. Bundgaard, Elsevier, 1985; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, each of which are hereby incorporated by reference in their entirety.
2. Compounds
[0060] Provided herein are compounds that are activators of TRPML1. In certain embodiments, provided is a compound of Formula I: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, X1, X2, X3, R1, R3, R4, and q are each independently as defined herein.
[0061] In certain embodiments, provided is a compound of Formula I: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
X1 is N or CR7;
X2 is N or CR6; and the dashed bond is a double bond; or
X2 is NR2; R1 is oxo; and the dashed bond is a single bond;
X3 is N or CR5;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -XR11CfOiN(R11 )2. -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R2 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl;
R3 is: ; wherein X is N or CH; each R4 is independently hydroxy, halo, cyano, -CD3, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C, 6 alkyl, -C(O)OC, 6 alkyl, -C(O)NRaRb, -S(O)2 C1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
R5, R6 and R7 are each independently hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, -NR12aC(O)OR12, or heteroaryl optionally substituted with one to five substituents independently selected from halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, or C3-6 cycloalkyl; provided that when X is N, then R10 is not -NR12aC(O)R12 or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1- C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N( C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl. [0062] In certain embodiments, provided is a compound of Formula I: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
X1 is N or CR7;
X2 is N or CR6; and the dashed bond is a double bond; or
X2 is NR2; R1 is oxo; and the dashed bond is a single bond;
X3 is N or CR5;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -XR11C(O)N(R11)2. -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R2 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O) C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy; R5, R6 and R7 are each independently hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0063] In certain embodiments, provided is a compound of Formula I: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
X1 is N or CR7;
X2 is N or CR6; and the dashed bond is a double bond; or
X2 is NR2; R1 is oxo; and the dashed bond is a single bond;
X3 is N or CR5; L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -0-, -S-, -C(0)-, -S(0)-, -S(0)2-, -C(0)0-*, -0C(0)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -NR11C(0)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R2 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, or C1-6 alkoxy;
R5, R6 and R7 are each independently hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2- C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a; R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -0-, -NH-, -S-, -S(0)-, -S(0)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(0)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(0)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0064] In certain embodiments, X1 is N or CR7. In certain embodiments, X1 is N. In certain embodiments, X1 is CR7.
[0065] In certain embodiments, X2 is N or CR6; and the dashed bond, i.e., the bond between X2 and C(R’), is a double bond. In certain embodiments, X2 is N; and the dashed bond is a double bond. In certain embodiments, X2 is CR6; and the dashed bond is a double bond.
[0066] In certain embodiments, X2 is NR2; R1 is oxo; and the dashed bond, i.e., the bond between X2 and C(R’), is a single bond.
[0067] In certain embodiments, X3 is N or CR5. In certain embodiments, X3 is N. In certain embodiments, X3 is CR5.
[0068] In certain embodiments, provided is a compound of Formula IA: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, X2, X3, R1, R3, R4, and q are each independently as defined herein.
[0069] In certain embodiments, provided is a compound of Formula IB: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R1, R3, R4, and q are each independently as defined herein. [0070] In certain embodiments, provided is a compound of Formula IB: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein:
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -NR11C(O)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2.6 alkenyl)-, -C(O)N(C2.6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0071] In certain embodiments, provided is a compound of Formula IC: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R1, R3, R4, and q are each independently as defined herein.
[0072] In certain embodiments, provided is a compound of Formula IC: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein:
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -NR11C(O)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2- C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1- C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(0)N( C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0073] In certain embodiments, provided is a compound of Formula ID: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R2, R3, R4, and q are each independently as defined herein.
[0074] In certain embodiments, provided is a compound of Formula ID: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein:
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -NR11C(O)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R2 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0075] In certain embodiments, provided is a compound of Formula IE: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein Ring A, L, R1, R3, R4, and q are each independently as defined herein.
[0076] In certain embodiments, provided is a compound of Formula IE: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein:
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -XR11C(O )N( R11)2. -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R3 is: each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O) C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2.6 alkenyl)-, -C(O)N(C2.6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0077] In certain embodiments, Ring A is C3-10 cycloalkyl.
[0078] In certain embodiments, Ring A is heterocyclyl.
[0079] In certain embodiments, Ring A is aryl.
[0080] In certain embodiments, Ring A is heteroaryl.
[0081] In certain embodiments, Ring A is monocyclic C3-7 cycloalkyl, monocyclic heterocyclyl, phenyl, or monocyclic heteroaryl.
[0082] In certain embodiments, Ring A is monocyclic C3-7 cycloalkyl.
[0083] In certain embodiments, Ring A is monocyclic heterocyclyl.
[0084] In certain embodiments, Ring A is bicyclic heterocyclyl.
[0085] In certain embodiments, Ring A is phenyl.
[0086] In certain embodiments, Ring A is monocyclic heteroaryl.
[0087] In certain embodiments, Ring A is bicyclic heteroaryl.
[0088] In certain embodiments, Ring A is monocyclic C3-7 cycloalkyl, 5- or 6-membered monocyclic heterocyclyl, phenyl, or 5- or 6-membered monocyclic heteroaryl. [0089] In certain embodiments, Ring A is 5- or 6-membered monocyclic heterocyclyl.
[0090] In certain embodiments, Ring A is 5- or 6-membered monocyclic heteroaryl.
[0091] In certain embodiments, Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl.
[0092] In certain embodiments, Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl; and at least one R4 is cyano.
[0093] In certain embodiments, Ring A is 4- or 5 -membered heterocyclyl or 5- or 6-membered membered heteroaryl.
[0094] In certain embodiments, Ring A is 4- or 5 -membered heterocyclyl or 5- or 6-membered membered heteroaryl; and at least one R4 is cyano.
[0095] In certain embodiments, Ring A is 4- or 5 -membered heterocyclyl or 5- or 6-membered membered heteroaryl; q is 1 or 2; and at least one R4 is cyano.
[0096] In certain embodiments, Ring A is 5-membered heterocyclyl.
[0097] In certain embodiments, Ring A is 5-membered heterocyclyl; and at least one R4 is cyano.
[0098] In certain embodiments, Ring A is 5- or 6-membered monocyclic heteroaryl having 1 or 2 ring nitrogen atoms.
[0099] In certain embodiments, Ring A is 5-membered heteroaryl.
[0100] In certain embodiments, Ring A is 5-membered heteroaryl; and at least one R4 is cyano.
[0101] In certain embodiments, Ring A is cyclopropyl, cyclobutyl, azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
[0102] In certain embodiments, Ring A is cyclopropyl, cyclobutyl, azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl; and at least one R4 is cyano.
[0103] In certain embodiments, Ring A is cyclopropyl, azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, pyridyl, triazolo[4,5-c]pyridinyl, 4,6-dihydropyrrolo[3,4-d]pyrazolyl, or 4,6- dihydrofuro[3,4-c]pyrazolyl.
[0104] In certain embodiments, Ring A is azetidinyl, pyrrolidinyl, phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
[0105] In certain embodiments, Ring A is phenyl, pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
[0106] In certain embodiments, Ring A is pyrazolyl, triazolyl, oxadiazolyl, or pyridyl.
[0107] In certain embodiments, Ring A is pyrazolyl, triazolyl, oxadiazolyl, or pyridyl; and at least one
R4 is cyano.
[0108] In certain embodiments, q is 0, 1, or 2. In certain embodiments, q is 0. In certain embodiments, q is 1. In certain embodiments, q is 2.
[0109] In certain embodiments, each R4 is independently selected from halo, cyano, -CD3, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, -C(O)OC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -S(O)2C1-6 alkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy. [0110] In certain embodiments, each R4 is independently halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, -C(O)OC1-6 alkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy.
[0111] In certain embodiments, each R4 is independently selected from cyano, C1-6 alkyl, or -C(O)OC1 6 alkyl.
[0112] In embodiments, each R4 at least one R4 is cyano.
[0113] In certain embodiments, q is 1, 2, or 3; and at least one R4 is cyano.
[0114] In certain embodiments, q is 1 or 2; and at least one R4 is cyano.
[0115] In certain embodiments, L is a bond.
[0116] In certain embodiments, L is -C(O)NR11-*.
[0117] In certain embodiments, provided is a compound of Formula IF: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein R1, R3, and each R4 are independently as defined herein. [0118] In certain embodiments, provided is a compound of Formula IGG: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein q, R1, R3, and each R4 are independently as defined herein. [0119] In certain embodiments, provided is a compound of Formula IH: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein R1, R3, and each R4 are independently as defined herein. [0120] In certain embodiments, R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is independently optionally substituted with one to five Z1a.
[0121] In certain embodiments, R11 is hydrogen or C1-6 alkyl optionally substituted with one to five independently selected halo.
[0122] In certain embodiments, R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C3 10 cycloalkyl is independently optionally substituted with one to five Z1a.
[0123] In certain embodiments, R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C310 cycloalkyl is independently optionally substituted with one to five Z1a. In certain embodiments, R1 is hydrogen.
[0124] In certain embodiments, R1 is hydrogen or halo. In certain embodiments, R1 is hydrogen, halo, C3-10 cycloalkyl, -N(R11)2, or -OR11; wherein the C310 cycloalkyl is independently optionally substituted with one to five Z1a.
[0125] In certain embodiments, R3 is
[0126] In certain embodiments, m is 1. In certain embodiments, m is 2.
[0127] In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, m is 3.
[0128] In certain embodiments, m + n is 2. In certain embodiments, m and n are both 1.
[0129] In certain embodiments, R3 is
[0130] In certain embodiments, ml is 0. In certain embodiments, ml is 1. In certain embodiments, ml is
2. In certain embodiments, ml is 1 or 2.
[0131] In certain embodiments, nl is 0. In certain embodiments, nl is 1. In certain embodiments, nl is 2.
In certain embodiments, nl is 3. In certain embodiments, nl is 1, 2, or 3. In certain embodiments, nl is 1 or 2.
[0132] In certain embodiments, m2 is 0. In certain embodiments, m2 is 1. In certain embodiments, m2 is
2. In certain embodiments, m2 is 1 or 2.
[0133] In certain embodiments, n2 is 0. In certain embodiments, n2 is 1. In certain embodiments, n2 is 2.
In certain embodiments, n2 is 3. In certain embodiments, n2 is 1, 2, or 3. In certain embodiments, n2 is 1 or 2. [0134] In certain embodiments, R3 is
[0135] In certain embodiments, m3 is 0. In certain embodiments, m3 is 1.
[0136] In certain embodiments, n3 is 0. In certain embodiments, n3 is 1. In certain embodiments, n3 is 2. In certain embodiments, n3 is 0 or 1. In certain embodiments, n3 is 1 or 2.
[0137] In certain embodiments, m3 is 0, and n3 is 0.
[0138] In certain embodiments, R10 is -NR12aC(O)R12.
[0139] In certain embodiments, R10 is heteroaryl optionally substituted with one to five substituents independently selected from halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, or C3-6 cycloalkyl
[0140] In certain embodiments, each R12 is independently C1-6 alkyl or C3-7 cycloalkyl; wherein the C1-6 alkyl, or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a.
[0141] In certain embodiments, each R11 is independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-10 cycloalkyl.
[0142] In certain embodiments, each R11 is independently hydrogen, C1-6 alkyl, or C3-10 cycloalkyl.
[0143] In certain embodiments, R12a is C1-6 alkyl.
[0144] In certain embodiments, p is 0. In certain embodiments, p is 1. In certain embodiments, p is 2. In certain embodiments, p is 1 or 2. In certain embodiments, p is 0, 1, or 2.
[0145] In certain embodiments, m3 is 0, n3 is 0, and p is 0, 1, or 2.
[0146] In certain embodiments, m3 is 0 or 1; n3 is 0 or 1; and p is 0, 1, or 2.
[0147] In certain embodiments, each R8 is independently C1-6 alkyl.
[0148] In certain embodiments, p is 1 or 2; and each R8 is methyl.
[0149] In certain embodiments, p is 2; and each R8 is methyl.
[0150] In certain embodiments, R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five substituents independently selected from hydroxy, halo, cyano, C1-6 alkyl, -O-C1-6 alkyl, C3-7 cycloalkyl, or -S(O)2C1-6 alkyl; wherein each C1-6 alkyl, -O-C1-6 alkyl, C3-7 cycloalkyl, or -S(O)2C1-6 alkyl is independently optionally substituted with one to five independently selected halo.
[0151] In certain embodiments, R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z1a.
[0152] In certain embodiments, R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z1a; and each Z1a is independently halo, cyano, C1-6 alkyl, or -OR13. [0153] In certain embodiments, R9 is C1-6 alkyl, C1-6 alkoxy, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 alkoxy, or C3-7 cycloalkyl is independently optionally substituted with one to five halo, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, or C3-7 cycloalkyl.
[0154] In certain embodiments, R9 is C1-6 alkyl optionally substituted with one to five Z1a; and each Z1a is independently halo, cyano, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, or C3-7 cycloalkyl.
[0155] In certain embodiments, R9 is C1-6 alkyl.
[0156] In certain embodiments, R9 is C1-6 alkyl optionally substituted with one to five halo, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, or C3-7 cycloalkyl. In certain embodiments, R9 is C1-6 alkyl.
[0157] In certain embodiments, R9 is C1-6 alkoxy optionally substituted with one to five halo, C1-6 alkyl, C1-6 alkoxy, C1-6 haloalkyl, or C3-7 cycloalkyl. In certain embodiments, R9 is C1-6 alkoxy optionally substituted with one to five halo.
[0158] In certain embodiments, R9 is C3-7 cycloalkyl optionally substituted with one to five halo, cyano, C1-6 alkyl, or C1-6 haloalkyl.
[0159] In certain embodiments, R9 is C1-6 alkyl, C1-6 alkoxy, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 alkoxy, or C3-7 cycloalkyl is independently optionally substituted with one to five halo or cyano. ^
[0160] In certain embodiments, R9 is , , [0161] In certain embodiments, R9 is
[0162] In certain embodiments, R9 is
[0163] In certain embodiments, R9 is
[0164] In certain embodiments, provided is a compound of Formula II: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein each X2, X3, L, Ring A, R1, R4, R8, R9, m, n, p, and q are independently as defined herein.
[0165] In certain embodiments, provided is a compound of Formula II:
or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein:
X2 is N or CR6;
X3 is N or CR5;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(0)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(0)N(R11)2, -S(O)2N(R11)2, -NR11C(O)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a; each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, or C1-6 alkoxy;
R6 and R7 are each independently hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl; each Ra and Rb are independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; m is 1 or 2; n is 1, 2, or 3; wherein m + n is 2, 3, or 4; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0166] In certain embodiments, X1 is N.
[0167] In certain embodiments, X2 is N; and the dashed bond is a double bond.
[0168] In certain embodiments, X2 is CR6; and the dashed bond is a double bond. [0169] In certain embodiments, R6 is hydrogen, hydroxy, halo, or C3-7 cycloalkyl.
[0170] In certain embodiments, X2 is NR2; R1 is oxo; and the dashed bond is a single bond.
[0171] In certain embodiments, X3 is N.
[0172] In certain embodiments, X3 is CR7.
[0173] In certain embodiments, R7 is hydrogen.
[0174] In certain embodiments, X1 is N; and X3 is CH.
[0175] In certain embodiments, m and n are each 1.
[0176] In certain embodiments, provided is a compound of Formula IIA: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein each X2, Ring A, R1, R4, R8, R9, m, n, p, and q are independently as defined herein.
[0177] In certain embodiments, provided is a compound of Formula IIB: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein Ring A, R1, R4, R8, R9, and q are each independently as defined herein.
[0178] In certain embodiments, provided is a compound of Formula IIC: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein Ring A, R1, R4, R8, R9, and q are each independently as defined herein.
[0179] In certain embodiments, provided is a compound of Formula IID: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein p, R1, R9, each R4, and each R8 are independently as defined herein.
[0180] In certain embodiments, provided is a compound of Formula IIE: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein R9 and each R4 are independently as defined herein; and ring B is R1 is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is independently optionally substituted with one to five Z1a.
[0181] In certain embodiments, provided is a compound of Formula III: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein each X2, L, Ring A, R1, R4, R8, R9, ml, nl, m2, n2, p, and q are independently as defined herein.
[0182] In certain embodiments, provided is a compound of Formula III: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein:
X2 is N or CR6;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -NR11C(O)N(R11)2, -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a; each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
R6 is hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; wherein ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2.6 alkenyl)-, -C(O)N(C2.6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
[0183] In certain embodiments, provided is a compound of Formula IV : or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, mixture of stereoisomers thereof, wherein each X2, L, Ring A, R1, R4, R8, R9, m3, n3, p, and q are independently as defined herein.
[0184] In certain embodiments, provided is a compound of Formula IV:
or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein:
X2 is N or CR6;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -XR11C(O )N( R11)2. -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -0C(0)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a; each R4 is independently hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C1-6 alkyl, -C(O)OC1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
R6 is hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy; R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m3 is 0 or 1 ; n3 is 0, 1, or 2; wherein m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2-6 alkenyl)-, -C(O)N(C2-6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl. [0185] In certain embodiments, L is a bond; Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl; at least one R4 is cyano; and each R8 is independently C1-6 alkyl.
[0186] In certain embodiments, L is a bond; Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl; R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C3-10 cycloalkyl is independently optionally substituted with one to five Z1a; at least one R4 is cyano; and each R8 is independently C1-6 alkyl.
[0187] In certain embodiments, L is a bond; Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl; R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C3-10 cycloalkyl is independently optionally substituted with one to five Z1a; at least one R4 is cyano; each R8 is independently C1-6 alkyl; and R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z1a.
[0188] In certain embodiments, L is a bond; Ring A is C3-4 cycloalkyl, phenyl, 4- or 5-membered heterocyclyl, or 5- or 6-membered membered heteroaryl; R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C3-10 cycloalkyl is independently optionally substituted with one to five Z1a; at least one R4 is cyano; each R8 is independently C1-6 alkyl; and R9 is C1-6 alkyl, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C3-7 cycloalkyl, or heterocyclyl is independently optionally substituted with one to five Z1a.
[0189] In certain embodiments, provided is a compound selected from Table 1, or a pharmaceutically acceptable salt, isotopically enriched analog, prodrug, stereoisomer, or a mixture of stereoisomers thereof:
Table 1
74
75
78
[0190] In certain embodiments, provided is a compound selected from Table 2 or a pharmaceutically acceptable salt thereof.
Table 2
[0191] TRP (Transient receptor potential) channels are integral membrane proteins consisting of a superfamily of cation channels that allow permeability of both monovalent and divalent cations. TRP channels are subdivided into six subfamilies: TRPC, TRPV, TRPM, TRPP, TRPML, and TRPA, and are expressed in almost every cell and tissue. TRPs play an instrumental role in the regulation of various physiological processes. TRP channels are extensively represented in brain tissues and are present in both prokaryotes and eukaryotes, exhibiting responses to several mechanisms, including physical, chemical, and thermal stimuli. TRP channels are involved in the perturbation of Ca2+ homeostasis in intracellular calcium stores, both in neuronal and non-neuronal cells, and its discrepancy leads to several neuronal disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), and Amyotrophic lateral sclerosis (ALS). TRPs participate in neurite outgrowth, receptor signaling, and excitotoxic cell death in the central nervous system.
[0192] “Treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. Beneficial or desired clinical results may include one or more of the following: a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and/or diminishing the extent of the disease or condition); b) slowing or arresting the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and/or preventing or delaying the spread (e.g., metastasis) of the disease or condition); and/or c) relieving the disease, that is, causing the regression of clinical symptoms (e.g., ameliorating the disease state, providing partial or total remission of the disease or condition, enhancing effect of another medication, delaying the progression of the disease, increasing the quality of life, and/or prolonging survival.
[0193] “Prevention” or “preventing” means any treatment of a disease or condition that causes the clinical symptoms of the disease or condition not to develop. Compounds may, in certain embodiments, be administered to a subject (including a human) who is at risk or has a family history of the disease or condition.
[0194] “Subject” refers to an animal, such as a mammal (including a human), that has been or will be the object of treatment, observation, or experiment. The methods described herein may be useful in human therapy, and/or veterinary applications. In certain embodiments, the subject is a mammal. In certain embodiments, the subject is a human.
[0195] The term “therapeutically effective amount” or “effective amount” of a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof means an amount sufficient to effect treatment when administered to a subject, to provide a therapeutic benefit such as amelioration of symptoms or slowing of disease progression. For example, a therapeutically effective amount may be an amount sufficient to decrease a symptom of a disease or condition of as described herein. The therapeutically effective amount may vary depending on the subject, and disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the manner of administering, which can readily be determined by one of ordinary skill in the art.
[0196] The methods described herein may be applied to cell populations in vivo or ex vivo. “In vivo” means within a living individual, as within an animal or human. In this context, the methods described herein may be used therapeutically in an individual. “Ex vivo” means outside of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples including fluid or tissue samples obtained from individuals. Such samples may be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein may be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein may be used ex vivo to determine the optimal schedule and/or dosing of administration of a compound of the present disclosure for a given indication, cell type, individual, and other parameters. Information gleaned from such use may be used for experimental purposes or in the clinic to set protocols for in vivo treatment. Other ex vivo uses for which the compounds and compositions described herein may be suited are described below or will become apparent to those skilled in the art. The compounds may be further characterized to examine the safety or tolerance dosage in human or non- human subjects. Such properties may be examined using commonly known methods to those skilled in the art.
[0197] In some embodiments, modulation of TRPML1 comprises activation of TRPML1.
[0198] Provided herein, in certain embodiments, is a method of treating a disease or disorder that can be treated by activation of TRPML1, the method comprising administering to a subject in need thereof a compound described herein (e.g., a compound of Formula I, or subformula thereof) or pharmaceutically acceptable salts, solvates, hydrates, tautomers, stereoisomers, isotopically labeled derivatives thereof, or a composition described herein. [0199] In certain embodiments, provided are compounds, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, that activate TRPML1. In certain embodiments, the compounds provided herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, activates TRPML1.
[0200] In certain embodiments, provided is a method of activating TRPML1 activity comprising contacting a cell with an effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. The activating can be in vitro or in vivo. [0201] In certain embodiments, provided is a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, for use in activating TRPML1 activity (e.g., in vitro or in vivo).
[0202] In certain embodiments, the present disclosure provides use of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, in the manufacture of a medicament for activating TRPML1 activity (e.g., in vitro or in vivo).
[0203] In certain embodiments, the present disclosure provides use of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, in the manufacture of a medicament for treating a disease or disorder mediated, at least in part, by TRPML1. [0204] In certain embodiments, provided is a method for treating a disease or disorder mediated, at least in part, by TRPML1, comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof to a subject in need thereof.
[0205] In certain embodiments, provided is a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, for use in treating a disease or disorder mediated, at least in part, by TRPML1 in a subject in need thereof.
[0206] In certain embodiments, provided is a method of treating a disease or disorder which can be treated by modulation of lysosomes, comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof to a subject in need thereof.
[0207] In certain embodiments, provided is a method of treating a disease or disorder selected from the group consisting of a ciliopathy, neurodegenerative or neurological disease or disorder, lysosomal storage disease or disorder, lysosomal transport disease or disorder, glycogen storage disease or disorder, cholesteryl ester storage disease or disorder, a muscular disease (e.g., muscular dystrophy), a disease related to aging (e.g., photo aging of the skin), macular degeneration (e.g., Stargardt’s or age-related), and cancer (e.g., cancers of the blood, brain, bone, lung, liver, kidney, bladder, stomach, breast, prostate, ovary, testes, colon, pancreas, or skin), comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof to a subject in need thereof.
[0208] In certain embodiments, the disorder or disease is aging, bone diseases, cardiovascular diseases, congenital developmental disorders, eye diseases, hematological and solid malignancies, infectious diseases, inflammatory diseases, liver diseases, metabolic diseases, neurodegenerative or neurological disease or disorder, pancreatitis, renal diseases, skeletal muscle disorders, obesity, lysosomal storage diseases, hypertrophic cardiomyopathy, dilated cardiomyopathy, inclusion body myositis, Paget’s disease, or pulmonary diseases.
Neurodegenerative or Neurological diseases or disorders
[0209] In certain embodiments, the treating comprises reducing one or more symptoms or features of neurodegeneration.
[0210] In certain embodiments, provided is a method for treating a neurodegenerative or neurological disease or disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
[0211] The transient receptor potential channel mucolipin 1 (TRPML1) is a cation channel that can transport Ca2+, Fe2+ and Zn2+. The primary function of TRPML1 is to induce Ca2+ release from endolysosomal compartments, a vital process for endolysosomal function. Dysfunction in TRPML1- relevant endolysosomal pathways have been widely observed in preclinical and clinical samples for late- onset neurodegenerative diseases. The functions regulated by TRPML1 include lysosomal biogenesis dependent on TFEB family, maturation of late endosomes to lysosomes; endolysosomal trafficking; nutrient sensing and adaptation; positioning, exocytosis, fission, clearance, and reformation of lysosomes; autophagy, phagocytosis and clearance of aggregated proteins and pathogens. Loss-of- function mutations in the human TRPML1 gene MCOLN1 cause mucolipidosis type IV (MLIV), a rare recessive lysosomal storage disorder (LSD). MLIV is characterized by neurodegeneration, psychomotor impairment, ophthalmologic and gut defects. MLIV patient cells show multiple endolysosomal abnormalities including defective endolysosomal trafficking, vacuolation, altered positioning of the endolysosomal compartment, compromised maturation of lysosomes, dysregulated pH and autophagic defects. [0212] TRP channels are involved in the perturbation of Ca2+ homeostasis in intracellular calcium stores, both in neuronal and non-neuronal cells, and its dysfunction leads to several neuronal disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), and Amyotrophic lateral sclerosis (ALS). TRPs participate in neurite outgrowth, receptor signaling, and excitotoxic cell death in the central nervous system.
[0213] In some embodiments, the neurodegenerative or neurological disease or disorder is selected from the group consisting of Parkinson’s disease, GBA-Parkinson’s disease, LRRK2 Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis (ALS), Alzheimer’s disease, progressive supranuclear palsy, frontotemporal dementia, FTDP-17, corticobasal degeneration, Lewy body dementia, Pick’s disease, and multi system atrophy.
Alzheimer’s disease
[0214] Alzheimer’ s disease (AD) is a devastating neurodegenerative disorder characterized by severe memory loss and behavioral changes. The underlying pathology involves the accumulation of extracellular amyloid aggregates called senile plaques and intracellular neurofibrillary tangles, leading to the selective loss of synapses and neurons in the hippocampal and cerebral cortical regions. While the amyloid and tau hypotheses have been the traditional theories explaining AD, the dysregulation of calcium homeostasis has recently gained attention as a critical factor in AD pathogenesis.
[0215] Calcium is an essential intracellular messenger that binds to multiple proteins, receptors, and ion channels to regulate various physiological functions. During neurodegeneration, neurons become inefficient in regulating calcium levels. Enhanced pathological lesions induce neurotoxicity and cytokines, leading to dysregulation of calcium homeostasis and leaving neurons prone to excitotoxicity and apoptosis.
[0216] In certain embodiments, provided is a method for treating Alzheimer’s disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. Parkinson’s disease
[0217] Parkinson’ s disease (PD) is the second most predominant neurodegenerative disorder related to aging after AD. It is a movement disorder, depicted by the disintegration of dopaminergic neurons (DNs) in the substantia nigra (SN), that worsens over time. Eewy bodies are the cytoplasmic inclusions formed by a-synuclein protein and are considered the main pathological hallmark of PD. Several factors in PD can lead to the loss of DNs in SN, such as oxidative stress, mitochondrial dysfunction, protein aggregation, and alteration in calcium homeostasis. Studies reported that TRP channels could facilitate some of the mechanisms that advance the progression of the disease.
[0218] In certain embodiments, provided is a method for treating Parkinson’s disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. Huntington ’s disease
[0219] Huntington’s disease (HD) is an autosomal neurodegenerative disorder triggered by polyglutamine expansion in Huntingtin protein and represented by cognitive impairment, medium spiny neurons (MSN) loss in the striatum, and convulsive movements. Various channels have been reported to alter the K+ homeostasis, such as the Kir4.1 channel expressing striatal astrocytes in mutated HTT protein that disintegrates the extracellular K+ homeostasis hence provokes hyperexcitability in neurons, i.e., HD motor symptoms in striatal neurons. However, the normal Kir4.1 channel is a prevalent astrocytic K+ channel that plays a prominent role in balancing the cells resting membrane potential and buffering K+ ions in the brain. Furthermore, it is suggested that HD mHTT protein modifies the high voltage stimulated Ca2+ channels. Besides dysfunction in the Ca2+ channel, other ion channels have also exhibited reduced expression in several HD mouse models. Thus, modification in these ion channels disintegrates the ion homeostasis in cortical pyramidal neurons, due to which synaptic integration, neurotransmitter release, and genetic expression gets affected, which plays a central role in cortical dysfunction in HD. [0220] In certain embodiments, provided is a method for treating Huntington’s disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. Amyotrophic lateral sclerosis
[0221] Amyotrophic lateral sclerosis (ALS) is a neurological disorder characterized by the progressive loss of motor neurons in the brain stem, motor cortex, and spinal cord, resulting in muscle weakness, atrophy, and contraction. The disease is associated with the hyperexcitability of axons due to the continuous conduction of sodium ions (Na+) and subsequent reduction in potassium ion (K+) conduction. The lingual muscles, stimulated by motor neurons, are also susceptible to degeneration in ALS, linked to the differential expression of voltage-gated calcium channels (VGCCs).
[0222] In certain embodiments, provided is a method for treating amyotrophic lateral sclerosis, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof.
Lysosomal Storage Disorders
[0223] In certain embodiments, provided is a method for treating a lysosomal storage disease or disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. [0224] In some embodiments, the lysosomal storage disease or disorder is selected from the group consisting of Niemann-Pick disease, Gaucher’s disease, neuronopathic Gaucher’s disease, neuronal ceroid lipofuscinosis, sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, gangliosidoses, Gaucher Disease, Lysosomal acid lipase deficiency, sulfatidoses, mucopolysaccharidoses, mucolipidoses, lipidoses, and oligosaccharidoses. In some embodiments, the lysosomal storage disorder is selected from the group consisting of sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, Fabry disease, Schindler disease, beta-galactosidase disorder, GM1 gangliosidosis, GM2 gangliosidosis AB variant, GM2 gangliosidosis activator deficiency, Sandhoff disease, Tay- Sachs disease, Gaucher disease, lysosomal acid lipase deficiency, Niemann-Pick disease, metachromatic leukodystrophy, Saposin B deficiency, multiple sulfatase deficiency, Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome, Hunter syndrome, Sanfilippo syndrome, Morquio syndrome, Maroteaux-Lamy syndrome, Sly syndrome, hyaluronidase deficiency, sialidosis, I-cell disease, pseudo-Hurler polydystrophy, phosphotransferease deficiency, mucolipidin 1 deficiency, Santavuori-Haltia disease, Jansky-Bielchowsky disease, Batten- Spielmeyer- Vogt disease, Kufs disease, Finnish variant neuronal ceroid lipfuscinosis, late infantile variant neuronal ceroid lipfuscinosis, type 7 neuronal ceroid lipfuscinosis, northern epilepsy neuronal ceroid lipfuscinosis, Turkish late infantile neuronal ceroid lipfuscinosis, German/Serbian late infantile neuronal ceroid lipfuscinosis, congenital cathepsin D deficiency, Wolman disease, alpha-mannosidosis, beta-mannosidosis, aspartylgluosaminuria, and fucosidosis. In some embodiments, the lysosomal storage disorder is selected from the group consisting of Niemann-Pick disease, Gaucher’s disease, neuronopathic Gaucher’s disease, and neuronal ceroid lipofuscinosis.
[0225] In certain embodiments, provided is a method for treating a lysosomal transport disease or disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. In some embodiments, the lysosomal transport disease or disorder is selected from the group consisting of cystinosis, pycnodysostosis, Salla disease, sialic acid storage disease, and infantile free sialic acid storage disease.
[0226] In certain embodiments, provided is a method for treating a glycogen storage disease or disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound as disclosed herein (e.g., a compound of Formula I, or subformula thereof), or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. In some embodiments, the glycogen storage disease or disorder is selected from the group consisting of Pompe disease and Danon disease.
Ciliopathies
[0227] In certain embodiments, provided is a method of treating a ciliopathy, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound capable of modulating TRPML, or a therapeutically effective amount of a pharmaceutical composition comprising the compound and a pharmaceutically acceptable excipient.
[0228] In some embodiments, the ciliopathy is selected from the group consisting of polycystic kidney disease, pancreatic cysts in polycystic kidney disease, Bardet-Biedl syndrome, nephronophthisis, Joubert Syndrome, Meckel-Gruber Syndrome, oral-facial- digital syndrome, Senior Loken Syndrome, Birt-Hogg- Dube syndrome, Leber’s congenital amaurosis, Alstrom syndrome, Jeune asphyxiating thoracic dystrophy, Ellis van Creveld syndrome, Sensenbrenner syndrome, and primary ciliary dyskinesia. In some embodiments, the ciliopathy is autosomal dominant polycystic kidney disease, autosomal recessive polycystic kidney disease, or pancreatic cysts associated with autosomal dominant polycystic kidney disease. In some embodiments, the ciliopathy is polycystic kidney disease. In some embodiments, the ciliopathy is autosomal dominant polycystic kidney disease.
[0229] Other embodiments include use of the presently disclosed compounds in therapy.
Combination Therapy
[0230] In some embodiments of the methods and uses described herein, the method or use further comprises the use of one or more additional therapeutic agents.
