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WO2025106722A1 - Inhibitors of stk33, ret, clk, and/or related kinases and methods using same - Google Patents

Inhibitors of stk33, ret, clk, and/or related kinases and methods using same Download PDF

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Publication number
WO2025106722A1
WO2025106722A1 PCT/US2024/055990 US2024055990W WO2025106722A1 WO 2025106722 A1 WO2025106722 A1 WO 2025106722A1 US 2024055990 W US2024055990 W US 2024055990W WO 2025106722 A1 WO2025106722 A1 WO 2025106722A1
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indazol
methanone
biphenyl
ylethynyl
pyrrolidin
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French (fr)
Inventor
Martin Matzuk
Angela KU
Kiran Sharma
Kurt BOHREN
Choel Kim
Srinivas CHAMAKURI
Yong Wang
Feng Li
Mingxing TENG
Chandra MADASU
Ravikumar JIMMIDI
Courtney SUTTON
Zhi Tan
Qiuji YE
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Baylor College of Medicine
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Baylor College of Medicine
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A61K31/416—1,2-Diazoles condensed with carbocyclic ring systems, e.g. indazole
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/54—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings condensed with carbocyclic rings or ring systems
    • C07D231/56—Benzopyrazoles; Hydrogenated benzopyrazoles
    • 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
    • C07D401/02—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 two hetero rings
    • C07D401/12—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 two hetero rings linked by a chain containing hetero atoms as chain links
    • 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
    • C07D403/06—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 linked by a carbon chain containing only aliphatic carbon atoms
    • 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
    • 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
    • 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
    • C07D471/08—Bridged systems
    • 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
    • C07D471/10—Spiro-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
    • C07D487/08—Bridged 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
    • C07D491/08—Bridged 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
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/16—Masculine contraceptives

Definitions

  • Contraception is an important strategy for family planning and may help to curb this continued population explosion and the impact of humans on our environment.
  • birth control pill for women, there have been limited breakthroughs in contraception over the recent decades.
  • knockouts of testis-enriched genes have been created and about 250 of them have revealed a male fertility phenotype
  • few small-molecule inhibitors of these validated targets have been developed to date and even fewer of them have demonstrated a non-hormonal contraceptive effect in pre-clinical models.
  • the present disclosure addresses these needs.
  • the following exemplary' embodiments are provided, which do not limit the scope of the disclosure exemplified herein.
  • the disclosure provides in one aspect a compound of Formula (I): wherein L 1 , X, Y, Z 1 , Z 2 , Z 3 , R a and R b are defined elsewhere herein.
  • the disclosure provides in one aspect a pharmaceutical composition
  • a pharmaceutical composition comprising any compound of the disclosure and a pharmaceutically acceptable carrier.
  • the disclosure provides in one aspect a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the sterilization is temporary.
  • the disclosure provides in one aspect a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the infertility is temporary.
  • the disclosure provides in one aspect a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the disclosure provides in one aspect a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the disclosure provides in one aspect a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity', the method comprising administering the mammal a therapeutically’ effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the disclosure provides in one aspect a method of inhibiting at least one of CL1, CLK2, CLK3, and CLK4 in a mammal, the method comprising administering the mammal an effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • the disclosure provides in one aspect a method of treating, ameliorating, and/or preventing a disease or disorder caused by CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
  • FIG. 1 depicts DEC-Tec selection. Enrichment profile of BCM DNA-encoded chemical library (DECL) qDOS28_l against STK33 at 0.1 pM (x-axis, z-score) versus no target control (y-axis, z-score).
  • a series of hit compounds w ere identified with similar building block 1 (BB1 in blue; attached to the DNA), same building block 2 (BB2 in red), and same building block 3 (BB3 in black). The enrichment of each tri-synthon is shown as sequencing counts/z-score at 0.1 pM in the box.
  • FIGs. 2A-2D depict examples of STK33 hits, analogs, and biological characteristics.
  • FIG. 2A Chemical structure of CDD-2110, the hit with a short linker enriched in the STK33 selection. Blue, red, and black moieties correspond to BB1. BB2. and BB3, respectively.
  • FIG. 2B Chemical structures of CDD-221 1 , CDD-2212, and CDD-2807, the analogs of CDD-2110.
  • FIG. 2C Chemical properties, biochemical activity, cellular activity, and metabolism data of CDD-2110, CDD-2211, CDD-2212, and CDD-2807.
  • Dissociation constant (Ka) and inhibition constant (Ki) values were calculated from LanthaScreen binding assay and Z’-LYTE assay, respectively; half-maximal inhibitory concentration (ICso) values were calculated from NanoBRET assay.
  • Half-life (ti/2) was measured using either mouse liver microsomal (MLM) or human liver microsomal (HLM) stability assay; assay data >60 min is an extrapolated estimate and included for relative determination of half-life.
  • FIG. 2D Summary of CDD-2807 fractional occupancy values for kinases with >30% occupancy (red: 30-49.9%; yellow: 50-79.9%; green: 80-100%) from NanoBRET Target Engagement (TE) KI 92 assay performed at 1 pM.
  • FIGs. 3A-3E depict a crystal structure of a STK33/CDD-2211 complex.
  • FIG. 3A Overall structure of the STK33/CDD-2211 dimer complex. Chain A is shown with surface and chain B in cartoon. N and C termini are labeled. The bound CDD-2211 are shown in sticks with its carbon atoms in yellow, oxygens in red, and nitrogens in blue. The activation loop is colored in red.
  • FIG. 3B 2Fo-Fc density for CDD-2211 in the STK33/CDD-2211 complex contoured at lo.
  • FIG. 3C Electrostatic surface of the active site with CDD-2211.
  • FIG. 3D LIGPLOT diagram showing interactions between STK33 and CDD-2211.
  • FIG. 3E Detailed interaction between STK33 and CDD-2211. Hinge residue backbone atoms that form hydrogen bonds with the ligand are shown as spheres. Key interacting residues shown are shown as sticks.
  • FIGs. 4A-4H depict CDD-2807 treatment induces a reversible contraceptive effect.
  • FIG. 4C Testes from control and CDD-2807 treatment in mice from protocol 2 at 63 days are not significantly different.
  • FIGs. 5A-5B depict DEC-Tec selection and potent hits.
  • FIG. 5A Enrichment profile of BCM DECL qDOS28_l against STK33 at 0.5 pM (x-axis, z-score) versus no target control (y-axis. z-score).
  • FIG. 5B Enrichment profile of BCM DECL qDOS28_l against STK33 at 0.5 pM (x-axis, z-score) versus selection against STK33 at 0.5 pM performed in the presence of staurosporine (y-axis, z-score). Most hits are not observed in the presence of staurosporine, indicating competitive binding.
  • FIG. 6 provides a superimposition of the two chains of the STK33/CDD-2211 dimer complex.
  • a zoomed in panel on the right shows the bound CDD-2211 molecules with residues within 5 A of CDD-2211.
  • Chain A is colored in tan and chain B in magenta.
  • Two chains are nearly identical showing an RMSD of 0.61 A between shared 252 CA atoms.
  • Chains A and B are colored in magenta and tan respectively. Only CA atoms are used in aligning two chains
  • FIG. 7 provides a structure based sequence alignment of STK33, RET, and CLK4.
  • the CDD-2211 contact residues.
  • El 99 and M245, which are unique to STK33 are marked with arrows.
  • FIG. 8 provides a model of CDD-2807 docked to the active site of human STK.33. The same color theme is used as in FIG. 3E.
  • FIG. 9 provides a sequence alignment between human and mouse STK33 proteins. Sequence alignment was performed using the CLUSTAL W program and plotted with the ENDscript program. Identical amino acids as identified by the default ESPript parameters (Risler. global score 0.7) are highlighted with white in red background, similar residues are in red. The secondary structures of human STK33 kinase domain are indicated above the sequence. CDD-2211 contacting residues in the human STK33/CDD-2211 complex are marked with arrows
  • FIG. 10 provides evaluations of CDD-2807 pharmacokinetics in mice.
  • FIG. 11 shows protocol timelines of mouse breeding and reversibility study
  • FIGs. 12A-12D provide data showing no body weight changes of mice treated longterm with CDD-2807.
  • FIGs. 13A-13F provide data showing CDD-2807 crosses the BTB to alter sperm parameters but testis weight in mice.
  • FIG. 13B Despite the contraceptive effect of CDD-
  • FIGs. 14A-14B provide histological analysis of (FIG. 14A) testis and (FIG. 14B) epididymis segments (Caput & Corpus) of control and CDD-2807 -treated male mice from protocol 1.
  • Bouins fixed, 4 pm-longitudinal sections of the testis and epididymis were stained with PAS-hematoxylin. All 12 stages of spermatogenesis are represented in both control and CDD-2807 -treated mice and show normal tissue morphology and acrosome development. Epididymis from the control and CDD-2807-treated mice were similar histologically. Scale bars are included for reference.
  • the present disclosure provides a method of altering cellular signaling, such as kinase signaling, in a subject.
  • the subject is male.
  • the subject is female.
  • the method comprises administering to the subject a therapeutically effective amount of a compound of the disclosure.
  • the compound of the disclosure is orally administered to the subject.
  • the compound of the disclosure is administered to the male subject at a dosage and dosage timing necessary 7 to provide a contraceptive effect in the male subject.
  • provided herein is a method of altering cellular signaling in a subject. In some embodiments, provided herein is a method of altering kinase function in a subject. In some embodiments, provided herein is a method of altering STK signaling in a subject. In some embodiments, provided herein is a method of altering RET signaling in a subject. In some embodiments, provided herein is a method of altering CLK signaling in a subject. In some embodiments, the method comprises administering to the subject an effective amount of a compound of the disclosure, such as but not limited to a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), and/or formula (VII).
  • a compound of the disclosure such as but not limited to a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), and/or formula (VII).
  • a method of treating, ameliorating, and/or preventing a kinase-mediated disease and/or disorder comprising administering a compound of the disclosure, such as but not limited to a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), and/or formula (VII).
  • values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited.
  • a range of ‘'about 0.1% to about 5%” or '‘about 0. 1% to 5%” should be interpreted to include notjust about 0.1% to about 5%. but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range.
  • the acts can be carried out in any order, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.
  • acyl refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom.
  • the carbonyl carbon atom is bonded to a hydrogen forming a “formyl” group or is bonded to another carbon atom, which can be part of an alkyl, aryl, aralkyl cycloalkyl, cycloalkydalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group or the like.
  • An acyl group can include 0 to about 12, 0 to about 20, or 0 to about 40 additional carbon atoms bonded to the carbonyl group.
  • An acyl group can include double or triple bonds within the meaning herein.
  • An acryloyl group is an example of an acyl group.
  • An acyl group can also include heteroatoms within the meaning herein.
  • a nicotinoyl group (pyridyl-3-carbonyl) is an example of an acyl group within the meaning herein.
  • Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like.
  • the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a “haloacyl” group.
  • An example is a trifluoroacetyl group.
  • alkyl refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms.
  • straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups.
  • branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups.
  • alkyl encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl.
  • Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.
  • alkenyl refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms.
  • alkoxy refers to an oxygen atom connected to an alkyd group, including a cycloalkyl group, as are defined herein.
  • linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like.
  • branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopenty loxy, isohexyloxy, and the like.
  • cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.
  • An alkoxy group can include about 1 to about 12, about 1 to about 20. or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms.
  • an allyloxy group or a methoxy ethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.
  • alkynyl refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms.
  • amine refers to primary, secondary, and tertiary amines having, e.g, the formula N(group)? wherein each group can independently be H or non-H, such as alkyl, aryl, and the like.
  • Amines include but are not limited to R-NH 2 , for example, alkylamines, arylamines, alkylarylamines; R 2 NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like: and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like.
  • the term “amine” also includes ammonium ions as used herein.
  • amino group refers to a substituent of the form -NH 2 , - NHR, -NR2, -NR 3 + , wherein each R is independently selected, and protonated forms of each, except for -NR3 . which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine.
  • An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group.
  • alkylamino includes a monoalkylamino, dialkylamino, and trialkylamino group.
  • aminoalkyl refers to amine connected to an alkyl group, as defined herein.
  • the amine group can appear at any suitable position in the alkyd chain, such as at the terminus of the alkyl chain or anywhere within the alkyl chain.
  • aralkyl refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined herein.
  • Representative aralkyl groups include benzyl and phenylethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl.
  • Aralkenyl groups are alkenyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an ary l group as defined herein.
  • aryl refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring.
  • aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenyleny l, pyrenyl, naphthacenyl, chrysenyl. biphenylenyl, anthracenyl, and naphthyl groups.
  • aryl groups contain about 6 to about 14 carbons in the ring portions of the groups.
  • Aryl groups can be unsubstituted or substituted, as defined herein.
  • Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof.
  • C 6-10 - C 6-10 biaryl means a C 6-10 aryl moiety covalently bonded through a single bond to another C6-10 aryl moiety.
  • the C6-10 aryl moiety can be any of the suitable aryl groups described herein.
  • Non-limiting example of a C 6-10 - C 6-10 biaryl include biphenyl and binaphthyl.
  • composition or “pharmaceutical composition” refers to a mixture of at least one compound described herein with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary and topical administration.
  • cycloalkyl refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups.
  • the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7.
  • Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein.
  • Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.
  • cycloalkenyl alone or in combination denotes a cyclic alkenyl group.
  • a “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate.
  • a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's state of health.
  • a disease or disorder is "alleviated” if the severity of a symptom of the disease or disorder, the frequency with which such a symptom is experienced by a patient, or both, is reduced.
  • the terms “effective amount,” “pharmaceutically effective amount” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.
  • halo means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
  • haloalkyl group includes mono-halo alkyl groups, polyhalo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro.
  • haloalkyl include trifluoromethyl, 1,1 -di chloroethyl, 1,2-dichloroethyl, l,3-dibromo-3,3- difluoropropyl, peril uorobutyl. and the like.
  • heteroaryl refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to. N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members.
  • a heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure.
  • a heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth.
  • a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth.
  • Heteroaryl groups include, but are not limited to, groups such as pyrrolyl. pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl. indolyl. azaindolyl.
  • heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed herein. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein.
  • ary l and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1 -naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N- hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3- anthracenyl), thiophenyl (2 -thieny l, 3-thienyl), furyl (2 -furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl.
  • 2,3-dihydro-benzo[b]thiophenyl (2-(2,3- dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro- benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro- benzo[b]thiophenyl). 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl,
  • heteroarylalkyl refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined herein.
  • C6-10-5-6 membered heterobiaryl means a C6-10 aryl moiety covalently bonded through a single bond to a 5- or 6-membered heteroaryl moiety.
  • the C 6-10 aryl moiety and the 5-6-membered heteroaryl moiety can be any of the suitable aryl and heteroaryl groups described herein.
  • Non-limiting examples of a C 6-10 -5-6 membered heterobiaryl include: . When the C6-10-5-6 membered heterobiaryl is listed group), the C 6-10 -5-6 membered heterobiaryl is bonded to the rest of the molecule through the C6-10 moiety.
  • heterocyclyl refers to aromatic and non-aromatic ring compounds containing three or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S.
  • a heterocyclyl can be a cycloheteroalkyl, or a heteroaryl, or if polycyclic, any combination thereof.
  • heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members.
  • a heterocyclyl group designated as a C 2 -heterocyclyl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth.
  • a C 4 -heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth.
  • heterocyclyl group includes fused ring species including those that include fused aromatic and non-aromatic groups.
  • a dioxolanyl ring and a benzdioxolanyl ring system are both heterocyclyl groups within the meaning herein.
  • the phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl.
  • Heterocyclyl groups can be unsubstituted or can be substituted as discussed herein.
  • Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl. triazolyl.
  • oxazolyl isoxazolyl, thiazolyl, pyridinyL thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl.
  • substituted heterocyclyl groups can be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6- substituted, or disubstituted with groups such as those listed herein.
  • heterocyclylalkyl refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group as defined herein is replaced with a bond to a heterocyclyl group as defined herein.
  • Representative heterocyclyl alkyl groups include, but are not limited to, furan-2-yl methyl, furan-3-yl methyl, pyridine-3-yl methyl, tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl.
  • X 1 , X 2 , and X 3 are independently selected from noble gases” would include the scenario where, for example, X 1 , X 2 , and X 3 are all the same, wherein X 1 , X 2 , and X 3 are all different, wherein X 1 and X 2 are the same but X 3 is different, and other analogous permutations.
  • ⁇ refers to a substituent connecting via a single bond to a substituted molecule.
  • a substituent is monovalent, such as, for example, F or Cl, it is bonded to the atom it is substituting by a single bond.
  • organic group refers to any carbon-containing functional group. Examples can include an oxygen-containing group such as an alkoxy group, aryloxy group, aralkyloxy group, oxo(carbonyl) group; a carboxyl group including a carboxylic acid, carboxylate, and a carboxylate ester; a sulfur-containing group such as an alkyl and aryl sulfide group; and other heteroatom-containing groups.
  • Non-limiting examples of organic groups include OR, OOR, OC(O)N(R)2, CN, CF3, OCF3, R, C(O), methylenedioxy, ethylenedioxy, N(R) 2 , SR, SOR, SO2R, SO 2 N(R) 2 , SOsR, C(O)R, C(O)C(O)R, C(O)CH 2 C(O)R, C(S)R, C(O)OR, OC(O)R.