[0231] In some embodiments, the additional therapeutic agent is selected from the group consisting of an mTOR inhibitor, V2 receptor antagonist, tyrosine kinase inhibitor, somatostatin analog, glucosylceramide synthase inhibitor, microRNA-17 inhibitor, siRNA against p53, KEAP1-Nrf2 activator, xanthine oxidase inhibitor, PPARγ agonist, metformin, and beta hydroxybutyrate.
[0232] In some embodiments, the additional therapeutic agent is selected from the group consisting of tolvaptan, lixivaptan, mozavaptan, satavaptan, sirolimus, tacrolimus, everolimus, bosutinib, tesavatinib, imatinib, gefitinib, erlotinib, dasatinib, octreotide, pasireotide, venglustat, eliglustat, miglustat, microRNA-17 inhibitor, bardoxolone methyl, allopurinol, oxypurinol, pioglitazone, rosiglitazone, lobeglitazone, metformin, and beta by dr oxy butyr ate.
[0233] In some embodiments, the additional therapeutic agent is selected from the group consisting of an immunomodulator, a calcineurin inhibitor, a renin angiotensin aldosterone system inhibitor, an antiproliferative agent, an alkylating agent, a corticosteroid, an angiotensin converting enzyme inhibitor, an adrenocorticotropic hormone stimulant, an angiotensin receptor blocker, a sodium- glucose transport protein 2 inhibitor, a dual sodium- glucose transport protein 1/2 inhibitor, a nuclear Factor- 1 (erythroid- derived 2)-like 2 agonist, a chemokine receptor 2 inhibitor, a chemokine receptor 5 inhibitor, an endothelin 1 receptor antagonist, a beta blocker, a mineralocorticoid receptor antagonist, a loop or thiazide diuretic, a calcium channel blocker, a statin, a short- intermediate or long-acting insulin, a dipeptidyl peptidase 4 inhibitor, a glucagon-like peptide 1 receptor agonist, a sulfonylurea, an apoptosis signal-regulating kinase- 1 , a chymase inhibitor, a selective gly cation inhibitor, a renin inhibitor, an interleukin-33 inhibitor, a farnesoid X receptor agonist, a soluble guanylate cyclase stimulator, a thromboxane receptor antagonist, a xanthine oxidase inhibitor, an erythropoietin receptor agonist, a cannabinoid receptor type 1 inverse agonist, a NADPH oxidase inhibitor, an anti-vascular endothelial growth factor B, an anti-fibrotic agent, a neprilysin inhibitor, a dual CD80/CD86 inhibitor, a CD40 antagonist, a cellular cholesterol and lipid blocker, a PDGFR antagonist, a Slit guidance ligand 2, an AP0L1 inhibitor, an Nrl2 activator/NF-kB inhibitor, a somatostatin receptor agonist, a PPAR gamma agonist, a AMP activated protein kinase stimulator, a tyrosine kinase inhibitor, a glucosylceramide synthase inhibitor, an arginine vasopressin receptor 2 antagonist, a xanthine oxidase inhibitor, a vasopressin receptor 2 antagonist, anti-amyloid beta antibodies, anti-Tau antibodies, anti- synuclein antibodies, dopamine precursors (e.g. L-DOPA), dopamine agonists (e.g. bromocriptine, cabergoline, pergolide, pramipexole and apomorphine), MAO-B inhibitors (e.g. rasagiline and selegiline), anticholinergics (e.g. orphenadrine, procyclidine and trihexyphenidyl), enhancers of b- glucocerebrosidase activity (e.g. ambroxol and afegostat), amantadine, and agents capable of treating Alzheimer’s (e.g., acetylcholinesterase inhibitors such as tacrine, rivastigmine, galantamine, donepezil, and NMD A receptor antagonists such as memantine).
[0234] In some embodiments, the additional therapeutic agent is selected from the group consisting of COX inhibitors including arylcarboxylic acids (e.g. salicylic acid, acetylsalicylic acid, diflunisal, choline magnesium trisalicylate, salicylate, benorylate, flufenamic acid, mefenamic acid, meclofenamic acid, or triflumic acid), arylalkanoic acids (e.g., diclofenac, fenclofenac, alclofenac, fentiazac, ibuprofen, flurbiprofen, ketoprofen, naproxen, fenoprofen, fenbufen, suprofen, indoprofen, tiaprofenic acid, benoxaprofen, pirprofen, tolmetin, zomepirac, clopinac, indomethacin, or sulindac), an enolic acid (e.g., phenylbutazone, oxyphenbutazone, azapropazone, feprazone, piroxicam, or isoxicam), a treatment for pulmonary hypertension, including prostanoids (e.g., epoprostenol, iloprost, or treprostinil), endothelin receptor antagonist (e.g., bosentan, ambrisentan, or macitentan), phosphodiesterase-5 inhibitor (e.g., sildenafil or tadalafil), sGC stimulator (e.g., riociguat), rho-kinase inhibitor (e.g., Y-27632, fasudil, or H- 1152P), epoprostenol derivative (e.g., prostacyclin, treprostinil, beraprost, or iloprost), serotonin blocker (e.g., sarpogrelate), endothelin receptor antagonist (besentan, sitaxsentan, ambrisentan, or TBC3711), PDF inhibitor (e.g., sildenafil, tadalafil, udenafil, or vardenafil), soluble gunaylate cyclase inhibitor (e.g., riociguat or vericiguat), calcium channel blocker (e.g., amlodipine, bepridil, clentiazem, diltiazem, fendiline, gallopamil, mibefradil, prenylamine, semotiadil, terodiline, verapamil, aranidipine, bamidipine, benidipine, cilnidipine, efonidipine, elgodipine, felodipine, isradipine, lacidipine, lercanidipine, manidipine, nicardipine, nifedipine, nilvadipine, nimodipine, nisoldipine, nitrendipine, cinnarizine, flunarizine, lidoflazine, lomerizine, bencyclane, etafenone, or perhexiline), tyrosine kinase inhibitor (e.g., imatinib), inhaled nitric oxide and nitric oxide-donating agent (e.g., inhaled nitrite), IKB inhibitor (e.g., IMD 1041), prostacyclin receptor agonist (e.g., selexipag), stimulator of hematopoiesis (e.g., TXA 127 (angiotensin (1-7)), darbepoetin alfa, erythropoietin, or epoetin alfa), anticoagulant and platelet-inhibiting agent, diuretic, dietary and nutritional supplement (e.g., acetyl-L-carnitine, octacosanol, evening primrose oil, vitamin B6, tyrosine, phenylalanine, or vitamin C, L-dopa), immunosuppressant (for transplants and autoimmune-related RKD), anti-hypertensive drug (for high blood pressure -related RKD, e.g., angiotensin-converting enzyme inhibitors and angiotensin receptor blockers), insulin (for diabetic RKD), lipid/cholesterol-lowering agents (e.g., HMG- CoA reductase inhibitors such as atorvastatin or simvastatin), and treatments for hyperphosphatemia or hyperparathyroidism associated with CKD (e.g., sevelamer acetate, cinacalcet).
4. Kits
[0235] Provided herein are also kits that include a compound of the disclosure, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and suitable packaging. In certain embodiments, a kit further includes instructions for use. In one aspect, a kit includes a compound of the disclosure, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, and a label and/or instructions for use of the compounds in the treatment of the indications, including the diseases or conditions, described herein.
[0236] Provided herein are also articles of manufacture that include a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof in a suitable container. The container may be a vial, jar, ampoule, preloaded syringe, or intravenous bag.
5. Pharmaceutical Compositions and Modes of Administration
[0237] Compounds provided herein are usually administered in the form of pharmaceutical compositions. Thus, provided herein are also pharmaceutical compositions that contain one or more of the compounds described herein, or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers, or prodrug thereof, and one or more pharmaceutically acceptable vehicles selected from carriers, adjuvants, and excipients. Suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents, including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers, and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical art. See, e.g., Remington’s Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G.S. Banker & C.T. Rhodes, Eds.).
[0238] The pharmaceutical compositions may be administered in either single or multiple doses. The pharmaceutical composition may be administered by various methods including, for example, rectal, buccal, intranasal, and transdermal routes. In certain embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenously, intraperitoneally, parenterally, intramuscularly, subcutaneously, orally, topically, or as an inhalant.
[0239] One mode for administration is parenteral, for example, by injection. The forms in which the pharmaceutical compositions described herein may be incorporated for administration by injection include, for example, aqueous or oil suspensions, or emulsions, with sesame oil, corn oil, cottonseed oil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterile aqueous solution, and similar pharmaceutical vehicles. [0240] Oral administration may be another route for administration of the compounds described herein. Administration may be via, for example, capsule or enteric coated tablets. In making the pharmaceutical compositions that include at least one compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, the active ingredient is usually diluted by an excipient and/or enclosed within such a carrier that can be in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be in the form of a solid, semi-solid, or liquid material, which acts as a vehicle, carrier or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solid or in a liquid medium), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.
[0241] Some examples of suitable excipients include, e.g., lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup, and methyl cellulose. The formulations can additionally include lubricating agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preserving agents such as methyl and propylhydroxy- benzoates; sweetening agents; and flavoring agents.
[0242] The compositions that include at least one compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof can be formulated so as to provide quick, sustained or delayed release of the active ingredient after administration to the subject by employing procedures known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolutional systems containing polymer-coated reservoirs or drug-polymer matrix formulations. Another formulation for use in the methods disclosed herein employ transdermal delivery devices (“patches”). Such transdermal patches may be used to provide continuous or discontinuous infusion of the compounds described herein in controlled amounts. The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.
[0243] For preparing solid compositions such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogeneous mixture of a compound described herein or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof. When referring to these preformulation compositions as homogeneous, the active ingredient may be dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules.
[0244] The tablets or pills of the compounds described herein may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action, or to protect from the acid conditions of the stomach. For example, the tablet or pill can include an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer that serves to resist disintegration in the stomach and permit the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
[0245] Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable, aqueous, or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described herein. In certain embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. In other embodiments, compositions in pharmaceutically acceptable solvents may be nebulized by use of inert gases. Nebulized solutions may be inhaled directly from the nebulizing device, or the nebulizing device may be attached to a facemask tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered, in one embodiment, orally or nasally, from devices that deliver the formulation in an appropriate manner.
[0246] The amount of the compound in a pharmaceutical composition or formulation can vary within the full range employed by those skilled in the art. Typically, the formulation will contain, on a weight percent (wt %) basis, from about 0.01-99.99 wt % of a compound of this disclosure based on the total formulation, with the balance being one or more suitable pharmaceutical excipients. In one embodiment, the compound is present at a level of about 1-80 wt %. Representative pharmaceutical formulations are described below.
Formulation Example 1 - Tablet formulation
[0247] The following ingredients are mixed intimately and pressed into single scored tablets.
Formulation Example 2 - Capsule formulation
[0248] The following ingredients are mixed intimately and loaded into a hard-shell gelatin capsule Formulation Example 3 - Suspension formulation
[0249] The following ingredients are mixed to form a suspension for oral administration.
Formulation Example 4 - Injectable formulation
[0250] The following ingredients are mixed to form an injectable formulation.
Formulation Example 5 - Suppository Formulation
[0251] A suppository of total weight 2.5 g is prepared by mixing the compound of this disclosure with Witepsol® H-15 (triglycerides of saturated vegetable fatty acid; Riches-Nelson, Inc., New York), and has the following composition:
6. Dosing
[0252] The specific dose level of a compound of the present application for any particular subject will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease in the subject undergoing therapy. For example, a dosage may be expressed as a number of milligrams of a compound described herein per kilogram of the subject’s body weight (mg/kg). Dosages of between about 0.1 and 150 mg/kg may be appropriate. In certain embodiments, about 0.1 and 100 mg/kg may be appropriate. In other embodiments a dosage of between 0.5 and 60 mg/kg may be appropriate. In certain embodiments, a dosage of from about 0.0001 to about 100 mg per kg of body weight per day, from about 0.001 to about 50 mg of compound per kg of body weight, or from about 0.01 to about 10 mg of compound per kg of body weight may be appropriate. Normalizing according to the subject’s body weight is particularly useful when adjusting dosages between subjects of widely disparate size, such as occurs when using the drug in both children and adult humans or when converting an effective dosage in a non-human subject such as dog to a dosage suitable for a human subject.
7. Synthesis of the Compounds
[0253] The compounds may be prepared using the methods disclosed herein and routine modifications thereof, which will be apparent given the disclosure herein and methods well known in the art. Conventional and well-known synthetic methods may be used in addition to the teachings herein. The synthesis of typical compounds described herein may be accomplished as described in the following examples. If available, reagents and starting materials may be purchased commercially, e.g., from Sigma Aldrich or other chemical suppliers.
[0254] It will be appreciated that where typical process conditions (i.e., reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures. [0255] Additionally, conventional protecting groups (“PG”) may be necessary to prevent certain functional groups from undergoing undesired reactions. Suitable protecting groups for various functional groups as well as suitable conditions for protecting and deprotecting particular functional groups are well known in the art. For example, numerous protecting groups are described in Wuts, P. G. M., Greene, T. W., & Greene, T. W. (2006). Greene’s protective groups in organic synthesis. Hoboken, N.J., Wiley- Interscience, and references cited therein. For example, protecting groups for alcohols, such as hydroxy, include silyl ethers (including trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso- propylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ethers), which can be removed by acid or fluoride ion, such as NaF, TBAF (tetra-n-butylammonium fluoride), HF-Py, or HF-NEt3. Other protecting groups for alcohols include acetyl, removed by acid or base, benzoyl, removed by acid or base, benzyl, removed by hydrogenation, methoxyethoxymethyl ether, removed by acid, dimethoxytrityl, removed by acid, methoxymethyl ether, removed by acid, tetrahydropyranyl or tetrahydrofuranyl, removed by acid, and trityl, removed by acid. Examples of protecting groups for amines include carbobenzyloxy, removed by hydrogenolysis p-methoxybenzyl carbonyl, removed by hydrogenolysis, tert-butyloxycarbonyl, removed by concentrated strong acid (such as HC1 or CF3COOH), or by heating to greater than about 80 °C, 9-fluorenylmethyloxycarbonyl, removed by base, such as piperidine, acetyl, removed by treatment with a base, benzoyl, removed by treatment with a base, benzyl, removed by hydrogenolysis, carbamate group, removed by acid and mild heating, p-methoxybenzyl, removed by hydrogenolysis, 3,4-dimethoxybenzyl, removed by hydrogenolysis, p-methoxyphenyl, removed by ammonium cerium(IV) nitrate, tosyl, removed by concentrated acid (such as HBr or H2SO4) and strong reducing agents (sodium in liquid ammonia or sodium naphthalenide), troc (trichloroethyl chloroformate), removed by Zn insertion in the presence of acetic acid, and sulfonamides (Nosyl & Nps), removed by samarium iodide or tributyltin hydride.
[0256] Furthermore, the compounds of this disclosure may contain one or more chiral centers. Accordingly, if desired, such compounds can be prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or diastereomers or as stereoisomer-enriched mixtures. All such stereoisomers
(and enriched mixtures) are included within the scope of this disclosure, unless otherwise indicated. Pure stereoisomers (or enriched mixtures) may be prepared using, for example, optically active starting materials or stereoselective reagents well-known in the art. Alternatively, racemic mixtures of such compounds can be separated using, for example, chiral column chromatography, chiral resolving agents, and the like.
[0257] The starting materials for the following reactions are generally known compounds or can be prepared by known procedures or obvious modifications thereof. For example, many of the starting materials are available from commercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA), Bachem (Torrance, California, USA), Emka-Chemce or Sigma (St. Louis, Missouri, USA). Others may be prepared by procedures or obvious modifications thereof, described in standard reference texts such as Fieser and Fieser’s Reagents for Organic Synthesis, Volumes 1-15 (John Wiley, and Sons, 1991), Rodd’s Chemistry of Carbon Compounds, Volumes 1-5, and Suppiementals (Elsevier Science Publishers, 1989) organic Reactions, Volumes 1-40 (John Wiley, and Sons, 1991), March’s Advanced Organic Chemistry, (John Wiley, and Sons, 5th Edition, 2001), and Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., 1989).
General Synthesis
[0258] Scheme I illustrates general methods which can be employed for the synthesis of compounds described herein (e.g., Formula I), where Ring A, L, X1, X2, X3, the dashed line, R1, R3, R4, R8, R9, R10, p, m, n, ml, m2, m3, nl, n2, n3, and q are each independently as defined herein, and LG is a leaving group (e.g., halo, etc.). Scheme I
[0259] In Scheme I, compounds of Formula I can be prepared by contacting compound 1-1 with an amine 1-2 under suitable coupling reaction conditions, followed by optional functionalization or deprotection when required. Upon reaction completion, compounds of Formula I can be recovered and purified by conventional techniques such as neutralization, extraction, precipitation, chromatography, filtration and the like.
[0260] Further derivatization of the compound provided by the steps outlined in Scheme I, or any intermediate, provides additional compounds. It should be understood that any of the compounds or intermediates shown in Scheme I may be prepared using traditional methods or purchased from commercial sources. In addition, any of the intermediates or any product obtained by the process outlined in Scheme I can be derivatized at any step to provide various compounds of Formula I. In certain embodiments, the various substituents of the compounds or intermediates as used in Scheme I are as defined herein (e.g., for Formula I).
[0261] Scheme II shows an exemplary methods for further derivatization of R3, for Example. In Scheme III, each R8, R9, p, m, n, ml, m2, nl, and n2 are independently as defined herein, and LG is a leaving group (e.g., halo, alkoxy, etc.). Scheme II
[0262] In Scheme II, the BOC protected R3 moiety is deprotected to liberate the free amine, which can then be functionalized with compound III- 1 to provide the desired product.
[0263] In certain embodiments, provided is a process for providing a compound of Formula I: comprising contacting a compound of Formula 1-1 : with a compound of Formula 1-2: under conditions sufficient to provide the compound of Formula I; wherein Ring A, L, X1, X2, X3, the dashed line, R1, R3, R4, R8, and q are each independently as defined herein. EXAMPLES
[0264] The following examples are included to demonstrate specific embodiments of the disclosure. It should be appreciated by those skilled in the art that the techniques disclosed in the examples which follow represent techniques to function well in the practice of the disclosure, and thus can be considered to constitute specific modes of its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.
General Experimental Methods
[0265] Reagents and solvents obtained from commercial suppliers were used without purification or drying unless otherwise noted. 1H NMR were recorded using a Broker NMR tuned to 400 MHz with TMS used as an internal standard. LC-MS analysis was performed on a Waters UPLC with SQD-2 mass detector (Single quadruple). HPLC analysis was performed on a WATERS Acquity UPLC H CLASS or Acquity Arc system (with PDA & ELSD Detector). Purifications with p-HPLC were conducted using the following columns: Waters Xbridge Prep OBD 150 x 40 mm x 10 μm, Waters Xbridge BEH 100 x 30 mm x 10 μm, or Phenomenex Luna C18 100 x 40 mm x 3 μm.
Intermediates la & lb
[0266] [l,l'-biphenyl]-4-ylmethyl (R)-2-cyano-3-fluoro-2-methylpropanoate (Intermediate la) & [1,1'- biphenyl]-4-ylmethyl (S)-2-cyano-3-fluoro-2-methylpropanoate (intermediate lb)
[0267] (4-phenylphenyl)methyl 2-cyanopropanoate: To a solution of 2-cyanopropanoic acid (4 g, 40.4 mmol) and (4-phenylphenyl)methanol (8.92 g, 48.44 mmol) in DCM (60 mL) was added EDCI (15.48 g, 80.74 mmol) and DMAP (2.47 g, 20.18 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into H2O (80 mL) and extracted with DCM (3 x 50 mL). The combined organic phase was washed with brine (150 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to afford (4-phenylphenyl)methyl 2-cyanopropanoate (8.8 g, 82%).
[0268] (4-phenylphenyl)methyl 2-cyano-3-hydroxy-2-methyl-propanoate: To a solution of (4- phenylphenyl)methyl 2-cyanopropanoate (6.8 g, 25.6 mmol) in CH3 CN (68 mL) was added TEA (129.7 mg, 1.28 mmol) and formaldehyde (2.31 g, 76.9 mmol). The mixture was stirred at 50 °C for 16 h. The reaction mixture was filtered and concentrated under reduced pressure. The remaining mixture was poured into H2O (100 mL) and extracted with EtOAc (3 x 80 mL). The combined organic phase was washed with brine (150 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to afford (4-phenylphenyl)methyl 2-cyano-3-hydroxy-2-methyl-propanoate (6.6 g, 87%).
[0269] [l,l'-biphenyl]-4-ylmethyl 2-cyano-3-fluoro-2-methylpropanoate: To a solution of (4- phenylphenyl)methyl 2-cyano-3-hydroxy-2-methyl-propanoate (6.6 g, 22.35 mmol) in DCM (120 mL) was added DAST (18.01 g, 111.74 mmol) dropwise at 0 °C. The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into aqueous NaHCO3 (150 mL) and extracted with DCM (3 x 80 mL). The combined organic phase was washed with brine (200 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to afford [l,l'-biphenyl]-4-ylmethyl 2-cyano-3-fluoro-2-methylpropanoate (6.1 g, 92%). The racemic mixture was separated by chiral SEC as described below.
[0270] Chiral Separation: [l,l'-biphenyl]-4-ylmethyl 2-cyano-3-fluoro-2-methylpropanoate (Intermediate la & lb): The product [l,l'-biphenyl]-4-ylmethyl 2-cyano-3-fluoro-2-methylpropanoate (8.1 g) was used for SEC separation (column: DAICEL CHIRALPAK IG (250 mm*50 mm, 10 μm); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 25%, isocratic elution mode) to give [l,l'-biphenyl]- 4-ylmethyl (R)-2-cyano-3-fluoro-2-methylpropanoate (4.02 g, 49%, peak 1 (intermediate la)) and [1,1'- biphenyl]-4-ylmethyl (S)-2-cyano-3-fluoro-2-methylpropanoate (3.56 g, 44%, peak 2 (intermediate lb)). The absolute stereochemistry was determined using X-ray crystallography.
(R)-2-cyano-3-fluoro-2-methylpropanoic acid (intermediate 2a) & (S)-2-cyano-3-fluoro-2- methylpropanoic acid (intermediate 2b)
[0271] Intermediate 2a: To a solution of Intermediate la (4.00 g, 13.45 mmol) in THF (40 mL) and H2O (13 mL) was added LiOH.H2O (1.41 g, 33.63 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture concentrated under reduced pressure to remove the THF. The remaining aqueous mixture was diluted with water (50 mL), extracted with MTBE (2 x 20 mL), and acidified with HC1 (1 M) to pH~l. The mixture was extracted with EtOAc (3 x 30 mL). The combined organic phase was dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give Intermediate 2a (1.6 g, 91%).
[0272] Intermediate 2b: To a solution of Intermediate lb (3.56 g, 11.97 mmol) in THF (35 mL) and H2O (11 mL) was added LiOH.H2O (1.26 g, 29.93 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture filtered and concentrated under reduced pressure to remove THF. The remained aqueous layer was diluted with water (50 mL), extracted with MTBE (2x 20 mL), and acidified with HC1 (1 N) to pH~l. The mixture was extracted with EtOAc (3 x 30 mL). The combined organic phase was dried with anhydrous Na2SO4, filtered, and concentrated to give Intermediate 2b (1.3 g, 82%). (4-phenylphenyl)methyl (23)-2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate (intermediate 3a) and (4-phenylphenyl)methyl (2 R)-2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl- propanoate (intermediate 3b)
[0273] (4-phenylphenyl)methyl 2-cyanopropanoate: To a solution of 2-cyanopropanoic acid (45 g, 454.14 mmol) and (4-phenylphenyl)methanol (87.85 g, 476.85 mmol) in DCM (900 mL) was added EDCI (95.77 g, 499.56 mmol) and DMAP (5.55 g, 45.41 mmol) with stirring at 0 °C. The mixture was warmed to 25 °C and stirred for 12 h under N2 (g). I L saturated NH4CI aqueous solution was added to the reaction mixture. The resulting mixture was extracted with DCM (600 mL x 3). The combined organic phase was washed with brine (IL) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 1:0 to 1:1) to give (4-phenylphenyl)methyl 2-cyanopropanoate (164 g, 69%).
[0274] (4-phenylphenyl)methyl 2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate: To a solution of (4-phenylphenyl)methyl 2-cyanopropanoate (45 g, 169.6 mmol) in THE (450 mL) was added n-BuLi (2.5 M, 74.6 mL) at -78 °C and stirred for 10 min under Ni(g). TMSCF3 (72.4 g, 508.9 mmol) was added to the reaction mixture at -78 °C. The resulting mixture was warmed to 20 °C and stirred for 2 h under N2 (g). The reaction mixture was added to 1 L saturated NH4CI aqueous solution at 0 °C, the resulting mixture was extracted with EtOAc (800 mL x 3). The combined organic phase was washed with brine (1 L) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 1:0 to 1:1) to give (4-phenylphenyl)methyl 2-cyano-3,3-difluoro-2- methyl-3-trimethylsilyl-propanoate (40 g, 61%). The racemic mixture was separated by chiral SEC as described below to obtain enantiopure product.
[0275] (4-phenylphenyl)methyl (2R)-2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate and [(4-phenylphenyl)methyl (2S)-2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate : phenylphenyl)methyl 2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate (75 g, 193.55 mmol) was used for SEC separation (column: DAICEL CHIRALPAK AY (250 mm*50 mm, lOum); mobile phase: [CO2-IPA]; B%:25%, isocratic elution mode) to give isomer 1 (4-phenylphenyl)methyl (2S)-2- cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate (31 g, 41% yield, peak 1 (intermediate 3a)) and isomer 2 (4-phenylphenyl)methyl (2R)-2-cyano-3,3-difluoro-2-methyl-3-trimethylsilyl-propanoate (30 g, 40% yield, peak 2 (intermediate 3b)). The absolute stereochemistry was determined using X-ray crystallography. (S)-2-cyano-3,3-difluoro-2-methylpropanoic acid (Intermediate 4a) & (R)-2-cyano-3,3-difluoro-2- methylpropanoic acid (Intermediate 4b)
[0276] (S)-2-cyano-3,3-difluoro-2- methylpropanoic acid (Intermediate 4a): To a solution of Intermediate 3a (5 g, 12.90 mmol) in MeCN (60 mL) and H2O (20 mL) was added CsF (3.92 g, 25.81 mmol) at 20 °C and stirred for 1 h. NaOH (1.03 g, 25.81 mmol) in H2O (8 mL) was added to the reaction and stirred at 20 °C for 1 h. Seven batches were combined and the combined reaction mixture was poured into water (I L). The reaction mixture was extracted with DCM (600 mL x 4). The aqueous phase was adjusted to pH~2 with HC1 (2N) and then extracted with EtOAc (600 mL x 3). The combined organic phase was concentrated under reduced pressure to give (5)-2-cyano-3,3-difluoro-2- methylpropanoic acid (Intermediate 4a) (11 g, 69%). The absolute stereochemistry was determined using X-ray crystallography.
[0277] (R)-2-cyano-3,3-difluoro-2-methylpropanoic acid (Intermediate 4b): To a solution of Intermediate 3b (5 g, 12.90 mmol) in H2O (20 mL) and MeCN (60 mL) was added CsF (3.92 g, 25.81 mmol) at 20 °C and stirred for 1 h. NaOH (1.03 g, 25.81 mmol) in H2O (8 mL) was added to the reaction and stirred at 20 °C for 1 h. Nine batches were combined and the combined reaction mixture was poured into water (1.2 L) and extracted with DCM (600 mL x 4). The aqueous phase was adjust to pH~2 with HC1 (2N) and extracted with EtOAc (600 mL x 3). The combined organic phase was concentrated under reduced pressure to give (R)-2-cyano-3,3-difluoro-2-methylpropanoic acid (Intermediate 4b) (11.3 g, 60%). The absolute stereochemistry was determined using X-ray crystallography.
Example 1 (2R,5A)- tert -butyl 4-(5-cyclopropyl-8-phenylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate
[0278] 5-bromo-8-chloroquinazolin-4(3H)-one: 2-amino-6-bromo-3-chlorobenzoic acid (0.8 g, 3.99 mmol), trimethoxymethane (1.02 g, 9.58 mmol) and ammonium acetate (738 mg, 9.58 mmol) were taken up into a microwave tube in MeOH (20 mL). The sealed tube was heated at 120 °C for 3 h under microwave. The reaction mixture was concentrated under reduced pressure. Three batches were combined. The crude product was triturated with H2O (10 mL) and filtered to give 5 -bromo- 8- chloroquinazolin-4(3H)-one (2 g, 80%). [0279] (2R,5S)-tert -butyl 4-(5-bromo-8-chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 5-bromo-8-chloroquinazolin-4(3H)-one (1 g, 3.85 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (908 mg, 4.24 mmol) in MeCN (10 mL) was added DBU (2.93 g, 19.27 mmol) and PYBOP (3.01 g, 5.78 mmol). The mixture was stirred at 20 °C for 12 h. The mixture was quenched with H2O (30 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EtOAc = 5:1 to 4:1) to give (2R, 5S)- tert-butyl 4-(5-bromo-8-chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (500 mg, 28%). [0280] (2R,5S)-tert -butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5-bromo-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (350 mg, 0.77 mmol) and cyclopropylboronic acid (66 mg, 0.77 mmol) in dioxane (3 mL) was added Pd(dppf)Ch (56 mg, 0.07 mmol) and K3PO4 (489 mg, 2.30 mmol), Ag2O (89 mg, 0.38 mmol) under N2. The mixture was stirred at 90 °C for 3h. The mixture was quenched with H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE:EtOAc = 4:1 to 3:1) to give (2R,5S)-tert-butyl4-(8-chloro-5- cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (300 mg, 94%).
[0281] (2R ,5S)-tert -butyl 4-(5-cyclopropyl-8-phenylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (220 mg, 0.53 mmol) and phenylboronic acid (320 mg, 2.64 mmol) in dioxane (5 mL) and H2O (0.2 mL) was added Pd(dppf)Ch (77 mg, 0.11 mmol) and CS2CO3 (516 mg, 1.58 mmol). The mixture was stirred at 120 °C for 16 h. The mixture was quenched with H2O (15 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: Xbridge BEH C18 100 x 30 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 60%-90%, 8 min) to give (2R,5S)-tert-butyl 4-(5-cyclopropyl-8-phenylquinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (82.9 mg, 34%).
Example 2 (2R,5S)-tert -butyl 4-[6-bromo-8-(4-cyano-2-pyridinyl)quinazolin-4-yl]-2,5-dimethylpiperazine-l- carboxylate [0282] 6-bromo-8-iodoquinazolin-4(3H)-one: A solution of methyl 2-amino-5-bromo-3-iodobenzoate (500 mg, 1.40 mmol) and ammonium formate (177 mg, 2.81 mmol) in formamide (2 mL) was heated in a microwave reactor for 30 min at 200 °C. The mixture was then diluted with H2O (20 mL) and briefly sonicated. The resulting solid was isolated by filtration, washed with H2O (20 mL x 2) to give 6-bromo- 8-iodoquinazolin-4(3H)-one (420 mg, 43%).
[0283] (2R ,5.S)-tert -buty I 4-(6-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 6-bromo-8-iodoquinazolin-4(3H)-one (320 mg, 0.91 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (215 mg, 1.00 mmol) in MeCN (11 mL) was added DBU (694 mg, 4.56 mmol) and PYBOP (712 mg, 1.37 mmol) at 20 °C. The mixture was stirred at 20 °C for 12 h. The reaction mixture was poured into H2O (30 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to afford (2R,5S)-tert-butyl 4-(6-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (180 mg, 36%).
[0284] 2-(trimethylstannyl)isonicotinonitrile: To a solution of 2-bromopyridine-4-carbonitrile (2 g, 10.93 mmol) in toluene (50 mL) was added Pd(PPh3)4 (631 mg, 0.555 mmol) and 1, 1,1, 2,2,2- hexamethyldistannane (8.77 g, 26.77 mmol) at 20 °C under N2. The mixture was stirred at 110 °C for 16 h. The reaction mixture was cooled to 20 °C and concentrated under reduced pressure. The residue was purified by column chromatography (AI2O3, PE:EA = 5:1 to 1:1) to afford 2- (trimethylstannyl)isonicotinonitrile (1.2 g, 41%).
[0285] (2R ,5.S)-tert -butyl 4-(6-bromo-8-(4-cyanopyridin-2-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(6-bromo-8-iodoquinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.15 mmol) and 2-(trimethylstannyl)isonicotinonitrile (94 mg, 0.18 mmol) in DMF (1 mL) was added Pd(PPh3)4 (34 mg, 0.029 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100 x 30 mm xlO μm; mobile phase: [A: 10 mM NH4HCO3 in water, B: CH3CN; B% in A: 50%-80%], 8 min) to give (2R,5S)-tert-butyl 4-(6-bromo-8-(4-cyanopyridin-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (7.25 mg, 9%). Example 3
(2R,53)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)-6-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0286] (2R ,5.S )-tert -hutyl 4-(8-(4-cyanopyridin-2-yl)-6-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(6-bromo-8-(4-cyanopyridin- 2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (70 mg, 0.13 mmol) and cyclopropylboronic acid (34 mg, 0.40 mmol) in 1,4-dioxane (1 mL) and H2O (0.1 mL) was added Pd(dppf)Ch (10 mg, 0.013 mmol) and K2CO3 (55 mg, 0.40 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layer was washed with brine (3 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by p-TLC (PE:EtOAc = 3:1) to afford (2R,5S)-tert-butyl 4-(8-(4- cyanopyridin-2-yl)-6-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (19.33 mg, 30%).