  • patient refers to any animal, or cells thereof whether in vitro or in situ, amenable to the methods described herein.
  • the patient, subject or individual is a human.
  • the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non- toxic, z.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.
  • pharmaceutically acceptable salt refers to a salt of the administered compounds prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids or bases, organic acids or bases, solvates, hydrates, or clathrates thereof.
  • Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid.
  • inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate).
  • Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2- hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic.
  • Suitable pharmaceutically acceptable base addition salts of compounds described herein include, for example, ammonium salts, metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts.
  • Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N’-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.
  • the term '‘pharmaceutically acceptable carrier’ or “pharmaceutically acceptable excipient” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound described herein within or to the patient such that it may perform its intended function. Typically, such compounds are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound(s) described herein, and not injurious to the patient.
  • materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc: excipients, such as cocoa butter and suppository’ waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic cellulose,
  • “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound(s) described herein and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions.
  • the “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound(s) described herein.
  • Other additional ingredients that may be included in the pharmaceutical compositions used with the methods or compounds described herein are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.
  • solvent refers to a liquid that can dissolve a solid, liquid, or gas.
  • Non-limiting examples of solvents are silicones, organic compounds, water, alcohols, ionic liquids, and supercritical fluids.
  • substantially refers to a majority of, or mostly, as in at least about 50%. 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%.
  • substantially free of can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that the composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5. 3, 2.5, 2. 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0. 1, 0.01, or about 0.001 wt% or less.
  • substantially free of can mean having a trivial amount of, such that a composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4. 3.5, 3, 2.5. 2, 1.5, 1, 0.9. 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%.
  • substituted refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms.
  • functional group or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group.
  • substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups.
  • a halogen e.g., F, Cl, Br, and I
  • an oxygen atom in groups such as hydroxy groups, al
  • Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CFs, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R) 2 , SR, SOR, SO2R, SO 2 N(R) 2 , SO3R, C(O)R, C(O)C(O)R, C(O)CH 2 C(O)R, C(S)R, C(O)OR, OC(O)R.
  • a ‘"therapeutic’' treatment is a treatment administered to a subject who exhibits signs of pathology, for the purpose of diminishing or eliminating those signs.
  • thioalkyd refers to a sulfur atom connected to an alky 1 group, as defined herein.
  • the alky l group in the thioalky 1 can be straight chained or branched.
  • linear thioalkyl groups include but are not limited to thiomethyl, thioethyl, thiopropyl, thiobutyl, thiopentyl, thiohexyl, and the like.
  • branched alkoxy include but are not limited to iso-thiopropyl, sec-thiobutyl, tert-thiobutyl. iso-thiopentyl. isothiohexyl, and the like.
  • the sulfur atom can appear at any suitable position in the alkyl chain, such as at the terminus of the alkyl chain or anywhere within the alkyl chain.
  • treat means reducing the frequency or severity with which symptoms of a disease or condition are experienced by a subject by virtue of administering an agent or compound to the subject.
  • range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3. from 1 to 4, from 1 to 5. from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.
  • the present disclosure relates to an inhibitor of STK33.
  • the STK33 inhibitor is selective for STK33 over one or more members of related kinases.
  • the present disclosure relates to an inhibitor of RET.
  • the RET inhibitor is selective for RET over one or more members of related kinases.
  • the present disclosure relates to an inhibitor of CLK.
  • the CLK is CLK1.
  • the CLK is CLK2.
  • the CLK is CLK3.
  • the CLK is CLK4.
  • the CLK inhibitor is selective for CLK over one or more members of related kinases.
  • the selectivity is such that the pICso of the compound for the specific kinase (STK33, RET, and/or one or more CLK’s) is about 0.1, 0.2, 0,3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8.1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, or higher than the pIC50 of the compound for the other kinase in question.
  • the compound of formula (I) is the compound of formula (Ia): a).
  • X is -C(CH3)2-.
  • X is -CF2-.
  • X is -CHF-.
  • X is - CH(CH 3 )-.
  • X is -CH 2 -.
  • Z 1 , Z 2 , and Z 3 are CH.
  • Z 1 and Z 2 are CH, and Z 3 is N.
  • Z 1 and Z 3 are CH, and Z 2 is N.
  • Z 2 and Z 3 are CH, and Z 1 is N.
  • Y is optionally substituted phenyl. In some embodiments, Y is optionally substituted naphthyl. In some embodiments, Y is optionally substituted isoquinolinyl. In some embodiments, Y is optionally substituted quinolinyl. In some embodiments, Y is optionally substituted 2-pyridyl. In some embodiments, Y is optionally substituted 3-pyridyl. In some embodiments, Y is optionally substituted 4-pyridyl. In some embodiments, Y is optionally substituted pyrimidinyl. In some embodiments, Y is optionally substituted pyrazinyl. In some embodiments, Y is optionally substituted triazinyl.
  • R e1 and R e2 are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally substituted 3-to-8 membered heterocyclyl, or optionally substituted C3-C10 heteroaryl;
  • one of R a or R b is hydrogen or methyl.
  • R a and R b combine with the nitrogen atom to which they are bound to form optionally substituted 5-, 6-, or-7 membered heterocyclyl.
  • R a and R b combine with the nitrogen atom to which they are bound to form optionally substituted pyrrolidinyl.
  • NR a R b is . In some embodiments, . In some embodiments, NR a R b is . In some embodiments, NR a R b is . In some embodiments, NR a R b is . In some embodiments, NR a R b is . In some embodiments, NR a R b is i . In some embodiments, NR a R b is . In
  • NR a R b is —• NH . In some embodiments, NR a R b is ,
  • the group is selected from the group consisting of:
  • the compound of Formula (I) is a compound of Formula (II). In some embodiments, the compound of Formula (I) is a compound of Formula (III). In some embodiments, the compound of Formula (I) is a compound of Formula (IV). In some embodiments, the compound of Formula (I) is a compound of Formula (V). In some embodiments, the compound of Formula (I) is a compound of Formula (VI). In some embodiments, the compound of Formula (I) is a compound of Formula (VII).
  • provided herein is a compound selected from the group consisting of: (S)-3-([l,r-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-l-phenylethyl)-lH-indazole- 5-carboxamide,
  • Table A Compound reference code Molecular structure - 65 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2134 - 66 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2530 - 67 - 53366970.3 Attorney Docket.
  • compounds described herein are present in optically active or racemic forms. It 5 is to be understood that the compounds described herein encompass racemic, optically-active, regioisomeric and stereoisomeric forms, or combinations thereof that possess the therapeutically useful properties described herein. Preparation of optically active forms is achieved in any suitable manner, including by way of non-limiting example, by resolution of the racemic form with recrystallization techniques, synthesis from optically-active starting 10 materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. In certain embodiments, a mixture of one or more isomer is utilized as the therapeutic compound described herein. In other embodiments, compounds described herein contain one or more chiral centers.
  • These compounds are prepared by any means, including stereoselective synthesis, enantioselective synthesis and/or separation of a mixture of 15 enantiomers and/ or diastereomers. Resolution of compounds and isomers thereof is achieved by any means including, by way of non-limiting example, chemical processes, enzymatic processes, fractional crystallization, distillation, and chromatography.
  • the methods and formulations described herein include the use of N-oxides (if appropriate), crystalline forms (also known as polymorphs), solvates, amorphous phases, 20 and/or pharmaceutically acceptable salts of compounds having the structure of any compound(s) described herein, as well as metabolites and active metabolites of these compounds having the same type of activity.
  • Solvates include water, ether (e.g., tetrahydrofuran, methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetates and the like.
  • the compounds described herein exist in solvated forms with 25 pharmaceutically acceptable solvents such as water, and ethanol.
  • the compounds described herein exist in unsolvated form.
  • the compound(s) described herein can exist as tautomers. All tautomers are included within the scope of the compounds presented herein. - 97 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159)
  • compounds described herein are prepared as prodrugs.
  • a “prodrug” refers to an agent that is converted into the parent drug in vivo.
  • a prodrug upon in vivo administration, is chemically converted to the biologically, pharmaceutically or therapeutically active form of the compound.
  • a prodrug is enzymatically metabolized by one or more steps or processes to the biologically, pharmaceutically or therapeutically active form of the compound.
  • sites on, for example, the aromatic ring portion of compound(s) described herein are susceptible to various metabolic reactions. Incorporation of appropriate substituents on the aromatic ring structures may reduce, minimize or eliminate 10 this metabolic pathway.
  • the appropriate substituent to decrease or eliminate the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a deuterium, a halogen, or an alkyl group.
  • Compounds described herein also include isotopically-labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic 15 mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes suitable for inclusion in the compounds described herein include and are not limited to 2 H, 3 H, 11 C, 13 C, 14 C, 36 Cl, 18 F, 123 I, 125 I, 13 N, 15 N, 15 O, 17 O, 18 O, 32 P, and 35 S.
  • isotopically-labeled compounds are useful in drug and/or substrate tissue distribution studies.
  • substitution with heavier isotopes such as 20 deuterium affords greater metabolic stability (for example, increased in vivo half-life or reduced dosage requirements).
  • substitution with positron emitting isotopes such as 11 C, 18 F, 15 O, and 13 N, is useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy.
  • Isotopically-labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically-labeled 25 reagent in place of the non-labeled reagent otherwise employed.
  • the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.
  • the compounds described herein, and other related compounds having different 30 substituents are synthesized using techniques and materials described herein and as described, for example, in Fieser & Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., - 98 - 53366970.3 Attorney Docket.
  • Groups such as trityl, dimethoxytrityl, acetal and t-butyldimethylsilyl are acid labile and are used to 20 protect carboxy and hydroxy reactive moieties in the presence of amino groups protected with Cbz groups, which are removable by hydrogenolysis, and Fmoc groups, which are base labile.
  • Carboxylic acid and hydroxy reactive moieties are blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl, in the presence of amines that are blocked with acid labile groups, such as t-butyl carbamate, or with carbamates that are both 25 acid and base stable but hydrolytically removable.
  • carboxylic acid and hydroxy reactive moieties are blocked with hydrolytically removable protective groups such as the benzyl group, while amine groups capable of hydrogen bonding with acids are blocked with base labile groups such as Fmoc.
  • Carboxylic acid reactive moieties are protected by conversion to simple ester 30 compounds as exemplified herein, which include conversion to alkyl esters, or are blocked with oxidatively-removable protective groups such as 2,4-dimethoxybenzyl, while co- existing amino groups are blocked with fluoride labile silyl carbamates. Allyl blocking groups are useful in the presence of acid- and base- protecting groups since the former are stable and are subsequently removed by metal or pi-acid catalysts.
  • an allyl-blocked carboxylic acid is deprotected with a palladium-catalyzed reaction in the presence of acid labile t-butyl carbamate or base-labile acetate amine protecting groups.
  • Yet another form of protecting group is a resin to which a compound or intermediate is attached. As long as the residue is attached to the resin, that functional group is blocked 5 and does not react. Once released from the resin, the functional group is available to react.
  • blocking/protecting groups may be selected from: .
  • compositions 15 the present disclosure provides a pharmaceutical composition comprising at least one compound of the present disclosure and at least one pharmaceutically acceptable carrier.
  • the compositions containing the compound(s) described herein include a pharmaceutical composition comprising at least one compound as described herein and at 20 least one pharmaceutically acceptable carrier.
  • the pharmaceutical composition comprises Kolliphor EL, and aqueous buffer, or a combination thereof.
  • the aqueous buffer comprises phosphate buffered saline (PBS). In some embodiments, the aqueous buffer comprises 1x PBS. In certain embodiments, the - 100 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) pharmaceutical composition comprises about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, or about 40% Kolliphor EL. In some embodiments, the pharmaceutical composition comprises about 20% Kolliphor EL in 1x PBS.
  • the composition is formulated for an administration route 5 such as oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal, intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration.
  • the composition is formulated 10 as a pill, tablet, gelcap, or capsule for oral administration.
  • the present disclosure relates to a method of promoting male contraception in a male subject.
  • the present disclosure relates to a method 15 of promoting male sterility in a male subject.
  • the present disclosure relates to a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject.
  • the present disclosure relates to a method of reducing testes size in a male subject.
  • the present disclosure relates to a method of modulating a RET, STK33, and/or CLK kinase function and/or activity in a subject.
  • the method comprises administering to the subject a therapeutically effective amount of a compound of the disclosure. In certain embodiments, the method comprises administering to the male subject a therapeutically effective amount of a compound of the disclosure. In certain embodiments, the method comprises administering to the subject a 25 therapeutically effective amount of a compound of Formula (I). In certain embodiments, the method comprises administering to the male subject a therapeutically effective amount of a compound of Table A. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound 30 disclosed herein and/or the pharmaceutical composition thereof.
  • provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof.
  • a method of inhibiting STK33 in a mammal the method comprising administering the mammal an - 101 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) effective amount of a compound of Formula (II) and/or the pharmaceutical composition thereof.
  • provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some 5 embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (V) and/or the pharmaceutical composition 10 thereof.
  • provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VII) 15 and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound disclosed herein, and/or a pharmaceutical composition thereof.
  • provided herein is a method of sterilizing a male subject, the method 20 comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (II), and/or a pharmaceutical composition thereof. In some 25 embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III), and/or a pharmaceutical composition thereof.
  • provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 30 compound of Formula (IV), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (V), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method - 102 - 53366970.3 Attorney Docket.
  • No.046641-7059WO1(00159) comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI), and/or a pharmaceutical composition thereof.
  • a method of sterilizing a male subject the method comprising administering to the male subject a therapeutically effective amount of a 5 compound of Formula (VII), and/or a pharmaceutical composition thereof.
  • a method of promoting male contraception and/or infertility in a male subject the method comprising administering to the male subject a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof.
  • provided herein is a method of 10 promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof.
  • a method of promoting male contraception and/or infertility in a male subject the method comprising administering to the male subject a therapeutically effective amount of a 15 compound of Formula (II) and/or the pharmaceutical composition thereof.
  • provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof.
  • a method of promoting male contraception 20 and/or infertility in a male subject the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof.
  • provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (V) and/or the 25 pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof.
  • provided herein is a method of promoting male contraception and/or infertility in a male 30 subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VII) and/or the pharmaceutical composition thereof. In some embodiments, the method provides a contraceptive effect in the male subject. In some embodiments, the compound is administered orally to the male subject. In some embodiments, provided herein is a method of minimizing and/or reducing - 103 - 53366970.3 Attorney Docket.
  • No.046641-7059WO1(00159) spermatozoa number and/or motility in a male subject comprising administering to the male subject a therapeutically effective amount of a compound disclosed herein and/or the pharmaceutical composition thereof.
  • a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof.
  • provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (II) and/or the 10 pharmaceutical composition thereof.
  • a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof.
  • provided herein is a method of minimizing and/or reducing spermatozoa 15 number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 20 compound of Formula (V) and/or the pharmaceutical composition thereof.
  • provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof.
  • a method of 25 minimizing and/or reducing spermatozoa number and/or motility in a male subject the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VII) and/or the pharmaceutical composition thereof.
  • the method provides a contraceptive effect in the male subject.
  • the compound is administered orally to the male subject.
  • provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (I) and/or the - 104 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (II) and/or the pharmaceutical composition thereof.
  • provided herein is a method of inhibiting RET in a mammal, the method 5 comprising administering the mammal an effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, 10 the method comprising administering the mammal an effective amount of a compound of Formula (V) and/or the pharmaceutical composition thereof.
  • RET inhibitors can be used to treat, ameliorate, and/or prevent cancer, including but limited to medullary thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A).
  • MEN2A Multiple Endocrine Neoplasia Type IIa
  • a method of 20 treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression and/or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof.
  • the method comprises treating a RET mediated cancer.
  • the RET mediated cancer comprises medullary 25 thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A).
  • the disease or disorder comprises Hirschprung Disease, central hypoventilation syndrome, or renal agenesis.
  • provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the 30 method comprising administering the mammal a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof.
  • a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity the method comprising administering the mammal a therapeutically effective amount of a compound of - 105 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Formula (II) and/or the pharmaceutical composition thereof.
  • provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (III) and/or the 5 pharmaceutical composition thereof.
  • a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof.
  • provided herein is a method of treating, ameliorating, and/or 10 preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (V) and/or the pharmaceutical composition thereof.
  • a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity the method 15 comprising administering the mammal a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof.
  • a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity comprising administering the mammal a therapeutically effective amount of a compound of Formula (VII) and/or the 20 pharmaceutical composition thereof.
  • CLKs family comprises four homologous proteins, CLK1, CLK2, CLK3, and CLK4. They contribute to cell cycle progression, cell growth and disease occurrence via the regulation of splicing.
  • human CLK2 may play a role in fatty liver disease through participating in fatty acid oxidation and ketogenesis.
  • a method of treating, ameliorating, and/or preventing a disease or disorder caused by - 106 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CLK overexpression or CLK hyperactivity comprising administering the mammal a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof.
  • the disease or disorder comprises Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and 5 renal cancer, breast cancer, or a melanoma.
  • the compound of formula (I) is a compound of formula (II). Exemplary compounds of formula (II) are described elsewhere herein.
  • the compound of formula (I) is a compound of formula (III). Exemplary compounds of formula (III) are described elsewhere herein.
  • the compound of 10 formula (I) is a compound of formula (IV). Exemplary compounds of formula (IV) are described elsewhere herein.
  • the compound of formula (I) is a compound of formula (V). Exemplary compounds of formula (V) are described elsewhere herein.
  • the compound of formula (I) is a compound of formula (VI). Exemplary compounds of formula (VI) are described elsewhere herein.
  • the compound of formula (I) is a compound of formula (VII). Exemplary compounds of formula (VII) are described elsewhere herein.
  • the compound of formula (I) is a compound of formula (VII). Exemplary compounds of formula (VII) are described elsewhere herein.
  • the methods described herein include administering to the subject a therapeutically effective amount of at least one compound described herein, which is optionally formulated in a pharmaceutical composition.
  • a therapeutically effective amount 20 of at least one compound described herein present in a pharmaceutical composition is the only therapeutically active compound in a pharmaceutical composition.
  • the method further comprises administering to the subject an additional agent that treats, ameliorates, and/or prevents the contemplated condition.
  • the compound of the disclosure can be administered to the subject using any 25 administration route known to a person of skill in the art. Exemplary routes of administration are described elsewhere herein.
  • a composition comprising a compound of the disclosure is orally administered to the subject.
  • a pill, tablet, gelcap, or capsule comprising a compound of the disclosure is orally administered to the subject.
  • the compound of the disclosure can be administered to the subject in any dosage with any timing of dosage administration necessary to inhibit STK, or a combination thereof and to provide a desired therapeutic effect.
  • the compound of the disclosure is administered to the male subject in order to provide a contraceptive effect in the subject.
  • the subject is a mammal. In other embodiments, the mammal is a human. 5 Administration/Dosage/Formulations
  • the regimen of administration may affect what constitutes an effective amount.
  • the therapeutic formulations may be administered to the subject either prior to or after the onset of the disease or disorder. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a 10 bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.
  • compositions described herein to a patient may be carried out using known procedures, at dosages 15 and for periods of time effective to treat the disease or disorder in the patient.
  • An effective amount of the therapeutic compound necessary to achieve a therapeutic effect may vary according to factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat the disease or disorder in the patient.
  • Dosage regimens may be adjusted to provide the optimum therapeutic 20 response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
  • a non- limiting example of an effective dose range for a therapeutic compound described herein is from about 1 and 5,000 mg/kg of body weight/per day.
  • One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective 25 amount of the therapeutic compound without undue experimentation.
  • Actual dosage levels of the active ingredients in the pharmaceutical compositions described herein may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the selected dosage level depends upon a variety of factors including the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in - 108 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) the medical arts.
  • a medical doctor e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required.
  • Dosage unit form as 10 used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle.
  • compositions described herein are formulated using one or more pharmaceutically acceptable excipients or carriers.
  • the pharmaceutical compositions described herein comprise a therapeutically effective amount of 20 a compound described herein and a pharmaceutically acceptable carrier.
  • the carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, C3-C7 aliphatic or aryl n-hydroxyl compound, wherein n is 1 to 10, e.g.
  • glycerol propylene glycol, and liquid polyethylene glycol, and the like
  • suitable mixtures thereof and vegetable oils.
  • the proper fluidity may be maintained, for example, by 25 the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
  • isotonic agents for example, sugars, sodium chloride, or polyalcohols such as 30 mannitol and sorbitol, in the composition.
  • Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.
  • the compositions described herein are administered to the patient in dosages that range from one to five times per day or more.
  • - 109 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) the compositions described herein are administered to the patient in range of dosages that include, but are not limited to, once every day, every two, days, every three days to once a week, and once every two weeks.
  • a composition as described herein is a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound described herein, alone or in combination with a second pharmaceutical agent; and instructions for using the compound to treat, or reduce one or more 15 symptoms of a disease or disorder in a patient.
  • Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art.
  • the pharmaceutical preparations may be sterilized and if 20 desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic agents.
  • Routes of administration of any of the compositions described herein include oral, 25 nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical.
  • the compounds for use in the compositions described herein can be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, 30 subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration.
  • compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, - 110 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions described herein are not limited to the particular formulations and compositions that are 5 described herein.
  • compositions intended for oral use may be prepared according to any method known in the art and such compositions may contain 10 one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients that are suitable for the manufacture of tablets.
  • excipients include, for example an inert diluent such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate.
  • the tablets may be uncoated or they may be coated by known techniques for elegance or to 15 delay the release of the active ingredients.
  • Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert diluent.
  • the compound(s) described herein can be in the form of tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., polyvinylpyrrolidone, hydroxypropylcellulose or 20 hydroxypropyl methylcellulose); fillers (e.g., cornstarch, lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrates (e.g., sodium starch glycollate); or wetting agents (e.g., sodium lauryl sulphate).
  • binding agents e.g., polyvinylpyrrolidone, hydroxypropylcellulose or 20 hydroxypropyl methylcellulose
  • fillers e.g., cornstarch, lactose, microcrystalline cellulose or calcium phosphate
  • the tablets may be coated using suitable methods and coating materials such as OPADRYTM film coating systems available from Colorcon, West Point, Pa. (e.g., OPADRYTM OY Type, OYC 25 Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and OPADRYTM White, 32K18400).
  • OPADRYTM film coating systems available from Colorcon, West Point, Pa. (e.g., OPADRYTM OY Type, OYC 25 Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and OPADRYTM White, 32K18400).
  • Liquid preparation for oral administration may be in the form of solutions, syrups or suspensions.
  • the liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agent (e.g., 30 lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbic acid).
  • suspending agents e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats
  • emulsifying agent e.g., 30 lecithin or acacia
  • non-aqueous vehicles e.g., almond oil, oily esters or ethyl alcohol
  • preservatives e.g., methyl or propyl p-hydroxy benzoates or sorbic acid.
  • parenteral Administration the compounds as described herein may be formulated for injection
  • No.046641-7059WO1(00159) infusion or for administration in a bolus dose and/or continuous infusion.
  • Suspensions, solutions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing and/or dispersing agents may be used.
  • Sterile injectable forms of the compositions described herein may be aqueous or 5 oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol.
  • a non-toxic parenterally-acceptable diluent or solvent for example as a solution in 1, 3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer’s 10 solution and isotonic sodium chloride solution.
  • Sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or di-glycerides.
  • Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their 15 polyoxyethylated versions.
  • Additional dosage forms suitable for use with the compound(s) and compositions described herein include dosage forms as described in U.S. Patents Nos.6,340,475; 20 6,488,962; 6,451,808; 5,972,389; 5,582,837; and 5,007,790. Additional dosage forms suitable for use with the compound(s) and compositions described herein also include dosage forms as described in U.S. Patent Applications Nos.20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and 20020051820.
  • Additional dosage forms suitable for use with the compound(s) and compositions described herein also include dosage forms as 25 described in PCT Applications Nos. WO 03/35041; WO 03/35040; WO 03/35029; WO 03/35177; WO 03/35039; WO 02/96404; WO 02/32416; WO 01/97783; WO 01/56544; WO 01/32217; WO 98/55107; WO 98/11879; WO 97/47285; WO 93/18755; and WO 90/11757.
  • Dosing 30 The therapeutically effective amount or dose of a compound described herein depends on the age, sex and weight of the patient, the current medical condition of the patient and the progression of the disease or disorder in the patient being treated. The skilled artisan is able to determine appropriate dosages depending on these and other factors.
  • a suitable dose of a compound described herein can be in the range of from about - 112 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1,000 mg, for example, from about 1 mg to about 500 mg, such as about 5 mg to about 250 mg per day.
  • the dose may be administered in a single dosage or in multiple dosages, for example from 1 to 4 or more times per day.
  • the amount of each dosage may be 5 the same or different.
  • a dose of 1 mg per day may be administered as two 0.5 mg doses, with about a 12-hour interval between doses.
  • the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days.
  • a 5 mg per day dose may 10 be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on.
  • the administration of the compound(s) described herein is optionally given continuously; alternatively, the dose of drug being administered is temporarily reduced or temporarily 15 suspended for a certain length of time (i.e., a “drug holiday”).
  • the length of the drug holiday optionally varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days.
  • the dose reduction during a drug holiday includes 20 from 10%-100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
  • a maintenance dose is administered if necessary.
  • the dosage or the frequency of administration, or both is reduced to a level at which the improved disease is retained.
  • 25 patients require intermittent treatment on a long-term basis upon any recurrence of symptoms and/or infection.
  • the compounds described herein can be formulated in unit dosage form.
  • unit dosage form refers to physically discrete units suitable as unitary dosage for patients undergoing treatment, with each unit containing a predetermined quantity of active material 30 calculated to produce the desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier.
  • the unit dosage form may be for a single daily dose or one of multiple daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose. Toxicity and therapeutic efficacy of such therapeutic regimens are optionally - 113 - 53366970.3 Attorney Docket.
  • No.046641-7059WO1(00159) determined in cell cultures or experimental animals including, but not limited to, the determination of the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population).
  • the dose ratio between the toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD 50 and 5 ED50.
  • the data obtained from cell culture assays and animal studies are optionally used in formulating a range of dosage for use in human.
  • the dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity.
  • the dosage optionally varies within this range depending upon the dosage form employed and the route of administration utilized.
  • reaction conditions including but not limited to reaction 15 times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, are within the scope of the present application.
  • NMR spectra were recorded at room temperature using a Bruker Avance III HD 600 MHz 15 spectrometer ( 1 H NMR at 600 MHz and 13 C NMR at 150 MHz) or Bruker Avance 800 MHz spectrometer ( 1 H NMR at 800 MHz and 13 C NMR at 200 MHz). Chemical shifts ( ⁇ ) are reported in parts per million (ppm) with reference to solvent signals [ 1 H-NMR: CDCl3 (7.26 ppm), CD3OD (3.31 ppm), and DMSO-d6 (2.50 ppm); 13 C-NMR: CDCl3 (77.00 ppm), CD 3 OD (49.15 ppm), and DMSO-d 6 (39.51 ppm)].
  • the mixture was vacuum-purged and refilled with nitrogen for three cycles and then heated by microwave reactor at 80 o C for 1 h (or at indicated temperature for indicated period of time). After completion of the reaction (monitored by TLC and LC-MS), the reaction mixture was filtered through a pad of celite, and the filtrate was extracted between EtOAc and 1N HCl aqueous solution. The organic layer was washed with saturated 10 aqueous NaHCO 3 and brine, dried over anhydrous Na 2 SO 4 , filtered, and concentred under reduced pressure. The residue was purified by flash chromatography on silica with EtOAc/hexanes or CH 3 OH/CH 2 Cl 2 gradient elution to afford the desired product.
  • IC 50 values and corresponding K d values of biochemical activity against STK33 were calculated from Lanthascreen binding assay, and IC50 values of cellular activity against STK33 were calculated from NanoBRET Target Engagement intracellular 15 kinase assay.
  • ND means not determined either for compounds not passing kinase assay screening against STK33 at 500 nM or for compounds with IC50 >300 nM in the NanoBRET assay against STK33.
  • Example 1 Discovery of potent STK33 inhibitors from DNA-encoded chemical library 20 screening and hit optimization To identify small-molecule binders to the kinase domain (KD) of STK33, DNA- Encoded Chemistry Technology (DEC-Tec) was used to uncover potent and selective kinase inhibitors.
  • DEC-Tec DNA- Encoded Chemistry Technology
  • NanoBRETTM Target Engagement intracellular kinase assay HEK293T cells were cultured in DMEM (Gibco11965-092) with 10% FBS and 1% Pen-Strep (Gibco 15140-122) in 5% CO2, 37°C. The cell line underwent authentication by short tandem repeat (STR) profiling at Cytogenetics and Cell Authentication Core (CCAC) MD Anderson Cancer Center. NanoBRETTM TE intracellular kinase assay, K-10 assay (Cat.
  • HEK293 cells were transiently transfected with NanoLuc® (STK33, RET, CLK1, CLK2, and CLK4) fusion vectors. After 36 hours of transfection, cells were treated with fixed concentration of tracer (recommended by Promega) followed by compounds (inhibitors) for 2 hours of incubation. To determine the test compound affinity, 20 cells were titrated with varying concentrations of test compounds ranging from 40 ⁇ M to 0.4 nM at a fixed tracer concentration. Staurosporine was measured in parallel as a positive control. Freshly prepared NanoBRET Nano-Glo substrate plus extracellular NanoLuc inhibitor was then added to initiate the subsequent bioluminescence resonance energy transfer (BRET) measurements using a CLARIOstar Plus BMG LABTECH plate reader.
  • BRET bioluminescence resonance energy transfer
  • NanoBRET Target Engagement K192 assay HEK293 cells (ATCC) were cultured in DMEM (Gibco) + 10% FBS (Seradigm), and incubated in a humidified 37°C/5% CO2 incubator.
  • the NanoBRET Target Engagement K192 Kinase Selectivity System (Promega) 30 contains 192 unique transfection-ready NanoLuc/Kinase fusions, pre-diluted in either transfection carrier DNA or relevant cyclin DNA.
  • NanoLuc/Kinase fusion constructs were transfected into HEK293 cells using Fugene HD (Promega) according to the manufacturer’s protocol. Briefly, 10 ⁇ L of a 20 ⁇ g/mL DNA solution was added to white, TC-treated 96 well assay plates (Corning), followed by 30 ⁇ L of a diluted Fugene HD - 119 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) solution (20 ⁇ L Fugene/1 mL Opti-MEM).
  • HEK293 cells were resuspended at 3.3 ⁇ 10 5 c/mL in Opti-MEM + 1% FBS and added at a volume of 60 ⁇ L per well, to result in approximately 20000 cells/well.
  • Cells were incubated in a humidified 37°C/5% CO 2 incubator for 20 hours.
  • Tracer K-10 Promega was prepared at a 5 100 ⁇ concentration in DMSO (Sigma) and diluted to a working concentration of 10 ⁇ in tracer dilution buffer (12.5 mM HEPES, 31.25% PEG400, pH 7.5) (Promega).
  • Test compounds were prepared as concentrated stock solutions in DMSO and diluted in Opti-MEM to prepare 10 ⁇ working stocks.
  • Tracer K-10 was added 10 to cells at one of 4 different concentrations, ranging from 25 nM to 1 ⁇ M, based on target affinity, according to manufacturer’s protocol.
  • To measure BRET NanoBRET NanoGlo Substrate and Extracellular NanoLuc Inhibitor (Promega) were added according to the manufacturer’s protocol, and filtered luminescence was measured on a GloMax Discover luminometer equipped with 450 nm BP filter (donor) and 600 nm LP filter (acceptor), using 15 0.5s integration time. Milli-BRET units are calculated by multiplying the raw BRET units by 1000.
  • % Occupancy [1-(X-Z)/(Y-Z)] ⁇ 100
  • X BRET in the presence of the test compound and Tracer K10
  • Y BRET in the presence of Tracer K10 only
  • Z Full occupancy, or BRET in the absence of the test 20 compound and Tracer K10.
  • an untagged NanoLuc construct Promega
  • DEC-Tec affinity selections For these studies, two independent DEC-Tec selections of our in-house libraries were performed using full length recombinant human STK33 protein with an N-terminal His6 tag 25 (Eurofins #14-671).
  • SF9 cells were cultured in SF900III medium (Gibco) and infected with STK33 KD baculovirus in an optimal virus/ insect cells ratio.
  • Cells were harvested 48 h after - 120 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) infection; pellets were kept at ⁇ 80°C.
  • frozen cell pellet was resuspended in lysis buffer (25 mM Tris, pH 8.0, 150 mM NaCl, and 1mM TCEP) and lysed using homogenizer (Avestin).
  • both CDD-2110 and CDD-3348 demonstrated prominent potency toward STK33 in both biochemical and cellular assays.
  • Table B Chemical properties, biochemical activity, cellular activity, and metabolism data of the hit with a long linker, CDD-3348, and two STK33 inhibitors, namely compound 1 and ML281.
  • K d and K i values were calculated from LanthaScreen binding assay and Z’-LYTE assay, respectively, as described in the method section; IC50 values were calculated from NanoBRET assay; t 1/2 was measured using either mouse liver microsomal or human liver 30 microsomal stability assay.
  • Reported IC50 values of compound 1 and ML281 are 7 and 14 nM, respectively. ND means not determined. - 121 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) . . .
  • CDD-2211 R-isomer
  • CDD-2212 S-isomer
  • Synthetic route 1 (2) Synthetic route 2: 15 1-(1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide (A1, CDD-2110) 20 .
  • methyl 3-bromo-1H-indazole-5-carboxylate and 2-ethynyl-1,1’-biphenyl were utilized in the general procedure for Sonogashira coupling - 123 - 53366970.3 Attorney Docket.
  • step 2 the cross- coupling intermediate was subjected to the general procedure for hydrolysis without THP- 5 protected amine to yield 3-([1,1’-biphenyl]-2-ylethynyl)-1H-indazole-5-carboxylic acid.
  • step 3 the resulting acid and N-methyl-1-(pyrrolidin-3-yl)piperidine-4-carboxamide were utilized in the general procedure for amide coupling to afford the title compound (49% over two steps) as a light yellow oil;
  • step 2 the general procedure for Sonogashira coupling was employed using 2-ethynyl-1,1'-biphenyl.