Example 4 (2R,5S)-tert-butyl 2,5-dimethyl-4-(8-phenylquinazolin-4-yl)piperazine-l-carboxylate
[0287] (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of 8-bromo-4-chloroquinazoline (1 g, 4.11 mmol) and tert-butyl (2R,5S)-2,5- dimethylpiperazine-1 -carboxylate (968 mg, 4.52 mmol) in NMP (10 mL) was added DIEA (1.33 g, 10.27 mmol) at 20 °C. The mixture was stirred at 100 °C for 8 h. The reaction mixture was poured into H2O (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to afford (2R,5S)-tert-butyl 4-(8- bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (680 mg, 39%). [0288] (2R,5S)-tert -butyl 2,5-dimethyl-4-(8-phenylquinazolin-4-yl)piperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.19 mmol) and phenylboronic acid (116 mg, 0.95 mmol) in dioxane (1 mL) and H2O (0.2 mL) was added Pd(dppf)Ch (28 mg, 0.038 mmol) and CS2CO3 (186 mg, 0.57 mmol) at 20 °C under N2. The mixture was stirred at 120 °C for 16 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layer was washed with brine (3 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100 x 30 mm xlO μm; mobile phase: [A: 10 mM NH4HCO3 in water, B: CH3 CN; B% in A: 52%-82%], 8 min) to give (2R,5S)-tert-butyl 2,5-dimethyl-4-(8-phenylquinazolin-4- yl)piperazine-l -carboxylate (72.12 mg, 90%).
Example 5 (2R,5S)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate
[0289] (2R ,5.S )-tert -butyl 4-(8-(4-cyanopyridin-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (200 mg, 0.47 mmol) and 2-(trimethylstannyl)isonicotinonitrile (507 mg, 0.95 mmol) in DMF (2 mL) was added Pd(PPh3)4 (110 mg, 0.095 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layer was washed with brine (3 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to afford the product (160 mg, 75%) and 50 mg of the product was further purified by p-TLC (SiO2, PE:EtOAc = 1:1) to afford the pure (2R,5S)-tert-butyl 4-(8-(4-cyanopyridin-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (19.42 mg). Example 6
2-(4-((2S,5R)-2,5-dimethyl-4-pivaloylpiperazin-l-yl)quinazolin-8-yl)isonicotinonitrile
[0290] 2-(4-((2,S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)isonicotinonitrile hydrochloride: To a solution of (2R,5S)-tert-butyl 4-(8-(4-cyanopyridin-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (120 mg, 0.27 mmol) in EtOAc (1 mL) was added HCl/EtOAc (4 N, 5 mL) at 20 °C. The mixture was stirred at 20 °C for 2 h. The reaction mixture concentrated under reduced pressure to give the 2-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)isonicotinonitrile hydrochloride (110 mg, crude).
[0291] 2-(4-((2S,5R)-2,5-dimethyl-4-pivaloylpiperazin-l-yl)quinazolin-8-yl)isonicotinonitrile: To a solution of 2-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)isonicotinonitrile hydrochloride (50 mg, 0.13 mmol) in DCM (1 mL) was added TEA (40 mg, 0.39 mmol) and pivaloyl chloride (32 mg, 0.26 mmol) at 0 °C under N2. The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into H2O (3 mL) and extracted with DCM (3 x 2 mL). The combined organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by p-TLC (SiO2, PE:EtOAc = 1:1) to afford 2-(4-((2S,5R )-2,5-dimethyl-4-pivaloylpiperazin-l- yl)quinazolin-8-yl)isonicotinonitrile (28.47 mg, 50%).
Example 7
(2R,5S)-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate
[0292] (2R ,5.S )-tert -hutyl 4-(8-(4-cyano-1H -pyrazol-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (60 mg, 0.14 mmol) in DMF (2 mL) was added 1H-pyrazole-4-carbonitrile (40 mg, 0.43 mmol), K2CO3 (49 mg, 0.36 mmol), (1R,2R)-N1,N2-dimethylcyclohexane-l,2-diamine (6 mg, 0.04 mmol) and Cui (3 mg, 0.02 mmol) at 25 °C under N2. The mixture was stirred at 120 °C for 2 h. The reaction mixture was diluted with H2O (10 mL). The aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (Waters Xbridge BEH C18 100 x 30 mm x 10 um; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B in A: 50%-80%, over 8.0 min) to give (2R,5>S)-tert butyl 4-(8-(4-cyano-1H-pyrazol-l-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (35.4 mg, 28%).
Example 8 (2R,5S)-tert -butyl 4-(8-(cyclobutylcarbamoyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate
[0293] 2,4,6-trichlorophenyl 4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l- yl)quinazoline-8-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (100 mg, 0.23 mmol) and (2,4,6-trichlorophenyl) formate (81 mg, 0.35 mmol) in Toluene (1 mL) was added TEA (73 mg, 0.71 mmol), Pd(OAc)2 (6 mg, 0.02 mmol) and Xantphos (28 mg, 0.05 mmol) at 25 °C under N2. The mixture was stirred at 110°C for 2 h in sealed tube. The reaction solution was concentrated under reduce pressure. The residue was purified by prep-TLC (SiO2, PE:EA = 3:1) to give 2,4,6-trichlorophenyl 4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)quinazoline-8-carboxylate (70 mg, 52%).
[0294] (2R ,5.S)-tert -butyl 4-(8-(cyclobutylcarbamoyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2,4,6-trichlorophenyl) 4-[(2S,5R)-4-tert-butoxycarbonyl-2,5-dimethyl- piperazin-l-yl]quinazoline-8-carboxylate (60 mg, 0.11 mmol) and cyclobutylamine (23 mg, 0.32 mmol) in THE (2 mL) was added DMAP (7 mg, 0.06 mmol) and TEA (54 mg, 0.53 mmol) at 25 °C. The mixture was stirred at 50 °C for 2 h. The reaction mixture was diluted with H2O (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: Waters Xbridge BEH Cl 8 100 x 30 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 45%-75%, 8 min) to give (2R,5S)-tert-butyl 4-(8- (cyclobutylcarbamoyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (8.81 mg, 18%). Example 9 (2R,5S)-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)pyrido[3,4- d|pyrimidin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0295] 3-amino-2-chloroisonicotinic acid: To a solution of methyl 3-amino-2-chloroisonicotinate (2 g, 10.72 mmol) in MeOH (10 mL) and H2O (10 mL) was added NaOH (1.16 g, 28.94 mmol). The mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove MeOH. The aqueous solution was adjusted pH to 3~4 by 3N HC1. The precipitate was filtered and dried under reduced pressure to give 3-amino-2-chloroisonicotinic acid (1.7 g, 92%).
[0296] 8-chloropyrido[3,4-d]pyrimidin-4(3H)-one : To a solution of 3-amino-2-chloroisonicotinic acid (500 mg, 2.90 mmol) in n-BuOH (10 mL) was added formamidine acetate (1.51 g, 14.49 mmol) at 20 °C. The mixture was stirred at 140 °C for 40 h. The reaction mixture was filtered and the solid was collect o give 8-chloropyrido[3,4- d]pyrimidin-4(3H)-one (600 mg, 57%).
[0297] (2R ,5S)-tert -butyl 4-(8-chloropyrido[3,4-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 8-chloropyrido|3.4-d |pyrimidin-4(3/7)-onc (600 mg, 3.30 mmol) and tert- butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (779 mg, 3.63 mmol) in MeCN (10 mL) was added DBU (2.52 g, 16.52 mmol) and PYBOP (2.58 g, 4.96 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into H2O (30 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-TLC (SiO2, PE:EA = 1:1) to give (2R,5S)-tert- butyl 4-(8-chloropyrido[3,4-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 12%).
[0298] (2R,5S)-tert-butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)pyrido[3,4-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloropyrido[3,4- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (70 mg, 0.19 mmol) and 1H-pyrazole-4- carbonitrile (51.73 mg, 0.55 mmol) in DMF (1.5 mL) was added K2CO3 (64 mg, 0.46 mmol), (15,25)- N1,N2-dimcthylcyclohcxanc- l ,2-diaminc (8 mg, 0.06 mmol), and Cui (4 mg, 0.02 mmol) at 20 °C under N2. The mixture was stirred at 110 °C for 16 h. The reaction mixture was poured into H2O (10 mL), and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1 to 0:1) and further purified by prep-HPLC (column: Waters Xbridge Prep OBD C18: 150 x 40mm x 10 um; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 30%-67%, 8 min) to give tert-butyl (2R,5S)-tert-butyl 4-(8-(4-cyano-1H-pyrazol-l- yl)pyrido[3,4-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (6.06 mg, 7 %).
Example 10
(2R,5S)-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0299] (2R ,5.S )-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5- cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (300 mg, 0.72 mmol) in DMF (2 mL) was added 1H-pyrazole-4-carbonitrile (201 mg, 2.16 mmol), K2CO3 (249 mg, 1.80 mmol), (1R, 2R)- N1,N2-dimcthylcyclohcxanc-l ,2-diaminc (31 mg, 0.22 mmol) and Cui (14 mg, 0.07 mmol) at 25 °C under N2. The mixture was stirred to 110 °C for 16 h. The reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was first purified by silica gel column chromatography (PE:EtOAc = 4:1 to 3:1) to give the crude product (150 mg), which was further purified by prep-HPLC with the following conditions: (column: Waters Xbridge Prep OBD C18 150 x 40 mm x 10 μm; mobile phase: A: 10 mm NH4HCO3 in water, B: MeCN; B in A: 45%-75%, over 8.0 min) to give (2R,5S)-tert-butyl 4-(8-(4-cyano-1H-pyrazol-l-yl)-5-cyclopropylquinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (14.67 mg, 4%).
Example 11 (2R,5S)-tert -butyl 4-(8-(3-cyanopyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate
[0300] (2R ,5.S)-tert -butyl 4-(8-(3-cyanopyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (100 mg, 0.23 mmol) in dioxane (1 mL) and pyrrolidine-3-carbonitrile hydrochloride (31 mg, 0.23 mmol) was added DIEA (92 mg, 0.71 mmol), XantPhos (27 mg, 0.04 mmol), t-BuONa (57 mg, 0.59 mmol) and Pd2(dba)3 (22 mg, 0.02 mmol) at 20 °C. The mixture was stirred at 100 °C for 16 h. The reaction mixture was diluted with water (5 mL), extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna Cl 8 100 x 30mm x 3μm; Mobile phase: A: 0.2% FA in water and B: MeCN; B% in A: 20%-60%, 8.0 min) to give (2R,5S)-tert-butyl 4-(8-(3-cyanopyrrolidin- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (16.32 mg, 15%).
Example 12 (2R,5A)-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)pyrido[4,3-d] pyrimidin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0301] (2R ,5.S)-tert -butyl 4-(8-bromopyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 8-bromo-4-chloropyrido[4,3-d]pyrimidine (0.2 g, 0.82 mmol) and tert- butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (351 mg, 1.64 mmol) in NMP (5 mL) was added DIEA (529 mg, 4.09 mmol). The mixture was stirred at 140 °C for 16 h. The reaction mixture was poured into H2O (15 mL) at 20 °C, and then extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 3:1 to 1:1) to give (2R,5S)-tert-butyl 4-(8-bromopyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (300 mg, 87 %).
[0302] (2R ,5.S )-tert -butyl 4-(8-(4-cyano-1H -pyrazol-l-yl)pyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromopyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (150 mg, 0.36 mmol) and 1H-pyrazole-4- carbonitrile (66 mg, 0.71 mmol) in dioxane (3 mL) was added CS2CO3 (231 mg, 0.71 mmol), (Bu4NICuI)2 (60 mg, 0.07 mmol) and (1R,2R)-N1,N2-dimethylcyclohexane-l,2-diamine (20 mg, 0.14 mmol) at 20 °C under N2. The mixture was stirred at 120 °C for 16 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EtOAc = 3:1 to 1:1) and further purified by prep-TLC (SiO2, PE: EtOAc = 2:1) to give pure (2R,5S )-tert-butyl 4-(8-(4-cyano-1H-pyrazol-l-yl)pyrido[4,3-d]pyrimidin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (21.14 mg, 13 %).
Example 13 l-(5-cyclopropyl-4-((2S,5R)-4-(l-fluorocyclopropanecarbonyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-1H -pyrazole-4-carbonitrile
[0303] l-(5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-1H -pyrazole-4- carbonitrile hydrochloride: A solution of (2R,5S)-tert-butyl 4-(8-(4-cyano-1H-pyrazol-l-yl)-5- cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (60 mg, 0.13 mmol) in IM HCl/MeOH (5 mL) was stirred at 20°C for 4 h. The reaction mixture was concentrated under reduced pressure to give l-(5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-1H-pyrazole- 4-carbonitrile hydrochloride (80 mg, crude).
[0304] l-(5-cyclopropyl-4-((2S,5R)-4-(l-fluorocyclopropanecarbonyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-1H -pyrazole-4-carbonitrile: To a solution of l-(5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-1H-pyrazole-4-carbonitrile hydrochloride (50 mg, 0.12 mmol) in DMF (1 mL) was added 1 -fluorocyclopropanecarboxylic acid (19 mg, 0.18 mmol), DIEA (79 mg, 0.61 mmol), and HATU (70 mg, 0.18 mmol) at 20 °C. The mixture was stirred at 20 °C for 6 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with the following conditions: column: Phenomenex luna Cl 8 100 x 40 mm x 5 μm; mobile phase: A: 10 mm 0.2% FA in water, B: MeCN; B in A: 5%-45% B, 8.0 min) to give l-(5-cyclopropyl-4-((2S,5R)-4- ( 1 -fluorocyclopropanecarbonyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1H-pyrazole-4-carbonitrile (2 mg, 27% yield).
Example 14 (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0305] l-methyl-3-(4,4,5,5-tetramethyl- 1,3,2-dioxaborolan-2-yl )-1H -pyrazole-5-carbonitrile: To a solution of 3-bromo-l -methyl- 1H-pyrazole-5-carbonitrile (200 mg, 1.08 mmol) in 1,4-dioxane (10 mL) was added bis(pinacolato)diboron (819 mg, 3.23 mmol), KO Ac (317 mg, 3.23 mmol) and Pd(dppf)Ch (79 mg, 0.11 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)- 1H pyrazole-5-carbonitrile (280 mg, 80% purity, 89%).
[0306] (2R ,5.S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-cyclopropylquinazolin-4-yl)- 2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5- cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (100 mg, 0.24 mmol) in 1,4-dioxane (5 mL) and H2O (1 mL) was added l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H pyrazole-5-carbonitrile (112 mg, 0.48 mmol), XPhos (23 mg, 0.048 mmol), K3PO4 (102 mg, 0.48 mmol), KOAc (47 mg, 0.48 mmol) and Pd2(dba)3 (22 mg, 0.024 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (10 mL), extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC with the following conditions: (column: Waters Xbridge Prep OBD Cl 8 150 x 40 mm x 10 μm; mobile phase: A: 10 mm NH4HCO3 in water, B: MeCN; B in A: 55%-95%, 8.0 min) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-l- methyl- 1 H-pyrazol-3-y l )-5-cyc lopropy Iquinazol in-4-y l )-2,5-dimcthy I pi pcrazinc- 1 -carboxylate (25 mg, 21%). Example 15 (2R,5S)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0307] 8-bromo-4-chloropyrido[4,3-d]pyrimidine: To a solution of 8-bromopyrido[4,3-d]pyrimidin- 4(3H)-one (1.6 g, 7.08 mmol) in SOCh (14 mL) was added DMF (517 mg, 7.08 mmol) at 20 °C under N2. The mixture was stirred at 85 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was stirred in (PE:EtOAc = 1: 1) (40 mL) at 20 °C for 30 min, filtered, and concentrated under reduced pressure to give 8-bromo-4-chloropyrido|4,3-d|pyrimidinc (600 mg, 18%).
[0308] (2R ,5.S)-tert -butyl 4-(8-bromopyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 8-bromo-4-chloropyrido[4,3-d]pyrimidine (1.5 g, 6.14 mmol) and tert- butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (2.63 g, 12.27 mmol) in NMP (20 mL) was added DIEA (3.96 g, 30.68 mmol) at 20 °C. The mixture was stirred at 140 °C for 16 h. The reaction mixture was poured into H2O (100 mL) at 20 °C, extracted with EtOAc (3 x 40 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 3:1 to 1:1) to give (2R,5S)-tert-butyl 4-(8-bromopyrido| 4,3-d| pyrimidin-4-y l )-2,5-dimcthy 1 pi perazine- 1 -carboxylate (2.5 g, 96%).
[0309] 8-bromo-4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide: To a solution of (2R,5S)-tert-butyl 4-(8-bromopyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (2.1 g, 4.97 mmol) in DCM (30 mL) was added m-CPBA (3.03 g, 14.92 mmol, 85%) at 0 °C under N2. The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into 10% Na2S2O3 (30 mL) at 20 °C, extracted with DCM (3 x 15 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc:MeOH = 10:1) to give 8-bromo-4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide (1.3 g, 60%).
[0310] (2R ,5.S)-tert -buty I 4-(8-bromo-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of 8-bromo-4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide (1.3 g, 2.97 mmol) in MeOH (15 mL) was added TEA (900 mg, 8.90 mmol) and TosCl (735 mg, 3.86 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into H2O (50 mL), extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 4:1 to 1:1) to give (2R,5S)-tert butyl 4-(8-bromo-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (900 mg, 67%).
[0311] (2R ,5.S)-tert -buty I 4-(8-(4-cyanopyridin-2-yl)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-bromo-5- methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (50 mg, 0.11 mmol) and 2- (trimethylstannyl)isonicotinonitrile (59 mg, 0.22 mmol) in DMF (1 mL) was added Pd(PPh3)4 (13 mg, 0.01 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (10 mL), extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18: 100 x 30mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 35%-65%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(4- cyanopyridin-2-yl)-5-methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (5.63 mg, 10 %).
Example 16 (2R,5S)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)-5-hydroxypyrido[4,3-dlpyrimidin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0312] (2R ,5.S)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)-5-hydroxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-(4-cyanopyridin-2-yl)-5- methoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (110 mg, 0.23 mmol) in MeCN (2 mL) was added TMSC1 (50 mg, 0.46 mmol) and Nal (69 mg, 0.46 mmol) at 20 °C. The mixture was stirred at 80 °C for 2 h. The reaction mixture was poured into H2O (5 mL) at 20 °C, extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (2 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18: 150 x 40mm x 10 μm; Mobile phase: A: 10 mM NH4HCO3 in water and B: MeCN; Gradient: B% = 20%-75%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(4-cyanopyridin- 2-yl)-5-hydroxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (11.33 mg, 11%). Example 17
(2R,5S)-tert -butyl 4-(8-(4-cyanopyridin-2-yl)-6-methyl-5-oxo-5,6-dihydropyrido[4,3-d]pyrimidin-4- yl)-2,5-dimethylpiperazine-l-carboxylate
[0313] (2R ,5.S )-tert -hutyl 4-(8-(4-cyanopyridin-2-yl)-6-methyl-5-oxo-5,6-dihydropyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-(4- cyanopyridin-2-yl)-5-hydroxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (110 mg, 0.12 mmol) in DMF (3 mL) was added NaH (10 mg, 0.24 mmol, 60% purity) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. Mel (22 mg, 0.15 mmol) in DMF (0.3 mL) was added to the reaction mixture. The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into sat. NH4CI (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: Waters Xbridge BEH C18: 100 x 30mm x 10 μm; Mobile phase: A: 10 mM NH4HCO3 in water and B: MeCN; Gradient: B% = 40%-74%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(4- cyanopyridin-2-yl)-6-methyl-5-oxo-5,6-dihydropyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (26 mg, 45%).
Example 18 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- cvclopropylqumazolin-8-yl )-l -methyl- 1H -pyrazole-5-carbonit rile
[0314] 3-(5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H - pyrazole-5-carbonitrile hydrochloride: To a solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H- pyrazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (40 mg, 0.08 mmol) in MeOH (1.5 mL) was added HCl/MeOH (0.5 mL). The mixture was stirred at 25 °C for 4 h. The mixture was concentrated under reduced pressure to give 3-(5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile hydrochloride (30 mg, 94%, crude).
[0315] 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- cyclopropylquinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: A mixture of 3-(5-cyclopropyl-4- ((2S,5R)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile hydrochloride (30 mg, 0.07 mmol), 1 -cyanocyclobutanecarboxylic acid (13 mg, 0.1 mmol), CMPI (27 mg, 0.1 mmol), DIEA (46 mg, 0.35 mmol) in THE (1 mL) was degassed and purged with N2 three times. The mixture was stirred at 50 °C for 2 h under N2 atmosphere. The mixture was poured into water (6 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (3 mL), dried over anhydrous Na2SO4 , filtered, and concentrated under reduced pressure. The mixture was purified by p-HPLC (column: Waters Xbridge BEH Cl 8 100 x 30 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 30%-75%, 8 min) to give 3-(4-((2S,5R)-4-(l- cyanocyclobutane- 1 -carbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-cyclopropylquinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile (10.46 mg, 30%).
Example 19 (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-ethoxypyrido[4,3-d]pyrimidin-4-yl)- 2,5-dimethylpiperazine- 1 -carboxylate
[0316] (2R,5S)-tert-butyl 4-(8-bromo-5-ethoxypyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of 8-bromo-4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide (200 mg, 0.46 mmol) in EtOH (4 mL) was added TEA (139 mg, 1.37 mmol) and TosCl (113 mg, 0.59 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated under reduced pressure to give a residue, diluted with H2O (20 mL), extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica column chromatography (PE:EtOAc = 4:1 to 1:1) to give (2R,5S)-tert-butyl 4-(8-bromo-5- ethoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (80 mg, 38%).
[0317] (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-ethoxypyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8- bromo-5-ethoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (90 mg, 0.20 mmol) and l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (67 mg, 0.29 mmol) in dioxane (9 mL) and H2O (1.5 mL) was added K3PO4 (82 mg, 0.39 mmol), KO Ac (38 mg, 0.39 mmol), XPhos (18 mg, 0.04 mmol) and Pd2(dba)3 (18 mg, 0.02 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18: 100 x 30mm x 3 μm; Mobile phase: A: 0.2% FA in water and B: MeCN; Gradient: B% = 25%-55%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(5- cyano- 1 -methyl- 1H-pyrazol-3-yl)-5-ethoxypyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (50.60 mg, 53%).
Example 20 trans -(2R,5S)-tert-butyl 4-(8-(5-cvano-l-inethyl-1H -pyraz(»l-3-yl)-5-(2- (ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate
[0318] trans-(2R,5S)-tert-butyl 4-(8-chloro-5-(2-(ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (300 mg, 0.66 mmol) and trans-2- (ethoxycarbonyl)cyclopropyl)boraneylidene)-λ3-fluoraneyl)potassium(III) fluoride (174 mg, 0.79 mmol) in H2O (0.6 mL) and toluene (2.4 mL) was added K3PO4 (279 mg, 1.32 mmol) and [2-(2- aminophenyl)phenyl]-chloro-palladium;bis(l-adamantyl)-butyl-phosphane (44 mg, 0.07 mmol) under N2. The mixture was stirred at 100 °C for 16 h. The mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1) to give trans-(2R,5S)-tert-butyl 4-(8-chloro-5-(2- (ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (115 mg, 36%).
[0319] trans-(2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(2- (ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of trans-(2R,5S)-tert-butyl 4-(8-chloro-5-(2-(ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (85 mg, 0.17 mmol) and l-methyl-3-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (49 mg, 0.21 mmol) in 1,4-dioxane (1.2 mL) and H2O (0.2 mL) was added K3PO4 (74 mg, 0.35 mmol), Pd(dba)2 (10 mg, 0.02 mmol), XPhos (17 mg, 0.03 mmol) and KOAc (34.12 mg, 0.35 mmol). The mixture was stirred at 100 °C for 16 h. The mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100x30 mmxlO um; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 60%-100%, 8 min) to give trans-(2R,5S)- tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(2-(ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (51 mg, 52%) as a mixture of diastereomers.
Example 21
(2R,5S)-tert -butyl 4-(5-cvan()-8-(5-cvano- l-inethyl-1H -pyrazol-3-yl )(|uinazolin-4-yl )-2,5- dimethylpiperazine- 1 -carboxylate
[0320] (2R,5S)-tert -butyl 4-(8-chloro-5-cyanoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5-bromo-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (300 mg, 0.66 mmol) in DMA (5 mL) was added Zn(CN)2 (31 mg, 0.26 mmol) and Pd(OAc)2 (7 mg, 0.033 mmol) and DPPF (36 mg, 0.066 mmol) at 20 °C under N2. The mixture was stirred at 80 °C for 16 h. The reaction mixture was poured into water (10 mL) and extracted with DCM (3x 5 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude was purified by p-TLC (PE:EtOAc = 3:1) to afford (2R,5S)-tert-butyl 4-(8-chloro-5-cyanoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (70 mg, 26%).
[0321] (2R,5S)-tert -butyl 4-(5-cyano-8-(5-cyano-l-methyl-1H -pyrazol-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-cyanoquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (70 mg, 0.17 mmol) and l-methyl-3-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (49 mg, 0.21 mmol) in dioxane (1.2 mL) and H2O (0.2 mL) was added K3PO4 (74 mg, 0.35 mmol), KOAc (34 mg, 0.35 mmol), Pd(dba)2 (10 mg, 0.017 mmol) and XPhos (17 mg, 0.035 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 3 h. The reaction mixture was poured into H2O (4 mL) and extracted with EtOAc (3 x 2 mL). The combined organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH Cl 8 75 x 30 mm x3 μm; mobile phase: A: 0.2% FA in water, B: MeCN; B% in A: 50%-80%, 8 min) to give (2R,5S)-tert- butyl 4-(5-cyano-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (36 mg, 44%). Example 22 (2R,5A)-tert -butyl 4-(8-(5-cvano- 1-methyl- 1H -l,2,4-triazol-3-yl )-5-cvcl()propyl(piinaz()lin-4-yl )-2,5- dimethylpiperazine- 1 -carboxylate
[0322] 3-bromo-l-methyl-1H -l,2,4-triazole-5-carbonitrile: To a solution of 5-bromo-4H- 1,2,4- triazole-3-carbonitrile (1.4 g, 8.09 mmol) in DMF (30 mL) was added K2CO3 (3.36 g, 24.28 mmol) and Mel (1.72 g, 12.14 mmol) at 20 °C. The mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc (3 x 40 mL). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give 3-bromo-l- methyl-1H-l,2,4-triazole-5-carbonitrile (300 mg, 19%).
[0323] (2R,5S)-tert-butyl 4-(5-cyclopropyl-8-(4,4,5,5-tetramethyl-L3,2-dioxahorolan-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8- chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 0.36 mmol) in 1,4- dioxane (5 mL) was added Bis(pinacolato)diboron (274 mg, 1.08 mmol), PCy3 (10 mg, 0.04 mmol), KO Ac (141 mg, 1.44 mmol) and Pd2(dba)3 (33 mg, 0.04 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 3 h. The reaction mixture was filtered and concentrated under reduced pressure to give (2R,5S)-tert-butyl 4-(5-cyclopropyl-8-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (150 mg, crude), which was used directly in the next step.
[0324] (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H -l,2,4-triazol-3-yl)-5-cyclopropylquinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5-cyclopropyl-8- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 0.30 mmol) in 1,4-dioxane (5 mL) and H2O (0.5 mL) was added 5-bromo-2-methyl-l,2,4-triazole-3- carbonitrile (110 mg, 0.59 mmol), K2CO3 (122 mg, 0.89 mmol) and Pd(dppf)Ch (22 mg, 0.03 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 16 h. The reaction mixture was quenched by addition of H2O (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (Phenomenex Luna Cl 8 100 x 30 mm x 10 μm; mobile phase; mobile phase: [A: 10 mM 0.2% FA in water, B: MeCN; B% in A: 10%-40% 8.0 min) to give (2R,5S)-tert-butyl 4-(8-(5- cyano- 1 -methyl- 1H- 1 ,2,4-triazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (22 mg, 15%). Example 23 (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-cyclopropylpyrido|4,3v/]pyrimidin- 4-yl)-2,5-dimethylpiperazine-l-carboxylate
[0325] 4-((2S,5R)-4-(tert -butoxycarbonyl )-2,5-dhnethylpiperazin- 1-yl )-8-(5-cyano-l -methyl- 1 II- pyrazol-3-yl)pyrido[4,3-d]pyrimidine 6-oxide: To a solution of 8-bromo-4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide (200 mg, 0.46 mmol) and l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (112 mg, 0.48 mmol) in 1,4-dioxane (4 mL) and H2O (0.5 mL) was added K3PO4 (194 mg, 0.91 mmol), KO Ac (90 mg, 0.91 mmol), XPhos (44 mg, 0.09 mmol) and Pd2(dba)3 (42 mg, 0.05 mmol) under N2 at 20 °C. The mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (EtOAc:MeOH = 10:1 to 5:1) to give 4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)pyrido[4,3- d]pyrimidine 6-oxide (170 mg, 80%).
[0326] (2R,5S)-tert -butyl 4-(5-chloro-8-(5-cyano-l-methyl-1H -pyrazol-3-yl)pyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of 4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)pyrido[4,3- d]pyrimidine 6-oxide (170 mg, 0.37 mmol) in toluene (4 mL) and DMF (1.5 mL) was added a solution of POCI3 (112 mg, 0.73 mmol) in toluene (0.5 mL). The mixture was stirred at 20 °C for 3 h. The reaction mixture was quenched by water (3 mL) at 20 °C, adjusted pH to 7~8 using sat. NaHCO3. extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc:MeOH = 10:1 to 5:1) to give (2R,5S)-tert-butyl 4-(5-chloro-8-(5-cyano-l- methyl-1H-pyrazol-3-yl)pyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (70 mg, 40%).
[0327] (2R,5S)-tert-buty\ 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-cyclopropylpyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5- chloro-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)pyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (40 mg, 0.08 mmol) and cyclopropylboronic acid (9 mg, 0.11 mmol) in dioxane (1 mL) was added K3PO4 (53 mg, 0.25 mmol) and Pd(dppf)Ch (6 mg, 0.01 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 12 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18: 100 x 30mm x 10 μm; Mobile phase: A: 10 mM NH4HCO3 in water and B: MeCN; Gradient: B% = 55%-85%, 8 min) to give (2R,5S)- tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-cyclopropylpyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate (8.15 mg, 20%).
Example 24
(2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-( trans-2- cyanocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate
[0328] frans-2-(4-((2S,5R)-4-(terf-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-8-(5-cyano-l- methyl-1H -pyrazol-3-yl)quinazolin-5-yl)cyclopropanecarboxylic acid: To a solution of trans- (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(2- (ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (120 mg, 0.21 mmol) in THF (2 mL) and H2O (2 mL) was added LiOH.l-kO (22 mg, 0.54 mmol). The mixture was stirred at 20 °C for 16 hr. The reaction mixture concentrated under reduced pressure to remove THF. Water (5 mL) was added to the remaining mixture and extracted with MTBE (5 ml). The aqueous phase was adjusted to pH 4-5 by IN HC1 and extracted with EtOAc (3 x 10 mL). The combined organic phase was dried with anhydrous Na2SO4, filtered, and concentrated to give tran.s-2-(4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-5- yl)cyclopropanecarboxylic acid (60 mg, crude).
[0329] (2R,5S)-tert -butyl 4-(5-(trans-2-carbamoylcyclopropyl)-8-(5-cyano-l-methyl-1H -pyrazol-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of traiis-2-(4-((2S.5R)-4- (tert-butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-5- yl)cyclopropanecarboxylic acid (60 mg, 0.11 mmol) in DMF (2 mL) was added NH4CI (30 mg, 0.56 mmol), DIEA (44 mg, 0.34 mmol) and HATU (52 mg, 0.14 mmol). The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into water (10 ml) and extracted with EtOAc (3 x10 mL). The combined organic phase was dried with anhydrous Na2SO4, filtered, and concentrated to give (2R,5S)- tert-butyl 4-(5-(trans-2-carbamoylcyclopropyl)-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (60 mg, crude).
[0330] (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(frans-2- cyanocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)- tert-butyl 4-(5-(trans-2-carbamoylcyclopropyl)-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.15 mmol) in THE (2 mL) was added methoxycarbonyl- (triethylammonio)sulfonyl-azanide (108 mg, 0.45 mmol). The mixture was stirred at 60 °C for 16 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100x30 mmxlO um; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 40%-85%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(trans-2-cyanocyclopropyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (34 mg, 44%) as a mixture of diastereomers.
Example 25 (2R ,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-methoxyquinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0331] (2R,5S)-tert -hutyl 4-(8-chloro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5-bromo-8- chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (380 mg, 0.83 mmol) and 4,4,5,5- tetramethyl-2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (233 mg, 0.92 mmol) in dioxane (8 mL) was added Pd2(dba)3 (77 mg, 0.084 mmol) and PCy3 (24 mg, 0.084 mmol) and KO Ac (245 mg, 2.50 mmol). The mixture was stirred at 80 °C for 12 h under N2. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phases were washed with brine (10 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (PE:EtOAc = 4:1 to 3:1) to give (2R,5S)-tert-butyl 4-(8-chloro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate (350 mg, 83%).
[0332] (2R,5S)-tert butyl 4-(8-chloro-5-hydroxyquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan- 2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (350 mg, 0.696 mmol) in THE (4 mL) was added 1 M NaOH (55.68 mg, 1.39 mmol) at 0°C, followed by addition of H2O2 (276 mg, 2.44 mmol, 30% in water). The mixture was stirred at 0 °C for 20 min. The reaction was quenched with 10% Na2SC>3 aqueous (10 mL) and acidified with IN HC1 to pH = 2. The aqueous layers were extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (PE:THF = 1:1 to 0 : 1) to give (2R,5S)-tert-butyl 4-(8-chloro-5-hydroxyquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (250 mg, 91%).