  • step 3 the cross-coupling intermediate was subjected to the 5 general procedure for removal of Boc-protecting group to afford the title compound (41% over three steps) as a light yellow oil;
  • 1 H NMR 600 MHz, DMSO-d 6 , two rotamers
  • step 1 methyl 3-iodo-1-(tetrahydro-2H-pyran-2-yl)- 1H-indazole-5-carboxylate, derived from methyl 3-iodo-1H-indazole-5-carboxylate via the 10 general procedure for THP protection, and 2-ethynyl-1,1'-biphenyl were utilized in the general procedure for Sonogashira coupling.
  • step 2 the cross-coupling intermediate was subjected to the general procedure for hydrolysis with THP-protected amine to yield 3-([1,1'- biphenyl]-2-ylethynyl)-1-(tetrahydro-2H-pyran-3-yl)-1H-indazole-5-carboxylic acid (75% over two steps) as a pale yellow solid;
  • step 3 the acid and tert- butyl 4,7-diazaspiro[2.5]octane-7-carboxylate were utilized in the general procedure for 20 amide coupling.
  • step 4 the general procedure for removal of THP and Boc-protecting group was employed to afford the title compound (35% over two steps) as a yellow oil;
  • 13 C NMR (150 MHz, DMSO-d6) ⁇ 143.2, 140.7, 139.9, 132.9, 129.7, 129.4, 129.1, 128.4, 128.2, 25 127.7, 125.9
  • No.046641-7059WO1(00159) 143.3, 140.2, 140.0, 135.7, 132.8, 131.9, 129.7, 129.4, 129.1, 129.1, 128.3, 128.2, 127.9, 127.7, 127.7, 126.0, 125.4, 123.7, 120.2, 118.6, 115.2, 111.2, 92.8, 84.0, 48.6, 36.5, 30.9, 25.5;
  • a oven dried microwave vial equipped with magnetic stir bar was charged with the amide (0.05 mmol, 1.0 equiv.), [1,1'-biphenyl]-2- ylboronic acid (0.1 mmol, 2.0 equiv.), Pd(PPh 3 ) 4 (0.005 mmol, 0.1 equiv.), and K 2 CO 3 (0.1 mmol, 2.0 equiv.).
  • the vial was sealed with a microwave cap, evacuated and backfilled with nitrogen, followed by addition of anhydrous DMF (0.3 mL) and water (0.1 mL, purged with nitrogen) via syringe. The mixture was vacuum-purged and refilled with nitrogen for three cycles and then heated by microwave reactor at 100 o C for 1 h.
  • a oven dried microwave vial equipped with magnetic stir bar was charged with the amide (0.1 mmol, 1.0 equiv.), potassium ([1,1’- biphenyl]-2-ylmethyl)trifluoroborate (0.25 mmol, 2.5 equiv.), Pd(OAc) 2 (0.01 mmol, 0.1 equiv.), RuPhos (0.02 mmol, 0.2 equiv.), and Cs2CO3 (0.3 mmol, 3.0 equiv.).
  • the vial was sealed with a microwave cap, evacuated and backfilled with nitrogen, followed by addition of anhydrous toluene (3 mL) and water (0.3 mL, purged with nitrogen) via syringe.
  • Table C Validation of STK33/CDD-2211 interactions by structure-activity-relationship (SAR) study of CDD-2211 analogs.
  • ND means not determined for compounds not passing kinase assay screening against STK33 at 500 ⁇ M.
  • each CDD compound 2.0 ⁇ M was incubated in the mouse or human liver microsomes (0.5 mg protein/mL) at 37 °C in 1 ⁇ PBS fortified with NADPH (1.0 mM). The samples were collected at specific time-points 0, 30, and 60 min in duplicate. The reactions were terminated by adding equivalent volume of ice-cold CH 3 OH and vortexed.
  • CDD- 2807 was dissolved in 10% Captisol containing 5% DMSO to prepare the solutions of 0.5 mg/mL and 2.5 mg/mL.
  • One group of mice were administered with 5 mg/kg CDD-2807 (i.p., 10 ⁇ L/g of 0.5 mg/mL solution) and the second group with 25 mg/kg CDD-2807 (i.p., 10 ⁇ L/g of 2.5 mg/mL solution).
  • the blood samples were centrifuged at 2000 g for 3 min at 4 oC, and the plasma samples were transferred to tubes and stored at ⁇ 80°C before analysis. All the samples were analyzed by the Thermo TSQ Quantis MS coupled with a Thermo Vanquish UHPLC (San Jose, CA). CDD-2807 was separated on a Luna C18 column (1 mm ⁇ 50 mm, 1.6 ⁇ m, Phenomenex, Torrance, CA), and eluted by a water-acetonitrile mobile phase system (both containing 0.1% formic acid, v/v) at the flow rate of 0.15 mL/min.
  • CDD-2807 was monitored under the selected reaction monitoring (SRM) mode coupled with a positive electrospray ionization (ESI) source.
  • SRM reaction monitoring
  • ESI positive electrospray ionization
  • the SRM ion pairs were 447 ⁇ 321 for CDD-2807.
  • the ion spray voltage was set at 3500 V.
  • High-purity nitrogen was used as the sheath gas (35 arbitrary unit), auxiliary gas (7 arbitrary unit), and high-purity argon was used as the collision gas.
  • the temperatures of the ion transfer tube and the vaporizer were set at 300°C and 275°C, respectively.
  • the concentration of CDD-2807 in plasma was quantified using individual calibration curves. The calibration curve was regressed with a weight of 1/x 2 with high linearity (r 2 >0.99).
  • PK parameters such as half-time (t1/2), area under the plasma concentration–time curve during the period of observation (AUC 0–t ), area under the plasma concentration–time curve from zero to infinity (AUC0– ⁇ ), clearance normalized by the bioavailability (CL/F), volume of distribution normalized by the bioavailability (Vd/F) and the mean residence time (MRT) were calculated by WinNonlin software (Certara, Princeton, NJ) by noncompartmental analysis.
  • the plasma concentration–time curves were plotted in Prism 9 (GraphPad, San Diego, CA) as mean ⁇ S.D. CDD-2807 concentrations in testis and brain.
  • testis and brain harvested from day 45 of protocol 1 and day 63 of protocol 2 were weighed and snap-frozen in liquid nitrogen. The samples were stored at ⁇ 80°C before analysis. Twenty-five mg of testis or brain tissue was homogenized in 150 ⁇ L of 50% methanol, and 50 ⁇ L of homogenate was added to 200 ⁇ L of ice-cold CH3OH containing 0.1 ⁇ M of agomelatine (internal standard). The mixture was vortexed and centrifuged at 15000 g for 15 min, and supernatants were transferred into sample vials for analysis.
  • CDD-2807 was analyzed by the same LC-MS/MS method used in the pharmacokinetic study The concentration of CDD-2807 in the testis and brain was determined by using individual calibration curves established with the corresponding tissue homogenates from untreated mice. The calibration curve was regressed with a weight of 1/x 2 with high linearity (r 2 >0.99). Alanine transaminase (ALT) and aspartate transaminase (AST) assays. On day 45 of protocol 1 and day 63 of protocol 2, three mice from each protocol were euthanized, and blood was collected through cardiac puncture. The serum was isolated by centrifuging the whole blood samples at 1500 g for 10 min and then maintained on wet ice before analysis.
  • ALT Alanine transaminase
  • AST aspartate transaminase
  • CDD-2807 was subjected to NanoBRET Target Engagement K192 assay (Promega) at a dose of 1 ⁇ M against 192 full- length protein kinases in HEK293 cells.
  • STK33 was the most potent binder (95.9% occupancy) although other kinases were engaged by CDD-2807 (FIG.2D).
  • ThermoFisher dose response assays and NanoBRET cellular assays were used to evaluate potency of CDD-2807 against off-target kinases possessing >80% occupancy, including CDC-like kinase 4 (CLK4) (94.7% occupancy), CLK2 (89.7% occupancy), rearranged during transfection (RET) (85.8% occupancy), and CLK1 (83.5% occupancy).
  • CLK4 CDC-like kinase 4
  • RET rearranged during transfection
  • CLK1 83.5% occupancy
  • CDD-2807 was the most potent ligand of STK33 (FIG.2C). Taken together, CDD-2807 is a potent, stable, and relatively selective STK33 inhibitor, showing >9- fold selectivity versus the other kinases (FIG.2C).
  • CDD-2110 compared to CDD-3348 (e.g., 96-fold to 437-fold selectivity of STK33 over CLK4 in biochemical assays and 14-fold to 23-fold selectivity of STK33 over CLK4 in the NanoBRET assay; FIG.2C and Table B) also suggested that the selectivity toward STK33 could be improved by modification of the linker at BB1.
  • Table B A summary of the data in relation to other molecular scaffolds is shown in Table B.
  • a summary of the pharmacokinetic (PK) parameters is shown in Table G.
  • frozen cell pellet was resuspended in lysis buffer (25 mM Tris, pH 8.0, 150 mM NaCl, and 1mM TCEP) and lysed using homogenizer (Avestin). After centrifugation at 48,000g for 1 h, the soluble fraction was loaded onto pre-equilibrated Strep-Tactin Superflow resin (IBA Lifesciences GmbH) and eluted with elution buffer (25 mM Tris, pH 8.0, 150 mM NaCl, 1 mM TCEP, and 2 mM desthibiotin).
  • the protein-inhibitor mixture was concentrated using Amicon Ultra- 15 centrifugal filters (Millipore Sigma) to 14 mg/mL.
  • Mosquito SPT labtech
  • the data was integrated and scaled by using iMosflm and SCALA, respectively.
  • the crystal structures of the STK33/CDD-2211 complex was determined by molecular replacement using a monomeric model of STK33 KD (residues 99 ⁇ 383) generated by Alpha fold as a search model.
  • CDD-2211 was traced and fitted manually into electron density by using COOT.
  • the final models have gone through several rounds of refinement using phenix.refine (46) followed by manual model building using COOT (47).
  • the visualization program PyMOL was used (48). A summary of data collection is shown herein. Table H. Data collection and refinement statistics.
  • Each monomer shows a nearly identical pose for CDD-2211 (FIG.3B) bound in the ATP pocket (FIG.6). All residues in the construct are ordered except for a segment of the activation loop (residues 273-283 in chain A and 274-284 in chain B are disordered).
  • the STK33 dimer is formed by the activation loop segment and the following two-turn helix with the APE motif from one monomer docking onto a surface formed between ⁇ E and ⁇ G helices of the other monomer (FIG.3A).
  • CDD-2211 binds in the active site formed between the small and large lobes (FIGs. 3A-3E and FIG.6).
  • the biphenyl (BB3) binds to the outer lip of the active site near the hinge (FIG.3C). It interacts with L122 and G123 on top and G198, E199, and M245 at the bottom through van der Waals interactions (FIGs.3D and 3E).
  • the indazole (BB2) binds the ribose pocket near the hinge region. It forms hydrogen bonds with backbones of E193 and C195 at the hinge (FIG.3E).
  • the pyrrolidine with dimethyl amine (BB1) contacts the Mg 2+ positioning loop, glycine-rich loop, and activation loop. The dimethyl amine hydrogen bonds with D265 that normally interacts with Mg 2+ .
  • BB1 is shielded by F127 at the Gly rich-loop (top) and E242 and N243 at the catalytic loop (bottom), and it interacts with these residues through van der Waals interactions.
  • This cocrystal structure is consistent with structure-activity-relationship (SAR) studies from a set of CDD-2211 analogs (Table B).
  • SAR structure-activity-relationship
  • a computational model of STK33 bound with CDD-2807 was generated based on the crystal structure of the STK33/CDD-2211 complex (FIG.8).
  • the model suggests that the nitrogen of the piperdine moiety at BB1 would interact with the side chain of E199 through a hydrogen bond while its interactions with the active site at BB2 and BB3 would be similar as in the STK33/CDD-2211 complex.
  • CDD-2211 similar to CDD-2807, demonstrated some (although low) inhibition of off-target kinases CLK1, CLK2, CLK4, and RET, as shown in FIG.2C.
  • crystal structures of RET and CLK4 were aligned as representatives with the STK33/CDD-2211 complex and compared the binding site residues.
  • Example 5 Mouse study of contraceptive effects Based on molecular modeling, mouse and human STK33 have identical ATP binding pockets, consistent with evolutionary conservation and function of STK33.
  • the human STK33 amino acids that are in contact with CDD-2211 are identical in mouse STK33 (FIG. 9), suggesting that CDD-2807 would equally inhibit mouse and human STK33.
  • reproductive outcomes in male were evaluated in mice dosed with CDD-2807, a potent and stable compound. Fertility analysis Sexually mature male C57/129 hybrid mice were randomly assigned to either the vehicle control treatment or the CDD-2807 treatment groups.
  • Drug formulation and injections were as follows: mice receiving the 15 mg/kg CDD-2807 twice daily dosing had the compound dissolved in DMSO, which was injected at 5% of the final volume in 10% Captisol and water. For mice receiving a single daily dose of 50 mg/kg CDD-2807 once daily, the compound was dissolved in DMSO and injected at 5% in 20% Captisol and water. All injections were administered as an intraperitoneal (IP) injection, and for the 15 mg/kg CDD-2807 and DMSO control mice, the injections were delivered in the morning and the evening. Mice in the 50 mg/kg dosing received IP injections once in the evening. Weights were collected daily prior to drug administration and monitored throughout the entirety of the injection period.
  • IP intraperitoneal
  • Injections were given for 21 days prior to introduction of female mice.
  • males were housed continuously with two C57/129 sexually mature females for 45 days to generate approximately two months of fertility data.
  • the total number of pups born per litter sired by each male were counted and the total number of litters and pups sired per male over the entire trial was calculated.
  • Mean pup number and SEM were used to determine differences in average number of pups sired using a t-test using Prism 9. Histological analysis Briefly, tissue was fixed in Bouin’s fixative, washed with 70% ethanol, embedded in paraffin, sectioned at 4- ⁇ m thickness, and stained by periodic acid-Schiff (PAS)- hematoxylin.
  • PAS periodic acid-Schiff
  • sperm were then collected and moved to a separate tube and then sperm were spun down at 300 x g for 5 minutes at room temperature, allowing sperm to collect at the bottom without any epididymis segments being present.
  • Media was removed and sperm cells were resuspended in DPBS to wash.
  • sperm were spun down a second time and then resuspended in fresh 2.5% glutaraldehyde/PBS and fixed for 30 minutes at room temperature on a rocker. After fixation, sperm were dehydrated in increasing concentrations of ethanol form 20%, 30%, 50%, 60%, 70%, 90%, and 100%. During each dehydration step, sperm were incubated at room temperature for 15 minutes.
  • the density of the sperm changes and requires higher speeds for each subsequent spin down; when sperm are in 100% ethanol, it requires ⁇ 1200 x g to pellet them.
  • Sperm were incubated overnight in 100% ethanol at 4°C, and the following day the samples were split and then spun down at 1500 x g for 5 minutes at room temperature and then resuspended in 50% tert-butanol/ethanol and incubated for 15 minutes. Samples were then imaged at the Houston Cincinnati Research Institute Microscopy SEM/AFM core on a FEI Nova NanoSEM 230 instrument.
  • CDD-2807 has a plasma t 1/2 of over 11 hours with dose-dependent increase of the maximum concentration (Cmax) and area under the curve (AUC) when delivered intraperitoneally (FIG.10 and Table G)
  • adult male mice were subjected to two protocols (FIGs.11A-11B).
  • the six control treated males in protocol 1 sired an average of 1.83 ⁇ 0.07 litters/male and 6.75 ⁇ 0.19 pups/female/litter in month 1, four of the six CDD-2807-treated males sired litters (either one or two litters; 1.0 ⁇ 0.37 litters/male) and at a lower 0.92 pups/female/litter (FIG.4A).
  • litters were born to all females housed with the six control males (2.0 ⁇ 0 litters male; 7.25 ⁇ 0.17 pups/female/litter) (FIG.4A).
  • no litters were produced by females housed with males who received CDD-2807 (FIG.4A).
  • CDD-2807 For males in protocol 1 evaluated at day 45 of CDD-2807 treatment or day 63 of protocol 2 mice, CDD-2807 easily crossed the blood-testis barrier (89.6 ⁇ 14.1 ng or 114.3 ⁇ 7.9 ng of CDD-2807/mg tissue, respectively), whereas it was extremely low in the brains of the protocol 1 males (0.28 ⁇ 0.09 ng of CDD-2807/mg tissue) and undetectable in the protocol 2 males (FIG.13A). Despite the contraceptive effect of CDD-2807, testis size of CDD-2807-treated mice was not changed versus control males (FIG.4C; FIG.13B).
  • CDD-2807-treated males showed no difference in body weight versus controls (FIG.12C) or alterations in serum ALT (control: 47.7 ⁇ 5.3 IU/L; CDD-2807: 20.3 ⁇ 5.3 IU/L) or AST (control: 48.2 ⁇ 23.4 IU/L; CDD-2807: 35.8 ⁇ 3.1 IU/L).
  • CDD-2807-treated males showed no statistical difference in body weight versus controls (FIG.12D) or alterations in serum ALT (control: 37.7 ⁇ 13.6 IU/L; CDD- 2807: 46.0 ⁇ 11.0 IU/L) or AST (control: 42.1 ⁇ 14.9 IU/L; CDD-2807: 29.8 ⁇ 6.8 IU/L), confirming the relative safety of CDD-2807.
  • sperm from the cauda epididymides of each mouse were analyzed using Computer Assisted Sperm Analysis (CASA).
  • CASA Computer Assisted Sperm Analysis
  • sperm counts were lower in the controls in protocol 1 mice versus CDD-2807-treated mice (FIG.13C), sperm motility (FIG.13D), progressive sperm (FIG.13E), and hyperactivated sperm (FIG.13F) were reduced in the CDD-2807-treated mice.
  • protocol 2 sperm counts (FIG.4D), sperm motility (FIG.4E), progressive sperm (FIG.4F), and hyperactivated sperm (FIG.4G) were all statistically reduced in the CDD- 2807-treated mice versus control mice.
  • CDD-2807 a hit from DEC-Tec screening was identified that was developed into a potent inhibitor of STK33, showed that our inhibitor (CDD-2807; molecular weight of 447 Da) easily crossed the BTB to cause infertility, and demonstrated that the contraceptive effects were quickly reversible. Because CDD-2807 is more metabolically stable ( ⁇ 10 hours) in HLM versus MLM ( ⁇ 1 hour) (FIG.2C), CDD-2807 could be metabolically more stable in men than mice, allowing us to translate our findings to men. Additionally, a proteolysis-targeting chimera (PROTAC)-based chemical knockdown approach is possible for testis-specific degradation of STK33.
  • PROTAC proteolysis-targeting chimera
  • CDD-2807 represents an excellent chemical probe that can be used to investigate STK33 signaling in various contexts other than male contraception. This work illuminates that kinases can serve as beneficial targets for treating human conditions beyond oncology indications.
  • Sequence Listing STK33 (Human) (SEQ ID NO:1) MADSGLDKKSTKCPDCSSASQKDVLCVCSSKTRVPPVLVVEMSQTSSIGSAESLISLERKKE KNINRDITSRKDLPSRTSNVERKASQQQWGRGNFTEGKVPHIRIENGAAIEEIYTFGRILGK GSFGIVIEATDKETETKWAIKKVNKEKAGSSAVKLLEREVNILKSVKHEHIIHLEQVFETPK KMYLVMELCEDGELKEILDRKGHFSENETRWIIQSLASAIAYLHNNDIVHRDLKLENIMVKS SLIDDNNEINLNIKVTDFGLAVKKQSRSEAMLQATCGTPIYMAPEVISAHDYSQQCDIWSIG VVMY
  • Embodiment 2 provides the compound of Embodiment 1, wherein the compound of formula (I) is the compound of formula (Ia): a).
  • Embodiment 4 provides the compound of any one of Embodiments 1-3, wherein Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolinyl, optionally substituted quinolinyl, optionally substituted 2-pyridyl, optionally substituted 3-pyridyl, optionally substituted 4-pyridyl, optionally substituted pyrimidinyl, optionally substituted pyrazinyl, or optionally substituted triazinyl.
  • Embodiment 6 provides the compound of any one of Embodiments 1-5, wherein one of R a or R b is hydrogen or methyl.
  • Embodiment 7 provides the compound of any one of Embodiments 1-6, wherein R a and R b combine with the nitrogen atom to which they are bound to form optionally substituted 5-, 6-, or-7 membered heterocyclyl.
  • Embodiment 8 provides the compound of any one of Embodiments 1-7, wherein R a and R b combine with the nitrogen atom to which they are bound to form optionally substituted pyrrolidinyl.
  • Embodiment 9 provides the compound of any one of Embodiments 1-3 and 5-8, which is a compound of Formula (II), (III), (IV), or (V):
  • Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein NR a R b is selected from the group consisting of: , , , , , , herein the group is selected from the group consisting of: , , R c5 is optionally substituted phenyl.
  • Embodiment 13 provides the compound of any one of Embodiments 5-10 and 12, wherein R c1 , R c2 , R c3 , and R c4 are each independently hydrogen.
  • Embodiment 14 provides the compound of Embodiment 5, which is a compound of formula (VI): d3 R d4 R R d2 d1 H d 5 R N R N I).
  • Embodiment 16 prov Embodiments 1-15, which is selected from the group consisting of: 3-([1,1'-biphenyl]-2-ylethynyl)-N-(4-methoxyphenyl)-1H-indazole-5-carboxamide, (S)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, (R)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, 3-([1,1'-biphenyl]-2-ylethynyl)-
  • Embodiment 17 provides a compound selected from Table A, or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof.
  • Embodiment 18 provides a pharmaceutical composition comprising the compound of any one of Embodiments 1-17 and a pharmaceutically acceptable carrier.
  • Embodiment 19 provides a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17, and/or the pharmaceutical composition of Embodiment 18, optionally wherein the sterilization is temporary.
  • Embodiment 20 provides a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18, optionally wherein the infertility is temporary.
  • Embodiment 21 provides a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18.
  • Embodiment 22 provides the method of any one of Embodiments 19-21, wherein the compound provides a contraceptive effect in the male subject.
  • Embodiment 23 provides the method of any one of Embodiments 19-22, wherein the compound is administered orally to the male subject.
  • Embodiment 24 provides a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of Embodiments 1-16 and/or the pharmaceutical composition of Embodiment 18.
  • Embodiment 25 provides a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18.
  • Embodiment 26 provides the method of Embodiment 25, wherein the disease or disorder comprises a RET-mediated cancer.
  • Embodiment 27 provides the method of Embodiment 26, wherein the RET-mediated cancer comprises medullary thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A).
  • MEN2A Multiple Endocrine Neoplasia Type IIa
  • Embodiment 28 provides the method of Embodiment 25, wherein the disease or disorder comprises Hirschprung Disease, central hypoventilation syndrome, and/or renal agenesis.
  • Embodiment 29 provides a method of inhibiting at least one of CL1, CLK2, CLK3, and CLK4 in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18.
  • Embodiment 30 provides a method of treating, ameliorating, and/or preventing a disease or disorder caused by CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18.
  • Embodiment 31 provides a method of Embodiment 30, wherein the disease or disorder comprises Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and renal cancer, breast cancer, and/or melanoma.

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Abstract

In one aspect, the present disclosure provides compounds that inhibit STK33, RET, CLK family kinases, and/or related kinases.

Description

TITLE OF THE INVENTION Inhibitors of STK33, RET, CLK, and/or Related Kinases and Methods Using Same CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No.63/598,802, filed November 14, 2023, which is incorporated herein by reference in its entirety. SEQUENCE LISTING The XML file named “046641-7059WO1 - Sequence Listing.xml” created on November 13, 2024, comprising 7.2 Kbytes, is hereby incorporated by reference in its entirety. BACKGROUND OF THE DISCLOSURE Although women have had small-molecule contraceptive pill options for over 50 years, there is no effective, non-hormonal oral contraceptive pill for men. Thus, there is a need to identify and validate contraceptives that target novel proteins and structures that are not based on endocrine interventions. The population on our planet is increasing at an extremely rapid rate. In the last 60 years, the world’s population has increased by more than 2.6-fold, growing from 3 billion people in 1960 to 8 billion in 2022, with projections indicating it will reach 9 billion by 2037. With this high rate of population growth, it is unclear whether every child born will have sufficient food, water, and education in the near future. Contraception is an important strategy for family planning and may help to curb this continued population explosion and the impact of humans on our environment. However, following the introduction of the birth control pill for women, there have been limited breakthroughs in contraception over the recent decades. Despite this clear need for more affordable, long-acting, reversible, and consequently safe contraceptives, there is no effective oral contraceptive pill available for men. Although over 800 knockouts of testis-enriched genes have been created and about 250 of them have revealed a male fertility phenotype, few small-molecule inhibitors of these validated targets have been developed to date and even fewer of them have demonstrated a non-hormonal contraceptive effect in pre-clinical models. The present disclosure addresses these needs. The following exemplary' embodiments are provided, which do not limit the scope of the disclosure exemplified herein.
The disclosure provides in one aspect a compound of Formula (I):
Figure imgf000004_0001
wherein L1, X, Y, Z1, Z2, Z3, Ra and Rb are defined elsewhere herein.
The disclosure provides in one aspect a pharmaceutical composition comprising any compound of the disclosure and a pharmaceutically acceptable carrier.
The disclosure provides in one aspect a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure. In certain embodiments, the sterilization is temporary.
The disclosure provides in one aspect a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure. In certain embodiments, the infertility is temporary.
The disclosure provides in one aspect a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
The disclosure provides in one aspect a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
The disclosure provides in one aspect a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity', the method comprising administering the mammal a therapeutically’ effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
The disclosure provides in one aspect a method of inhibiting at least one of CL1, CLK2, CLK3, and CLK4 in a mammal, the method comprising administering the mammal an effective amount of at least one compound and/or pharmaceutical composition of the disclosure. The disclosure provides in one aspect a method of treating, ameliorating, and/or preventing a disease or disorder caused by CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound and/or pharmaceutical composition of the disclosure.
BRIEF DESCRIPTION OF THE DRAWINGS
The following detailed description of exemplary embodiments of the disclosure will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosure, non-limiting embodiments are show n in the drawings. It should be understood, how ever, that the disclosure is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.
FIG. 1 depicts DEC-Tec selection. Enrichment profile of BCM DNA-encoded chemical library (DECL) qDOS28_l against STK33 at 0.1 pM (x-axis, z-score) versus no target control (y-axis, z-score). A series of hit compounds w ere identified with similar building block 1 (BB1 in blue; attached to the DNA), same building block 2 (BB2 in red), and same building block 3 (BB3 in black). The enrichment of each tri-synthon is shown as sequencing counts/z-score at 0.1 pM in the box.
FIGs. 2A-2D depict examples of STK33 hits, analogs, and biological characteristics. (FIG. 2A) Chemical structure of CDD-2110, the hit with a short linker enriched in the STK33 selection. Blue, red, and black moieties correspond to BB1. BB2. and BB3, respectively. (FIG. 2B) Chemical structures of CDD-221 1 , CDD-2212, and CDD-2807, the analogs of CDD-2110. (FIG. 2C) Chemical properties, biochemical activity, cellular activity, and metabolism data of CDD-2110, CDD-2211, CDD-2212, and CDD-2807. Dissociation constant (Ka) and inhibition constant (Ki) values were calculated from LanthaScreen binding assay and Z’-LYTE assay, respectively; half-maximal inhibitory concentration (ICso) values were calculated from NanoBRET assay. Half-life (ti/2) was measured using either mouse liver microsomal (MLM) or human liver microsomal (HLM) stability assay; assay data >60 min is an extrapolated estimate and included for relative determination of half-life. (FIG. 2D) Summary of CDD-2807 fractional occupancy values for kinases with >30% occupancy (red: 30-49.9%; yellow: 50-79.9%; green: 80-100%) from NanoBRET Target Engagement (TE) KI 92 assay performed at 1 pM.
FIGs. 3A-3E depict a crystal structure of a STK33/CDD-2211 complex. (FIG. 3A) Overall structure of the STK33/CDD-2211 dimer complex. Chain A is shown with surface and chain B in cartoon. N and C termini are labeled. The bound CDD-2211 are shown in sticks with its carbon atoms in yellow, oxygens in red, and nitrogens in blue. The activation loop is colored in red. (FIG. 3B) 2Fo-Fc density for CDD-2211 in the STK33/CDD-2211 complex contoured at lo. (FIG. 3C) Electrostatic surface of the active site with CDD-2211. (FIG. 3D) LIGPLOT diagram showing interactions between STK33 and CDD-2211. (FIG. 3E) Detailed interaction between STK33 and CDD-2211. Hinge residue backbone atoms that form hydrogen bonds with the ligand are shown as spheres. Key interacting residues shown are shown as sticks.
FIGs. 4A-4H depict CDD-2807 treatment induces a reversible contraceptive effect. (FIG. 4A, FIG. 4B) Average litter size from the two-month fertility' assessment showed that CDD-2807-treated mice had a significant (*P<0.0001) reduction in fertility in months one and two in protocol 1 (FIG. 4A; n = 6) and protocol 2 (FIG. 4B; n = 7). Fertility was restored to equal levels as controls after halting CDD-2807 treatment in protocol 1. (FIG. 4C) Testes from control and CDD-2807 treatment in mice from protocol 2 at 63 days are not significantly different. (FIG. 4D- FIG. 4G) CDD-2807-treated mice (n = 3) from protocol 2 at 63 days had a statistically significant (*P<0.005) decrease in sperm counts (FIG. 4D), motility (FIG. 4E), progressive sperm (FIG. 4F), and hyperactivated sperm (FIG. 4G) compared to control mice (n = 3). (FIG. 4H) By SEM analysis, 94.7 ± 2.7% of the sperm in the CDD-2807 -treated mice (n = 3) in protocol 2 at 63 days had morphologic defects, including head and tail defects as shown, compared to only 10.9 ± 2.7% abnormal sperm for the controls (n = 3).
FIGs. 5A-5B depict DEC-Tec selection and potent hits. (FIG. 5A) Enrichment profile of BCM DECL qDOS28_l against STK33 at 0.5 pM (x-axis, z-score) versus no target control (y-axis. z-score). (FIG. 5B) Enrichment profile of BCM DECL qDOS28_l against STK33 at 0.5 pM (x-axis, z-score) versus selection against STK33 at 0.5 pM performed in the presence of staurosporine (y-axis, z-score). Most hits are not observed in the presence of staurosporine, indicating competitive binding.
FIG. 6 provides a superimposition of the two chains of the STK33/CDD-2211 dimer complex. A zoomed in panel on the right shows the bound CDD-2211 molecules with residues within 5 A of CDD-2211. Chain A is colored in tan and chain B in magenta. Two chains are nearly identical showing an RMSD of 0.61 A between shared 252 CA atoms. Chains A and B are colored in magenta and tan respectively. Only CA atoms are used in aligning two chains
FIG. 7 provides a structure based sequence alignment of STK33, RET, and CLK4. The CDD-2211 contact residues. El 99 and M245, which are unique to STK33 are marked with arrows.
FIG. 8 provides a model of CDD-2807 docked to the active site of human STK.33. The same color theme is used as in FIG. 3E.
FIG. 9 provides a sequence alignment between human and mouse STK33 proteins. Sequence alignment was performed using the CLUSTAL W program and plotted with the ENDscript program. Identical amino acids as identified by the default ESPript parameters (Risler. global score 0.7) are highlighted with white in red background, similar residues are in red. The secondary structures of human STK33 kinase domain are indicated above the sequence. CDD-2211 contacting residues in the human STK33/CDD-2211 complex are marked with arrows
FIG. 10 provides evaluations of CDD-2807 pharmacokinetics in mice.
FIG. 11 shows protocol timelines of mouse breeding and reversibility study
FIGs. 12A-12D provide data showing no body weight changes of mice treated longterm with CDD-2807. Body weight changes during treatment in protocol 1 (FIG. 12A; n = 3- 6 mice) and body weight of protocol 1 mice sacrificed at day 45 in protocol 1 (FIG. 12C; n = 2-3 mice). Body weight changes during treatment in protocol 2 (FIG. 12B; n = 4-7 mice) and body weight of protocol 2 mice sacrificed at day 63 (FIG. 12D; n = 3 mice).
FIGs. 13A-13F provide data showing CDD-2807 crosses the BTB to alter sperm parameters but testis weight in mice. (FIG. 13A) Levels of CDD-2807 in the testes and brains of protocol 1 mice (n = 3) at day 45 and protocol 2 mice (n = 3) at day 63; no CDD-2807 was observed in control mouse tissues. (FIG. 13B) Despite the contraceptive effect of CDD-2807, testis size of CDD-2807 -treated mice from protocol 1 (n = 3) at day 45 and protocol 2 (n = 3) at day 63 were not changed versus protocol 1 control males (n = 2) and protocol 2 control males (n = 3), respectively. (FIG. 13C) Vehicle control mice (n = 2) in protocol 1 unexpectedly had a low sperm count. (FIG. 13D- FIG. 13F) CDD-2807-treated mice (n = 3) had decreased sperm motility (FIG. 13D), progressive sperm (FIG. 13E), and hyperactivated sperm (FIG. 13F) compared to vehicle control mice (n = 2).
FIGs. 14A-14B provide histological analysis of (FIG. 14A) testis and (FIG. 14B) epididymis segments (Caput & Corpus) of control and CDD-2807 -treated male mice from protocol 1. Bouins fixed, 4 pm-longitudinal sections of the testis and epididymis were stained with PAS-hematoxylin. All 12 stages of spermatogenesis are represented in both control and CDD-2807 -treated mice and show normal tissue morphology and acrosome development. Epididymis from the control and CDD-2807-treated mice were similar histologically. Scale bars are included for reference. DETAILED DESCRIPTION OF THE DISCLOSURE
In one aspect, the present disclosure provides a method of altering cellular signaling, such as kinase signaling, in a subject. In some embodiments, the subject is male. In some embodiments, the subject is female. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound of the disclosure. In some embodiments, the compound of the disclosure is orally administered to the subject. In some embodiments, the compound of the disclosure is administered to the male subject at a dosage and dosage timing necessary7 to provide a contraceptive effect in the male subject.