[0333] (2R,5S)-tert butyl 4-(8-chloro-5-methoxyquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-hydroxyquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (200 mg, 0.51 mmol) in THF (4 mL) was added K2CO3 (140 mg, 1.02 mmol) and Mel (108 mg, 0.763 mmol). The mixture was stirred at 40 °C for 4 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-TLC (silica gel, PE:EtOAc = 1:2) to give (2R,5S)-tert-butyl 4-(8-chloro-5-methoxyquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (50 mg, 24%).
[0334] (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-methoxyquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5- methoxyquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (45 mg, 0.11 mmol) in 1,4-dioxane (2 mL) and H2O (0.2 mL) was added 2-methyl-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole-3- carbonitrile (52 mg, 0.22 mmol), Pd2(dba)3 (11 mg, 0.011 mmol), XPhos (11 mg, 0.022 mmol) and K3PO4 (47 mg, 0.22 mmol) and KOAc (22 mg, 0.22 mmol) under N2. The mixture was stirred at 100 °C for 6 h. The reaction mixture was poured into water (5 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 80 x 30mm x 3um; mobile phase: A: 0.2% FA in water, B: MeCN; B% in A: 20%-55%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-methoxyquinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (17 mg, 32%).
Example 26 (2R,5S)-tert -butyl 4-(8-(5-cyano-1H -L2,4-triazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0335] 3-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-1H -l,2,4-triazole-5-carbonitrile: To a solution of 3-bromo-1H-l,2,4-triazole-5-carbonitrile (500 mg, 2.89 mmol) in DCM (5 mL) was added TEA (438 mg, 4.34 mmol) and SEM-C1 (530 mg, 3.18 mmol) at 20°C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into water (10 mL) and extracted with DCM (3 xlO mL).The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 5:1) to give 3-bromo-l-((2-(trimethylsilyl)ethoxy)methyl)-1H-l,2,4-triazole-5-carbonitrile (600 mg, 68%).
[0336] (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-5-cyclopropylquinazolin-8- yl)boronic acid: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (0.8 g, 1.92 mmol) and bis(pinacolato)diboron (1.46 g, 5.76 mmol) in dioxane (40 mL) was added Pd2(dba)3 (176 mg, 0.19 mmol) and PCy3 (54 mg, 0.19 mmol) and KOAc (753 mg, 7.67 mmol) under N2. The mixture was stirred at 110 °C for 16 h. The reaction mixture was filtered, the filtrate was used directly in the next step.
[0337] (2R,5S)-tert -butyl 4-(8-(5-cyano-l-((2-(trimethylsilyl)ethoxy)methyl)-1H -l,2,4-triazol-3-yl)- 5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (4-((2S,5R)-4- (tert-butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-cyclopropylquinazolin-8-yl)boronic acid (400 mg, 0.79 mmol) in dioxane (20 mL) and H2O (2 mL) was added 3-bromo-l-((2- (trimethylsilyl)ethoxy)methyl)-1H-l,2,4-triazole-5-carbonitrile (286 mg, 0.944 mmol), Pd(dppf)Ch (58 mg, 0.078 mmol) and K2CO3 (217 mg, 1.57 mmol). The mixture was stirred at 100 °C for 6 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (3 x20 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (PE:EtOAc = 4:1 to 3:1) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-l-((2-(trimethylsilyl)ethoxy)methyl)-1H-l,2,4-triazol-3- yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (300 mg, 63%).
[0338] (2R,5S)-tert -butyl 4-(8-(5-cyano-1H -l,2,4-triazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-(5-cyano-l-((2- (trimethylsilyl)ethoxy)methyl)-1H-l,2,4-triazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (100 mg, 0.165 mmol) in THE (1 mL) was added TBAF (0.83 mL, 0.83 mmol, 1 M in THE). The mixture was stirred at 20 °C for 12 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 8 mL). The combined organic layer was washed with brine (5 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna Cl 8 80 x 30mm x 3um; mobile phase: A: 0.2% FA in water, B: MeCN; B% in A: 60%-100%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-1H-l,2,4- triazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (30.25 mg, 40%). Example 27 (2R,53)-tert -butyl 4-(8-(5-cvano-l -methyl-1H -pyraz()l-3-yl)-5-(diinethylainino)pyrid()| 4,3- d ]pyrirnidin-4-yl)-2,5-dirnethylpiperazine-l -carboxylate
[0339] (2R,5S)-tert -hutyl 4-(5-amino-8-bromopyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of 8-bromo-4-((2S,5R)-4-(tert -butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)pyrido[4,3-d]pyrimidine 6-oxide (500 mg, 1.14 mmol) in pyridine (18.05 g, 228.14 mmol) was added TosCl (261 mg, 1.37 mmol) at 20 °C. After addition, the mixture was stirred at 20°C for 1 h, and then 2-aminoethanol (1.74 g, 28.52 mmol) was added 20 °C. The resulting mixture was stirred at 20 °C for 16 h. The mixture was poured into H2O (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (PE:EtOAc = 1:1 to 0:1) to give (2R,5S)-tert-butyl 4-(5-amino-8-bromopyrido[4,3-d]pyrimidin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (220 mg, 44%).
[0340] (2R,5S)-tert -butyl 4-(8-bromo-5-(dimethylamino)pyrido[4,3-d]pyrimidin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (2R,5S)-tert-butyl 4-(5-amino-8-bromopyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (100 mg, 0.23 mmol) in DMF (2 mL) was added NaH (22 mg, 0.57 mmol) at 0 °C under N2. After addition, the mixture was stirred at 0 °C for 30 min, and then Mel (97 mg, 0.69 mmol) was added. The resulting mixture was stirred at 20 °C for 2 h. The reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-TLC (SiO2, EtOAc) to give (2R,5S)-tert-butyl 4-(8-bromo-5- (dimethylamino)pyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (70 mg, 65%).
[0341] (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(dimethylamino)pyrido[4,3- d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8- bromo-5-(dimethylamino)pyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (30 mg, 0.064 mmol) in 1,4-dioxane (2 mL) and H2O (0.4 mL) was added 2-methyl-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)pyrazole-3-carbonitrile (22 mg, 0.097 mmol), K3PO4 (27 mg, 0.13 mmol), KOAc (13 mg, 0.13 mmol), XPhos (6 mg, 0.013 mol) and Pd2(dba)3 (6 mg, 0.006 mmol) under N2 atmosphere. The suspension was degassed and purged with N2 for 3 times. The mixture was stirred at 80 °C for 2 h under N2. Two batches were combined. The mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100 x 30 mm x 3 um; mobile phase: A: 0.2% FA in water, B: MeCN; B% in A: 30%-65%, 8 min) to give (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5- (dimethylamino)pyrido[4,3-d]pyrimidin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (21 mg, 33%).
Example 28
(2R,5S) -tert - butyl 4-(5-cyclopropyl-8-( trans-2-(ethoxycarbonyl)cyclopropyl)quinazolin-4-yl)-2,5- dimethylpiperazine- 1 -carboxylate
[0342] (2R,5S)-tert -butyl 4-(5-cyclopropyl-8-( trans-2-(ethoxycarbonyl)cyclopropyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8-chloro-5- cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (50 mg, 0.12 mmol) and ethyl trans- 2-(trifluoro-14-boraneyl)cyclopropane-l -carboxylate, potassium salt (32 mg, 0.14 mmol) in toluene (1.6 mL) and H2O (0.4 mL) was added K3PO4 (25 mg, 0.12 mmol) and [2-(2-aminophenyl)phenyl]-chloro- palladium;bis(l-adamantyl)-butyl-phosphane (8 mg, 0.01 mmol). The mixture was stirred at 100 °C for 16 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH Cis 100x30 mmxlO um; mobile phase: A: lOmM NH4HCO3 in water, B: CH3 CN; B% in A: 35%-85%, 8 min) to give (2R,5S)-tert-butyl 4-(5-cyclopropyl-8-(trans-2-(ethoxycarbonyl)cyclopropyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (20 mg 33%) as a mixture of diastereomers.
Example 29 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- trans-2- cyanocyclopropylquinazolin-8-yl)-l-methyl-lh-pyrazole-5-carbonitrile [0343] 3-(5-(trans-2-cyanocyclopropyl)-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l- methyl-1H -pyrazole-5-carbonitrile hydrochloride: A solution of tert-butyl (2R,5S)-4-[5-[trans-2- cyanocyclopropyl] -8-(5-cyano- 1 -methyl-pyrazol-3-yl)quinazolin-4-yl]-2,5-dimethyl-piperazine- 1 - carboxylate (15 mg, 0.029 mmol) in EtOAc (0.2 mL) was added 4N HCl/EtOAc (1 mL) .The mixture was stirred at 20 °C for 7 h . The reaction mixture was concentrated under reduced pressure to give 3-(5- (trans-2-cyanocyclopropyl)-4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile hydrochloride (15 mg, crude).
[0344] 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- trans-2- cyanocyclopropylquinazolin-8-yl)-l-methyl-lh-pyrazole-5-carbonitrile: To a solution of 1- cyanocyclobutanecarboxylic acid (5.02 mg, 0.04 mmol), DIEA (14.39 mg, 0.11 mmol,) and HATU (10.16 mg, 0.026 mmol) in DMF (1 mL) was added 3-(5-(trans-2-cyanocyclopropyl)-4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile hydrochloride (10 mg, 0.022 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into NH4CI (2 mL) and extracted with EtOAc (3 x 2 mL). The organic phase was separated and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100 x 30mm x 3um; mobile phase: A: 0.2% FA in water, B: CH3CN; B% in A: 15%-55%, 8 min) to give 3-(4- ((2S,5R)-4-( 1 -cyanocyclobutane- 1 -carbonyl)-2,5-dimethylpiperazin- 1 -y l)-5- trans-2- cyanocyclopropylquinazolin-8-yl)-l -methyl- lh-pyrazole-5-carbonitrile (5 mg, 46% yield) as a mixture of diastereomers.
Example 30 l-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- cvclopropylquinazolin-8-yl )-3-methyl- 1H -pyrazole-4-carbonitrile
[0345] 8-chloro-5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazoline hydrochloride: (2R,5S)-tert-butyl 4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (200 mg, 0.48 mmol) in MeOH (1 mL) was added HCl/MeOH (4N, 2 mL).The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give 8-chloro-5-cyclopropyl-4- ((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazoline hydrochloride (200 mg, crude).
[0346] l-((2R,5S)-4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carbonyl)cyclobutane-l-carbonitrile: To a solution of 8-chloro-5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin-l-yl)quinazoline hydrochloride (180 mg, 0.51 mmol) and 1 -cyanocyclobutane- 1- carboxylic acid (127 mg, 1.02 mmol) in THF (5 mL) was added DIEA (329 mg, 2.55 mmol) and CMPI (260 mg, 1.02 mmol). The mixture was stirred at 50 °C for 2 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 3 mL). The organic phase was concentrated under reduced pressure. The residue was purified by silica gel chromatography column (PE:EA = 10:1 to 5:1) to give 1- ((2R,5S)-4-(8-chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carbonyl)cyclobutane- 1 - carbonitrile (200 mg, 93%).
[0347] l-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- cyclopropylquinazolin-8-yl)-3-methyl-1H -pyrazole-4-carbonitrile: To a mixture of l-((2R,5S)-4-(8- chloro-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carbonyl)cyclobutane-l -carbonitrile (150 mg, 0.35 mmol) and 3-methyl-1H-pyrazole-4-carbonitrile (189 mg, 1.77 mmol) in DMF (5 mL) was added N1 ,N2 -dimethy Icy clohexane-l,2-diamine (40 mg, 0.28 mmol), K2CO3 (147 mg, 1.06 mmol) and Cui (27 mg, 0.14 mmol) at 20°C under N2. The mixture was stirred at 180 °C for 8 h . The mixture was poured into H2O (20 mL). The aqueous phase was extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100 x 30mm x 3um; mobile phase: A: 0.2% FA in water, B: MeCN; B% in A: 30%-70%, 8 min) to give l-(4- ((2S,5R)-4-(l -cyanocyclobutane- l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5-cyclopropylquinazolin-8-yl)- 3-methyl-1H-pyrazole-4-carbonitrile (6.7 mg, 4%).
Example 31 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- (methoxymethyl)quinazolin-8-yl)-l-methyl- 1H-pyrazole-5-carbonitrile
[0348] (2R,5S)-tert butyl 4-(8-chloro-5-(hydroxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (200 mg, 438.81 μmol) and (tributylstannyl)methanol (211 mg, 0.66 mmol) in dioxane (8 mL) was added Pd(PPh3)4 (51 mg, 0.044 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 16 h. Then (tributylstannyl)methanol (56 mg, 0.18 mmol) and Pd(PPh3)4 (51 mg, 0.044 mmol) were added at 20 °C under N2, the mixture was stirred at 90 °C for 20 h. Two batches were combined. The reaction mixture was poured into H2O (20 mL) and extracted with EtOAc (3 x 8 mL). The combined organic layer was washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to afford (2R,5S)-tert-butyl 4-(8-chloro-5-(hydroxymethyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (110 mg, 31%).
[0349] (2R,5S)-tert butyl 4-(8-chloro-5-(methoxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5-(hydroxymethyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (90 mg, 0.22 mol) in DMF (1 mL) was added NaH (13 mg, 0.33 mmol, 60% purity) at 0 °C under N2. The mixture was stirred at 0 °C for 30 min, then Mel (47 mg, 0.33 mmol) was added. The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into NH4CI (3 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (5mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by p-TLC (silica gel, PE:EtOAc = 1:1) to afford (2R,5S)-tert-butyl 4-(8-chloro-5- (methoxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (70 mg, 75%).
[0350] (2R,5S)-tert -butyl 4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(methoxymethyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (2R,5S)-tert-butyl 4-(8-chloro-5- (methoxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (70 mg, 0.17 mmol) and 1- methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (58 mg, 0.25 mmol) in dioxane (1 mL) and H2O (0.2 mL) was added K3PO4 (71 mg, 0.33 mmol) and KOAc (33 mg, 0.33 mmol) and XPhos (16 mg, 0.033 mmol) and Pd2(dba)3 (15 mg, 0.017 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 16 h. The reaction mixture was poured into H2O (3 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (5mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep- TLC (silica gel, PE:EtOAc = 1:1) to afford the (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H-pyrazol-3- yl)-5-(methoxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 97%).
[0351] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(methoxymethyl)quinazolin-8-yl)-l-methyl-1H - pyrazole-5-carbonitrile hydrochloride: To a solution of (2R,5S)-tert-butyl 4-(8-(5-cyano-l-methyl-1H- pyrazol-3-yl)-5-(methoxymethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (80 mg, 0.16 mmol) in 1,4-dioxane (0.5 mL) was added 4N HCl/dioxane (1 mL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give 3-(4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)-5-(methoxymethyl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile hydrochloride (100 mg, crude).
[0352] 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5-dimethylpiperazin-l-yl)-5- (methoxymethyl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 3-(4- ((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(methoxymethyl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5- carbonitrile hydrochloride (50 mg, 0.12 mmol) and 1 -cyanocyclobutane- 1 -carboxylic acid (22 mg, 0.18 mmol) in THE (1 mL) was added CMPI (45 mg, 0.18 mmol) and DIEA (76 mg, 0.58 mmol). The mixture was stirred at 50 °C for 16 h. The reaction mixture was poured into H2O (3 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH Cl 8 100 x 30 mm x3 μm; mobile phase: [A: 10 mM FA in water, B: CH3 CN; B% in A: 20%-50%, 8 min) to give 3-(4-((2S,5R)-4-(l-cyanocyclobutane-l-carbonyl)-2,5- dimethylpiperazin-l-yl)-5-(methoxymethyl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (19 mg, 33%).
Example 32A & 32B
3-(4-((2S,5R)-4-((R)-2-cyano-3-fluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-5- cvcl(»propylquinazol in-8-yl )-l-inethyl- 1H -pyraz(»le-5-carb(»nitrile and 3-(4-((2S,5R)-4-((3)-2-cyano-
3-fluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-5-cyclopropylquinazolin-8-yl)-l-methyl- 1H -pyrazole-5-carbonitrile
[0353] 3-(5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H - pyrazole-5-carbonitrile hydrochloride: A solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H- pyrazol-3-yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (130 mg, 0.27 mmol) in 1,4-dioxane (2 mL) was added 4N HCl/l,4-dioxane (4 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give 3-(5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile hydrochloride (100 mg, 88%) as a white solid.
[0354] 3-(4-((2S,5R)-4-((R)-2-cyano-3-fluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-5- cyclopropylquinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 3-(5-cyclopropyl- 4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (150 mg, 0.39 mmol) and 2-cyano-3-fluoro-2-methylpropanoic acid (102 mg, 0.77 mmol) in THE (2 mL) was added CMPI (148 mg, 0.58 mmol) and DIEA (250 mg, 1.94 mmol). The mixture was stirred at 50 °C for 2 h. The mixture was diluted with water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography column (PE:EtOAc = 3:1) to give (90 mg, diastereomers) as white solid, which was further separated by SEC (column: DAICEL CHIRALCEL OX (250mm*30mm,10um); mobile phase: [CO2- MeOH(0.1%NH3H2O)]; B%:45%, isocratic elution mode) to give 3-(4-((2S,5R)-4-((R)-2-cyano-3- fluoro-2-methylpropanoyl)-2,5-dimethylpiperazin- 1 -yl)-5-cyclopropylquinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile (19 mg, 9%, peak 1 (32A), 19 mg, 9%, peak 2 (32B)).
[0355] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 39 tert-butyl N-[(2r,3s)-l-[8-(5-cyano-l-methyl-pyrazol-3-yl)-5-cyclopropyl-quinazolin-4-yl]-2-methyl- azetidin-3-yl]-N-methyl-carbamate
[0356] Tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2-methylazetidin-3-yl)carbamate:
To a solution of 5-bromo-4,8-dichloroquinazoline (1g, 3.60 mmol) and tert-butyl ((2r,3s)-2- methylazetidin-3-yl)carbamate hydrochloride (962 mg, 4.32 mmol) in NMP (30 mL) was added DIEA (2.33 g, 17.99 mmol). The mixture was stirred at 100 °C for 12 h. The reaction mixture was poured into water (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2-methylazetidin-3-yl)carbamate (1.5 g, 98%).
[0357] Tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2-methylazetidin-3- yl)(methyl)carbamate: To a solution of tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2- methylazetidin-3-yl)carbamate (1.5 g, 3.51 mmol) in DMF (15 mL) was added NaH (169 mg, 4.21 mmol, 60% in dispersion oil) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h, then CH3I (598 mg, 4.21 mmol) was added. The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (3 x 15 mL). The combined organic phase was washed with brine (15 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 7:1 to 5:1) to give tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2-methylazetidin-3-yl)(methyl)carbamate (1.5 g, 97%).
[0358] Tert-butyl ((2r,3s)-l-(8-chloro-5-cyclopropylquinazolin-4-yl)-2-methylazetidin-3- yl)(methyl)carbamate: To a solution of tert-butyl ((2r,3s)-l-(5-bromo-8-chloroquinazolin-4-yl)-2- methylazetidin-3-yl)(methyl)carbamate (1.4 g, 3.17 mmol) and cyclopropylboronic acid (545 mg, 6.34 mmol) in dioxane (20 mL) was added K3PO4 (2.02 g, 9.51 mmol), Ag2O (220 mg, 0.95 mmol) and Pd(dppf)Cl2-CH2Cl2 (259 mg, 0.32 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 20 h. The reaction mixture was poured into water (40 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give tert-butyl ((2R,35)-l-(8-chloro-5-cyclopropylquinazolin-4-yl)-2- methylazetidin-3-yl)(methyl)carbamate (130 mg, 10%).
[0359] tert-butyl N-[(2r,3s)-l-[8-(5-cyano-l-methyl-pyrazol-3-yl)-5-cyclopropyl-quinazolin-4-yl]-2- methyl-azetidin-3-yl]-N-methyl-carbamate (trans stereochemistry): tert-butyl (( 2r.3s )- l -( 8-cliloro-5- cyclopropylquinazolin-4-yl)-2-methylazetidin-3-yl)(methyl)carbamate (130 mg, 0.32 mmol) 2-methyl-5- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole-3-carbonitrile (150 mg, 0.65 mmol) in 1,4-dioxane (5 mL) and H2O (1 mL) was added K3PO4 (137 mg, 0.65 mmol,) KOAc (63 mg, 0.65 mmol), XPhos (31 mg, 0.064 mmol) and Pd2(dba)3 (30 mg, 0.032 mmol) under N2. The suspension was degassed and purged with N2 for 3 times. The mixture was stirred at 100 °C for 2 h. The reaction mixture was poured into water (5mL) and extracted with EtOAc (3 x 3 mL). The combined organic phase was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH Cl 8 100 x 30 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 65%-85%, 8.0 min) to give tert-butyl N-[(2r,3s)-l-[8-(5-cyano-l-methyl-pyrazol-3-yl)-5-cyclopropyl-quinazolin-4-yl]-2- methyl-azetidin-3-yl]-N-methyl-carbamate as an enantiomeric mixture of trans diastereomers (24 mg, 16%). m/z = 474.3 [M+H]+.
[0360] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 42 3-[4-[(2S,5R)-4-(2-cyano-3-fluoro-2-methyl-propanoyl)-2,5-dimethyl-piperazin-l-yl]-5-pyrazol-l-yl- quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0361] 5-bromo-8-chloro-4-methoxyquinazoline: A solution of 5-bromo-4,8-dichloroquinazoline (2 g, 7.14 mmol) in MeOH (15 mL) was stirred at 20 °C for 0.5 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (3 x 10 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 8:1 to 5:1) to give 5-bromo-8-chloro-4- methoxyquinazoline (450 mg, 23%).
[0362] 8-chloro-5-( 1H-pyrazol- l-yl)quinazolin-4-ol: To a mixture of 5-bromo-8-chloro-4- methoxyquinazoline (700 mg, 2.56 mmol) and 1H-pyrazolc (350 mg, 5.12 mmol) in DMF (10 mL) was added K2CO3 (1.1 g, 7.68 mmol), Ni,N2-dimethylcyclohexane-l,2-diamine (300 mg, 2.05 mmol) and Cui (200 mg, 1.02 mmol) at 20°C under N2. The mixture was stirred at 140 °C for 4 h. The reaction mixture was poured into water (50 mL) and extracted with EtOAc (3 x 10 mL). The aqueous phase was dried under reduced pressure and purified by silica gel column chromatography (PE: EtOAc = 1:1 to 0:1) to give 8-chloro-5-(1H-pyrazol-l-yl)quinazolin-4-ol (270 mg, 14%).
[0363] tert-butyl (2R,5S)-4-(8-chloro-5-(1H -pyrazol-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-
1 -carboxylate: To a solution of 8-chloro-5-( 1H-pyrazol- l -yl)quinazolin-4-ol (160 mg, 0.65 mmol), tert- butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (153 mg, 0.71 mmol) in CH3CN (3 mL) was added DBU (495 mg, 3.24 mmol) and PyBOP (506 mg, 0.97 mmol). The mixture was stirred at 20 °C for 3 h. The reaction mixture was poured into water (15 mL) and extracted with EtOAc (3 x 5 mL). The organic phase was washed with brine (5 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 2:1 to 1:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-( 1H-pyrazol- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate
(200 mg, 70%).
[0364] tert-butyl (2R ,5.S)-4-(8-(5-cvano-l-methyl-1H -pyrazol-3-yl)-5-(1H -pyrazol-l-yl)quinazolin- 4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8-chloro-5-(1H- pyrazol-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (20 mg, 0.045 mmol) in dioxane (1 mL) and H2O (0.2 mL) was added l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H- pyrazole-5-carbonitrile (21 mg, 0.09 mmol), K3PO4 (19 mg, 0.09 mmol), KOAc (9 mg, 0.09 mmol), XPhos (5 mg, 0.009 mmol), Pd2(dba)3 (4 mg, 0.004 mmol) under N2. The suspension was degassed and purged with N23 times. The mixture was stirred at 80 °C for 2 h under N2. The reaction mixture was poured into water (2 mL) and extracted with EtOAc (3 x 1 mL). The combined organic phase was concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100 x 30 mm x 3 μm; mobile phase: A: 0.2% FA in water, B: ACN; B% in A: 20%-55%, 8.0 min) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l -methyl- 1H-pyrazol-3-yl)-5-( 1H-pyrazol- l -yl)quinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (22 mg, 10%).
[0365] 3-(4-((2.S,5R )-2,5-diinethylpiperazin-l-yl )-5-( 1H-pyraz(»l- l-ylppiinaz(»lin-8-yl )- 1-inethyl-1H- pyrazole-5-carbonitrile: To a solution of tert-butyl (2R, 5S)-4-(8-(5-cyano-l -methyl- l rt-pyrazol-3-yl)-5- ( 1H-pyrazol- 1 -yl )quinazolin-4-yl )-2,5-dimcthylpipcrazinc- 1 -carboxylate (250 mg, 0.49 mmol) in DCM (5 mL) was added trimethylsilyl trifluoromethanesulfonate (379 mg, 1.70 mmol) and 2,6- dimethylpyridine (261 mg, 2.43 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give crude 3-(4-((2S,5R)-2,5-dimethylpiperazin-l- yl)-5-(1H-pyrazol-l-yl)quinazolin-8-yl)-l -methyl-1H-pyrazole-5-carbonitrile (600 mg, 33% purity) which was used directly in the next step.
[0366] 3-[4-[(2S,5R)-4-(2-cyano-3-fluoro-2-methyl-propanoyl)-2,5-dimethyl-piperazin-l-yl]-5- pyrazol-l-yl-quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)-5-( 1 H-pyrazol- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1 H-pyrazole-5-carbonitrile (201 mg, 0.48 mmol) and 2-cyano-3-fluoro-2-methylpropanoic acid (127 mg, 0.97 mmol) in DMF (20 mL) was added BOP-CI (248 mg, 0.98 mmol) and DIEA (315 mg, 2.43 mmol). The mixture was stirred at 50 °C for 16 h. The reaction mixture was poured into water (40 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phase was concentrated under reduced pressure. The residue was purified by prep- HPLC (column: Waters Xbridge BEH Cl 8 100 x 30 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: CH3CN; B% in A: 45%-75%, 8.0 min) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2- methylpropanoyl)-2,5-dimethylpiperazin- 1 -yl)-5-(1H-pyrazol- 1 -yl)quinazolin-8-yl)- 1 -methyl-1H- pyrazole-5-carbonitrile (31 mg, 12%). m/z = 527.3 [M+H]+.
[0367] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 53A & 53B 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lS)- 2,2-difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4-[2- cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lR)-2,2- difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile [0368] 3-((2R,5S)-4-(5-bromo-8-iodo-quinazolin-4-yl)-2,5-dimethyl-piperazin-l-yl]-2,2- bis(fluoromethyl)-3-oxopropanenitrile: To a solution of 5-bromo-4-chloro-8-iodoquinazoline (1.06 g, 2.88 mmol) and 3-((2R,5S)-2,5-dimethylpiperazin-l-yl)-2,2-bis(fluoromethyl)-3-oxopropanenitrile hydrochloride (810 mg, 2.88 mmol) in NMP (30 mL) was added DIEA (1.86 g, 14.38 mmol, 2.50 mL), the mixture was stirred at 90 °C for 1 h. The reaction mixture was poured into H2O (60 ml) and extracted with EtOAc (3 x 30 mL). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 3-((2R,5S)-4-(5-bromo-8-iodo-quinazolin-4-yl)-2,5- dimethyl-piperazin-l-yl]-2,2-bis(fluoromethyl)-3-oxopropanenitrile (1.2 g, 72%).
[0369] 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a solution of 3- ((2R,5S)-4-(5-bromo-8-iodo-quinazolin-4-yl)-2,5-dimethyl-piperazin-l-yl]-2,2-bis(fluoromethyl)-3- oxopropanenitrile (1.2 g, 2.08 mmol) and l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H- pyrazole-5-carbonitrile (411 mg, 1.76 mmol) in dioxane (15 mL) and H2O (3 mL) was added Pd(dppf)Ch (152 mg, 0.21 mmol) and K3PO4 (881 mg, 4.15 mmol) under N2. The mixture was stirred at 60 °C for 1 hour.
[0370] The residue was poured into H2O (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phase was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2- (fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5- carbonitrile (650 mg, 56%).
[0371] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-vinylquinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 3-(5-bromo-4-((2S,5R)-
4-(2-cyano-3-fhioro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (200 mg, 0.35 mmol) and Potassium vinyltrifluoroborate (72 mg, 0.53 mmol) in dioxane (4 mL) was added Pd(dppf)Cl2-CH2Cl2 (30 mg, 0.035 mmol) and Ag2O (25 mg, 0.11 mmol), K3PO4 (229 mg, 1.08 mmol). The mixture was stirred at 100 °C for 12 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 3 mL). The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 5:1 to 2:1) to give
3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- vinylquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (130 mg, 72%).
[0372] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-
5-[(lS)-2,2-difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-
4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lR)-2,2- difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 3-(4- ((25,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- vinylquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (130 mg, 0.25 mmol) in THF (2 mL) was added TMSCF3 (184 mg, 1.29 mmol) and Nal (20 mg, 0.13 mmol). The mixture was stirred at 60 °C for 16 h. The residue was poured into H2O (5mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromagraphy (PE:EtOAc = 5:1 to 3:1) to give 120 mg as mixture of diastereomers. Then the mixture was purified by SEC to give 3-[4- [(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[2,2- difluorocyclopropyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile as two separate isomers with unknown stereochemistry (24mg, 17%, peak 1, m/z = 555.2 [M+H]+ (53A), 5 mg, 3 %, peak 2, m/z = 555.2 [M+H]+ (53B)).
[0373] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 59 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- methoxyquinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile
[0374] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5- carbonitrile: To a solution of 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)- 2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (200 mg, 0.35 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2’-bi(l,3,2-dioxaborolane) (137 mg, 0.54 mmol) in dioxane (4 mL) was added KO Ac (105 mg, 1.08 mmol), PCy3 (10. mg, 0.035 mmol), Pd2(dba)3 (32 mg, 0.035 mmol) under N2, The mixture was stirred at 80 °C for 16 h. The reaction mixture was poured into water (8 mL) and extracted with EtOAc (3 x 3 mL). The combined organic phase was washed with brine (3 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 20:1 to 10:1) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro- 2-(fhioromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (120 mg, 55%).
[0375] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-hydroxyquinazolin-8-yl)- l-methyl- 1H-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)-4-(2- cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (100 mg, 0.16 mmol) in 1,4- dioxane (1 mL) and H2O (1 mL) was added oxone (28 mg, 0.18 mmol) at 0 °C. The mixture was stirred at 0 °C for 2 h. The reaction mixture was poured into water (4 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1 to 0:1) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)-5-hydroxyquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (60 mg, 73%). [0376] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-methoxyquinazolin-8-yl)- l-methyl- 1H-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)-4-(2- cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)-5-hydroxyquinazolin-8-yl)- 1 - methyl-1H-pyrazole-5-carbonitrile (50 mg, 0.1 mmol) in THE (1 mL) was added CH3I (21 mg, 0.15 mmol) and K2CO3 (28 mg, 0.2 mmol). The mixture was stirred at 40 °C for 4 h. The reaction mixture was poured into water (4 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (3 x 2 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100 x 30 mm x 10 μm; mobile phase: A:10 mM NH4HCO3 in water, B: ACN; B% in A: 35%-65%, 8.0 min) to give 3-(4- ((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- methoxyquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (14 mg, 27%). m/z = 509.2 [M+H]+.
[0377] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 62 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- cyclopropyl-6-fluoroquinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile
[0378] Tert-butyl (2.S, 5R )-4-(2-cvanoacety I )-2,5-dimethyl pi perazine- 1 -carboxy late: To a solution of tert-butyl (2S,5R)-2,5-dimethylpiperazine-l -carboxylate (5 g, 23.33 mmol) in THF (50 mL) was added 2- cyanoacetic acid (2.38 g, 28.00 mmol), DIEA (9.05 g, 69.99 mmol) and CMPI (7.15 g, 28.00 mmol) at 20 °C. The mixture was stirred at 50 °C for 4 h. The reaction mixture was added to H2O (50 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with brine (2 x 15 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1) to give tert-butyl (2S,5R)-4-(2- cyanoacetyl)-2,5-dimethylpiperazine-l -carboxylate (6.5 g, 99%).
[0379] Tert-butyl (2S,5R)-4-(2-cyano-3-hydroxy-2-(hydroxymethyl)propanoyl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2S,5R)-4-(2-cyanoacetyl)-2,5- dimethylpiperazine-1 -carboxylate (4.5 g, 15.99 mmol) and HCHO (2.86 g, 35.19 mmol, 37% in water) in dioxane (50 mL) was added a solution of TEA (32 mg, 0.32 mmol) in THF (0.1 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 12 h. The reaction mixture was quenched by addition of H2O (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (3 x 20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1) to give tert-butyl (2S,5R)-4-(2- cyano-3-hydroxy-2-(hydroxymethyl)propanoyl)-2,5-dimethylpiperazine-l-carboxylate (1.1 g, 20%).
[0380] (2.S,5R )-tert -butyl 4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazine- 1-carboxylate: To a solution of (2S,5R)-tert-butyl 4-(2-cyano-3-hydroxy-2-(hydroxymethyl)propanoyl)- 2,5-dimethylpiperazine-l -carboxylate (1.5 g, 4.39 mmol) in DCM (20 mL) was added dropwise DAST (2.12 g, 13.18 mmol) at -40 °C under N2. The mixture was stirred at 20 °C for 12 h. The reaction mixture was quenched by addition of sat. NaHCO3 (50 mL) and extracted with DCM (3 x 15 mL). The combined organic layers were washed with brine (3 x 15 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1) to give (2S,5R)-tert-butyl 4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazine-1 -carboxylate (750 mg, 49% yield).