In some embodiments, provided herein is a method of altering cellular signaling in a subject. In some embodiments, provided herein is a method of altering kinase function in a subject. In some embodiments, provided herein is a method of altering STK signaling in a subject. In some embodiments, provided herein is a method of altering RET signaling in a subject. In some embodiments, provided herein is a method of altering CLK signaling in a subject. In some embodiments, the method comprises administering to the subject an effective amount of a compound of the disclosure, such as but not limited to a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), and/or formula (VII).
In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a kinase-mediated disease and/or disorder, such as but not limited to cancer, the method comprising administering a compound of the disclosure, such as but not limited to a compound of formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), and/or formula (VII).
Reference will now be made in detail to certain embodiments of the disclosed subject matter, examples of which are illustrated in part in the accompanying drawings. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.
Throughout this document, values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a range of ‘'about 0.1% to about 5%” or '‘about 0. 1% to 5%” should be interpreted to include notjust about 0.1% to about 5%. but also the individual values (e.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.
In the methods described herein, the acts can be carried out in any order, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.
Definitions
The term “about” as used herein can allow for a degree of variability7 in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or of a stated limit of a range, and includes the exact stated value or range.
In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. The statement “at least one of A and B” or “at least one of A or B” has the same meaning as “A. B, or A and B.” In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting: information that is relevant to a section heading may occur within or outside of that particular section. All publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference.
The term “acyl” as used herein refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is bonded to a hydrogen forming a “formyl” group or is bonded to another carbon atom, which can be part of an alkyl, aryl, aralkyl cycloalkyl, cycloalkydalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl, heteroarylalkyl group or the like. An acyl group can include 0 to about 12, 0 to about 20, or 0 to about 40 additional carbon atoms bonded to the carbonyl group. An acyl group can include double or triple bonds within the meaning herein. An acryloyl group is an example of an acyl group. An acyl group can also include heteroatoms within the meaning herein. A nicotinoyl group (pyridyl-3-carbonyl) is an example of an acyl group within the meaning herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like. When the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a “haloacyl” group. An example is a trifluoroacetyl group.
The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some embodiments, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.
The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=C=CCH2, -CH=CH(CH3). - CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.
The term “alkoxy” as used herein refers to an oxygen atom connected to an alkyd group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like. Examples of branched alkoxy include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopenty loxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20. or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxy ethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.
The term “alkynyl” as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms. 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some embodiments, from 2 to 8 carbon atoms. Examples include, but are not limited to - C =CH. -OC(CH3), -C =C(CH2CH3). -CH2C=CH, -CH2C=C(CH3). and -CH2C^C(CH2CH3) among others.
The term "amine" as used herein refers to primary, secondary, and tertiary amines having, e.g, the formula N(group)? wherein each group can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NH2, for example, alkylamines, arylamines, alkylarylamines; R2NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, heterocyclylamines and the like: and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein.
The term “amino group” as used herein refers to a substituent of the form -NH2, - NHR, -NR2, -NR3 +, wherein each R is independently selected, and protonated forms of each, except for -NR3 . which cannot be protonated. Accordingly, any compound substituted with an amino group can be viewed as an amine. An “amino group” within the meaning herein can be a primary, secondary, tertiary, or quaternary amino group. An “alkylamino” group includes a monoalkylamino, dialkylamino, and trialkylamino group.
The term “aminoalkyl” as used herein refers to amine connected to an alkyl group, as defined herein. The amine group can appear at any suitable position in the alkyd chain, such as at the terminus of the alkyl chain or anywhere within the alkyl chain.
The term “aralkyl” as used herein refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined herein. Representative aralkyl groups include benzyl and phenylethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-ethyl-indanyl. Aralkenyl groups are alkenyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an ary l group as defined herein.
The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenyleny l, pyrenyl, naphthacenyl, chrysenyl. biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof. As used herein, the term “C6-10- C6-10 biaryl” means a C6-10 aryl moiety covalently bonded through a single bond to another C6-10 aryl moiety. The C6-10 aryl moiety can be any of the suitable aryl groups described herein. Non-limiting example of a C6-10- C6-10 biaryl include biphenyl and binaphthyl. As used herein, the term “composition” or “pharmaceutical composition” refers to a mixture of at least one compound described herein with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary and topical administration. The term “cycloalkyl” as used herein refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 4, 5, 6, or 7. Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “cycloalkenyl” alone or in combination denotes a cyclic alkenyl group. A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal's state of health.
A disease or disorder is "alleviated" if the severity of a symptom of the disease or disorder, the frequency with which such a symptom is experienced by a patient, or both, is reduced.
As used herein, the terms “effective amount,” “pharmaceutically effective amount” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.
The terms “halo,” “halogen,” or “halide” group, as used herein, by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.
The term “haloalkyl” group, as used herein, includes mono-halo alkyl groups, polyhalo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1,1 -di chloroethyl, 1,2-dichloroethyl, l,3-dibromo-3,3- difluoropropyl, peril uorobutyl. and the like.
The term “heteroaryl” as used herein refers to aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to. N, O, and S; for instance, heteroaryl rings can have 5 to about 8-12 ring members. A heteroaryl group is a variety of a heterocyclyl group that possesses an aromatic electronic structure. A heteroaryl group designated as a C2-heteroaryl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise a C4-heteroaryl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms sums up to equal the total number of ring atoms. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl. pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl. indolyl. azaindolyl. indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups can be unsubstituted, or can be substituted with groups as is discussed herein. Representative substituted heteroaryl groups can be substituted one or more times with groups such as those listed herein.
Additional examples of ary l and heteroaryl groups include but are not limited to phenyl, biphenyl, indenyl, naphthyl (1 -naphthyl, 2-naphthyl), N-hydroxytetrazolyl, N- hydroxytriazolyl, N-hydroxyimidazolyl, anthracenyl (1-anthracenyl, 2-anthracenyl, 3- anthracenyl), thiophenyl (2 -thieny l, 3-thienyl), furyl (2 -furyl, 3-furyl) , indolyl, oxadiazolyl, isoxazolyl, quinazolinyl, fluorenyl, xanthenyl. isoindanyl. benzhydryl, acridinyl, thiazolyl, pyrrolyl (2-pyrrolyl). pyrazolyl (3-pyrazolyl). imidazolyl (1 -imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl), triazolyl (1,2,3-triazol-l-yl, l,2,3-triazol-2-yl l,2,3-triazol-4-yl, l,2,4-triazol-3-yl), oxazolyl (2-oxazolyl, 4-oxazolyl, 5-oxazolyl), thiazolyl (2-thiazolyl, 4- thiazolyl. 5-thiazolyl), pyridyl (2-pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl), pyrazinyl, pyridazinyl (3- pyridazinyl. 4- pyridazinyl, 5-pyridazinyl), quinolyl (2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl, 6- quinolyl, 7-quinolyl, 8-quinolyl), isoquinolyl (1 -isoquinolyl, 3-isoquinolyl, 4-isoquinolyl, 5- isoquinolyl, 6-isoquinolyl, 7-isoquinolyl, 8-isoquinolyl), benzo[b]furanyl (2-benzo[b]furanyl, 3-benzo[b]furanyl, 4-benzo[b] furanyl. 5-benzo[b]furanyl. 6-benzo[b] furanyl. 7- benzofb] furanyl), 2,3-dihydro-benzo[b]furanyl (2-(2,3-dihydro-benzo[b]furanyl), 3-(2,3- dihydro-benzo[b] furanyl), 4-(2,3-dihydro-benzo[b]furanyl), 5-(2,3-dihydro-benzo[b]furanyl),
6-(2,3-dihydro-benzo[b]furanyl), 7-(2,3-dihydro-benzo[b] furanyl), benzo[b]thiophenyl (2- benzo[b]thiophenyl, 3-benzo[b]thiophenyl, 4-benzo[b] thiophenyl, 5-benzo[b]thiophenyl, 6- benzo[b]thiophenyl, 7-benzo[b]thiophenyl). 2,3-dihydro-benzo[b]thiophenyl, (2-(2,3- dihydro-benzo[b]thiophenyl), 3-(2,3-dihydro-benzo[b]thiophenyl), 4-(2,3-dihydro- benzo[b]thiophenyl), 5-(2,3-dihydro-benzo[b]thiophenyl), 6-(2,3-dihydro- benzo[b]thiophenyl). 7-(2,3-dihydro-benzo[b]thiophenyl), indolyl (1-indolyl, 2-indolyl,
3-indolyl, 4-indolyl, 5-indolyl. 6-indolyl, 7-indolyl), indazole (1-indazolyl, 3-indazolyl,
4-indazolyl, 5-indazolyL 6-indazolyl, 7-indazolyl), benzimidazolyl (1 -benzimidazolyl, 2-benzimidazolyl, 4-benzimidazolyl, 5-benzimidazolyl, 6-benzimidazolyl, 7-benzimidazolyl, 8-benzimidazolyl), benzoxazolyl (1-benzoxazolyl, 2-benzoxazolyl), benzothiazolyl (1- benzothiazolyl, 2-benzothiazolyl, 4-benzothiazolyl, 5 -benzothiazolyl, 6-benzothiazolyl,
7-benzothiazolyl), carbazolyl (1-carbazolyL 2-carbazolyl. 3-carbazolyl. 4-carbazolyl), 5H-dibenz[b,f|azepine (5H-dibenz[b,f| azepin- 1-yl, 5H-dibenz[b,f|azepine-2-yl, 5H-dibenz[b,f|azepine-3-yl, 5H-dibenz[b,f|azepine-4-yl, 5H-dibenz[b,f|azepine-5-yl),
10,1 l-dihydro-5H-dibenz[b,f| azepine (10,1 l-dihydro-5H-dibenz[b.f|azepine-l-yl,
10,1 l-dihydro-5H-dibenz[b,f]azepine-2-yl, 10,1 l-dihydro-5H-dibenz[b,f|azepine-3-yl.
10,1 l-dihydro-5H-dibenz[b,f|azepine-4-yl, 10,1 l-dihydro-5H-dibenz[b,f|azepine-5-yl), and the like. The term “heteroarylalkyl” as used herein refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined herein. As used herein, the term “C6-10-5-6 membered heterobiaryl” means a C6-10 aryl moiety covalently bonded through a single bond to a 5- or 6-membered heteroaryl moiety. The C6-10 aryl moiety and the 5-6-membered heteroaryl moiety can be any of the suitable aryl and heteroaryl groups described herein. Non-limiting examples of a C6-10-5-6 membered heterobiaryl include: . When the C6-10-5-6 membered heterobiaryl is listed group), the C6-10-5-6 membered
Figure imgf000015_0001
heterobiaryl is bonded to the rest of the molecule through the C6-10 moiety. As used herein, the term “5-6 membered- C6-10 heterobiaryl “ is the same as a C6-10-5- 6 membered heterobiaryl, except that when the 5-6 membered C6-10 heterobiaryl is listed as a substituent (e.g., as an “R” group), the 5-6 membered- C6-10 heterobiaryl is bonded to the rest of the molecule through the 5-6-membered heteroaryl moiety. The term “heterocyclyl” as used herein refers to aromatic and non-aromatic ring compounds containing three or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S. Thus, a heterocyclyl can be a cycloheteroalkyl, or a heteroaryl, or if polycyclic, any combination thereof. In some embodiments, heterocyclyl groups include 3 to about 20 ring members, whereas other such groups have 3 to about 15 ring members. A heterocyclyl group designated as a C2-heterocyclyl can be a 5-ring with two carbon atoms and three heteroatoms, a 6-ring with two carbon atoms and four heteroatoms and so forth. Likewise, a C4-heterocyclyl can be a 5-ring with one heteroatom, a 6-ring with two heteroatoms, and so forth. The number of carbon atoms plus the number of heteroatoms equals the total number of ring atoms. A heterocyclyl ring can also include one or more double bonds. A heteroaryl ring is an embodiment of a heterocyclyl group. The phrase “heterocyclyl group” includes fused ring species including those that include fused aromatic and non-aromatic groups. For example, a dioxolanyl ring and a benzdioxolanyl ring system (methylenedioxyphenyl ring system) are both heterocyclyl groups within the meaning herein. The phrase also includes polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. Heterocyclyl groups can be unsubstituted or can be substituted as discussed herein. Heterocyclyl groups include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl. triazolyl. tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyL thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl, dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl, azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl. and quinazolinyl groups. Representative substituted heterocyclyl groups can be mono-substituted or substituted more than once, such as, but not limited to, piperidinyl or quinolinyl groups, which are 2-, 3-, 4-, 5-, or 6- substituted, or disubstituted with groups such as those listed herein.
The term “heterocyclylalkyl” as used herein refers to alkyl groups as defined herein in which a hydrogen or carbon bond of an alkyl group as defined herein is replaced with a bond to a heterocyclyl group as defined herein. Representative heterocyclyl alkyl groups include, but are not limited to, furan-2-yl methyl, furan-3-yl methyl, pyridine-3-yl methyl, tetrahydrofuran-2-yl ethyl, and indol-2-yl propyl.
The term '‘independently selected from” as used herein refers to referenced groups being the same, different, or a mixture thereof, unless the context clearly indicates otherwise. Thus, under this definition, the phrase “X1, X2, and X3 are independently selected from noble gases” would include the scenario where, for example, X1, X2, and X3 are all the same, wherein X1, X2, and X3 are all different, wherein X1 and X2 are the same but X3 is different, and other analogous permutations.
The term “monovalent” as used herein refers to a substituent connecting via a single bond to a substituted molecule. When a substituent is monovalent, such as, for example, F or Cl, it is bonded to the atom it is substituting by a single bond.
The term '‘organic group” as used herein refers to any carbon-containing functional group. Examples can include an oxygen-containing group such as an alkoxy group, aryloxy group, aralkyloxy group, oxo(carbonyl) group; a carboxyl group including a carboxylic acid, carboxylate, and a carboxylate ester; a sulfur-containing group such as an alkyl and aryl sulfide group; and other heteroatom-containing groups. Non-limiting examples of organic groups include OR, OOR, OC(O)N(R)2, CN, CF3, OCF3, R, C(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SOsR, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R. C(0)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O- 2N(R)C(0)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)C0N(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(0)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, C(=NOR)R, and substituted or unsubstituted (Ci-Cioo)hydrocarbyl. wherein R can be hydrogen (in examples that include other carbon atoms) or a carbon-based moiety, and wherein the carbon-based moiety can be substituted or unsubstituted.
The terms “patient,” “subject,” or “individual” are used interchangeably herein, and refer to any animal, or cells thereof whether in vitro or in situ, amenable to the methods described herein. In a non-limiting embodiment, the patient, subject or individual is a human.
As used herein, the term “pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non- toxic, z.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.
As used herein, the language “pharmaceutically acceptable salt” refers to a salt of the administered compounds prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids or bases, organic acids or bases, solvates, hydrates, or clathrates thereof.
Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric (including sulfate and hydrogen sulfate), and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2- hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic. P-hydroxy butyric, salicylic, galactaric and galacturonic acid.
Suitable pharmaceutically acceptable base addition salts of compounds described herein include, for example, ammonium salts, metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N’-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.
As used herein, the term '‘pharmaceutically acceptable carrier’ or “pharmaceutically acceptable excipient” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound described herein within or to the patient such that it may perform its intended function. Typically, such compounds are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound(s) described herein, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc: excipients, such as cocoa butter and suppository’ waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound(s) described herein and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound(s) described herein. Other additional ingredients that may be included in the pharmaceutical compositions used with the methods or compounds described herein are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.
The term “solvent” as used herein refers to a liquid that can dissolve a solid, liquid, or gas. Non-limiting examples of solvents are silicones, organic compounds, water, alcohols, ionic liquids, and supercritical fluids. The term “substantially” as used herein refers to a majority of, or mostly, as in at least about 50%. 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or 100%. The term “substantially free of’ as used herein can mean having none or having a trivial amount of, such that the amount of material present does not affect the material properties of the composition including the material, such that the composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4, 3.5. 3, 2.5, 2. 1.5, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0. 1, 0.01, or about 0.001 wt% or less. The term “substantially free of’ can mean having a trivial amount of, such that a composition is about 0 wt% to about 5 wt% of the material, or about 0 wt% to about 1 wt%, or about 5 wt% or less, or less than, equal to, or greater than about 4.5 wt%, 4. 3.5, 3, 2.5. 2, 1.5, 1, 0.9. 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.01, or about 0.001 wt% or less, or about 0 wt%.
The term “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CFs, OCF3, R, O (oxo), S (thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R. C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O- 2N(R)C(O)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety; for example, R can be hydrogen, (Ci- Cioo)hydrocarbyl, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl.
A ‘"therapeutic’' treatment is a treatment administered to a subject who exhibits signs of pathology, for the purpose of diminishing or eliminating those signs.
The term “thioalkyd” as used herein refers to a sulfur atom connected to an alky 1 group, as defined herein. The alky l group in the thioalky 1 can be straight chained or branched. Examples of linear thioalkyl groups include but are not limited to thiomethyl, thioethyl, thiopropyl, thiobutyl, thiopentyl, thiohexyl, and the like. Examples of branched alkoxy include but are not limited to iso-thiopropyl, sec-thiobutyl, tert-thiobutyl. iso-thiopentyl. isothiohexyl, and the like. The sulfur atom can appear at any suitable position in the alkyl chain, such as at the terminus of the alkyl chain or anywhere within the alkyl chain.