[0381] 3-((2R,5S)-2,5-dimethylpiperazin-l-yl)-2,2-bis(fluoromethyl)-3-oxopropanenitrile hydrochloride: To a solution of (2S,5R)-tert -butyl 4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazine-1 -carboxylate (750 mg, 2.17 mmol) in EtOAc (5 mL) was added HCl/EtOAc (4 M, 5 mL) at 20 °C. The mixture was stirred at 20 °C for 30 min. The reaction mixture was concentrated under reduced pressure to give 3-((2R,5S)-2,5-dimethylpiperazin-l-yl)-2,2-bis(fluoromethyl)-3- oxopropanenitrile hydrochloride (500 mg, 82%).
[0382] Methyl 6-amino-2-bromo-3-fluorobenzoate: To a solution of 6-amino-2-bromo-3- fluorobenzoic acid (1 g, 4.27 mmol) in MeOH (20 mL) was added H2SO4 (9.20 g, 93.80 mmol) at 0 °C under N2. The mixture was stirred at 80 °C for 48 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (10 mL), adjusted to pH = 3 by 2 N NaOH and extracted with MTBE (3 x 20 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give methyl 6-amino-2- bromo-3-fluorobenzoate (850 mg).
[0383] Methyl 2-amino-6-bromo-5-fluoro-3-iodobenzoate: To a solution of methyl 6-amino-2-bromo- 3 -fluoro-benzoate (850 mg, 3.43 mmol) in AcOH (13 mL) was added NIS (732 mg, 3.26 mmol) at 20 °C under N2. The mixture was stirred at 50 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The reaction mixture was diluted with H2O (15 mL) and adjusted to pH = 7 by 1 N NaOH and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (3 x 5 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give methyl 2-amino-6-bromo-5-fluoro-3-iodobenzoate (900 mg, 70%).
[0384] 2-amino-6-bromo-5-fluoro-3-iodobenzoic acid: To a solution of methyl 2-amino-6-bromo-5- fluoro-3-iodobenzoate (700 mg, 1.87 mmol) in MeOH (7 mL) and H2O (7 mL) was added NaOH (300 mg, 7.49 mmol) at 20 °C under N2. The mixture was stirred at 80 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (20 mL) and adjusted to pH = 1 by 3 N HC1. The suspension was filtered and the filter cake was dried under reduced pressure to give 2- amino-6-bromo-5-fluoro-3-iodobenzoic acid (780 mg).
[0385] 5-bromo-6-fluoro-8-iodoquinazolin-4-ol: To a solution of 2-amino-6-bromo-5-fluoro-3-iodo- benzoic acid (700 mg, 1.94 mmol) in n-BuOH (10 mL) was added formamidine acetate (1.21 g, 11.67 mmol) at 20 °C. The mixture was stirred at 140 °C for 1 h in sealed tube. The residue was poured into H2O (20 mL). The suspension was filtered and the filter cake was dried under reduced pressure to give 5- bromo-6-fluoro-8-iodoquinazolin-4-ol (570 mg, 79%).
[0386] 5-bromo-4-chloro-6-fluoro-8-iodoquinazoline: To a solution of 5-bromo-6-fluoro-8- iodoquinazolin-4-ol (450 mg, 1.22 mmol) in toluene (9 mL) was added POCl3 (748 mg, 4.88 mmol) and DIEA (315 mg, 2.44 mmol) at 20 °C. The mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give 5-bromo-4-chloro-6-fluoro-8-iodoquinazoline (670 mg, crude), which was used directly in the next step.
[0387] 3-((2R,5S)-4-(5-bromo-6-fluoro-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazin-l-yl)-2,2- bis(fluoromethyl)-3-oxopropanenitrile: To a solution of 5-bromo-4-chloro-6-fluoro-8-iodo-quinazoline (470 mg, 1.21 mmol) in NMP (10 mL) was added DIEA (784 mg, 6.07 mmol) and 3-((2R,5S)-2,5- dimethylpiperazin-l-yl)-2,2-bis(fluoromethyl)-3-oxopropanenitrile hydrochloride (410 mg, 1.46 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 1 h. The reaction mixture was poured into H2O (20 ml) and extracted with EtOAc (3 x lOmL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 3-((2R,5S)-4-(5-bromo-6-fluoro-8- iodoquinazolin-4-yl)-2,5-dimethylpiperazin- 1 -yl)-2,2-bis(fluoromethyl)-3-oxopropanenitrile (750 mg, 86%).
[0388] 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)-6-fluoroquinazolin-8-yl)- l-methyl- 1H-pyrazole-5-carbonitrile: To a solution of 3-[(2R,5S)-4-(5-bromo-6-fluoro-8-iodo-quinazolin-4-yl)-2,5-dimethyl-piperazin-l-yl]-2,2- bis(fluoromethyl)-3-oxopropanenitrile (650 mg, 1.09 mmol) in 1,4-dioxane (10 mL) and H2O (2 mL) was added K3PO4 (463 mg, 2.18 mmol), l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H- pyrazole-5-carbonitrile (280 mg, 1.20 mmol) and Pd(dppf)Ch (80 mg, 0.11 mmol) at 20 °C under N2. The mixture was stirred at 60 °C for 1 h. The reaction mixture was poured into H2O (20 ml) and extracted with EtOAc (3 x lOmL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 3-(5-bromo-4-((25,5R)-4-(2-cyano-3- fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)-6-fluoroquinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile (540 mg, 86%).
[0389] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-cyclopropyl-6-fluoroquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a solution of 3-(5- bromo-4-((2S,5R )-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-6- fluoroquinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (200 mg, 0.35 mmol) in dioxane (4 mL) was added K3PO4 (221 mg, 1.04 mmol), cyclopropylboronic acid (150 mg, 1.74 mmol), Ag2O (24 mg, 0.11 mmol), and Pd(dppf)Cl2-CH2Cl2 (28 mg, 0.03 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 2 h. The reaction mixture was poured into H2O (5 ml) and extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with brine (2 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100 x 30 mm x3 μm; mobile phase: A: 0.2% FA in water, B: CH3CN; B% in A: 35%-70%, 8 min) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)- 5-cyclopropyl-6-fluoroquinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (15 mg, 8%). m/z. = 537.2 [M+H]+. [0390] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 69 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5-(l- fluorocyclopropyl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile
[0391] Tert-butyl 2-cyano-3-hydroxy-2-(hydroxymethyl)propanoate : To a solution of HCHO
(63.23 g, 779.22 mmol, 37% aqueous) in dioxane (500 mL) was added tert-butyl 2-cyanoacetate (50 g,
354.19 mmol) and TEA (716 mg, 7.08 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. Then TEA (716 mg, 7.08 mmol) in THE (5 mL) was added to the mixture. The mixture was stirred at 20 °C for 16 h. Two batches were carried out in parallel. The reaction mixture was poured into water (500 mL) and extracted with EtOAc (3 x 200 mL). The combined organic phase was washed with brine (200 mL), dried over with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 2:1 to 1:1) to give tert-butyl 2-cyano-3- hydroxy-2-(hydroxymethyl)propanoate (124 g, 43% purity).
[0392] Tert-butyl 2-cyano-3-fluoro-2-(fluoromethyl)propanoate: To a solution of tert-butyl 2-cyano- 3-hydroxy-2-(hydroxymethyl)propanoate (64 g, 318.06 mmol) in DCM (640 mL) was added DAST (153.80 g, 954.18 mmol) at -40 °C under N2. The mixture was stirred at 20 °C for 16 h. The reaction mixture was diluted with aq. NaHCO3 (600 mL), and then extracted with DCM (3 x 200 mL). The combined organic layers were washed with brine (200 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 5:1) to give tert-butyl 2-cyano-3-fhroro-2-(fluoromethyl)propanoate (24 g, 36%). [0393] 2-cyano-3-fluoro-2-(fluoromethyl)propanoic acid: To a solution of tert-butyl 2-cyano-3- fluoro-2-(fluoromethyl)propanoate (1 g, 4.87 mmol) in DCM (5 mL) was added TFA (7.68 g, 67.31 mmol). The mixture was stirred at 20 °C for 4 h. The reaction mixture was concentrated under reduced pressure to give 2-cyano-3-fluoro-2-(fluoromethyl)propanoic acid (0.78 g).
[0394] Tert-butyl (2R,5S)-4-(8-chloro-5-vinylquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a mixture of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (3 g, 6.58 mmol) and potassium vinyltrifluoroboratc (882 mg, 6.58 mmol) in dioxane (40 mL) was added K3PO4 (4.19 g, 19.75 mmol), Ag2O (458 mg, 1.97 mmol), and Pd(dppf)Ch. DCM (538 mg, 0.66 mmol) at 20°C under N2. The mixture was stirred at 100 °C for 16 h. The residue was diluted with H2O (90 mL) and extracted with EtOAc (3 x 30 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-vinylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (2.1 g, 79 %).
[0395] Tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluoro-2-iodoethyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-vinylquinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (900 mg, 2.23 mmol) in DCM (15 mL) was added NIS (1.01 g, 4.47 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then pyridine ;hydrofluoride (1.11 g, 11.17 mmol) was added at 0 °C under N2. The mixture was stirred at 20 °C for 12 h. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM (3 x 10 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4- (8-chloro-5-(l-fluoro-2-iodoethyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (300 mg, 37%). [0396] Tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluorovinyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluoro-2-iodoethyl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (300 mg, 0.55 mmol) in DCM (5 mL) was added DBU (166 mg, 1.09 mmol, 0.16 mL) at 20 °C under N2. The mixture was stirred at 20 °C for 16 h. The residue was diluted with H2O (30 mL) and the aqueous phase was extracted with DCM (3 x 10 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluorovinyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (200 mg, 87%).
[0397] Tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluorocyclopropyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (E)-N-(2,6-diisopropylphenyl)-l-[6-[(E)-N-(2,6- diisopropylphenyl)-C-methyl-carbonimidoyl]-2-pyridyl]ethanimine (40 mg, 0.08 mmol) in THE (3 mL) was added cobalt(II) bromide (18 mg, 0.08 mmol) at 20 °C under N2. The mixture was stirred at 20 °C for 12 h. Then a solution of tert-butyl (2R,5S)-4-(8-chloro-5-(l-fluorovinyl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (350 mg, 0.83 mmol) in THE (1 mL), dibromomethane (289 mg, 1.66 mmol, 0.12 mL) and Zn (109 mg, 1.66 mmol) were added into above solution at 20 °C under N2. The mixture was stirred at 20 °C for 2 h. The reaction mixture was quenched by sat. NH4CI (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give tert-butyl (2R,5S)-4-[8-chloro-5-(l- fluorocyclopropyl)quinazolin-4-yl]-2,5-dimethyl-piperazine-l-carboxylate (200 mg, 55%).
[0398] Tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(l- fluorocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-( 1 -fluorocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (180 mg, 0.41 mmol) and l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5- carbonitrile (193 mg, 0.83 mmol) in dioxane (3 mL) and H2O (0.6 mL) was added K3PO4 (176 mg, 0.83 mmol), KOAc (81 mg, 0.83 mmol), XPhos (40 mg, 0.08 mmol) and Pd2(dba)3 (38 mg, 0.04 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The residue was diluted with H2O (15 mL) and the aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl- 1H-pyrazol-3-yl)-5-( 1 -fluorocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (150 mg, 72%) .
[0399] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(l-fluorocyclopropyl)quinazolin-8-yl)-l-methyl- 1H -pyrazole-5-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3- yl)-5-(l-fluorocyclopropyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 0.3 mmol) in DCM (2 mL) was added trimethylsilyl trifluoromethanesulfonate (231mg, 1.04 mmol) and 2,6- dimethylpyridine (254 mg, 2.37 mmol, 0.228 mL) at 0 °C. The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give 3-(4-((2S,5R)-2,5-dimethylpiperazin-l- yl)-5-(l-fluorocyclopropyl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (480 mg, 25% purity). [0400] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-
5-(l-fluorocyclopropyl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 2- cyano-3-fluoro-2-(fluoromethyl)propanoic acid (88 mg, 0.59 mmol) in DCM (2 mL) was added 1 -chloro- N,N, 2-trimethyl-l -propenylamine (99 mg, 0.74 mmol, 0.10 mL). The mixture was stirred at 0 °C for 0.5 h. The above reaction mixture was added to a solution of 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(l- fluorocyclopropyl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (480 mg, 0.30 mmol, 25% purity) in DCM (2 mL) and DIEA (115 mg, 0.89 mmol, 0.15 mL) at 0 °C. The mixture was stirred at 20 °C for 2 h. The residue was diluted with H2O (6 mL) and the aqueous phase was extracted with EtOAc (2 x 3 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 x 7 μm; mobile phase: A: lOmM NH4HCO3 in water, B: ACN; B% in A: 45%-75%, 8.0 min) to give 3-(4-((2S,5R)-4-(2-cyano-3-fhroro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5-(l- fluorocyclopropyl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (29 mg, 15%). m/z = 537.2 [M+H]+.
[0401] The following compound was prepared according to the procedures described herein using the appropriate starting material.
Example 71 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)-5- (pyridin-3-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile
[0402] Methyl 2-amino-6-bromo-3-iodobenzoate: To a solution of methyl 2-amino-6-bromo-benzoate (60 g, 260.80 mmol) in AcOH (600 mL) was added NIS (58.68 g, 260.80 mmol) at 20 °C under N2. The mixture was stirred at 50 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (150 mL) and adjusted to pH = 7 by 1 N NaOH and extracted with EtOAc (3 x 50 mL). The combined organic phase was washed with brine (3 x 20 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give methyl 2-amino-6-bromo-3-iodobenzoate (18 g, 19%).
[0403] 2-amino-6-bromo-3-iodobenzoic acid: To a solution of methyl 2-amino-6-bromo-3-iodo- benzoate (18 g, 50.57 mmol) in MeOH (70 mL) and H2O (70 mL) was added NaOH (8.09 g, 202.27 mmol) at 20 °C. The mixture was stirred at 80 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (30 mL) and adjusted to pH = 1 by 3 N HC1. The suspension was filtered and the filter cake was dried under reduced pressure to give 2-amino-6-bromo-3- iodobenzoic acid (16 g, 93% yield).
[0404] 2-amino-6-bromo-3-iodobenzamide: To a solution of 2-amino-6-bromo-3-iodo-benzoic acid
(16 g, 46.79 mmol) in DMF (160 mL) was added HATU (21.35 g, 56.12 mmol), NH4CI (3.75 g, 70.19 mmol) and DIEA (30.24 g, 233.97 mmol). The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (450 mL) and extracted with EtOAc (3 x 100 mL). The combined organic phase was washed with brine (3 x 50 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 2-amino-6-bromo-3-iodobenzamide (17.9 g).
[0405] 5-bromo-8-iodoquinazolin-4-ol: To a solution of 2-amino-6-bromo-3-iodo-benzamide (17.9 g, 52.50 mmol) in diethoxymethoxyethane (110 mL) was added p-TsOH (904 mg, 5.25 mmol) at 20 °C. The mixture was stirred at 120 °C for 1 h. The residue was poured into H2O (60 mL). The suspension was filtered and the filter cake was dried under reduced pressure to give 5-bromo-8-iodoquinazolin-4-ol (14.5 g, 78% yield).
[0406] 5-bromo-4-chloro-8-iodoquinazoline: To a solution of 5-bromo-8-iodoquinazolin-4-ol (5 g, 14.25 mmol) in toluene (50 mL) was added POCl3 (8.74 g, 56.99 mmol) and DIEA (3.68 g, 28.50 mmol) at 20 °C. The mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give crude 5-bromo-4-chloro-8-iodoquinazoline (5.26 g), which was used into next step directly.
[0407] Tert butyl (2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a solution of 5-bromo-4-chloro-8-iodoquinazoline (5.3 g, 14.35 mmol) and tert-butyl (2R,5S)-2,5-dimethylpiperazine-l -carboxylate (4.61 g, 21.52 mmol) in NMP (80 mL) was added DIEA (9.27 g, 71.74 mmol). The mixture was stirred at 90 °C for 2 h. The reaction mixture was poured into H2O (150 mL) and extracted with EtOAc (3 x 50 mL). The combined organic phase was washed with brine (3 x 30 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (7 g, 89%).
[0408] 5-bromo-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-iodoquinazoline: To a solution of tert- butyl (2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (1 g, 1.83 mmol) in DCM (15 mL) was added trimethylsilyl trifluoromethanesulfonate (1.42 g, 6.40 mmol) and 2,6- dimethylpyridine (1.57 g, 14.62 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to remove solvent to give crude 5-bromo-4-((2S,5R)- 2,5-dimethylpiperazin-l-yl)-8-iodoquinazoline (0.81 g) which was used into next step directly.
[0409] 3-((2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazin-l-yl)-2,2- bis(fluoromethyl)-3-oxopropanenitrile: To a solution of 2-cyano-3-fluoro-2-(fluoromethyl)propanoic acid (400 mg, 2.68 mmol) in DCM (8 mL) was added l-chloro-N,N,2-trimethyl-l -propenylamine (430 mg, 3.22 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to 5-bromo-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-iodoquinazoline (800 mg, 1.79 mmol) and DIEA (693.75 mg, 5.37 mmol) in DCM (8 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (20 mL) and extracted with DCM (3 x 10 mL). The combined organic phase was washed with brine (5 mL), dried over with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 3-((2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazin-l- yl)-2,2-bis(fluoromethyl)-3-oxopropanenitrile (0.38 g). [0410] 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a solution of 3- ((2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazin-l-yl)-2,2-bis(fluoromethyl)-3- oxopropanenitrile (280 mg, 0.48 mmol) in dioxane (3 mL) and H2O (0.6 mL) was added K3PO4 (206 mg, 0.97 mmol), l-methyl-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (124 mg, 0.53 mmol) and Pd(dppf)Ch (35 mg, 0.05 mmol) at 20 °C under N2. The mixture was stirred at 60 °C for 1 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 2:1) to give 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-
2.5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile (0.18 g, 66%).
[0411] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(pyridin-3-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a solution of 3-(5-bromo-4- ((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 - methyl- 1H-pyrazole-5-carbonitrile (50 mg, 0.09 mmol) in dioxane (2 mL) was added Pd(PPh3)4 (10 mg, 0.009 mmol) and tributyl(3-pyridyl)stannane (127 mg, 0.18 mmol) at 20 °C under N2 -The mixture was stirred at 100 °C for 16 h. The reaction was diluted with H2O (5 mL) and then extracted with EtOAc (3 x 3 mL). The combined organic phase was washed with brine (2 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 xlO μm; mobile phase: A: 10 mM NH4HCO3 in water, B: CH3CN, B% in A 35%-65%, over 8 min) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-
2.5-dimethylpiperazin- 1 -yl)-5-(pyridin-3-yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (12 mg, 24%). m/z = 556.2 [M+H]+.
[0412] The following SFC and prep HPLC separation methods were employed for isolating certain compounds disclosed herein as single isomers. In the Table below, the peak number refers to the order of elution.
[0413] SFC Method: Column: DAICEL CHIRALPAK AD (250 mm x 30 mm x 10 μm): mobile phase: [CCE-MeOH (0.1%NH3H2O)]; B%: 50%, isocratic elution mode).
Prep HPLC Method: mobile phase: Eluent A: H2O (0.1% formic acid or TEA), Eluent B: MeCN (0.1% formic acid or TEA); Waters Xbridge Prep OBD 150 x 40 mm x 10 μm, Waters Xbridge BEH 100 x 30 mm x 10 μm, or Phenomenex Luna CIS 100 x 40 mm x 3 μm. [0414] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 118A and 118B 3-[4-[(2S,5R)-4-[(2R)-2-cyano-2-cyclopropyl-3-fluoro-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4-[(2S)-2- cyano-2-cyclopropyl-3-fluoro-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0415] ethyl 2-cyano-2-cyclopropyl-3-fluoropropanoate: To a mixture of ethyl 2-cyano-2- cyclopropylacetate (2 g, 13.06 mmol) in DMF (20 mL) was added NaH (783 mg, 19.59 mmol, 60% purity) at 0 °C under N2. The mixture was stirred at 0 °C for 30 min, then was added fluoroiodomethane (2.30 g, 14.36 mmol) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into ice saturated NH4CI solution (60 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (SiO2, PE:EtOAc = 10: 1 to 5: 1) to give ethyl 2-cyano-2-cyclopropyl-3-fluoropropanoate (1.5 g, 62% yield).
[0416] 2-cyano-2-cyclopropyl-3-fluoropropanoic acid: To a solution of ethyl 2-cyano-2-cyclopropyl- 3-fluoropropanoate (150 mg, 0.81 mmol) in THE (2 mL) and H2O (1 mL) was added LiOH.H2O (169 mg, 4.05 mmol) at 20 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was diluted with water (5 mL) and extracted with MTBE (2 x 2 mL). The water phase was adjusted pH = 5 by IN HC1. The mixture was extracted with EtOAc (2 x 3 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 2-cyano- 2-cyclopropyl-3-fluoropropanoic acid (100 mg).
[0417] 3-[4-[(2S,5R)-4-[(2R)-2-cyano-2-cyclopropyl-3-fluoro-propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile and 3-|4-|(2.S',5R )-4- [(2S)-2-cyano-2-cyclopropyl-3-fluoro-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 2-cyano-2-cyclopropyl-3- fluoropropanoic acid (71 mg, 0.45 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l- en-1 -amine (90 mg, 0.68 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8- yl)-l-methyl-1H-pyrazole-5-carbonitrile (100 mg, 0.23 mmol) and DIEA (70 mg, 0.54 mmol) with DCM (2 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into ice water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 mm x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: CH3 CN; B% in A: 50%-80% over 8.0 min) to diastereomeric mixture (29 mg, 22% yield) which was further purified by SFC (column: DAICEL CHIRALPAK IC(250 mm x 30 mm, 10 μm); mobile phase: [CO2-MeOH(0.1%NH3H2O)]; B%:50%, isocratic elution mode) to give and 3-[4-[(2S,5R)-4-[2-cyano-2-cyclopropyl-3-fluoro-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile as two separate diastereomers (6.90 mg, 5% yield, peak 1; 7.01 mg, 5% yield, peak 2). m/z = 582.3 [M+H]+ and m/z = 582.3 [M+H]+.
[0418] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 129 A & 129B 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4-[(2S)-2- cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0419] tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(5- bromo-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (8 g, 15.20 mmol) and (5-fluoropyridin-3-yl)boronic acid (6.42 g, 45.59 mmol) in dioxane (100 mL) and H2O (20 mL) was added K3PO4 (9.68 g, 45.59 mmol), Ag20 (1.06 g, 4.56 mmol) and Pd(dppf)Cl2-CH2Cl2 (1.24 g, 1.52 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 3 h. The reaction mixture was quenched by water (300 mL) and extracted with EtOAc (3 x 100 mL). The combined organic phase was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexane:EtOAc = 1:1) to afford tert-butyl (2R, 5S)-4-(8-(5-cyano-l -methyl- 1H-pyrazol-3-yl)-5-(5- fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (3.4 g, 41%). [0420] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-l- methyl- 1H-pyrazole-5-carbonitrile: To a mixture of tert-butyl (2R,5S)-4-(8-(5-cyano-l -methyl- 1H- pyrazol-3-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (3.2 g, 5.90 mmol) in DCM (50 mL) was added TMSOTf (4.59 g, 20.64 mmol) and 2,4-dimethylpyridine (3.16 g, 29.49 mmol) at 20 °C under N2. The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by silica gel column chromatography (EtOAc) to afford 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (5.2 g, 50% purity).
[0421] 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile and 3-[4-[(2S,5R)-4- [(2S)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 2-cyano-3,3-difluoro-2- methylpropanoic acid (54 mg, 0.36 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l- en-1 -amine (72 mg, 0.54 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8- yl)-l -methyl- 1H-pyrazole-5-carbonitrile (80 mg, 0.18 mmol) and DIEA (56 mg, 0.43 mmol) in DCM (2 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into ice water (10 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: l_WePure Biotech XP tC18 75 x 40 mm x 10 μm; mobile phase: [A: 10 mM NH4HCO3 in water, B: CH3CN; B% in A: 45%-75%] over 8.0 min) to diasteromeric mixture (28 mg, 27% yield) which was further purified SEC separation (column: ChiralPak IH, 250 x 30mm, 10μm; mobile phase: [CC>2-EtOH(0.1% NH3H2O)]; B%:30%, isocratic elution mode) to give 3- [4-[(2S,5R)-4-[2-cyano-3,3-difhroro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile as two separate diastereomers (6.78 mg, 10 % yield, peak 1 (129 A); 9.46 mg, 8 % yield, peak 2 (129B)). m/z = 574.2 [M+H]+ and m/z = 574.2 [M+H]+.
[0422] The following compound was prepared according to the procedures described herein using the appropriate starting material. z =
Example 131 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- tetrahydrofuran-3-yl-quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0423] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(2,5-dihydrofuran-3-yl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 3- (5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (150 mg, 0.27 mmol) in dioxane (1 mL) and H2O (0.2 mL) was added 2-(2,5-dihydrofuran-3-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (63 mg, 0.32 mmol), K2CO3 (74 mg, 0.54 mmol) and Pd(dppf)Ch (20 mg, 0.03 mmol) under N2. The mixture was stirred at 100 °C for 2 h. The reaction mixture was poured into H2O (3 mL) and extracted with EtOAc (3 x 1 mL). The combined organic phase was washed with brine (1 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 4:1 to 3:1) to give 3-(4-((25,5R)-4-(2-cyano-3-fluoro-2- (fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)-5-(2,5-dihydrofuran-3-yl)quinazolin-8-yl)- 1 - methyl-1H-pyrazole-5-carbonitrile (90 mg, 61%).
[0424] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(tetrahydrofuran-3-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a mixture of Pd/C (134 mg, 0.13 mmol, 10% purity) in THE (10 mL) was added 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2- (fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)-5-(2,5-dihydrofuran-3-yl)quinazolin-8-yl)- 1 - methyl- 1H-pyrazole-5-carbonitrile (230 mg, 0.42 mmol) under N2. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred under H2 (30 Psi) at 50 °C for 32 h. The reaction mixture was filtered and concentrated under reduced pressure to give a residue. The residue was dissolved in CHCI3 (10 mL), to which was added MnO2 (73 mg, 0.84 mmol). The mixture was stirred at 70 °C for 4 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with the following condition (column: WePure Biotech XP tC 18 150 x 40 mm x 7μm; mobile phase: A: 10 mM NH4HCO3 in water, B: MeCN; B% in A: 53%-73%, 8.0 min) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)- 5-(tetrahydrofuran-3-yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (9 mg, 4%). m/z = 549.2 [M+H]+.
[0425] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 134 reZ-3-(4-((lS,6R)-5-((S)-2-cyano-3-fluoro-2-methylpropanoyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-5- (5-fluoropyridin-3-yl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile
[0426] 5-bromo-4-chloro-8-iodoquinazoline: To a solution of 5-bromo-8-iodoquinazolin-4-ol (1 g, 2.85 mmol) in Toluene (10 mL) was added POCI3 (1.75 g, 11.40 mmol, 1.06 mL) and DIEA (737 mg, 5.70 mmol, 1 mL) at 20 °C under N2. The reaction mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give 5-bromo-4-chloro-8-iodoquinazoline (2.2 g, 45% purity) which was used into next step directly.
[0427] tert-butyl (1R ,6S)-5-(5-bromo-8-iodoquinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2- carboxylate: To a solution of 5-bromo-4-chloro-8-iodoquinazoline (2 g, 2.44 mmol, 45% purity) in DIEA (20 mL) was added rel- tert-butyl ( 1 R ,6.S')-2, 5-diazabicyclo| 4.2.0 |octanc-2-carboxy late (500 mg, 2.36 mmol) at 20 °C. The reaction mixture was stirred at 140 °C for 1 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 5:1) to give tert-butyl (1R,6S)-5-(5-bromo-8-iodoquinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate (1 g, 78% yield). [0428] Tert-butyl ( lR ,6.S)-5-(5-bromo-8-(5-cvano-l -methyl- 1H -pyrazol-3-yl)quinazolin-4-yl)-2,5- diazabicyclo[4.2.0]octane-2-carboxylate: To a solution of tert-butyl (1R,6S)-5-(5-bromo-8- iodoquinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate (800 mg, 1.47 mmol) and l-methyl-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H pyrazole-5-carbonitrile (308 mg, 1.32 mmol) in 1,4- dioxane (10 mL) and H2O (2 mL) was added K3PO4 (623 mg, 2.93 mmol) and Pd(dppf)Ch (107 mg, 0.15 mmol) at 20 °C under N2. The reaction mixture was stirred at 60 °C for 1 h. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (1R,6S)-5-(5-bromo-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)quinazolin-4-yl)-2,5- diazabicyclo[4.2.0]octane-2-carboxylate (330 mg, 43%).
[0429] tert-butyl (1R,6S)-5-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate: To a solution of tert-butyl (1R,65)-5- (5-bromo-8-(5-cyano-l-methyl-1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2- carboxylate (280 mg, 0.53 mmol) and (5-fluoropyridin-3-yl)boronic acid (226 mg, 1.60 mmol) in 1,4- dioxane (5 mL) was added K3PO4 (340 mg, 1.60 mmol), Ag2O (37 mg, 016 mmol) and Pd(dppf)Cl2*CH2Cl2 (44 mg, 0.05 mmol) at 20 °C under N2. The reaction mixture was stirred at 100 °C for 12 h. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1 to 1:1) to give tert-butyl (1R,6S)-5-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(5- fluoropyridin-3-yl)quinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate (160 mg, 55 % yield).
[0430] 3-(4-(( LS,6R )-2,5-diazabicyclo|4.2.0|octan-2-yl)-5-(5-fluoropyridm-3-yl)qumazolin-8-yl)-l- methyl- 1H-pyrazole-5-carbonitrile: To a solution of tert-butyl ( l R .6.S')-5-(8-(5-cyano- l -mcthyl- l /7- pyrazol-3-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate (160 mg, 0.30 mmol) in DCM (3 mL) was added 2,6-lutidine (159 mg, 1.48 mmol) and TMSOTf (230 mg, 1.04 mmol) at 0 °C under N2, the reaction mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give 3-(4-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)-5-(5- fluoropyridin-3-yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (370 mg, crude).
[0431] reZ-3-(4-((lS,6R)-5-((S)-2-cyano-3-fluoro-2-methylpropanoyl)-2,5-diazabicyclo[4.2.0]octan- 2-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-l-methyl-1H-pyrazole-5-carbonitrile: To a solution of CS>2-cyano-3-fluoro-2-mcthyl propanoic acid (41 mg, 0.31 mmol) in DCM (1 mL) was added 1-chloro- MM2-trimcthyl-prop- l -cn- l -amine (50 mg, 0.37 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. The mixture was then added to a solution of 3-(4-((1S,6R)-2,5-diazabicyclo[4.2.0]octan-2-yl)- 5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- l -mcthyl-1H-pyrazolc-5-carbonitrilc (260 mg, 0.21 mmol) and DIEA (53 mg, 0.41 mmol) in DCM (2 mL) at 0 °C, followed by stirring at 0 °C under N2 for 1 h. The reaction mixture was diluted with water (5 mL) and extracted with DCM (3 x 3 mL). The combined organic layers were washed with brine (3 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Waters xbridge 150 x 25 mm x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: CH3CN; B% in A: 45%-75%, 9 min) to give reZ-3-(4-((1S,6R)-5-((S)-2-cyano-3-fluoro-2-methylpropanoyl)-2,5-diazabicyclo[4.2.0]octan-2-yl)-5-(5- fluoropyridin-3-yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (47.05 mg, 41% yield), m/z = 554.1 [M+H]+.
[0432] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 145 l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-5-methoxy-pyrazole-4-carbonitrile
[0433] tert-butyl (2R,5S)-4-(5-bromo-8-(4-(ethoxycarbonyl)-5-methoxy-1H -pyrazol-l- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2A,,5S’)-4-(5- bromo-8-iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 0.92 mmol) and ethyl 3- methoxy-1H-pyrazole-4-carboxylate (311 mg, 1.83 mmol) in DMF (6 mL) was added K2CO3 (253 mg, 1.83 mmol) and Cui (69 mg, 0.37 mmol) A^l ,N2-dimcthylcyclohcxanc-l ,2-diaminc (103 mg, 0.73 mmol). The mixture was stirred at 100 °C for 5 h. Two parallel batches were combined. The reaction mixture was poured into water (25 mL) and extracted with EtOAc (3 x 15 mL). The combined organic phase was washed with brine (15 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give tert-butyl (2R,5S)-4-(5-bromo-8-(4-(ethoxycarbonyl)-5-methoxy-1H-pyrazol-l-yl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (860 mg, 80% yield).
[0434] tert-butyl (2R,5S)-4-(8-(4-(ethoxycarbonyl)-5-methoxy-1H -pyrazol-l-yl)-5-(5-fluoropyridin- 3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R.5S)-4-(5- bromo-8-(4-(ethoxycarbonyl)-5-methoxy- 1H-pyrazol-l -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (860 mg, 1.46 mmol) and (5-fluoro-3-pyridyl)boronic acid (411 mg, 2.92 mmol) in 1,4- dioxane (10 mL) was added Pd(dppf)Cl2-CH2Cl2 (119 mg, 0.15 mmol), Ag2O (101 mg, 0.45 mmol) and K3PO4 (929 mg, 4.38 mmol). The mixture was stirred at 100 °C for 36 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-(4-(ethoxycarbonyl)-5-methoxy-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (480 mg, 54% yield).