The terms “treat,” “treating” and “treatment.” as used herein, means reducing the frequency or severity with which symptoms of a disease or condition are experienced by a subject by virtue of administering an agent or compound to the subject.
Throughout this disclosure, various aspects of the disclosure can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3. from 1 to 4, from 1 to 5. from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.
Compounds
In one aspect, the present disclosure relates to an inhibitor of STK33. In some embodiments, the STK33 inhibitor is selective for STK33 over one or more members of related kinases. In some embodiments, the present disclosure relates to an inhibitor of RET. In some embodiments, the RET inhibitor is selective for RET over one or more members of related kinases. In some embodiments, the present disclosure relates to an inhibitor of CLK. In some embodiments, the CLK is CLK1. In some embodiments, the CLK is CLK2. In some embodiments, the CLK is CLK3. In some embodiments, the CLK is CLK4. In some embodiments, the CLK inhibitor is selective for CLK over one or more members of related kinases.
In some embodiments, the selectivity is such that the pICso of the compound for the specific kinase (STK33, RET, and/or one or more CLK’s) is about 0.1, 0.2, 0,3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8.1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, or higher than the pIC50 of the compound for the other kinase in question. In some embodiments, provided herein is a compound of Formula (I), or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof. In some embodiments, the compound of Formula (I) is: I), wherein: 1
Figure imgf000021_0001
L is selected from the group consisting of -C≡C-,-CH=CH- , -CH2-, and a bond; X is selected from the group consisting of -C(=O)-, -C(CH3)2-, -CF2-, -CHF-, - CH(CH3)-, and -CH2; Z1, Z2, or Z3 are independently CH or N, wherein 0 or 1 of Z1, Z2, and Z3 is N; Ra and Rb are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, and optionally substituted 3-to-8 membered heterocyclyl, or Ra and Rb may combine with the nitrogen atom to which they are bound to form optionally substituted 3-to-8 membered heterocyclyl; and Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C3-C10 heteroaryl, or CN; or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof. In certain embodiments, the compound of formula (I) is the compound of formula (Ia): a). In some embodiments,
Figure imgf000021_0002
iments, X is -C(CH3)2-. In some embodiments, X is -CF2-. In some embodiments, X is -CHF-. In some embodiments, X is - CH(CH3)-. In some embodiments, X is -CH2-. In some embodiments, Z1, Z2, and Z3 are CH. In some embodiments, Z1 and Z2 are CH, and Z3 is N. In some embodiments, Z1 and Z3 are CH, and Z2 is N. In some embodiments, Z2 and Z3 are CH, and Z1 is N. In some embodiments, Y is optionally substituted phenyl. In some embodiments, Y is optionally substituted naphthyl. In some embodiments, Y is optionally substituted isoquinolinyl. In some embodiments, Y is optionally substituted quinolinyl. In some embodiments, Y is optionally substituted 2-pyridyl. In some embodiments, Y is optionally substituted 3-pyridyl. In some embodiments, Y is optionally substituted 4-pyridyl. In some embodiments, Y is optionally substituted pyrimidinyl. In some embodiments, Y is optionally substituted pyrazinyl. In some embodiments, Y is optionally substituted triazinyl. In some embodiments, Y is substituted with at least one substituent independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, -C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3-C10 heteroaryl; or two adjacent substituents may combine with the atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl. In some embodiments, Re1 and Re2 are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally substituted 3-to-8 membered heterocyclyl, or optionally substituted C3-C10 heteroaryl; In some embodiments, one of Ra or Rb is hydrogen or methyl. In some embodiments, Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted 5-, 6-, or-7 membered heterocyclyl. In some embodiments, Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted pyrrolidinyl. In some embodiments , wherein Rc1, Rc2, Rc3, Rc4, and Rc5 are independently selected from t
Figure imgf000022_0001
of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, -C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3-C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl; wherein Re1 and Re2 are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally substituted 3-to-8 membered heterocyclyl, or optionally substituted C3-C10 heteroaryl. In some embodiments, the compound of Formula (I) is a compound of Formula (II), (III), (IV), or (V): V), w
Figure imgf000023_0001
Rc1, Rc2, Rc3, Rc4, and Rc5 are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, - C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3- C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally naphthyl, optionally 3-to-8 membered heterocyclyl, and optionally substituted 5-to-8- membered heteroaryl, optionally wherein two of Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, can combine to form an optionally substituted 2-to-7-atom alkylene or heteroalkylene group (e.g., spiro- or fused-bicyclic ring system); X1 is C(Rd3)(Rd4), O, or NRf, wherein Rf is selected from the group consisting of optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl; and X2 is C(Rd5)(Rd6), O, or NRf, wherein Rf is selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl. In some embodiments, NRaRb is selected from the group consisting of: , ,
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
some embodiments,
H n some embodiments, NRaRb is
Figure imgf000026_0002
. In some embodiments,
Figure imgf000026_0003
. In some embodiments, NRaRb is
Figure imgf000026_0004
. In some embodiments, NRaRb is
Figure imgf000026_0005
. In some embodiments, NRaRb is . In some
Figure imgf000026_0006
embodiments, NRaRb is i . In some embodiments, NRaRb is
Figure imgf000026_0007
. In
O,
Figure imgf000026_0008
some embodiments, NRaRb is —• NH . In some embodiments, NRaRb is
Figure imgf000026_0009
,
In some embodiments, the
Figure imgf000026_0010
group is selected from the group consisting of:
Figure imgf000026_0011
Figure imgf000027_0001
Figure imgf000028_0001
In some embodiments, provided herein is a compound of Formula (VI):
Figure imgf000029_0001
In some embodiments, provided herein is a compound of formula (VII):
Figure imgf000029_0002
In some embodiments, the compound of Formula (I) is a compound of Formula (II). In some embodiments, the compound of Formula (I) is a compound of Formula (III). In some embodiments, the compound of Formula (I) is a compound of Formula (IV). In some embodiments, the compound of Formula (I) is a compound of Formula (V). In some embodiments, the compound of Formula (I) is a compound of Formula (VI). In some embodiments, the compound of Formula (I) is a compound of Formula (VII). In some embodiments, provided herein is a compound selected from the group consisting of:
Figure imgf000029_0003
(S)-3-([l,r-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-l-phenylethyl)-lH-indazole- 5-carboxamide,
(R)-3-([l,l'-biphenyl] -2-ylethyny l)-N-(2-(dimethylamino)-2-oxo-l -phenylethyl)- 1H- mdazole -5 -carboxamide,
3-([l,r-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-l-phenylethyl)-lH-indazole-5- carboxamide,
Figure imgf000030_0001
(3-([l ,T-biphenyl]-2-ylethynyl)-l H-indazol-5-yl)(pyrrolidin-l -yl)methanone,
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-hydroxypyrrolidin-l-yl)methanone,
Figure imgf000030_0002
(R)-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-hydroxypyrrolidin-l-yl)methanone,
(S)-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-liydroxypyrrolidin-l-yl)methanone,
Figure imgf000030_0003
l-(3-([l.r-biphenylJ-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-one,
Figure imgf000030_0004
l-(l-(3-([l,r-biphcnyl]-2-ylcthynyl)-lH-indazolc-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide,
(R)- 1 -( l-(3 -([ 1 , 1 '-biphenyl] -2-ylethyny 1)- 1 H-indazole-5 -carbony l)py rrolidin-3 -y 1)-N- methylpiperidine-4-carboxamide,
(S)-l-(l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide.
Figure imgf000031_0001
l-(l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethylpiperidine-4-carboxamide,
(R)- 1 -( 1 -(3 -([ 1 , 1 ’-biphenyl] -2-ylethynyl)- 1 H-indazole-5 -carbonyl)pyrrolidin-3 -y 1)-N,N- dimethylpiperidine-4-carboxamide,
(S)-l-(l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethy lpiperidine-4-carboxamide,
Figure imgf000031_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2.5-diazabicyclo[2.2.2]octan-2- yl)methanone.
Figure imgf000031_0003
(3-([l.r-biphcnyl]-2-ylcthynyl)-lH-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone,
Figure imgf000032_0001
4-(2-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)-2.6-diazaspiro[3.5]nonane-6- carbonyl)-N-methylcyclohexane-l -carboxamide,
(R)-4-(2-(3 -([1,1 '-biphenyl] -2-ylethynyl)- 1 H-indazole-5 -carbony l)-2.6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-l-carboxamide,
(S)-4-(2-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)-2.6-diazaspiro[3.5]nonane- 6-carbonyl)-N-methylcyclohexane-l -carboxamide,
(3 -( [ 1 , 1 '-bipheny 1] -2 -y lethyny 1) - 1 H-indazol-5 -y 1) (3 -(dimethy lamino)pyrrolidin- 1 - yl)methanone,
Figure imgf000032_0002
(R)-(3-([ 1 , 1 ’ -biphenyl] -2-ylethynyl)- lH-indazol-5 -y l)(3-(dimethylamino)pyrrolidin- 1 - yl)methanone,
Figure imgf000032_0003
(S)-(3 -([ 1.1 ’-biphenyl] -2-ylethynyl)- 1 H-indazol-5 -y 1) (3 -(dimethy lamino)pyrrolidin- 1 - yl)methanone.
Figure imgf000032_0004
(3-(phenylethynyl)-lH-indazol-5-yl)(pyrrolidin-l-yl)methanone,
Figure imgf000033_0001
3-([l. l'-biphenyl] -2 -ylethynyl)-N-(2-(dimethylamino)ethyl)-N -methyl- lH-indazole-5- carboxamide.
Figure imgf000033_0003
(R)-(3-([ 1 , 1 ’-biphenyl] -2-y lethynyl)- 1 H-pyrazolo [3 ,4-b ] pyridin-5 -y 1)(3 - (dimethylamino)pyrrolidin-l-yl)methanone,
(S)-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-l -yl)methanone,
(3-([l, l'-biphenyl] -2-y lethynyl)-l H-pyrazolo[3, 4-b] pyridin-5 -y 1)(3- (dimethylamino)pyrrolidin-l-yl)methanone,
Figure imgf000033_0002
(R)-(3-([ 1 , 1 '-biphenyl] -2-y lethynyl)- 1 H-indazol-5 -y l)(3-(methy lamino)pyrrolidin- 1 - yl)methanone,
(S)-(3 -([ 1 , 1 '-biphenyl] -2-y lethy nyl)- 1 H-indazol-5 -y 1)(3 -(methylamino)py rrolidin- 1 - yl)methanone,
(3 -([ 1 , 1 '-biphenyl] -2-y letliyny 1) - 1 H-indazol-5 -y 1)(3 -(methylamino) py rrolidin- 1 - yl)methanone,
Figure imgf000034_0002
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -ipheny lethy ny 1)- 1 H-indazol-5-y l)methanone,
(S)-(3-(dimethylamino)pyrrolidin-l-yl)(3-(phenylethynyl)-lH-indazol-5-yl)methanone,
(3-(dimethylamino)pyrrolidin-l-yl)(3-(phenylethynyl)-lH-indazol-5-yl)methanone,
Figure imgf000034_0001
(R)-(3-((2.6-dichlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone. (S)-(3-((2.6-dichlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone. (3-((2,6-dichlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone.
Figure imgf000035_0001
(R)-(3-((2-chlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidm-l- yl)methanone.
(S)-(3-((2-chlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone.
(3-((2-chlorophenyl)ethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l-yl)methanone,
Figure imgf000035_0002
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(trifluoromethoxy)pheny l)ethyny 1)- 1 H-indazol- 5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-l-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-lH-indazol-
5-yl)methanone, (3-(dimethylamino)pyrrolidin-l-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-lH-indazol-5- yl)methanone,
Figure imgf000035_0003
(R)-(3-(dimethy lamino)py rrolidin- 1 -y 1)(3 -((2-fluoropheny l)ethy ny 1)- 1 H-indazol-5- yl)methanone,
(S)-(3 -(dimethy lamino)pyrrolidin- 1 -y 1) (3 -((2-fluoropheny l)ethynyl)- 1 H-indazol-5- yl)methanone. (3-(dimethylamino)pyrrolidin-l-yl)(3-((2-fluorophenyl)ethynyl)-lH-indazol-5- yl)methanone.
Figure imgf000036_0001
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-morpholinopheny l)ethynyl)- 1 H-indazol-5 - yl)methanone.
(S)-(3 -(dimethy lamino)pyrrolidin- 1 -y 1) (3 -((2-morpholinopheny l)ethyny 1)- 1 H-indazol-5 - yl)methanone.
(3-(dimethylamino)pyrrolidin-l-yl)(3-((2-morpholinophenyl)ethynyl)-l H-indazol-5- yl)methanone.
Figure imgf000036_0002
(R)-(3-([lH'-biphenyl]-3-ylethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl (methanone.
(S)-(3-([ LI ’-biphenyl] -3 -ylethynyl)-l H-indazol-5 -yl)(3 -(dimethy lamino)pyrrolidin-l- yl)methanone,
((3-([Ll'-biphenyl]-3-ylethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
Figure imgf000036_0003
(R)-(3-([l,r-biphenyl]-4-ylethynyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
(S)-(3 -([ 1, 1 ’-biphenyl] -4-y lethynyl)- 1 H-indazol-5 -y 1)(3 -(dimethy lamino)pyrrolidin- 1 - yl)methanone,
(3-([ l.l'-bipheny l]-4-ylethynyl)-lH-indazol-5-yl)(3 -(dimethy lamino)pyrrolidin-l- yl)methanone,
Figure imgf000037_0001
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(pyridin-3 -y l)pheny l)ethy ny 1)- 1 H-indazol-5 - yl)methanone,
(S)-(3 -(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(pyridin-3 -y l)phenyl)ethy ny 1)- lH-indazol-5 - yl)methanone,
(3-(dimethylamino)pyrrolidin-l-yl)(3-((2-(pyridin-3-yl)phenyl)etliynyl)-lH-indazol-5- yl)methanone,
Figure imgf000037_0002
(R)-(3-(dimethylamino)pyrrolidin-l-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-lH-indazol-5- yl)methanone,
(S)-(3 -(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(pyridin-4-y l)phenyl)ethyny 1)- 1 H-indazol-5 - yl)methanone,
(3-(dimethy lamino)pyrrolidin- 1 -yl)(3-((2-(pyridin-4-yl)pheny l)ethyny 1)- 1 H-indazol-5 - yl)methanone,
Figure imgf000037_0003
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -(isoquinolin-5 -y lethynyl)- 1 H-indazol-5 - yl)methanone.
(S)-(3-(dimethylamino)pyrrolidin-l-yl)(3-(isoquinolin-5-ylethynyl)-lH-indazol-5- yl)methanone.
(3-(dimethy lamino)pyrrolidin- 1 -y 1) (3 -(isoquinolin-5 -y lethynyl)- 1 H-indazol-5 -y l)methanone,
Figure imgf000038_0003
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3.6-diazabicyclo[3.1.1]heptan-3- yl)methanone.
Figure imgf000038_0001
(3-([Ll'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- yl)methanone,
Figure imgf000038_0002
(3-([l.r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone,
Figure imgf000039_0001
(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(8-oxa-3-azabicyclo[3.2.1]octan-3- yl)methanone.
Figure imgf000039_0002
(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-phenylpiperazin-l-yl)methanone,
(R)- 3-([l J ’-biphenyl] -2-ylethynyl)-lH-indazol-5-yl)(3-phenylpiperazin-l-yl)methanone,
(S)- 3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-phenylpiperazin-l-yl)methanone,
Figure imgf000039_0003
(R)-(3-([ 1.1 ’-biphenyl] -2-y lethynyl)- 1 H-indazol-5 -y 1) (3 -aminopyrrolidin- 1 -yl)methanone.
(S)-(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-aminopyrrolidin-l-yl)methanone, (3-([l J ’-biphenyl] -2-y lethynyl)-! H-indazol-5-yl)(3-aminopyrrolidin-l-yl)methanone, ( l)- 1 H-indazol-5 -y 1)( 1.4-diazepan- 1 -y l)methanone,
Figure imgf000040_0001
(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(4,7-diazaspiro[2.5]octan-4-yl)methanone,
Figure imgf000040_0003
(3-([l.l'-biphenyl] -2-ylethynyl)-lH-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-8-yl)methanone,
Figure imgf000040_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-methyl-2,7-diazaspiro[3.5]nonan-7- yl)methanone,
Figure imgf000041_0001
(3-([l J'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000041_0002
(7-methyl-2,7-diazaspiro[3.5]nonan-2-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5- yl)mcthanonc,
Figure imgf000041_0003
(3 -( [ 1 , l'-bipheny 1] -2-y lethyny 1) - 1 H-indazol-4-y 1) (7-methy 1-2, 7-diazaspiro [3.5 ] nonan-2- yl)methanone.