[0435] l-(4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-8-yl)-5-methoxy- 1H-pyrazole-4-carboxylic acid: To a solution of tert-butyl (2R,5>S)-4- (8-(4-(ethoxycarbonyl)-5-methoxy-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (480 mg, 0.79 mmol) in THE (4 mL) H2O (1 mL) was added NaOH (127 mg, 3.17 mmol). The mixture was stirred at 50 °C for 48 h. The reaction mixture was diluted with H2O (10 mL) and extracted with MTBE (10 ml). The aqueous phase was adjusted pH = 1 by 3N HC1. Then the mixture extracted with EtOAc (3 x 5 ml) and concentrated under reduced pressure to give l-(4- ((25,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-5- methoxy- 1 H-pyrazolc-4-carboxylic acid (330 mg, 72% yield).
[0436] tert-butyl (21f,5S)-4-(8-(4-carbamoyl-5-methoxy-1H -pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of l-(4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-5-methoxy-1H- pyrazole-4-carboxylic acid (330 mg, 0.57 mmol) in DMF (5 mL) was added NH4CI (61 mg, 1.14 mmol), HATU (261 mg, 0.69 mmol) and DIEA (221 mg, 1.71 mmol). The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 3:1) to give compound tert-butyl (2R,5S)-4-(8-(4-carbamoyl-5-methoxy-1H- pyrazol- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (270 mg, 82% yield).
[0437] tert-butyl (2R,5S)-4-(8-(4-cyano-5-methoxy-1H -pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8-(4- carbamoyl -5-methoxy-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (270 mg, 0.46 mmol) in THF (5 mL) was added methoxycarbonyl- (triethylammonio)sulfonyl-azanide (557 mg, 2.34 mmol). The mixture was stirred at 60 °C for 2 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-(4-cyano-5-methoxy-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (220 mg, 84% yield).
[0438] l-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-5- methoxy- 1H-pyrazole-4-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(4-cyano-5-methoxy-1H- pyrazol- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (220mg, 0.39 mmol) in DCM (3 mL) was added TMSOTf (306 mg, 1.38 mmol) and 2,6-lutidine (211 mg, 1.97 mmol) at 20°C. The reaction mixture stirred at 20°C for 2h and then concentrated under reduced pressure. The residue was purified by prep-TLC (SiO2, DCM:MeOH = 10:1) to give l-(4-((2S,5R)-2,5- dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-5-methoxy-1H-pyrazole-4-carbonitrile (120 mg, 66% yield).
[0439] l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-(5-fluoro-3-pyridyl)quinazolin-8-yl]-5-methoxy-pyrazole-4-carbonitrile: To a solution of 2-cyano- 3-fluoro-2-(fluoromethyl)propanoic acid (30 mg, 0.19 mmol) in DCM (0.25 mL) was added 1-chloro- N,N,2-trimethyl-prop-l-en-l -amine (31 mg, 0.23 mmol) at 0°C, the mixture was stirred at 0°C for 0.5 h. Then this mixture was added to a mixture of l-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5- fluoropyridin-3-yl)quinazolin-8-yl)-5-methoxy-1H-pyrazole-4-carbonitrile (60 mg, 0.13 mmol) and DIEA (85 mg, 0.65 mmol) in DCM (0.5 mL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into water (3 mL) and extracted with DCM (3 x 1 mL).The combined organic phase was concentrated under reduced pressure. The residue was purified by prep-HPLC with the following condition (column: Waters Xbridge BEH Cl 8 150 x 40 mm x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 57%-77%, 8.0 min) to give l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2- (fluoromethyl)propanoyl] -2,5 -dimethyl-piperazin- 1 -yl] -5 -(5 -fluoro-3 -pyridyl)quinazolin- 8-yl] -5- methoxy-pyrazole-4-carbonitrile (16 mg, 20% yield), m/z = 590.2 [M+H]+.
[0440] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 153 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- pyrrolidin-l-yl-quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0441] tert-butyl (2R,5S)-4-(8-chloro-5-(pyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a mixture of tert-butyl (2R,5S)-4-(5-amino-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (210 mg, 0.54 mmol) in DMF (3 mL) was added NaH (47 mg, 1.18 mmol, 60% purity) at 0 °C under N2. The mixture was stirred at 0 °C for 30 min, then was added 1,4- diiodobutane (199 mg, 0.64 mmol) in DMF (0.5 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. The reaction mixture was poured into ice saturated aqueous NH4CI (15 mL). The aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE: EtOAc = 3:1) to give tert- butyl (2R,5S)-4-(8-chloro-5-(pyrrolidin- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (200 mg, 84% yield).
[0442] tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(pyrrolidin-l-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-(pyrrolidin- l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (200 mg, 0.45mmol) and l-methyl-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (209 mg, 0.90 mmol) in 1,4- dioxane (5 mL) and H2O (1 mL) was added K3PO4 (190 mg, 0.90 mmol), KOAc (88 mg, 0.90 mmol), XPhos (43 mg, 0.09 mmol) and Pd2(dba)3 (41 mg, 0.04 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The residue was diluted with H2O (15 mL) and the aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)- 5-(pyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (100 mg, 43% yield). [0443] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(pyrrolidin-l-yl)quinazolin-8-yl)-l-methyl-1H- pyrazole-5-carbonitrile: To a mixture of tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5- (pyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (100 mg, 0.19 mmol) in DCM (2 mL) was added 2,4-lutidine (104 mg, 0.97 mmol, 0.11 mL) and TMSOTf (151 mg, 0.68 mmol, 0.12 mL) at 20 °C under N2. The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent to give 5-[4-[(2S,5R)-2,5-dimethylpiperazin-l-yl]-5-pyrrolidin-l-yl- quinazolin-8-yl]-2-methyl-pyrazole-3-carbonitrile (370 mg, 20% purity).
[0444] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-pyrrolidin-l-yl-quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 2-cyano-3- fluoro-2-(fluoromethyl)propanoic acid (29 mg, 0.19 mmol) in DCM (2 mL) was added 1-Chloro-N,N,2- trimethyl-1 -propenylamine (31 mg, 0.23mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to a mixture of was added to 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5- (pyrrolidin-l-yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (270 mg, 0.13 mmol) and DIEA (50 mg, 0.39 mmol) in DCM (1 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. The reaction mixture was poured into H2O (6 mL). The aqueous phase was extracted with DCM (3 x 2 mL). The combined organic layers were washed with brine (2 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 x 7 μm; mobile phase: A: lOmM NH4HCO3 in water, B: ACN; B% in A: 60%-80%, 8.0 min) to give 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2- (fluoromethyl)propanoyl] -2,5 -dimethyl-piperazin- 1 -yl] -5 -pyrrolidin- 1 -yl-quinazolin- 8 -yl] - 1 -methyl- pyrazole-5-carbonitrile (11 mg, 15% yield), m/z = 548.3 [M+H]+.
[0445] The following compound was prepared according to the procedures described herein using the appropriate starting material.
Examples 155A & 155B
3-[(2R,5S)-4-[5-cyclopropyl-8-[4-(trifluoromethyl)triazol-2-yl]quinazolin-4-yl]-2,5-dimethyl- piperazin-l-yl]-2,2-bis(fluoromethyl)-3-oxo-propanenitrile & 3-[(2R,5S)-4-[5-cyclopropyl-8-[4- (trifluoromethyl)triazol-l-yl]quinazolin-4-yl]-2,5-dimethyl-piperazin-l-yl]-2,2-bis(fluoromethyl)-3- oxo-propanenitrile
[0446] tert -butyl (2R,5S)-4-(5-cyclopropyl-8-(4-(trifluoromethyl)-lZM,2,3-triazol-l-yl)quinazolin- 4-yl)-2,5-dimethylpiperazine-l-carboxylate and tert -butyl (2R,5S)-4-(5-cyclopropyl-8-(4- (trifluoromethyl)-2H-l, 2, 3-triazol-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate: To a solution of (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-cyclopropylquinazolin-8- yl)boronic acid (200 mg, 0.47 mmol) in pyridine (3 mL) was added Cu(OAc)2 (170 mg, 0.94 mmol) and 4-(trifluoromethyl)-1H-l,2,3-triazole (129 mg, 0.94 mmol) at 20 °C under O2. The mixture was stirred at 80 °C for 4 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (5mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =5:1 to 4:1) to give a mixture of tert-butyl (2R,5S)-4-(5-cyclopropyl-8-(4-(trifluoromethyl)- \ H- 1 ,2,3-triazol- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate and tert-butyl (2R,5S)-4-(5- cyclopropyl-8-(4-(trifluoromethyl)-2H- 1 ,2,3-triazol-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (150 mg, 62% yield).
[0447] 5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-(4-(trifluoromethyl)-1H -l,2,3- triazol-l-yl)quinazoline and tert-butyl 5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-(4- (trifluoromethyl)-2H-l,2,3-triazol-2-yl)quinazoline: To a solution of tert- (2R,5S)-4-(5-cyclopropyl-8- (4-(trifluoromethyl)- \ H- 1 ,2,3-triazol- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate and tert-butyl (2R,5S)-4-(5-cyclopropyl-8-(4-(trifluoromethyl)-2H-1,2,3-triazol-2-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (150 mg, 0.29 mmol) in DCM (1 mL) was added TMSOTf (225 mg, 1.01 mmol) and 2,6-lutidine (155 mg, 1.45 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =1:1 to 0:1) to give a mixture of tert-5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)-8-(4-(trifluoromethyl)- \H- 1 ,2,3-triazol- 1 -yl)quinazoline and tert-butyl 5- cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)-8-(4-(trifluoromethyl)-2H- 1 ,2,3-triazol-2- yl)quinazoline (100 mg, 83% yield).
[0448] 3-[(2R,5S)-4-[5-cyclopropyl-8-[4-(trifluoromethyl)triazol-2-yl]quinazolin-4-yl]-2,5- dimethyl-piperazin-l-yl]-2,2-bis(fluoromethyl)-3-oxo-propanenitrile and 3-[(2R,5S)-4-[5- cyclopropyl-8-[4-(trifluoromethyl)triazol-l-yl]quinazolin-4-yl]-2,5-dimethyl-piperazin-l-yl]-2,2- bis(fluoromethyl)-3-oxo-propanenitrile: To a solution of 2-cyano-3-fluoro-2-(fluoromethyl)propanoic acid (57 mg, 0.38 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l-en-l -amine (64 mg, 0.48 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to a mixture of 5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-(4-(trifluoromethyl)-1H-1,2,3- triazol- 1 -yl)quinazoline and tert-butyl 5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)-8-(4-
(trifluoromethyl)-277-l,2,3-triazol-2-yl)quinazoline (100 mg, 0.24 mmol) and DIEA (124 mg, 0.96 mmol) in DCM (2 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (2 mL) and extracted with DCM (3 x 3 mL). The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with following condition (column: WePure Biotech XP tC18 150 x 40 mm x 7μm; mobile phase: A: lOmM NH4HCO3 in water, B: MeCN; B% in A: 55%-90%,
8.0 min) to give the title compounds as two separate isomers (21 mg, 16% yield, peak 1 (155A); 23 mg,
16% yield, peak 2 (155B)). m/z = 549.2 [M+H]+ & m/z = 549.2 [M+H]+.
[0449] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 189 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(2- cyclopropylpyrazol-3-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0450] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-(2-cyclopropylpyrazol-3-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 3- (5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (50 mg, 0.09 mmol) and l-cyclopropyl-5- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H pyrazole (42 mg, 0.18 mmol) in dioxane (1 mL) was added Pd(dppf)Ch CH2Cl2 (7.33 mg, 0.01 mmol) and Ag2O (6 mg, 0.02 mmol) and K3PO4 (57 mg, 0.26 mmol) under N2. The mixture was stirred at 100 °C for 16 h. The mixture was poured into by H2O (3 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with brine (3 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: WePure Biotech XP tC18 150 x 40 mm x 7 μm; mobile phase: A: lOmM NH4HCO3 in water, B: MeCN; B% in A: 55%-75%, 8 min) to give 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2- (fluoromethyl)propanoyl]-2,5-dimethyl-piperazin- 1 -yl]-5-(2-cyclopropylpyrazol-3-yl)quinazolin-8-yl] - 1 - methyl-pyrazole-5-carbonitrile (28 mg, 53%). m/z = 585.3 [M+H]+.
[0451] The following compound wase prepared according to the procedures described herein using the appropriate starting material. Example 191A & 191B
3-[4-[(2S,5R)-2,5-dimethyl-4-[(2R)-2-methyl-l,l-dioxo-thietane-2-carbonyl]piperazin-l-yl]-5-(3- fluorophenyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-2,5-dimethyl-4- [(2S)-2-methyl-l,l-dioxo-thietane-2-carbonyl]piperazin-l-yl]-5-(3-fluorophenyl)quinazolin-8-yl]-l- methyl-pyrazole-5-carbonitrile
[0452] tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(3-fluorophenyl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 0.95 mmol) and (3-fluorophenyl)boronic acid (399 mg, 2.85 mmol) in 1,4-dioxane (10 mL) was added Pd(dppf)Cl2 .CH2Cl2 (78 mg, 0.09 mmol), Ag2O (66 mg, 0.28 mmol) and K3PO4 (605 mg, 2.85 mmol) under N2. The mixture was stirred at 100 °C for 16 h. The mixture was poured into by H2O (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL) and dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Commercial hexanes:EtOAc = 4:1 to 3:1) to give tert-butyl (2R,5>S)-4-(8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)-5-(3-fluorophenyl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (500 mg, 97% yield).
[0453] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(3-fluorophenyl)quinazolin-8-yl)-l-methyl-1H- pyrazole-5-carbonitrile: To a solution of t tert-butyl (2R,5S)-4-(8-(5-cyano-l -methyl- 1H-pyrazol-3-yl)- 5-(3-fluorophenyl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 0.92 mmol) in DCM (5 mL) was added TMSOTf (718 mg, 3.23 mmol) and 2,6-lutidine (791 mg, 7.39 mmol). The mixture was concentrated under reduced pressure. The crude product was purified by silica gel chromatography (EtOAc: MeOH=20:l to 10:1) to give 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(3- fluorophenyl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (800 mg, 50% purity).
[0454] 3-(4-((2S,5R)-4-(l,l-dioxidothietane-2-carbonyl)-2,5-dimethylpiperazin-l-yl)-5-(3- fluorophenyl)quinazolin-8-yl)- l-methyl- 1H-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)- 2,5-dimethylpiperazin- 1 -yl)-5-(3-fluorophenyl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (400 mg, 0.45 mmol) and l,l-dioxothietane-2-carboxylic acid (71 mg, 0.47 mmol) in THF (5 mL) was added CMPI (174 mg, 0.68 mmol) and DIEA (293 mg, 2.26 mmol). The mixture was stirred at 50 °C for 2 h. The mixture was poured into by H2O (5 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL) and dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Commercial hexanes: EtOAc = 3:1 to 1:1) to give 3-(4-((2S,5R)-4-(l,l-dioxidothietane-2-carbonyl)-2,5-dimethylpiperazin-l-yl)-5-(3- fhrorophenyl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (187 mg, 72% yield).
[0455] 3-[4-[(2S,5R)-2,5-dimethyl-4-[(2R)-2-methyl-l,l-dioxo-thietane-2-carbonyl]piperazin-l-yl]- 5-(3-fluorophenyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile and 3-[4-[(2S,5R)-2,5- dimethyl-4- [(2S)-2-methyl- 1 , 1 -dioxo-thietane-2-carbonyl]piperazin- 1 -yl] -5- (3- fluorophenyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)-4- (1,1 -dioxidothietane-2-carbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-(3-fluorophenyl)quinazolin-8-yl)- 1 - methyl- 1 H-pyrazolc-5-carbonitrilc (100 mg, 0.17 mmol) in DMF (1 mL) was added CS2CO3 (114 mg, 0.35 mmol) and Mel (37 mg, 0.26 mmol) at 20 °C. The mixture was stirred at 20 °C for 16 h. The reaction mixture was poured into water (5 mL). The precipitated solid was filtered, the filter cake was washed with water (5 mL). The filter cake was dried under reduced pressure to give diastereomeric mixture (80 mg) as white solid which was purified by SEC (column: DAICEL CHIRALCEL OD (250mmx30mm, 10 um); mobile phase: [CO2 -IPA(0.1 % NH3H2O)]; B%:50%, isocratic elution mode) to give 3-(4-((2S,5R)-2,5-dimethyl-4-(2-methyl-l,l-dioxidothietane-2-carbonyl)piperazin-l-yl)-5-(3- fluorophenyl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (30 mg, 29% yield, peak 1 (191A); 25 mg, 25% yield, peak 2 (191B)). m/z = 588.3 [M+H]+ and m/z = 588.3 [M+H]+.
[0456] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 195 3-[4-[(2S,5R)-4-[(2S)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(2- oxopyrrolidin-l-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0457] tert-butyl (2R,5S)-4-(8-chloro-5-((diphenylmethylene)amino)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (1 g, 2.19 mmol) and diphenylmethanimine (477 mg, 2.63 mmol) in dioxane (20 mL) was added Pd2(dba)3 (201 mg, 0.22 mmol), Xantphos (254 mg, 0.44 mmol) and CS2CO3 (2.14 g, 6.58 mmol). The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into water (40 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-((diphenylmethylene)amino)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (1.2 g, 98% yield).
[0458] tert-butyl (2R,5S)-4-(5-amino-8-chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate: To a mixture of tert-butyl (2R,5S)-4-(8-chloro-5-((diphenylmethylene)amino)quinazolin-
4-yl)-2,5-dimethylpiperazine-l -carboxylate (1.2 g, 2.16 mmol) in THF (10 mL) was added HC1 (1 M, 21.58 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 2 h. The reaction mixture was poured into H2O (15 mL). The aqueous phase was extracted with MTBE (3 x 5 mL). Then the aqueous phase was neutralized with NaHCO3 aqueous (30 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with brine (15 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue to give tert-butyl (2R ,5.S’)-4- (5-amino-8-chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (840 mg, 99% yield).
[0459] tert-butyl (2R,5S)-4-(5-(4-bromobutanamido)-8-chloroquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-amino-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (840 mg, 2.14 mmol) and 4-bromobutanoyl chloride (596 mg, 3.22 mmol) in THF (10 mL) was added K2CO3 (740 mg, 5.36 mmol). The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (20 mL). The aqueous phase was extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (20 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 3:1) to give tert-butyl (2R ,5.S’)-4- (5-(4-bromobutanamido)-8-chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (860 mg, 74 % yield).
[0460] tert-butyl (2R,5,S)-4-(8-chloro-5-(2-oxopyrrolidin-l-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2R .5.S')-4-(5-(4-bromobutanamido)-8- chloroquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (860 mg, 1.59 mmol) in DMF (10 mL) was added NaH (287 mg, 7.17 mmol) at 0°C .The mixture was stirred at 20°C for 1 h. The reaction mixture was poured into ice saturated NH4CI solution (20 mL). The reaction mixture was poured into H2O (10 mL). The aqueous phase was extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1 to 0:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-(2-oxopyrrolidin-l-yl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l -carboxylate (380 mg, 52% yield).
[0461] tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H -pyrazol-3-yl)-5-(2-oxopyrrolidin-l- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2A',5S’)-4-(8- chloro-5-(2-oxopyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (180 mg, 0.4 mmol) and 2-methyl-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazole-3-carbonitrile (136 mg, 0.59 mmol) in dioxane (2 mL) H2O (0.4 mL) was added Pd2(dba)3 (36 mg, 0.04 mmol), XPhos (37 mg, 0.08 mmol), K3PO4 (166 mg, 0.78 mmol) and KOAc (76 mg, 0.78 mmol). The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with brine (3 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc=5:l to 3:1) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(2- oxopyrrolidin-l-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (180 mg, 87% yield).
[0462] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(2-oxopyrrolidin-l-yl)quinazolin-8-yl)-l- methyl-1H -pyrazole-5-carbonitrile: To a solution of tert-butyl (2R .5.S')-4-(8-(5-cyano- l -mcthyl- l /7- pyrazol-3-yl)-5-(2-oxopyrrolidin- 1 -yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (380 mg, 0.7 mmol) in DCM (5 mL) was added TMSOTf (557 mg, 2.51 mmol) and 2,6-dimethylpyridine (384 mg, 3.58 mmol). The mixture was stirred at 0 °C for 1 h. The reaction mixture concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA:MeOH =1:0 to 10:1) to give 3-(4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)-5-(2-oxopyrrolidin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile (300 mg, 97% yield).
[0463] 3-[4-[(2S,5R)-4-[(2S)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(2-oxopyrrolidin-l-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of intermediate 4B (69 mg, 0.46 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l-en-l- amine (65 mg, 0.48 mmol) at 0°C, the mixture was stirred at 0 °C for 0.5 h. Then this mixture was added to the mixture of 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(2-oxopyrrolidin-l-yl)quinazolin-8-yl)-l- methyl-1H-pyrazole-5-carbonitrile (100 mg, 0.23 mmol) and DIEA (150 mg, 1.16 mmol) in DCM (2 mL) .The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into water (3 mL) and extracted with DCM (3 x 1 mL). The combined organic phase was concentrated under reduced pressure. The residue was purified by prep-HPLC with the following condition (column: WePure Biotech XP tC18 150 x 40 mm x 7 μm; mobile phase: A: lOmM NH4HCO3 in water, B: ACN; B% in A: 35%-65%, 8.0 min) to give 3-[4-[(2S,5R)-4-[(2S)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(2-oxopyrrolidin-l-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile (23 mg, 18 % yield), m/z = 562.2 [M+H]+.
[0464] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 198 l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- cyclopropyl-quinazolin-8-yl]-4-methoxy-pyrazole-3-carbonitrile
[0465] tert -butyl (2R,5S)-4-(8-(3-bromo-4-methoxy-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (4-((25,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)-5-cyclopropylquinazolin-8-yl)boronic acid (500 mg, 1.17 mmol) and 3-bromo-4- methoxy-1H-pyrazole (415 mg, 2.35 mmol) in pyridine (10 mL) was added Cu(OAc)2 (426 mg, 2.35 mmol) under O2. The mixture was stirred at 70 °C for 16 h. The mixture was poured into H2O (20 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (10 mL) and dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Commercial hexanes:EtOAc = 3:1 to 1:1) to give tert-butyl (2R,5S)-4-(8-(3- bromo-4-methoxy- 1H-pyrazol- 1 -yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (320 mg, 49% yield).
[0466] tert-butyl (2R,5S)-4-(8-(3-cyano-4-methoxy-1H -pyrazol-l-yl)-5-cyclopropylquinazolin-4-yl)- 2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8-(3-bromo-4-methoxy- 1H-pyrazol- 1 -yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (320 mg, 0.57 mmol) in DMF (3 mL) was added Pd(dppf)Ch (42 mg, 0.06 mmol), Zn(CN)2 (67 mg, 0.57 mmol) and Zn (38 mg, 0.57 mmol). The mixture was stirred at 120 °C for 2 h. The mixture was poured into by H2O (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL) and dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography (Commercial hexanes:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(8- (3-cyano-4-methoxy- 1H-pyrazol- 1 -yl)-5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine- 1 - carboxylate (160 mg, 55% yield).
[0467] l-(5-cyclopropyl-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-4-methoxy-1H - pyrazole-3-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(3-cyano-4-methoxy-1H-pyrazol-l-yl)- 5-cyclopropylquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (160 mg, 0.32 mmol) in DCM (2 mL) was added TMSOTf (247 mg, 1.11 mmol) and 2,6-lutidine (272 mg, 2.54 mmol). The mixture was stirred at 20 °C for 1 h. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (EtOAc: MeOH = 20:1 to 10:1) to give l-(5-cyclopropyl-4-((2S,5R)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-4-methoxy-1H-pyrazole-3-carbonitrile (250 mg, 35% purity).
[0468] l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-cyclopropyl-quinazolin-8-yl]-4-methoxy-pyrazole-3-carbonitrile: To a solution of 2-cyano-3- fluoro-2-(fluoromethyl)propanoic acid (90 mg, 0.61 mmol) in DCM (2 mL) was added l-chloro-N,N,2- trimethyl-prop-l-en-1 -amine (27 mg, 0.20 mmol). The mixture was stirred at 0°C for 1 h. Then the mixture was added to a mixture of l-[5-cyclopropyl-4-[(2S,5R)-2,5-dimethylpiperazin-l-yl]quinazolin-8- yl]-4-methoxy-pyrazole-3-carbonitrile (233 mg, 0.20 mmol) and DIEA (26 mg, 0.20 mmol) in DCM (5 mL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into H2O (5 mL) and extracted with DCM (3 x 5 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: WePure Biotech XP tC18 150 x 30 mm x 7 μm; mobile phase: A: lOmM NH4HCO3 in water, B: MeCN; B% in A: 55%-75%, 8 min) to give l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2- (fluoromethyl)propanoyl] -2,5 -dimethyl-piperazin- 1 -yl] -5 -cyclopropyl-quinazolin- 8 -yl] -4-methoxy- pyrazole-3-carbonitrile (35 mg, 32% yield), m/z = 535.3 [M+H]+.
[0469] The following compounds were prepared according to the procedures described herein using the appropriate starting material. 199B m/z =
Example 200 4-chloro-2-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]thiazole-5-carbonitrile
[0470] tert-butyl (2R,5S)-4-(8-(4-chloro-5-cyanothiazol-2-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)boronic acid (380 mg, 0.56 mmol) in dioxane (5 mL) and H2O (1 mL) was added 2,4-dichlorothiazole-5-carbonitrile (151 mg, 0.85 mmol), Pd(dppf)Ch (41 mg, 0.06 mmol) and K2CO3 (155 mg, 1.12 mmol) under N2. The mixture was stirred at 100 °C for 2 h. The reaction mixture was poured into H2O (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =5:1 to 4:1) to give tert-butyl (2R,5S)-4-(8-(4-chloro-5-cyanothiazol-2-yl)- 5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (280 mg, 86% yield). [0471] 4-chloro-2-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8- yl)thiazole-5-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(4-chloro-5-cyanothiazol-2-yl)-5-(5- fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (280 mg, 0.48 mmol) in DCM (5 mL) was added TMSOTf (322 mg, 1.45 mmol) and 2,6-lutidine (259 mg, 2.41 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =1:1 to 0:1) to give 4-chloro-2-(4- ((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)thiazole-5-carbonitrile (160 mg, 69% yield). [0472] 4-chloro-2-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl- piperazin-l-yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]thiazole-5-carbonitrile: To a solution of 2- cyano-3-fluoro-2-(fluoromethyl)propanoic acid (37 mg, 0.25 mmol) in DCM (1 mL) was added 1-chloro- N,N,2-trimethyl-prop-1 -cn-1 -amine (45 mg, 0.33 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to a mixture of 4-chloro-2-(4-((2S,5R)-2,5-dimethylpiperazin- l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)thiazole-5-carbonitrile (80 mg, 0.17 mmol) and DIEA (65 mg, 0.50 mmol) in DCM (1 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (3 mL) and extracted with DCM (3 x 1 mL). The combined organic phase was washed with brine (1 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with following condition (column: Waters Xbridge BEH C18 100 x 30 mm x 10μm; mobile phase: A: lOmM NH4HCO3 in water, B: MeCN; B% in A: 50%- 80%, 8.0 min) to give 4-chloro-2-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5- dimethyl-piperazin- 1 -yl] -5 -(5 -fluoro-3 -pyridyl)quinazolin- 8 -yl] thiazole-5 -carbonitrile (18.66 mg, 18% yield), m/z = 611.1 [M+H]+.
[0473] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 205 & 206 5-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-3-carbonitrile & 3-[4-[(2S,5R)-4-[2- cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-5-carbonitrile
[0474] 3-bromo-l -(bromodifluoromethyl)-1H -pyrazole-5-carbonitrile and 5-bromo-l- (bromodifluoromethyl)-1H -pyrazole-3-carbonitrile: To a solution of 3-bromo- 1 H-pyrazolc-5- carbonitrile (2 g, 11.63 mmol) in DMF (20 mL) was added NaH (698 mg, 17.44 mmol, 60% purity). The mixture was stirred at 0 °C for 30 min, dibromo(difluoro)methane (4.88 g, 23.26 mmol) was added. The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into H2O (60 mL) and extracted with EtOAc (3 x 30 mL). The combined organic phase was washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =30:1 to 20:1) to give a mixture of 3-bromo-l- (bromodifluoromethyl)-1H-pyrazole-5-carbonitrile and 5-bromo-l-(bromodifluoromethyl)-1H-pyrazole- 3 -carbonitrile (1.1 g, 31% yield).
[0475] 3-bromo-l-(trifluoromethyl)-1H -pyrazole-5-carbonitrile and 5-bromo-l-(trifluoromethyl)- 1H -pyrazole-3-carbonitrile: To a solution of the mixture of 3-bromo-l -(bromodifluoromethyl)- \ H- pyrazole-5-carbonitrile and 5-bromo-l -(bromodifluoromethyl)- 1H-pyrazole-3-carbonitrile (1.1 g, 3.66 mmol) in DCE (10 mL) was added AgBF4 (1.42 g, 7.31 mmol) at 20 °C under N2. The mixture was stirred at 50 °C for 16 h. The reaction mixture was poured into H2O (10 mL) and extracted with DCM (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 50:1 to 40:1) to give a mixture of 3-bromo-l -(trifluoromethyl)- \ H- pyrazole-5-carbonitrile and 5-bromo-l-(trifluoromethyl)-1H-pyrazole-3-carbonitrile (580 mg, 66% yield) as colorless oil. The mixture was used into next step directly.
[0476] tert -butyl (2R,5S)-4-(8-chloro-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-chloroquinazolin- 4-yl)-2,5-dimethylpiperazine-l -carboxylate (8 g, 17.55 mmol) and (5-fluoro-3-pyridyl)boronic acid (3.71 g, 26.33 mmol) in dioxane (200 mL) and H2O (30 mL) was added Ag2O (1.22 g, 5.27 mmol), Pd(dppf)Cl2-CH2Cl2 (1.43 g, 1.76 mmol) and K3PO4 (11.18 g, 52.66 mmol) under N2. The mixture was stirred at 100 °C for 24 h. The reaction mixture was poured into H2O (200 mL) and extracted with EtOAc (3 x 200 mL). The combined organic phase was washed with brine (200 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 4:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (5.5 g, 66% yield).
[0477] (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-8-yl)boronic acid: To a solution of tert-butyl (2R,5S)-4-(8-chloro-5-(5-fhroropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (500 mg, 1.06 mmol) and 4,4,5,5-tetramethyl- 2-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-l,3,2-dioxaborolane (538 mg, 2.12 mmol) in toluene (20 mL) was added Catacxium A-Pd-G3 (77 mg, 0.10 mmol) and KO Ac (208 mg, 2.12 mmol) at 20 °C under N2. Four parallel batches were combined. The mixture was filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with following condition (column: Phenomenex luna C18 250 x 70 mm x 15μm; mobile phase: A: 10 mM TEA in water, B: MeCN; B% in A: 25%-55%, 20.0 min) to give (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3- yl)quinazolin-8-yl)boronic acid (1 g, 49% yield).
[0478] tert-butyl (2R,5S)-4-(8-(5-cyano-l-(trifluoromethyl)-1H -pyrazol-3-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate and tert-butyl (2R ,5.S)-4-(8-(3-cvano-l- (trifluoromethyl)-1H -pyrazol-5-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)boronic acid (400 mg, 0.59 mmol) in dioxane (5 mL) and H2O (1 mL) was added a mixture of 3-bromo-l-(trifluoromethyl)-1H-pyrazole-5- carbonitrile and 5-bromo-l-(trifluoromethyl)-1H-pyrazole-3-carbonitrile (212 mg, 0.89 mmol), Pd(dppf)Ch (43 mg, 0.06 mmol) and K2CO3 (163 mg, 1.18 mmol) under N2. The mixture was stirred at 90 °C for 2 h. The reaction mixture was poured into H2O (5 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =4:1 to 3:1) to give a mixture of tert-butyl (2R,5S)-4-(8-(5-cyano-l -(trifluoromethyl)- \H- pyrazol-3-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate and tert- butyl (2R,5S)-4-(8-(3-cyano-l -(trifluoromethyl)- 1H-pyrazol-5-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4- yl)-2,5-dimethylpiperazine-l -carboxylate (240 mg, 68% yield).
[0479] 5-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-l- (trifluoromethyl)-l£f-pyrazole-3-carbonitrile and 3-(4-((2.S,5R )-2,5-dimethylpiperazin-l-yl)-5-(5- fluoropyridin-3-yl)quinazolin-8-yl)-l-(trifluoromethyl)-1H -pyrazole-5-carbonitrile: To a mixture of tert-butyl (2R,5S)-4-(8-(5-cyano-l -(trifluoromethyl)- 1H-pyrazol-3-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate and tert-butyl (2R,5S)-4-(8-(3-cyano- 1 - (trifluoromethyl)-1H-pyrazol-5-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l- carboxylate (240 mg, 0.40 mmol) in DCM (3 mL) was added TMSOTf (268 mg, 1.21 mmol) and 2,6- lutidine (216 mg, 2.01 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc =1:1 to 0:1) to give the mixture of 5-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5- fhioropyridin-3-yl)quinazolin-8-yl)-l-(trifluoromethyl)-1H-pyrazole-3-carbonitrile and 3-(4-((2S,5R)- 2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1 -(trifluoromethyl)- 1H-pyrazole-5- carbonitrile (200 mg, 90% yield).
[0480] 5-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-(5-fluoro-3-pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-3-carbonitrile and 3-[4-[(2S,5R)- 4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-(trifluoromethyl)pyrazole-5-carbonitrile: To a solution of 2-cyano-3- fluoro-2-(fluoromethyl)propanoic acid (90 mg, 0.60 mmol) in DCM (1 mL) was added I -chloro-MM2- trimethyl-prop-l-en-1 -amine (108 mg, 0.81 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to a mixture of 5-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5- fhioropyridin-3-yl)quinazolin-8-yl)-l-(trifluoromethyl)-1H-pyrazole-3-carbonitrile and 3-(4-((2S,5R)- 2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1 -(trifluoromethyl)- 1H-pyrazole-5- carbonitrile (200 mg, 0.40 mmol) and DIEA (156 mg, 1.21 mmol) in DCM (1 mL) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into H2O (2 mL) and extracted with DCM (3 x 3 mL). The combined organic phase was washed with brine (3 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep- HPLC with following condition (column: Phenomenex Luna Cl 8 80 x 30 mm x 5 μm; mobile phase: A: 10 mM FA in water, B: MeCN; B% in A: 40%-70%, 8.0 min) to 5-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2- (fluoromethyl)propanoyl] -2,5 -dimethyl-piperazin- 1 -yl] -5 -(5 -fluoro-3 -pyridyl)quinazolin- 8 -yl] - 1 - (trifluoromethyl)pyrazole-3-carbonitrile (42 mg, 16% yield, peak 1 (205)) and 3-[4-[(2S,5R)-4-[2-cyano- 3-fluoro-2-(fluoromethyl)propanoyl] -2,5-dimethyl-piperazin- 1 -yl] -5-(5-fluoro-3-pyridyl)quinazolin-8- yl]-l-(trifhroromethyl)pyrazole-5-carbonitrile (8.5 mg, 3% yield, peak 2 (206)). m/z = 628.2 [M+H]+. [0481] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 210 3-[5-(3-chloro-2-pyridyl)-4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl- piperazin-l-yl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0482] 3-(5-(3-chloropyridin-2-yl)-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-l-methyl-1H -pyrazole-5-carbonitrile: To a solution of 3-(5- bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (150 mg, 0.27 mmol) tributyl-(3-chloro-2- pyridyl) stannane (162 mg, 0.4 mmol) in DMF (2 mL) was added a bis(tri-tert- butylphosphine)palladium(O) (27 mg, 0.053 mmol). The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into water (5 mL) and extracted with EtOAc (3 x 2 mL). The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC with the following condition (column: WePure Biotech XP tC18 150 x 40 x 10 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 50%-80%, 8.0 min) to give 3-(5-(3-chloropyridin-2-yl)-4-((2S,5R)-4-(2-cyano-3-fluoro-2- (fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5- carbonitrile (3 mg, 2% yield), m/z = 590.2 [M+H]+.
[0483] The following compounds were prepared according to the procedures described herein using the appropriate starting material. Example 214 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[2- (trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0484] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-[2-(trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a mixture of 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (100 mg, 179 mmol) and (2- (trifluoromethyl)pyridin-3-yl)boronic acid (103 mg, 0.58 mmol) in dioxane (2 mL) was added K3PO4 (114 mg, 0.54 mmol), Ag2O (13 mg, 0.05 mmol) and Pd(dppf)Cl2-CH2Cl2 (15 mg, 0.02 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (9 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with brine (3 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex luna C18 150 x 40 x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 60%-80%, 8.0 min) to give 3-[4- [(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[2- (trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile (29 mg, 26% yield), m/z = 624.2 [M+H]+.
Example 215 4-chloro-l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]pyrazole-3-carbonitrile
[0485] tert-butyl (2R ,5.S)-4-(8-(4-chloro-3-cyano-1H-pyrazol-l-yl)-5-(5-fhioropvridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of (4-((2S,5R)-4-(tert- butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)boronic acid (300 mg, 0.63 mmol) and 4-chloro-1H-pyrazole-3-carbonitrile (159 mg, 1.25 mmol) in pyridine (5 mL) was added Cu(OAc)2 (226 mg, 1.25 mmol). The mixture was stirred at 80 °C for 16 h under O2. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic phase was washed with brine (5 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexanes: EtOAc = 5:1 to 1:1) to give tert-butyl (2R,5S)-4-(8-(4-chloro-3-cyano-1H-pyrazol-l-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (150 mg, 43% yield).
[0486] 4-chloro-l-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1H-pyrazole-3-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(4-chloro-3-cyano-1H-pyrazol-l- yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (150 mg, 0.27 mmol) in DCM (3 mL) was added 2,6-lutidine (143 mg, 1.33 mmol) and TMSOTf (207 mg, 0.94 mmol) at 0 °C. The mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by prep-TLC (SiO2, DCM:MeOH = 10:1) to give 4-chloro-l-(4- ((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-1H-pyrazole-3- carbonitrile (120 mg, 97% yield).
[0487] 4-chloro- 1 - [4- [(2S,5R) -4- [2-cyano-3-fluoro-2-(fluoromethyl)propanoyl] -2,5-dimethyl- piperazin-l-yl]-5-(5-fluoro-3-pyridyl)quinazolin-8-yl]pyrazole-3-carbonitrile: To a solution of 2- cyano-3-fluoro-2-(fluoromethyl)propanoic acid (65 mg, 0.43 mmol) in DCM (1 mL) was added 1-chloro- N,N,2-trimethyl-prop-l-en-l -amine (61 mg, 0.45 mmol) at 0°C, the mixture was stirred at 0°C for 0.5 h. Then the mixture was added to a solution of 4-chloro-l-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5- fluoropyridin-3-yl)quinazolin-8-yl)-1H-pyrazole-3-carbonitrile (100 mg, 0.22 mmol) and DIEA (140 mg, 1.08 mmol) in DCM (1 mL) at 0°C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into water (5 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 mm x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 57%-77%, 8.0 min) to give 44- chloro- 1 -[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl] -2,5-dimethyl-piperazin- 1 -yl] -5-(5- fluoro-3-pyridyl)quinazolin-8-yl]pyrazole-3-carbonitrile (27 mg, 21% yield), m/z = 624.2 [M+H]+. [0488] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 219A & 219B 3-chloro-l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-[(lR,2R)-2-methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile & 3-chloro-l-[4- [(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[(lS,2S)-2- methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile
[0489] (5-bromo-4-((2S,5R)-4-(tertbutoxycarbonyl)-2,5-dimethylpiperazin-l-yl)quinazolin-8- yl)boronic acid: Solution 1: tert-butyl (2R,5S)-4-(5-bromo-8-iodoquinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (3.5 g, 6.40 mmol) in THF (70 mL). Solution 2: i-PrMgCl.LiCl, (1.3 M in THF, 14.76 mL). Solution 3: trimethyl borate (3.32 g, 31.98 mmol) in THF (70 mL). The solution 1 was pumped by Pump 1 (SI, Pl, 6.848 mL/min) to flow reactor 1 (FLR1, PFA, Dynamic mixer, 1.5 mL, 20 °C ; FLR1, PFA, Coils reactor, 3.175(1/8") mm, 14.96 mL, 20 °C). The solution 2 was pumped by Pump 2 (S2, P2, 1.382 mL/min) to flow reactor 1 (FLR1, PFA, Dynamic mixer, 1.5 mL, 20 °C ; FLR1, PFA, Coils reactor 3 175(1/8") mm 14 96 mL 20 °C) The residence time of flow reactor 1 was 2 min. The mixture was collected with a bottle (Solution 3). The Pump 1 and Pump 2 was started at the same time. The Pump 3 was started after. The reaction mixture was collected after running FLR1, 2 min. Stir for 2 min, quenched by aqueous NH4CI (150 mL). The aqueous phase was extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give (5-bromo-4-((2S,5R)-4-(ter/-butoxycarbonyl)- 2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)boronic acid (5.4 g, 55% purity).
[0490] tert -butyl (2R,5S)-4-(5-bromo-8-(3-chloro-4-cyano-1H -pyrazol-l-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of (5-bromo-4-((2S,5R)-4-(ter/-butoxycarbonyl)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)boronic acid (2 g, 4.30 mmol) and 3-chloro- l H-pyrazolc-4- carbonitrile (1.10 g, 8.60 mmol) in pyridine (20 mL) was added Cu(OAc)2 (1.56 g, 8.60 mmol) at 20 °C under O2. The reaction mixture was stirred at 80 °C under O2 (15 psi) for 12 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give tert-butyl (2R,5S)-4-(5-bromo-8-(3-chloro-4-cyano-1H-pyrazol-l-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (870 mg, 37%).
[0491] l-(5-bromo-4-((2.S,5R )-2,5-dhnethylpiperazin-l-yl )quinazolin-8-yl)-3-chloro-1H -pyrazole-4- carbonitrile: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8-(3-chloro-4-cyano-1H-pyrazol-l- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (800 mg, 1.46 mmol) in DCM (10 mL) was added 2,6-lutidine (784 mg, 7.31 mmol) and TMSOTf (1.14 g, 5.12 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 1:1 to 0:1) to give l-(5-bromo-4- ((25,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-3-chloro-1H-pyrazole-4-carbonitrile (730 mg, 95% yield).
[0492] l-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-3-chloro-1H-pyrazole-4-carbonitrile: To a solution of 2- cyano-3-fluoro-2-(fluoromethyl)propanoic acid (730 mg, 4.90 mmol) in DCM (10 mL) was added 1- chloro-N,N,2-trimethyl-prop-l-en-l -amine (546 mg, 4.09 mmol) at 0 °C under N2, the reaction mixture was stirred at 0 °C for 1 h. The mixture was added a solution of l-(5-bromo-4-((25,5R)-2,5- dimethylpiperazin-l-yl)quinazolin-8-yl)-3-chloro-1H-pyrazole-4-carbonitrile (730 mg, 1.63 mmol) and DIEA (845 mg, 6.54 mmol) in DCM (10 mL) at 0 °C, then the reaction mixture was stirred at 0 °C for 1 h. The reaction mixture was diluted with water (50 mL) and extracted with DCM (3 x 20 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 3:1) to give l-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)- 2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-3-chloro-1H-pyrazole-4-carbonitrile (440 mg, 23% yield). [0493] 3-chloro- 1 - [4- [(2S,5R) -4- [2-cyano-3-fluoro-2-(fluoromethyl)propanoyl] -2,5-dimethyl- piperazin-l-yl]-5-[(lR,2R)-2-methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile & 3- chloro-l-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-[(lS,2S)-2-methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile: To a solution of l-[5- bromo-4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l- yl]quinazolin-8-yl]-3-chloro-pyrazole-4-carbonitrile (100 mg, 0.12 mmol) and Potassium Trifluoro[(lR,2R)-rel-2-methylcyclopropyl]borate (39 mg, 0.24 mmol) in 1,4-dioxane (2 mL) was added K3PO4 (51 mg, 0.24 mmol) and Pd(dppf)Ch (8.8 mg, 0.012 mmol) at 20 °C under N2, then the reaction mixture was stirred at 100 °C under N2 for 16 h. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (FA) with the following conditions: (column: Phenomenex luna Cl 8 100 x 40 mm x 5 μm; mobile phase: A: 0.2% FA in water, B: ACN; B% in A: 45%-75%, 8.0 min) to give 25 mg of product as diastereomeric mixture (trans stereochemistry about cyclopropyl) which was further separated by chiral column [(s,s) WHELK-01 250 x 30 mm x 5 μm; mobile phase: A: CO2 , B: IPA(0.1% NH3H2O; B% in A: 20%-45% over 30 min, socratic elution mode) to give 3-chloro-l-[4-[(2S,5R)-4-[2- cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[2- methylcyclopropyl]quinazolin-8-yl]pyrazole-4-carbonitrile as two separate stereoisomers (9 mg, 21% yield, peak 1 (219A), 8 mg, 21% yield, peak 2 (219B). m/z = 553.2 [M+H]+ and m/z = 553.2 [M+H]+. [0494] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
220B m/z = Example 230 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[5- (trifluoromethyl)-1H -pyrazol-4-yl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0495] l-(tetrahydro-2H -pyran-2-yl )-4-(4,4,5,5-tetramethyl- L3,2-dioxahorolan-2-yl )-5-
(trifluoromethyl)-1H -pyrazole: To a mixture of 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-5- (trifluoromethyl)-1H-pyrazole (500 mg, 1.91 mmol) and 3,4-dihydro-2H-pyran (241 mg, 2.86 mmol) in THF (5 mL) was added TsOH (66 mg, 0.38 mmol) at 20 °C under N2-The mixture was stirred at 60 °C for 2 h. The reaction mixture was poured into H2O (15 mL). The aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (SiO2, Commercial hexanes: EtOAc = 3: 1) to give 1- (tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-5-(trifluoromethyl)-1H- pyrazole (280 mg, 42 % yield).
[0496] 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(l-(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)-1H-pyrazol-4-yl)quinazolin-8-yl)-l-methyl- 1H-pyrazole-5-carbonitrile: To a mixture of 3-(5-bromo-4-((2S,5R)-4-(2-cyano-3-fluoro-2- (fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5- carbonitrile (301 mg, 0.54 mmol) and l-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)-5-(trifluoromethyl)-1H-pyrazole (280 mg, 0.81 mmol) in dioxane (5 mL) was added K3PO4 (343 mg, 1.62 mmol), Ag2O (25 mg, 0.11 mmol) and Pd(dppf)Cl2-CH2Cl2 (44 mg, 0.06 mmol) at 20 °C under N2. The mixture was stirred at 90 °C for 16 h. The mixture was poured into H2O (9 mL) and extracted with EtOAc (3 x 3 mL). The combined organic layers were washed with brine (3 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Commercial hexanes: EtOAc = 1 : 1) to give 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin-l-yl)- 5-(l -(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)- 1 H-pyrazol-4-y l)quinazol in-8-y I)- 1 -methyl- 1H- pyrazole-5-carbonitrile (190 mg, 51% yield). [0497] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-[5-(trifluoromethyl)-1H -pyrazol-4-yl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a mixture of 3-(4-((2S,5R)-4-(2-cyano-3-fluoro-2-(fluoromethyl)propanoyl)-2,5-dimethylpiperazin- 1 -yl)-5- (1 -(tetrahydro-2H-pyran-2-yl)-5-(trifluoromethyl)- 1H-pyrazol-4-yl)quinazolin-8-yl)- 1 -methyl- 1H- pyrazole-5-carbonitrile (190 mg, 0.27 mmol) in dioxane (3 mL) was added 6 N HC1 (3 mL) at 20 °C under N2. The mixture was stirred at 50 °C for 1 h. The reaction mixture was poured into ice saturated aqueous NaHCO3 (10 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: Phenomenex luna C18 150 x 40 x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 50%-70%, 8.0 min) to give 3-[4-[(2S,5R)-4-[2-cyano-3-fhroro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin- l-yl]-5-[5-(trifluoromethyl)-1H-pyrazol-4-yl]quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile (48 mg, 29% yield), m/z = 613.2 [M+H]+.
[0498] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Examples 234A and 234B 3-[4-[(2S,5R)-4-[(2R)-2-cyano-2-cyclopropyl-2-fluoro-acetyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4-[(2S)-2- cyano-2-cyclopropyl-2-fluoro-acetyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0499] ethyl 2-cyano-2-cyclopropyl-2-fluoroacetate: To a solution of ethyl 2-cyano-2- cyclopropylacetate (1 g, 6.53 mmol) in THF (10 mL) was added dropwise LiHMDS (1 M in THF, 9.79 mL) at -78 °C. After addition, the mixture was stirred at -78 °C for 15 min, and then NFSI (3.09 g, 9.79 mmol) in THF (2 mL) was added dropwise at -78 °C. The resulting mixture was stirred at -78 °C for 4 h. The reaction mixture was poured into aqueous NH4CI (20 mL) and extracted with EtOAc (3 x 15 mL). The combined organic layers were washed with brine (15 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Commercial hexanes:EtOAc = 10:1) to give ethyl 2-cyano-2-cyclopropyl-2-fluoroacetate (1 g, 89% yield).
[0500] 2-cyano-2-cyclopropyl-2-fluoroacetic acid: To a solution of ethyl 2-cyano-2-cyclopropyl-2- fluoroacetate (500 mg, 2.92 mmol) in MeCN (5 mL) and H2O (5 mL) was added TBD (1.02 g, 7.30 mmol) at 20 °C. The mixture was stirred at 20 °C for 2 h. The reaction mixture was diluted with water (5 mL) and extracted with MTBE (2 x 2 mL). The water phase was adjusted pH = 5 by IN HC1 and extracted with EtOAc (3 x 2 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 2-cyano-2-cyclopropyl- 2-fluoroacetic acid (500 mg crude).
[0501] 3-[4-[(2S,5R)-4-[(2R)-2-cyano-2-cyclopropyl-2-fluoro-acetyl]-2,5-dimethyl-piperazin-l-yl]- 5-(5-fluoro-3-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile and 3-[4-[(2S,5R)-4-[(2S)- 2-cyano-2-cyclopropyl-2-fluoro-acetyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of 3-(4-((2S,5R)-2,5- dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (100 mg, 0.23 mmol) and 2-cyano-2-cyclopropyl-2-fluoroacetic acid (161 72 mg 0 45 mmol) in THF (5 mL) was added DIEA (146.04 mg, 1.13 mmol) and CMPI (86.61 mg, 0.34 mmol) at 20 °C. The mixture was stirred at 60 °C for 2 h. The reaction mixture was poured into water (10 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were washed with brine (15 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-TLC (SiO2, DCM:MeOH = 10:1) to give 3-(4-((2S,5R)-4-(2-cyano-2-cyclopropyl-2-fluoroacetyl)-2,5- dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5-carbonitrile (100 mg, 77% yield) as white solid. The mixture of diastereomers was isolated by prep-SFC (column: DAICEL CHIRALPAK IG 250 x 30mm x 10μm; mobile phase: A: [CO2-EtOH(0.1%NH3H2O), B: isocratic; B% in A: 50%, elution mode) to give 3-[4-[(2S,5R)-4-[2-cyano-2-cyclopropyl-2-fluoro-acetyl]- 2,5-dimethyl-piperazin- 1 -yl] -5-(5-fluoro-3-pyridyl)quinazolin-8-yl]- 1 -methyl-pyrazole-5-carbonitrile as two separate isomers (30 mg, 23% yield, peak 1 (234A), 33 mg, 26% yield, peak 2 (234B)). m/z = 568.2 [M+H]+ and m/z = 568.2 [M+H]+.
[0502] The following compound was prepared according to the procedures described herein using the appropriate starting material. z =
Example 236 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- methyl-l,3,4-oxadiazol-2-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0503] 2,4,6-trichlorophenyl 4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-8-(5- cyano-l-methyl-1H -pyrazol-3-yl)quinazoline-5-carboxylate: To a solution of tert-butyl (2R,5S)-4-(5- bromo-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (2 g, 3.80 mmol) and 2,4,6-trichlorophenyl formate (1.28 g, 5.70 mmol) in Toluene (20 mL) was added Pd(OAc)2 (85 mg, 0.38 mmol) and Xantphos (440 mg, 0.76mmol) and TEA (1.15 g, 11.40 mmol, 1.59 mL) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was quenched by water (20 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexanes: EtOAc = 3:1) to afford 2,4,6- trichlorophenyl 4-((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin- 1 -yl)-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazoline-5-carboxylate (2.47 g, 76% yield).
[0504] tert-butyl (2R,5S)-4-(5-(2-acetylhydrazine-l-carbonyl)-8-(5-cyano-l-methyl-1H -pyrazol-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of 2,4,6-trichlorophenyl 4- ((2S,5R)-4-(tert-butoxycarbonyl)-2,5-dimethylpiperazin-l-yl)-8-(5-cyano-l-methyl-1H-pyrazol-3- yl)quinazoline-5-carboxylate (2.47 g, 3.68 mmol) and acetohydrazide (2.73 g, 36.81 mmol) in DCM (50 mL) was added TEA (1.12 g, 11.04 mmol, 1.54 mL) and DMAP (450 mg, 3.68 mmol) and HOAt (1.00 g, 7.36 mmol, 1.03 mL) at 20 °C. The mixture was stirred at 50 °C for 16 h. The reaction mixture was quenched by water (150 mL) and extracted with DCM (3 x 50 mL). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc) to afford tert-butyl (2R,5S)-4-(5-(2-acetylhydrazine-l-carbonyl)-8-(5-cyano-l -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2, 5- dimethylpiperazine- 1 -carboxylate (0.52 g, 26%).
[0505] tert-butyl (2R ,5.S)-4-(8-(5-cvano-l-methyl-1H -pyrazol-3-yl)-5-(5-methyl-L3,4-oxadiazol-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-(2- acetylhydr azine- l-carbonyl)-8-(5-cyano-l -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2, 5- dimethylpiperazine-1 -carboxylate (0.52 g, 0.95 mmol) in MeCN (10 mL) was added p-TsCl (543 mg, 2.85 mmol) and DIEA (368 mg, 2.85 mmol) at 20 °C under N2. The mixture was stirred at 20 °C for 2 h. The reaction mixture was quenched by water (30 mL) and extracted with EtOAc (3 x 10 mL). The combined organic phase was washed with brine (10 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc) to afford tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(5-methyl-l,3,4- oxadiazol-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (0.43 g, 85% yield).
[0506] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-methyl-l,3,4-oxadiazol-2-yl)quinazolin-8-yl)- l-methyl- 1H-pyrazole-5-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H- pyrazol-3-yl)-5-(5-methyl-l,3,4-oxadiazol-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (0.43 g, 0.81 mmol) in DCM (5 mL) was added TMSOTf (361 mg, 1.62 mmol) and 2,6-lutidine (261 mg, 2.44 mmol) at 0 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (EtOAc) to afford 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-methyl-l,3,4-oxadiazol-2- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (0.3 g, 86% yield).
[0507] 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l- ylJ-5-(5-methyl-l,3,4-oxadiazol-2-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of intermediate 4B (23 mg, 0.15 mmol) in DCM (0.5 mL) was added l-chloro-N,N,2-trimethyl- prop- 1-en-l -amine (23 mg, 0.17 mmol) at 0 °C under N2. The mixture was stirred at 0 °C for 30 min. Then the mixture was added a solution of 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-methyl-l,3,4- oxadiazol-2-yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (50 mg, 0.12 mmol) and DIEA (45 mg, 0.35 mmol) in DCM (1 mL) at 0 °C. The mixture was stirred at 20 °C for 1 h. The reaction mixture was quenched by water (5 mL) and extracted with DCM (3 x 2 mL).The combined organic phase was washed with brine (2 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC(column: 2_Phenomenex Gemini Cl 8 75 x 40mm x 5 μm; mobile phase: A : 10 mM NH4HCO3 in water, B: CH3CN; B% in A: 35%-60% over 8.0 min) to afford 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- (5-methyl-l,3,4-oxadiazol-2-yl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile (7.1 mg, 10% yield), m/z = 561.2 [M+H]+.
[0508] The following compound was prepared according to the procedures described herein using the appropriate starting material. z =
Example 238 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l-yl]-5- (3,5-difluoro-2-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile
[0509] tert-butyl (2R ,5.S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(3,5-difhioropvridin-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a mixture of tert-butyl (2R,5S)-4-(5- bromo-8-(5-cyano- 1 -methyl- 1H-pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine- 1 -carboxylate (228 mg, 0.43 mmol) and 3,5-difluoro-2-(tributylstannyl)pyridine (350 mg, 0.87 mmol) in DMF (5 mL) was added Bis(tri-tert-butylphosphine)palladium(0) (44 mg, 0.09 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 3 h. The reaction mixture was poured into H2O (15 mL). The aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (5 mL). The combined organic phase was dried over anhydrous Na2SO4 filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Commercial hexanes: EtOAc = 1: 1) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l-methyl-1H-pyrazol-3-yl)-5-(3,5-difluoropyridin-2- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (210 mg, 87% yield).
[0510] 3-(5-(3,5-difluoropyridin-2-yl)-4-((2S,5R)-2,5-dimethylpiperazin-l-yl)quinazolin-8-yl)-l- methyl- 1H-pyrazole-5-carbonitrile: To a mixture of tert-butyl (2R .5.S')-4-(8-(5-cyano- l -mcthyl- l /7- pyrazol-3-yl)-5-(3,5-difluoropyridin-2-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (210 mg, 0.37 mmol) in DCM (3 mL) was added TMSOTf (291 mg, 1.31 mmol) and 2,4-dimethylpyridine (201 mg, 1.87 mmol) at 20 °C under N2-The mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to remove solvent. The residue was purified by silica gel column chromatography (Commercial hexanes: EtOAc = 1:1 to 0: 1) to give 3-(5-(3,5-difluoropyridin-2- yl)-4-((2S,5R)-2,5-dimethylpiperazin- 1 -yl)quinazolin-8-yl)- 1 -methyl- 1H-pyrazole-5 -carbonitrile (170 mg, 98% yield).
[0511] 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl-propanoyl]-2,5-dimethyl-piperazin-l- yl]-5-(3,5-difluoro-2-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5-carbonitrile: To a solution of intermediate 4B (41 mg, 0.28 mmol) in DCM (1 mL) was added l-chloro-N,N,2-trimethyl-prop-l-en-l- amine (44 mg, 0.33 mmol, 0.04 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 0.5 h. Then the mixture was added to a mixture of 3-(5-(3,5-difluoropyridin-2-yl)-4-((2S,5R)-2,5-dimethylpiperazin-l- yl)quinazolin-8-yl)-l -methyl- 1H-pyrazole-5-carbonitrile (85 mg, 0.18 mmol) and DIEA (72 mg, 0.55 mmol, 0.1 mL) in DCM (1 mL) at 0 °C under N2. The mixture was stirred at 0 °C for 1 h. The reaction mixture was poured into H2O (6 mL). The aqueous phase was extracted with DCM (3 x 2 mL). The combined organic layers were washed with brine (2 mL). The combined organic phase was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep- HPLC (column: Phenomenex luna C18 150 x 40 x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 50%-85%, 8.0 min) to give 3-[4-[(2S,5R)-4-[(2R)-2-cyano-3,3-difluoro-2-methyl- propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-(3,5-difluoro-2-pyridyl)quinazolin-8-yl]-l-methyl-pyrazole-5- carbonitrile (39 mg, 35% yield), m/z = 592.3 [M+H]+.
[0512] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
241B m/z = Example 249
3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[4-
(trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile
[0513] 8-chloro-4-methoxy-5-(4-(trifluoromethyl)pyridin-3-yl)quinazoline: To a solution of 5- bromo-8-chloro-4-methoxyquinazoline (1 g, 3.66 mmol), 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-4-(trifluoromethyl)pyridine (1.10 g, 4.02 mmol) in toluene (8 mL) was added Catacxium A-Pd-G2 (244 mg, 0.36 mmol) and K3PO4 (1.55 g, 7.31 mmol) in H2O (2 mL). The mixture was stirred at 100 °C for 4 h. The reaction mixture was poured into water (60 mL) and extracted with EtOAc (3 x 30 mL). The combined organic phases were washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexanes: EtOAc = 1:1) to give 8-chloro-4-methoxy-5-(4-(trifluoromethyl)pyridin-3-yl)quinazoline (3.65 g, 98% yield).
[0514] 8-chloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-4-ol: To a solution of 8-chloro-4- methoxy-5-(4-(trifluoromethyl)pyridin-3-yl)quinazoline (3.65 g, 10.74 mmol) in DMSO (30 mL) was added pyridine ;hydrochloride (6.21 g, 53.72 mmol). The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (50 ml) and extracted with EtOAc (3 x 20 mL). The combined organic phase was washed with brine (20 mL), dried with anhydrous Na2SO4, filtered and was concentrated under reduced pressure. The residue was triturated with EtOAc (10ml) and filtered to give 8-chloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-4-ol (1.8 g, 51% yield).
[0515] 4,8-dichloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazoline: To a solution of 8-chloro-5-(4- (trifluoromethyl)pyridin-3-yl)quinazolin-4-ol (1.6 g, 4.91 mmol) in toluene (20 mL) was added POCl3 (1.58 g, 10.32 mmol) and TEA (646 mg, 6.39 mmol) under N2. The mixture was stirred at 110 °C for 16 h. The reaction mixture was concentrated under reduced pressure. The crude product was triturated with EtOAc (10 ml) and filtered to give 4,8-dichloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazoline (2 g, crude) as white solid which was used into next step without further purification.
[0516] tert-butyl (2R,5S)-4-(8-chloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate: To a solution of 4,8-dichloro-5-(4-(trifluoromethyl)pyridin-3- yl)quinazoline (1.69 g, 4.91 mmol) and tert-butyl (2R ,5.S’)-2,5-dimcthylpipcrazinc- 1 -carboxylate (5.26 g, 24.56 mmol) in dioxane (30 mL) was added DIEA (3.17 g, 24.56 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into water (60 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic phase was washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexanes :EtOAc = 8:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-chloro-5-(4- (trifluoromethyl)pyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (1.1 g, 42% yield). [0517] tert-butyl (2R,5S)-4-(8-(5-cyano-l-(methyl- d3)-1H -pyrazol-3-yl)-5-(4- (trifluoromethyl)pyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(8-chloro-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (700 mg, 1.34 mmol) and 5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-2-(trideuteriomethyl)pyrazole-3-carbonitrile (633 mg, 2.68 mmol) in dioxane (7.5 mL) and H2O (1.5 mL) was added Pd2(dba)3 (122 mg, 0.13 mmol), XPhos (127 mg, 0.26 mmol) and K3PO4 (569 mg, 2.68 mmol), KOAc (263 mg, 2.68 mmol). The mixture was stirred at 100 °C for 2 h. The reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (3 x 10 mL). The combined organic phases were washed with brine (30 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexanes :EtO Ac = 8:1 to 3:1) to give tert-butyl (2R,5S)-4-(8-(5-cyano-l-(methyl-d3)-1H-pyrazol-3-yl)-5-(4-(trifluoromethyl)pyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (750 mg, 93% yield).
[0518] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-8- yl)-l-(methyl-d3)- 1H-pyrazole-5-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l- (methyl-d3)-1H pyrazol-3-yl)-5-(4-(trifluoromethyl)pyridin-3-yl)quinazolin-4-yl)-2,5- dimethylpiperazine-1 -carboxylate (750 mg, 1.26 mmol) in DCM (8 mL) was added TMSOTf (979 mg, 4.41 mmol) and 2,6-lutidine (674 mg, 6.30 mmol). The mixture was stirred at 15 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (MTBE:EtOAc = 1:0 to 0:1) to give 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(4- (trifluoromethyl)pyridin-3-yl)quinazolin-8-yl)-l-(methyl-d3)-1H pyrazole-5-carbonitrile (1 g, crude).
[0519] 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]- 5-[4-(trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile: To a solution of 2-cyano-3-fluoro-2-(fluoromethyl)propanoic acid (48 mg, 0.32 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l-en-l -amine (45 mg, 0.33 mmol) at 0 °C, the mixture was stirred at 0 °C for 0.5 h. Then this mixture and added to 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(4- (trifluoromethyl)pyridin-3-yl)quinazolin-8-yl)- 1 -(methy l- d3)- 1H-pyrazole-5-carbonitrile (160 mg, 0.16 mmol) and DIEA (104 mg, 0.81 mmol) in DCM (2 mL). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into water (2 mL) and extracted with DCM (3 x 1 mL). The combined organic phase was washed with brine (1 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by prep-HPLC (column: WePure Biotech XP tC18 150 x 40 x 5 μm; mobile phase: A: 10 mM NH4HCO3 in H2O, B: ACN; B% in A: 45%-75%, 8 min) to give 3-[4-[(2S,5R)-4-[2-cyano-3-fluoro-2-(fluoromethyl)propanoyl]-2,5-dimethyl-piperazin-l-yl]-5-[4- (trifluoromethyl)-3-pyridyl]quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile (25.51 mg, 25% yield), m/z = 627.2 [M+H]+.
[0520] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 266A & 266B 3-[4-[(2S,5R)-4-[(2S)-2-fluoro-l,l-dioxo-thietane-2-carbonyl]-2,5-dimethyl-piperazin-l-yl]-5-(5- fluoro-3-pyridyl)quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4- [(2R)-2-fluoro-l,l-dioxo-thietane-2-carbonyl]-2,5-dimethyl-piperazin-l-yl]-5-(5-fluoro-3- pyridyl)quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile
[0521] benzyl 2-fluorothietane-2-carboxylate 1,1-dioxide: To a solution of benzyl thietane-2- carboxylate 1,1-dioxide (1 g, 4.16 mmol) in THF (10 mL) was added dropwise LiHMDS (1 M in THF, 4.58 mL) at -78 °C under N2. The mixture was stirred at -78 °C for 15 min. Then NFSI (1.97 g, 6.24 mmol) was added at -78 °C. The mixture was stirred at -78 °C for 3 h. The reaction mixture was poured into aqueous NH4CI (20 mL) and extracted with EtOAc (3 xl5 mL). The combined organic layer were washed with brine (15 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexane:EtOAc = 1:1) to afford benzyl 2- fluorothietane-2-carboxylate 1,1-dioxide (0.62 g, 57%).
[0522] 2-fluorothietane-2-carboxylic acid 1,1-dioxide: To a solution of benzyl 2-fluorothietane-2- carboxylate 1,1-dioxide (0.92 g, 3.56 mmol) in MeCN (9 mL) and H2O (3 mL) was added 3, 4, 6, 7,8,9- hexahydro-2H-pyrimido[l,2-a]pyrimidine (992 mg, 7.12 mmol) at 20 °C. The mixture was stirred at 20 °C for 2 h. The mixture was added water (20 mL) and extracted with MTBE (3 x 5 mL). The aqueous phase was acidified with 3N HC1 to pH = 1 and extracted with EtOAc (3 x 10 mL). The combined organic phase was dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford 2-fluorothietane-2-carboxylic acid 1,1-dioxide (0.54 g, crude).
[0523] methyl 3-bromo- l -(methyl-d3)-1H -pyrazole-S-carboxylate: To a solution of methyl 3-bromo- 1H-pyrazole-5-carboxylate (80 g, 390.23 mmol) in DMF (1000 mL) was added K2CO3 (80.90 g, 585.34 mmol) and trideuterio(iodo)methane (113.13 g, 780.45 mmol). The mixture was stirred at 60 °C for 2 h. The reaction mixture was filtered and poured into H2O (3 L) and extracted with EtOAc (3 x 1 L). The combined organic phase was washed with brine (2 x 1 L), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 20:1 to 10:1) to give methyl 3-bromo-l -(methyl-d3)- 1H-pyrazole-5-carboxylate (74 g, 85 % yield).
[0524] 3-bromo-l-(methyl-d3)- 1H-pyrazole-5-carboxamide: A solution of methyl 3-bromo-l- (methyl-d3)-1H pyrazole-5-carboxylate (74 g, 333.25 mmol) in NH7McOH (7 M in MeOH, 714 mL) was stirred at 80 °C for 16 h in a 2 L of autoclave. The reaction mixture was concentrated under reduced pressure. The residue was triturated with MTBE (200 mL) at 20 °C for 30 min and filtered to give 3- brorno-l-(rnethyl-d3)-1H-pyrazole-5-carboxamide (61 g, 88% yield).
[0525] 3-bromo-l-(methyl-d3)-1H-pyrazole-5-carbonitrile: To a solution of 3-hromo- l -(methyl-d3)- 1H-pyrazole-5-carboxamide (60 g, 289.79 mmol) in DCM (500 mL) was added TFAA (213.03 g, 1.01 mol) and TEA (87.97 g, 869.38 mmol). The mixture was stirred at 20 °C for 2 h. The reaction mixture was poured into H2O (500 mL). The aqueous phase was extracted with DCM (3 x 500 mL). The combined organic phase was washed with brine (500 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 5:1 to 4:1) to give 3-bromo-l -(methyl-d3)- 1H-pyrazole-5-carbonitrile (51 g, 93% yield).
[0526] l-(inethyl-d3)-3-(4,4,5,5-tetrainethyl- 13,2-dioxab(»i(»lan-2-yl )-1H -pyraz(»le-5-carb(»nitrile:
To a mixture of 3-bromo-l -(methyl-d3)- 1H-pyrazole-5-carbonitrile (10 g, 52.90 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(l,3,2-dioxaborolane) (40.30 g, 158.71 mmol) in dioxane (200 mL) was added KO Ac (15.58 g, 158.71 mmol) and Pd(dppf)Ch (3.87 g, 5.29 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 16 h. The reaction mixture was poured into H2O (200 mL) and extracted with EtOAc (3 x 200 mL). The combined organic phase was washed with brine (200 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE:EtOAc = 10:1 to 8:1) to give l -(mcthyl-d;)-3-(4,4,5.5- tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (11.4 g, 91% yield).
[0527] tert-butyl (21f,5S)-4-(5-bromo-8-(5-cyano-l-(methyl-d3)-1H -pyrazol-3-yl)quinazolin-4-yl)- 2,5-dimethylpiperazine-l-carboxylate: To a solution of tert-butyl (2R,5S)-4-(5-bromo-8- iodoquinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (5 g, 9.14 mmol) and l -(mcthyl-d;)-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H pyrazole-5-carbonitrile (4.31 g, 18.27 mmol) in dioxane (100 mL) and H2O (20 mL) was added K3PO4 (4.85 g, 22.84 mmol) and Pd(dppf)Ch (669 mg, 0.91 mmol) at 20 °C under N2. The mixture was stirred at 60 °C for 2 h. The reaction mixture was quenched by water (300 mL) and extracted with EtOAc (3 x 100 mL). The combined organic phase was washed with brine (100 mL), dried with anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexane:EtOAc = 3:1) to afford tert-butyl (2R,5S)-4-(5-bromo-8-(5-cyano-l -(methyl-d3)- 1H-pyrazol-3-yl)quinazolin-4-yl)-2, 5- dimethylpiperazine-1 -carboxylate (2.7 g, 56%).
[0528] tert-butyl (2R,5S)-4-(8-(5-cyano-l -(methyl-d3)- 1H-pyrazol-3-yl)-5-(5-fluoropyridin-3- yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate: To a solution of tert-butyl (2R,5S)-4-(5- bromo-8-(5-cyano-l-(methyl-d3)-1H pyrazol-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l -carboxylate (1.2 g, 2.27 mmol) and (5-fluoropyridin-3-yl)boronic acid (639 mg, 4.53 mmol) in dioxane (10 mL) and H2O (2 mL) was added Ag2O (158 mg, 0.68 mmol), Pd(dppf)Cl2-CH2Cl2 (185 mg, 0.23 mmol) and K3PO4 (1.44 g, 6.80 mmol) at 20 °C under N2. The mixture was stirred at 100 °C for 3 h. The reaction mixture was quenched by water (30 mL), and then extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Hexane:EtOAc = 1:1) to afford tert-butyl (2R, 5S)-4-(8-(5-cyano-l -(methyl-d3)- 1H-pyrazol-3-yl)-5-(5- fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (0.65 g, 52% yield).
[0529] 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-l-
(methyl-d3)-1H -pyrazole-5-carbonitrile: To a solution of tert-butyl (2R,5S)-4-(8-(5-cyano-l-(methyl- d3)-1H-pyrazol-3-yl)-5-(5-fluoropyridin-3-yl)quinazolin-4-yl)-2,5-dimethylpiperazine-l-carboxylate (0.65 g, 1.19 mmol) in DCM (10 mL) was added TMSOTf (530 mg, 2.38 mmol) and 2,6-lutidine (383 mg, 3.57 mmol) at 0 °C under N2. The mixture was stirred at 20 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EtOAc) to afford 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)-l- (methyl-d3)-1H pyrazole-5-carbonitrile (0.39 g, 73%).
[0530] 3-[4-[(2S,5R)-4-[(2S)-2-fluoro-l,l-dioxo-thietane-2-carbonyl]-2,5-dimethyl-piperazin-l-yl]-5- (5-fhioro-3-pyridyl)quinazolin-8-yl]-l-(trideuteriomethyl)pyrazole-5-carbonitrile & 3-[4-[(2S,5R)-4- [(2R)-2-fluoro- 1 , 1 -dioxo-thietane-2-carbonyl] -2,5-dimethyl-piperazin- 1 -yl] -5-(5-fluoro-3- pyridyl)quinazolin-8-yl] - 1 -(trideuteriomethyl)pyrazole-5-carbonitrile [0531] : To a solution of 3-(4-((2S,5R)-2,5-dimethylpiperazin-l-yl)-5-(5-fluoropyridin-3-yl)quinazolin-
8-yl)- 1 -(methyl-d3 - 1 H-pyrazolc-5-carbonitrilc (150 mg, 0.34 mmol) and 2-fluorothietane-2-carboxylic acid 1,1-dioxide (170 mg, 1.01 mmol) in THE (4 mL) was added DIEA (218 mg, 1.68 mmol) and HATU
(192 mg, 0.5 mmol) at 20 °C. The mixture was stirred at 50 °C for 2 h. The reaction mixture was poured into water (10 mL), and extracted with EtOAc (3 x 5 mL). The combined organic phases were washed with brine (5 mL), dried with anhydrous Na2SO4 . filtered and concentrated under reduced pressure. The residue was purified by p-TLC (EtOAc) to afford 3-(4-((25,5R)-4-(2-fluoro-l,l-dioxidothietane-2- carbonyl)-2,5-dimethylpiperazin- 1 -yl)-5-(5-fluoropyridin-3-yl)quinazolin-8-yl)- 1 -(methyl-d3)- 1H- pyrazole-5-carbonitrile (46 mg, 23 % yield) as yellow solid. The mixture of diastereomers was separated by SFC (column: DAICEL CHIRALCEL OD(250mm x 30mm, lOum); mobile phase: [CO2 -
MeOH(0.1% NH3H2O)]; B%: 45%, isocratic elution mode) to afford 3-[4-[(2S,5R)-4-[2-fluoro-l,l- dioxo-thietane-2-carbonyl]-2,5-dimethyl-piperazin- 1 -yl] -5-(5-fluoro-3-pyridyl)quinazolin-8-yl]- 1 -
(trideuteriomethyl)pyrazole-5-carbonitrile as two separate stereoisomers (25 mg, 12% yield, peak 1
(266A), 14 mg, 7% yield, peak 2 (266B)). m/z = 596.2 [M+H]+ & m/z = 596.3 [M+H]+
[0532] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
Example 285
3-(l-((2S,5R)-4-((R)-2-cyano-3,3-difluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-8-(5- fluoropyridin-3-yl)isoquinolin-5-yl)-l-(methyl-d3)-1H -pyrazole-5-carbonitrile
[0533] 8-bromo-5-chloroisoquinoline 2-oxide: To a solution of 8 -bromo-5 -chloroisoquinoline (3 g, 12.37 mmol) in DCM (30 mL) was added m-CPBA (5.02 g, 24.74 mmol, 85% purity) at 0 °C under N2. The mixture was stirred at 25 °C for 1 h. The solid was collected by filtration, washed with hexanes (3 x 10 mL) and aq. NaHCO3 (20 mL) and dried under reduced pressure to give 8-bromo-5- chloroisoquinoline 2-oxide (2.8 g, 88% yield).
[0534] tert-butyl(2R,5S)-4-(8-bromo-5-chloroisoquinolin-l-yl)-2,5-dimethylpiperazine-l -carboxylate: To a solution of 8 -bromo-5 -chloroisoquinoline 2-oxide (3.80 g, 14.70 mmol) and tert-butyl (2R,5S)-2,5- dimethylpiperazine-1 -carboxylate (3.94 g, 18.38 mmol) in THF (25 mL) were added DIEA (7.22 g, 55.86 mmol) and PyBOP (8.91 g, 19.11 mmol). The mixture was stirred at 25 °C for 6 h. The reaction mixture was poured into water (30 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (2 x 10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (hexane: EtOAc = 5:1 to 3:1) to give tert-butyl(2R,5S)-4-(8-bromo-5-chloroisoquinolin-l-yl)-2,5-dimethylpiperazine-l- carboxylate (1.6 g, 24% yield).
[0535] tert-butyl (2R,5S)-4-(5-chloro-8-(5-fluoropyridin-3-yl)isoquinolin-l-yl)-2,5-dimethylpiperazine- 1 -carboxylate: A mixture of tert-butyl(2R,5S)-4-(8-bromo-5-chloroisoquinolin-l-yl)-2,5- dimethylpiperazine-1 -carboxylate (1.4 g, 2.15 mmol), (5-fhroropyridin-3-yl)boronic acid (607mg, 4.31 mmol), K3PO4 (1.37 g, 6.46 mmol), Ag2O (149 mg, 0.66 mmol) and Pd(dppf)C12.CH2C12 (175 mg, 0.22 mmol) in dioxane (10 mL) and H2O (2 mL) was degassed and purged with N2 for 3 times, and then it was stirred at 100 °C under N2 for 3 h. The reaction mixture was poured into water (10 mL) and extracted with EtOAc (3 x 10 mL). The combined organic layers were washed with brine (2 x 8 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel chromatography column (hexane:EtOAc = 4:1 to 2:1) to give tert-butyl (2R,5S)-4- (5-chloro-8-(5-fluoropyridin-3-yl)isoquinolin- 1 -yl)-2,5-dimethylpiperazine- 1 -carboxylate (900 mg, 89% yield). [0536] tert-butyl (2R,5S)-4-(5-(5-cyano-l-(methyl-d3)-1H-pyrazol-3-yl)-8-(5-fluoropyridin-3- yl)isoquinolin-l-yl)-2,5-dimethylpiperazine-l-carboxylate: A mixture of tert-butyl (2R,5S)-4-(5-chloro- 8-(5-fluoropyridin-3-yl)isoquinolin-l-yl)-2,5-dimethylpiperazine-l -carboxylate (200 mg, 0.43 mmol) 1- (methyl-d3)-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-1H-pyrazole-5-carbonitrile (150 mg, 0.64 mmol), K3PO4 (180 mg, 0.85 mmol), KOAc (83 mg, 0.85 mmol), XPhos (40.49 mg, 0.085 mmol) and Pd(dba)2 (24 mg, 0.042 mmol) in dioxane (5 mL) and H2O (0.5 mL) was degassed and purged with N2 for 3 times, and then the mixture was stirred at 100 °C under N2 for 2 h. Combined with another 100 mg batch. The reaction mixture was poured into water (5 mL) and extracted with EtOAc (3 x 8 mL). The combined organic layers were washed with brine (2 x 8 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (hexane: EtOAc = 4:1 to 2:1) to give tert-butyl (2R,5S)-4-(5-(5-cyano-l-(methyl-d3)-1H-pyrazol-3-yl)-8- (5-fluoropyridin-3-yl)isoquinolin-l-yl)-2,5-dimethylpiperazine-l -carboxylate (320 mg, 71% yield).
[0537] 3-(l-((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-(5-fluoropyridin-3-yl)isoquinolin-5-yl)-l-(methyl- d3)-1H-pyrazole-5-carbonitrile: A mixture of tert-butyl (2R,5S)-4-(5-(5-cyano-l-(methyl-d3)-1H- pyrazol-3-yl)-8-(5-fluoropyridin-3-yl)isoquinolin-l-yl)-2,5-dimethylpiperazine-l -carboxylate (320 mg, 0.59 mmol) in DCM (4 mL), 2,6-lutidine (251mg, 2.35 mmol) and TMSOTf (522 mg, 2.35 mmol) at 0 °C and then mixture was stirred at 25 °C under N2 for 1 h. The reaction mixture was poured into water (5 mL) and extracted with DCM (3 x 5 mL). The combined organic layers were washed with brine (2 x 8 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give 3-(l- ((2S,5R)-2,5-dimethylpiperazin-l-yl)-8-(5-fluoropyridin-3-yl)isoquinolin-5-yl)-l-(methyl-d3)-1H- pyrazole-5-carbonitrile (300 mg, 57% yield, 50% purity).
[0538] 3-(l-((2S,5R)-4-((R)-2-cyano-3,3-difluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-8- (5-fhioropyridin-3-yl)isoquinolin-5-yl)-l-(methyl-d3)-1H-pyrazole-5-carbonitrile: To a solution of intermediate 4B (40 mg, 0.27 mmol) in DCM (2 mL) was added l-chloro-N,N,2-trimethyl-prop-l-en-l- amine (40 mg, 0.30 mmol ) at 0 °C under N2. The mixture was stirred at 0 °C for 30 mins. Then the mixture was added to a solution of DIEA (65 mg, 0.51 mmol) and 3-(l-((2S,5R)-2,5-dimethylpiperazin- l-yl)-8-(5-fluoropyridin-3-yl)isoquinolin-5-yl)-l-(methyl-d3)-1H-pyrazole-5-carbonitrile (150 mg, 0.17 mmol) in DCM (2 mL). The mixture was stirred at 20 °C for 1 h. The reaction mixture was poured into water (8 mL) and extracted with EtOAc (3 x 5 mL). The combined organic layers were washed with brine (2 x 5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purification by prep-HPLC (column: CD24-WePure Biotech XPT Cl 8 150 x 25 x 7 μm; mobile phase: A: 10 mM NH4HCO3 in water, B: ACN; B% in A: 39%-69%, 15 min) to give 3-(l- ((2S,5R)-4-((R)-2-cyano-3,3-difluoro-2-methylpropanoyl)-2,5-dimethylpiperazin-l-yl)-8-(5- fluoropyridin-3-yl)isoquinolin-5-yl)-l-(methyl-d3)-1H-pyrazole-5-carbonitrile (48 mg, 49% yield), m/z =
576.2 [M+H]+.
[0539] The following compounds were prepared according to the procedures described herein using the appropriate starting material.
[0540] The following SFC separation method was employed for isolating certain compounds disclosed herein as single isomers: Column: DAICEL CHIRALPAK AD (250 mm x 30 mm x 10 pm): mobile phase: [CO2 -MeOH (0.1%NH3H2O)]; B%: 50%, isocratic elution mode).
BIOLOGICAL EXAMPLE 1
Biochemical Assay of the Compounds hTRPMLl FLIPR Assay
[0541] General Information: TRPML1 is a cation-permeable ion channel localized on the lysosomal membrane. The principal of the assay involves fluorescence measurement of a sensitive calcium dye as an indication of intracellular calcium concentration change in cells expressing mutant TRPML1 localized on the plasma membrane. Cellular calcium responses are measured in the presence or absence of test compounds to identify and characterize modulators of TRPML1 -dependent calcium conductance.
[0542] Cell Culture: HEK293 cells stably expressing HA conjugated human TRPML1 (HA- hTRPMLl) were generated by Lenti- virus infection (with CMV promoter and puromycin selection). Mutations were introduced to TRPML1 sequence (L15A L16A L577A L578A) to enable the plasma membrane expression of TRPML1. Cells were cultured in a humidified and air-controlled (5% CO2 ) incubator at 37 °C. Cells were cultured in DMEM (Thermo Fisher) supplemented with Fetal Bovine Serum (Thermo Fisher, 10% v/v), Penicillin/Streptomycin (Thermo Fisher, 1% v/v) and puromycin
(Thermo Fisher, 4 pg/ml).
[0543] Assay Protocol: The day before the assay, cells were lifted and dissociated using TrypLE Express Enzyme (Gibco) and were seeded at a density of 12k cells/well (30 pF) in poly-D-lysine coated 384-well microplates with black-wall (Greiner) in growth medium. The cell plates were placed in cell culture incubator (5% CO2 at 37 °C) until the next day.
[0544] On the day of the assay, cell plates were centrifuged at 300 rpm for 7s to remove the growth medium. FLIPR Calcium 6 dye (Molecular Devices) were diluted in assay buffer and were added to the cell plates at 20 μL/well. The assay buffer consists of Hank's Balanced Salt Solution (Gibco) supplemented with 1% of probenecid (Thermo Fisher) and 2% of IM HEPES (Gibco). The cell plates were then centrifuge at 300 rpm for 30 seconds and incubated with 5% CO2 at 37 °C for 2 h.
[0545] Compounds were dissolved at 10 mM with DMSO. Compound plates were prepared on the day of experiment at 3X of final concentrations. Compound titration was created using Echo acoustic dispenser (Beckman Coulter) in 384-well polypropylene plates (Greiner). Each compound plate included high controls (20 μM ML-SA5), low controls (1% DMSO) and different doses of up to 16 compounds. Each compound was titrated at 22 concentrations ranging from 300 pM to 143 pM, with final concentrations from 100 pM to 47.7 pM. DMSO concentration is maintained at 1% as final concentrations across the plate.
[0546] The fluorescent signals in each well from cell plates were determined with an excitation wavelength of 480 nm and an emission wavelength of 540 nm using FLIPR Tetra (Molecular Devices). The background fluorescence signals were first recorded for 60 seconds. Then, 10 pL of compound solution was transferred to each well of cell plates from compound plates using a FLIPR pipettor. Solutions were mixed three times using the pipettor. Fluorescence signals were measured for 290 seconds.
[0547] Data Analysis: Background-subtracted maximum signals were calculated as cellular responses. The cellular responses with compound treatment were expressed as % of assay window defined by average of high controls and average of low controls. The %-values were plotted as a function of test compound concentration in GraphPad Prism. The data were described by a 4-parameter logistic function that enabled estimation of each compound’s EC50 (potency).
[0548] Activity of the tested compounds is provided in Table 4 below as follows: A = EC50 < 0.1 pM; B = EC50 0.1 -0.5 pM; C = 0.5 pM < EC50 < 2 pM; D = EC50 > 2 pM.
Table 4
[0549] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
[0550] The disclosure illustratively described herein may suitably be practiced in the absence of any element or elements, limitation, or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claims.
[0551] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference individually. In case of conflict, the present specification, including definitions, will control. [0552] It is to be understood that while the disclosure has been described in conjunction with the above embodiments, that the foregoing description and examples are intended to illustrate and not limit the scope of the disclosure. Other aspects, advantages, and modifications within the scope of the disclosure will be apparent to those skilled in the art to which the disclosure pertains.

Claims

What is claimed is:
1. A compound of Formula I: or a pharmaceutically acceptable salt, isotopically enriched analog, tautomer, stereoisomer, or mixture of stereoisomers thereof, wherein: the dashed bond represents a single or double bond;
X1 is N or CR7;
X2 is N or CR6; and the dashed bond is a double bond; or
X2 is NR2; R1 is oxo; and the dashed bond is a single bond;
X3 is N or CR5;
L is a bond, C1-2 alkylene, C2 alkenylene, C2 alkynylene, -NR11-, -O-, -S-, -C(O)-, -S(O)-, -S(O)2-, -C(O)O-*, -OC(O)-*, -C(O)NR11-*, -NR11C(O)-*, -NR11S(O)-*, -NR11S(O)2-*, -S(O)NR11-*, -S(O)2NR11-*, -NR11C(O)NR11-, -NR11S(O)NR11-, -NR11S(O)2NR11-, -OC(O)NR11-*, or -NR11C(O)O-*; wherein the * bond is attached to Ring A;
Ring A is C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl;
R1 is hydrogen, halo, cyano, nitro, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, -OR11, -SR11, -C(O)R11, -C(O)OR11, -S(O)R11, -S(O)2R11, -C(O)N(R11)2, -NR11C(O)R11, -NR11S(O)R11, -NR11S(O)2R11, -S(O)N(R11)2, -S(O)2N(R11)2, -XR11CfOiN(R11 )2. -NR11S(O)N(R11)2, -NR11S(O)2N(R11)2, -OC(O)N(R11)2, or -NR11C(O)OR11; wherein the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl, is optionally substituted with one to five Z1a;
R2 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl;
R3 is: wherein X is N or CH; each R4 is independently hydroxy, halo, cyano, -CD3, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, -C(O)C, 6 alkyl, -C(O)OC, 6 alkyl, -C(O)NRaRb, -S(O)2C1-6 alkyl, C3-7 cycloalkyl, or 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy;
R5, R6 and R7 are each independently hydrogen, hydroxy, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkylthio, -NRaRb, or C3-7 cycloalkyl;
Ra and Rb are each independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, -(CH2)0-2-C3-7 cycloalkyl, or 3-7 membered heterocyclyl; each R8 is independently C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl, C1-6 haloalkyl, or C3-7 cycloalkyl is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkoxy, or C3-7 cycloalkyl; or two R8 together with the carbon atom(s) attached thereto can be taken together to form oxo, or a fused or spiro C3-7 cycloalkyl, or a fused or spiro 4-7 membered heterocyclyl; wherein the C3-7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted with halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, or C1-6 haloalkoxy;
R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, -O-C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is optionally substituted with one to five Z1a;
R10 is -C(O)OR12, -NR12aC(O)R12, -C(O)N(R12)2, -NR12aC(O)OR12, or heteroaryl optionally substituted with one to five substituents independently selected from halo, hydroxy, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, or C3-6 cycloalkyl; provided that when X is N, then R10 is not -NR12aC(O)R12 or -NR12aC(O)OR12; each R11 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a; each R12 is independently C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein the C1-6 alkyl, C3-7 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1a;
R12a is hydrogen, C1-6 alkyl, or C3-7 cycloalkyl; wherein the C1-6 alkyl or C3-7 cycloalkyl is independently optionally substituted with one to five Z1a; or two R12, R12 and R12a, or R12a and an R8, together with the atom to which each is attached form a heterocyclyl; wherein the heterocyclyl is independently optionally substituted with one to five Z1b; m is 1 or 2; n is 1, 2, or 3; ml is 0, 1, or 2; nl is 0, 1, 2, or 3; m2 is 0, 1 , or 2; n2 is 0, 1, 2, or 3; n3 is 0, 1, or 2; m3 is 0 or 1 ; wherein m + n is 2, 3, or 4; ml + nl is 0, 1, 2, 3, or 4; m2 + n2 is 0, 1, 2, 3, or 4; m3 + n3 is 0, 1, 2, or 3; p is 0, 1, 2, 3, 4, 5, 6, 7, or 8; q is 0, 1, 2, 3, or 4; each Z1a is independently halo, cyano, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R13)2, -OR13, -SR13, -C(O)R13, -C(O)OR13, -S(O)R13, -S(O)2R13, -C(O)N(R13)2, -NR13C(O)R13, -NR13S(O)R13, -NR13S(O)2R13, -S(O)N(R13)2, -S(O)2N(R13)2, -NR13C(O)N(R13)2, -NR13S(O)N(R13)2, -NR13S(O)2N(R13)2, -OC(O)N(R13)2, or -NR13C(O)OR13; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each R13 is independently hydrogen, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl is independently optionally substituted with one to five Z1b; each Z1b is independently halo, cyano, -OH, -SH, -NH2, -NO2, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -L1-C1-6 alkyl, -L1-C2-6 alkenyl, -L1-C2-6 alkynyl, -L1-C1-6 haloalkyl, -L1-C3 -10 cycloalkyl, -L1 -heterocyclyl, -L1-aryl, or -L1 -heteroaryl; and each L1 is independently -O-, -NH-, -S-, -S(O)-, -S(O)2-, -N(C1-6 alkyl)-, -N(C2-6 alkenyl)-, -N(C2-6 alkynyl)-, -N(C1-6 haloalkyl)-, -N(C3-io cycloalkyl)-, -N(heterocyclyl)-, -N(aryl)-, -N(heteroaryl)-, -C(O)-, -C(O)O-, -C(O)NH-, -C(O)N(C1-6 alkyl)-, -C(O)N(C2.6 alkenyl)-, -C(O)N(C2.6 alkynyl)-, -C(O)N(C1-6 haloalkyl)-, -C(0)N(C3 -10 cycloalkyl)-, -C(O)N(heterocyclyl)-, -C(O)N(aryl)-, -C(O)N(heteroaryl)-, -NHC(O)-, -NHC(O)O-, -NHC(O)NH-, -NHS(O)-, or -S(O)2NH-; wherein each C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C3-10 cycloalkyl, heterocyclyl, aryl, and heteroaryl of Z1b and L1 is further independently optionally substituted with one to five halo, cyano, -OH, -SH, -NH2, -NO2, -SF5, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C3-10 cycloalkyl, heterocyclyl, aryl, or heteroaryl.
2. The compound of claim 1 , wherein Ring A is monocyclic C3-7 cycloalkyl, monocyclic heterocyclyl, phenyl, or monocyclic heteroaryl.
3. The compound of claim 1, wherein Ring A is monocyclic C3-7 cycloalkyl, 5- or 6-membered monocyclic heterocyclyl, phenyl, or 5- or 6-membered monocyclic heteroaryl.
4. The compound of claim 1, wherein q is 0, 1, 2, or 3.
5. The compound of claim 1, wherein each R4 is independently selected from halo, cyano, -CD3, C1-6 alkyl, C1-6 haloalkyl, C1-6 alkoxy, -C(O)OC1-6 alkyl, -C(O)N(C1-6 alkyl)2, -S(O)2C1-6 alkyl, and 5-6 membered heteroaryl; wherein each is independently optionally substituted with halo, hydroxy, cyano, C1-6 alkyl, or C1-6 alkoxy.
6. The compound of claim 1 , wherein q is 1 , 2, or 3 ; and at least one R4 is cyano.
7. The compound of claim 1 , wherein L is a bond.
8. The compound of claim 1, wherein L is -C(O)NR11-*.
9. The compound of any preceding claim, wherein R3 is
10. The compound of claim 9, wherein m is 1.
11. The compound of claim 9 or 10, wherein n is 1.
12. The compound of any preceding claim, wherein p is 0, 1 , or 2.
13. The compound of any preceding claim, wherein each R8 is independently C1-6 alkyl.
14. The compound of any preceding claim, wherein p is 1, or 2; and each R8 is methyl.
15. The compound of any preceding claim, wherein X1 is N.
16. The compound of any preceding claim, wherein X2 is N; and the dashed bond is a double bond.
17. The compound of any one of claims 1-15, wherein X2 is CR6; and the dashed bond is a double bond.
18. The compound of claim 17, wherein R6 is hydrogen, hydroxy, halo, or C3-7 cycloalkyl.
19. The compound of any one of claims 1-15, wherein X2 is NR2; R1 is oxo; and the dashed bond is a single bond.
20. The compound of any preceding claim, wherein X3 is N.
21. The compound of any one of claims 1-20, wherein X3 is CR7.
22. The compound of claim 21, wherein R7 is hydrogen.
23. The compound of claim 1, wherein the compound is represented by Formula IIA: or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof.
24. The compound of any preceding claim, wherein R9 is C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl, or heterocyclyl; wherein the C1-6 alkyl, C1-6 alkoxy, C3-7 cycloalkyl or heterocyclyl is independently optionally substituted with one to five substituents independently selected from hydroxy, halo, cyano, C1-6 alkyl, -O-C1-6 alkyl C cycloalkyl or S(O) C alkyl; wherein each C alkyl O C alkyl, C3-7 cycloalkyl, or -S(O)2C1-6 alkyl is independently optionally substituted with one to five independently selected halo.
25. The compound of claim 18, wherein R1 is hydrogen, halo, cyano, C1-6 alkyl, C3-10 cycloalkyl, heterocyclyl, aryl, heteroaryl, -N(R11)2, or -OR11; wherein the C1-6 alkyl or C3-10 cycloalkyl is independently optionally substituted with one to five Z1a.
26. A compound selected from Table 1, or a pharmaceutically acceptable salt thereof.
27. A compound selected from Table 2, or a pharmaceutically acceptable salt thereof.
28. A pharmaceutical composition comprising a compound of any preceding claim, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, and a pharmaceutically acceptable carrier.
29. A method for activating TRPML1, the method comprising contacting a cell with an effective amount of a compound of any one of claims 1-27, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, the pharmaceutical composition of claim 28.
30. The method of claim 29, wherein the contacting is in vivo.
31. A method for treating a disease or disorder mediated, at least in part, by TRPML1, the method comprising administering to a subject in need thereof, an effective amount of a compound of any one of claims 1-20, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, or the pharmaceutical composition of claim 28.
32. The method of claim 31, wherein the disease or disorder is aging, bone diseases, cardiovascular diseases, congenital developmental disorders, eye diseases, hematological and solid malignancies, infectious diseases, inflammatory diseases, liver diseases, metabolic diseases, neurodegenerative or neurological disease or disorder, pancreatitis, renal diseases, skeletal muscle disorders, obesity, lysosomal storage diseases, hypertrophic cardiomyopathy, dilated cardiomyopathy, inclusion body myositis, Paget’s disease, or pulmonary diseases.
33. The method of claim 32, wherein the disease or disorder is a ciliopathy, ciliopathy, neurodegenerative or neurological disease or disorder, lysosomal storage disease or disorder, lysosomal transport disease or disorder, glycogen storage disease or disorder, cholesteryl ester storage disease or disorder, a muscular disease, a disease related to aging, macular degeneration, or cancer.
34. The method of claim 33, wherein the neurodegenerative or neurological disease or disorder is selected from the group consisting of Parkinson’s disease, GBA-Parkinson’s disease, LRRK2 Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis (ALS), Alzheimer’s disease, progressive supranuclear palsy, frontotemporal dementia, FTDP-17, corticobasal degeneration, Lewy body dementia, Pick’s disease, and multi system atrophy.
35. The method of claim 33, wherein the lysosomal storage disease or disorder is selected from the group consisting of Niemann-Pick disease, Gaucher’s disease, neuronopathic Gaucher’s disease, sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, gangliosidoses, Gaucher Disease, Lysosomal acid lipase deficiency, sulfatidoses, mucopolysaccharidoses, mucolipidoses, lipidoses, and oligosaccharidoses. In some embodiments, the lysosomal storage disorder is selected from the group consisting of sphingolipidoses, Farber disease, Krabbe disease, Galactosialidosis, Fabry disease, Schindler disease, beta-galactosidase disorder, GM1 gangliosidosis, GM2 gangliosidosis AB variant, GM2 gangliosidosis activator deficiency, Sandhoff disease, Tay- Sachs disease, Gaucher disease, lysosomal acid lipase deficiency, Niemann-Pick disease, metachromatic leukodystrophy, Saposin B deficiency, multiple sulfatase deficiency, Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome, Hunter syndrome, Sanfilippo syndrome, Morquio syndrome, Maroteaux-Lamy syndrome, Sly syndrome, hyaluronidase deficiency, sialidosis, I-cell disease, pseudo-Hurler polydystrophy, phosphotransferease deficiency, mucolipidin 1 deficiency, Santavuori-Haltia disease, Jansky-Bielchowsky disease, Batten- Spielmeyer- Vogt disease, Kufs disease, Finnish variant neuronal ceroid lipfuscinosis, late infantile variant neuronal ceroid lipfuscinosis, type 7 neuronal ceroid lipfuscinosis, northern epilepsy neuronal ceroid lipfuscinosis, Turkish late infantile neuronal ceroid lipfuscinosis, German/Serbian late infantile neuronal ceroid lipfuscinosis, congenital cathepsin D deficiency, Wolman disease, alpha-mannosidosis, beta-mannosidosis, aspartylgluosaminuria, and fucosidosis. In some embodiments, the lysosomal storage disorder is selected from the group consisting of Niemann-Pick disease, Gaucher’s disease, and neuronopathic Gaucher’s disease.
36. Use of a compound of any one of claims 1-27, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, or the pharmaceutical composition of claim 28, for treating a disease or disorder mediated, at least in part, by TRPML1.
37. A compound of any one of claims 1-27, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, or the pharmaceutical composition of claim 28, for use in therapy.
38. A compound of any one of claims 1-27, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, or the pharmaceutical composition of claim 28, for use in treating a disease or disorder mediated, at least in part, by TRPML1.
39. The use of a compound of any one of claims 1-27, or a pharmaceutically acceptable salt, isotopically enriched analog, stereoisomer, tautomer, or mixture of stereoisomers thereof, or the pharmaceutical composition of claim 28, for the manufacture of a medicament for treating a treating a disease or disorder mediated, at least in part, by TRPML1.
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