Figure imgf000041_0004
(3-([l, l'-bipheny l]-2-ylethynyl)-lH-indazol-5-yl)(5-mediy 1-2, 7-diazaspiro [3.5]nonan-2- yl)methanone,
(R)- (3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(5-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone,
(S)- (3-([l, l'-bipheny l]-2-y lethynyl)-lH-indazol-5-y l)(5-methyl-2, 7-diazaspiro[3.5]nonan-2- yl)mcthanonc,
Figure imgf000042_0001
7-(3-([ 1J '-biphenyl] -2 -yletiiynyl)-lH-indazole-5-carbonyl)-2,7-diazaspiro[3.5]nonan-l -one,
Figure imgf000042_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-6-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone,
Figure imgf000043_0001
(2.8-diazaspiro[4.5]decan-2-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-lH-indazol-5- yl)methanone,
Figure imgf000043_0004
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(6,9-diazaspiro[4.5]decan-6-yl)methanone,
Figure imgf000043_0002
(S)-(3-([l , 1 ’-biphenyl] -2-ylethynyl)-l H-indazol-5-yl)(2-phenylpiperazin-l -yl)methanone, (R)-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-phenylpiperazin-l-yl)methanone,
(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-phenylpiperazin-l-yl)methanone,
Figure imgf000043_0003
(S)-(2-phenylpiperazin-l-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-lH-indazol-5- yl)methanone,
(R)-(2-phenylpiperazin-l-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-lH-indazol-5- yl)methanone,
(2-pheny Ipiperazin- 1 -y 1)(3 -((2-(lrifluoromethoxy)pheny l)ethy ny 1)- 1 H-indazol-5- yl)methanone.
Figure imgf000044_0001
(R)-(3-(| 1. l'-bipheny 11 -2-y lethynyl)- 1 H-indazol-5 -yl)(2-(4-chloropheny l)piperazin- 1 - yl)methanone.
(S)-(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-(4-chlorophenyl)piperazin-l- yljmcthanonc.
(3-([L l'-bipheny l]-2-ylethynyl)-lH-indazol-5-yl)(2-(4-chlorophenyl)piperazin-l- yl)methanone,
Figure imgf000044_0002
(S)-(2-phenylpiperazin-l-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-lH-indazol-5-yl)methanone,
(R)-(2-phenylpiperazin-l-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-lH-indazol-5- yl)methanone.
(2-pheny Ipiperazin- l-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-l H-indazol-5 -yl)mcthanonc.
Figure imgf000045_0001
(R)-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-benzylpiperazin-l-yl)methanone,
(S)-(3-([ l,l'-biphenyl]-2-y lethynyl)-l H-indazol-5-yl)(2-benzy Ipiperazin- l-yl)methanone, (3-([ 1.1 '-biphenyl] -2-y lethynyl)- 1 H-indazol-5 -y l)(2-benzylpiperazin- 1 -y l)methanone,
Figure imgf000045_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)((2S,6S)-2,6-dimethylpiperazin-l- yl)methanone.
(3-(| 1. 1 '-biphenyl |-2-y lethynyl)- 1 H-indazol-5-yl)((2R.6S)-2.6-diincthy Ipiperazin- 1 - yl)methanone. (3-([ 1 , 1 '-biphenyl] -2-y lethynyl)- 1 H-indazol-5 -y l)((2R.6R)-2.6-dimethy Ipiperazin- 1 - yl)methanone.
Figure imgf000045_0003
(S)-(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-benzylpiperazin-l-yl)methanone, (R)-(3-(| 1. l'-bipheny l|-2 -y lethynyl)-! H-indazol-5 -y l)(2-benzy Ipiperazin- l-yl)methanone, (3-([ 1 , 1 '-biphenyl] -2-y lethynyl)- 1 H-indazol-5 -yl)(2-benzy Ipiperazin- 1 -yl)methanone,
(3-(| 1. 1 '-biphenyl |-2-y lethynyl)- 1 H-indazol-5-yl)(2-isopropy Ipiperazin- 1 -yl)mcthanonc.
Figure imgf000046_0001
(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2,9-diazaspiro[5.5]undecan-2- yl)methanone,
Figure imgf000046_0002
(3-([l,r-biphenyl]-2-yletliynyl)-lH-indazol-5-yl)(2,7-diazaspiro[4.5]decan-2-yl)methanone,
Figure imgf000047_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(8-oxa-l-azaspiro[4.5]decan-l- yl)methanone,
Figure imgf000047_0003
(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-oxa-6-azaspiro[3.4]octan-6- yl)methanone,
Figure imgf000047_0001
(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(8-oxa-2-azaspiro[4.5]decan-2- yl)methanone,
Figure imgf000048_0003
(3-((2-(pyridin-4-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2.7-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000048_0001
(3-((2-(pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone,
Figure imgf000048_0002
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[l,l'-biphenyl]-2-yl)ethynyl)-lH- indazol-5 -y l)methanone,
Figure imgf000049_0001
(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(6-methyl-2,6-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000049_0002
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(5-methyl-2,5-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000049_0003
(3-(| 1 J'-biphenyl|-2-ylethynyl)-lH-indazol-5-yl)(5-methyl-2,5-diazaspiro|3.5|nonan-2- yl)methanone.
Figure imgf000049_0004
(3-((2-(pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2.6-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000050_0001
(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(2-methyl-2,5-diazaspiro[3.5]nonan-5- yl)methanone,
Figure imgf000050_0002
(3-((2-(pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000050_0003
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((4-(trifluoromethoxy)-[l,l'-biphenyl]-2-yl)ethynyl)-lH- indazol-5 -y l)methanone,
Figure imgf000050_0004
(R)-2-((5-(3-(dimcthylammo)pyrrolidinc-l -carbonyl)- lH-indazol-3-yl)ctliynyl)-N- methylbenz amide,
(S)-2-((5-(3-(dimethylamino)pyrrolidine-l -carbonyl)- lH-indazol-3-y l)ethynyl)-N- methylbenz amide, 2-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)ethynyl)-N- methylbenzamide,
Figure imgf000051_0001
(R)-N-(2-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3- yl)ethynyl)phenyl)acetamide, (S)-N-(2-((5 -(3-(dimcthy lamino)py rrolidinc- 1 -carbonyl)- 1 H-indazol-3 - yl)ethynyl)phenyl)acetamide,
N-(2-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3- yl)ethynyl)phenyl)acetamide,
Figure imgf000051_0002
(2,6-diazaspiro|3.5|nonan-2-yl)(3-((5-(trifluoromethyl)-| 1 J '-biphenyl |-2-yl)ethynyl)-l H- indazol-5 -y l)methanone.
Figure imgf000051_0003
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[l,r-biphenyl]-2-yl)ethynyl)-lH- indazol-5 -y l)methanone,
Figure imgf000052_0003
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((2Xtrifluoromethoxy)-[l J'-biphenyl]-2-yl)ethynyl)-lH- indazol-5-yl)methanone,
Figure imgf000052_0001
(3-((2-(naphthalen-2-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone,
Figure imgf000052_0002
2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-N-methyl-[l,l'- bipheny 1] -2-carboxamide,
Figure imgf000053_0001
2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)etliynyl)-N-methyl-[l,r- bipheny 1] -3 -carboxamide,
Figure imgf000053_0002
N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-[l,r-biphenyl]-2- yl)acetamide,
Figure imgf000053_0003
2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-N-mediyl-[l,r- bipheny 1] -4-carboxamide,
Figure imgf000053_0004
(3-((2-(6-amino-4-methylpyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6- diazaspiro [3.5]nonan-2-y l)methanone,
Figure imgf000054_0001
(3-((2-(pyridin-4-yl)phenyl)ethynyl)-lH-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000054_0002
(3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yljmcthanonc.
Figure imgf000054_0003
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[l,r-biphenyl]-2-yl)etliynyl)-lH- indazol-5 -y l)metlianone,
Figure imgf000054_0004
(2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[l,r-biphenyl] -2-yl)ethynyl)-lH- indazol-5 -y l)metlianone,
Figure imgf000054_0005
(2,6-diazaspiro [3.5]nonan-2-yl)(3-((2'-(trifluorometlioxy )- [ 1 , l'-bipheny 1] -2-y l)ethyny 1)- 1 H-
Figure imgf000055_0001
(3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2.6-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000055_0002
(3-((2-(2-(pyrrolidin-l-yl)pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6- diazaspiro [3.5]nonan-2-yl)methanone.
Figure imgf000055_0003
(3-((2-(2-(piperazin- 1 -yl)pyridin-3 -yl)phenyl)ethy ny 1)- 1 H-indazol-5 -yl)(2,6- diazaspiro [3.5]nonan-2-y l)methanone,
Figure imgf000055_0004
(3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone,
Figure imgf000056_0001
(R)-2-((5-(3-(dimethylammo)pyrrolidine-l -carbonyl)- lH-indazol-3-yl)ethynyl)-N- pheny Ibenzamide . (S)-2-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)ethynyl)-N- pheny Ibenzamide , 2-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)ethynyl)-N- pheny Ibenzamide ,
Figure imgf000056_0002
(3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone.
Figure imgf000056_0003
(3-((2-(6-amino-4-mcthylpyridin-3-yl)phcnyl)cthynyl)-lH-indazol-5-yl)(2,6- diazaspiro [3.5 ] nonan-2 -y l)methanone ,
Figure imgf000056_0004
(R)-7-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)etliynyl)indolin-2-one,
(S)-7-((5-(3-(dmiethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)ethynyl)indolin-2-one,
7-((5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazol-3-yl)etliynyl)indolin-2-one,
Figure imgf000057_0001
(3-((2-(2-(pyrrolidin- 1 -y l)pyridin-3 -yl)pheny l)ethyny 1)- 1 H-indazol-5 -yl)(2,6- diazaspiro [3.5 ] nonan-2 -y l)mcthanonc ,
Figure imgf000057_0002
(3-((2-(2-(piperazin-l-yl)pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6- diazaspiro [3.5]nonan-2-y l)methanone.
Figure imgf000057_0003
(R)-(3-(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(4-methylpiperazine- 1 - carbonyl)phenyl)ethynyl)-lH-indazol-5-yl)methanone.
(S)-(3 -(dimethy lamino)pyrrolidin- 1 -y 1)(3 -((2-(4-methylpiperazine- 1 - carbonyl)phenyl)ethynyl)-lH-indazol-5-yl)methanone. (3-(dimethylamino)pyrrolidin-l-yl)(3-((2-(4-methylpiperazine-l-carbonyl)phenyl)ethynyl)- lH-mdazol-5-yl)methanone,
Figure imgf000058_0001
N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-[l,l'-biphenyl]-2- yl)acetamide,
Figure imgf000058_0002
2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-N-methyl-[l.r- bipheny 1] -3 -carboxamide,
Figure imgf000058_0003
2'-((5-(2.6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-N-methyl-[l,l'- biphenyl] -4-carboxamide,
Figure imgf000059_0001
(3-((2-(2-(4-methylpiperazin-l-yl)pyridin-3-yl)phenyl)ethynyl)-lH-indazol-5-yl)(2,6- diazaspiro [3.5]nonan-2-y l)methanone.
Figure imgf000059_0002
(S)-l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide,
(R)-l -(3-([ 1 , 1' -biphenyl] -2-ylethynyl)-l H-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide, l-(3-([l J '-biphenyl] -2-ylethynyl)-lH-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide,
Figure imgf000059_0003
N-(2'-((5-(2.6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-[l,r-biphenyl]-2- yl)pivalamide.
Figure imgf000059_0004
5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazole-3-carbonitrile,
(R)-5-(3-(dimethylamino)pyrrolidine-l-carbonyl)-lH-indazole-3-carbonitrile,
(S)-5-(3-(dimethylamino)pyrroli dine-1 -carbonyl)- lH-indazole-3-carbonitrileT
Figure imgf000060_0001
(3-([l,r-biphenyl]-2-yl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
(R)-(3-([l,r-biphenyl]-2-yl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
(S)-(3-([ 1 , 1 '-biphenyl] -2-y 1)- lH-indazol-5 -y 1) (3 -(dimethy lamino)pyrrolidin- 1 - yl)methanone,
Figure imgf000060_0002
(3-([l,r-biphenyl]-2-ylmethyl)-lH-indazol-5-yl)(3-(dimethylammo)pyrrolidin-l- yl)methanone,
(R)-(3-([l,r-biphenyl]-2-ylmethyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
(S)-(3-([l,r-biphenyl]-2-ylmethyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
Figure imgf000060_0003
(3-(2-([l,r-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin-l- yl)methanone,
(E)-(3-(2-([l,l,-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1 -yl)methanone, (R,E)-(3-(2-([l,l'-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3- (dimethy lamino)pyrrolidin- 1 -yl)methanone.
(S,E)-(3-(2-([l,l’-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3-
(dimethy lamin o)py rrolidin- 1 -yl)methanone,
(Z)-(3-(2-([l,l'-biphenyl]-2-yl)vinyl)-lEI-indazol-5-yl)(3-(dimethylamino)pyrrolidin- l-yl)methanone,
(R.Z)-(3-(2-([l,l'-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3-
(dimethy lamin o)py rrolidin- 1 -yl)methanone,
(S,Z)-(3-(2-([l,l'-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(3-
(dimethylamino)pyrrolidin-l-yl)methanone,
Figure imgf000061_0001
(3 -(2-([ 1, 1 ’-biphenyl] -2-yl)vinyl)- 1 H-indazol-5 -yl)(7-oxa-2-azaspiro|3.5 Jnonan-2- yl)methanone,
(E)-(3 -(2-([ 1 , l'-bipheny 1] -2-yl)viny 1)- 1 H-indazol-5-y l)(7-oxa-2-azaspiro [3.5]nonan- 2-yl)melhanone,
(Z)-(3-(2-([l.l'-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan- 2-yl)methanone,
Figure imgf000061_0002
(3 -(2-([ 1 , l'-biphenyl] -2-yl)vinyl)- 1 H-indazol-5 -yl)(7-methyl-2, 7- di azaspiro [3.5]nonan-2-yl)methanone.
(E)-(3-(2-([l.l'-biphenyl]-2-yl)vinyl)-lH-indazol-5-yl)(7-methyl-2.7- diazaspiro [3.5]nonan-2-yl)methanone,
(Z)-(3 -(2-([ 1 , l'-biphenyl] -2-yl)vinyl)- 1 H-indazol-5 -yl)(7-methyl-2, 7- di azaspiro [3.5]nonan-2-yl)methanone.
Figure imgf000062_0001
l-(l-(3-([l.l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea,
(R)-l-(l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea,
(S )- 1 -( 1 -(3 -([ 1. l'-bipheny 1] -2-ylethyny 1)- 1 H-indazole-5-carbony l)pyrrolidin-3-yl)-3- phenylurea.
Figure imgf000062_0002
3-(l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-3,4- dihydroquinazolin-2(lH)-one,
(R)-3-(l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-
3,4-dihydroquinazolin-2(lH)-one,
(S)-3-(l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-
3,4-dihydroquinazolin-2(lH)-one,
Figure imgf000062_0003
(3-([lJ'-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-((4,6-dimethylpyrimi din-2- y l)amino)pyrrolidin- 1 -y l)methanone, (R)-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-((4,6-dimethylpyrimi din-2- y l)amino)pyrrolidin- 1 -y l)methanone,
(S)-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin- 1 -yl)methanone,
Figure imgf000063_0001
N-(l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-2t3- dihydrobenzo[b] [ 1 ,4] dioxine-6-carboxamide,
(R)-N-(l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-
2,3-dihydrobenzo[b][l,4]dioxine-6-carboxamide, (S)-N-(l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3-yl)-
2,3-dihydrobenzo[b][l,4]dioxine-6-carboxamide,
Figure imgf000063_0002
N-(2-((l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (R)-N-(2-((l-(3-([l,l'-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluorornethyl)benzamide,
(S)-N-(2-((l-(3-([l,r-biphenyl]-2-ylethynyl)-lH-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide,
Figure imgf000064_0002
(3 -((2'-morpholino- [ 1 , l'-bipheny 1J -2-y l)ethy ny 1)- 1 H-indazol-5 -yl)(2,6- diazaspiro [3.5]nonan-2-yl)methanone,
Figure imgf000064_0001
7-(2-((5 -(2,6-diazaspiro [3.5]nonane-2-carbonyl)- 1 H-indazol-3 - yl)ethynyl)phenyl)isoindolin-l-one,
Figure imgf000065_0001
2'-((5-(2.6-diazaspiro[3.5]nonane-2-carbonyl)-lH-indazol-3-yl)ethynyl)-N.N- dimethyl-[ 1 , 1 '-biphenyl] -2-carboxamide,
Figure imgf000065_0002
(3-((2'-(pyrrolidin-l-ylmethyl)-[l,r-biphenyl]-2-yl)ethynyl)-lH-indazol-5-yl)(2,6- di azaspiro [3.5]nonan-2-yl)methanone.
Figure imgf000065_0003
(3-((2'-(morpholinomethyl)-[l,r-biphenyl]-2-yl)ethynyl)-lH-indazol-5-yl)(2,6- di azaspiro [3.5]nonan-2-yl)methanone.
Figure imgf000065_0004
Attorney Docket. No.046641-7059WO1(00159) (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(vinylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone, 1H-indazol-5-yl)(6-(ethylsulfonyl)-2,6-
Figure imgf000066_0001
5 diazaspiro[3.5]nonan-2-yl)methanone, yl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(2,6-
Figure imgf000066_0002
diazaspiro[3.5]nonan-2-yl)methanone, 10
Figure imgf000066_0003
thynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone,
Figure imgf000066_0004
ynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, - 64 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)meth
Figure imgf000067_0001
anone, 5 -2-
Figure imgf000067_0002
-2- yl)methanone. 10 In some aspects, provided herein, is a compound selected from those listed in Table A: Table A Compound reference code Molecular structure
Figure imgf000067_0003
- 65 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2134
Figure imgf000068_0001
- 66 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2530
Figure imgf000069_0001
- 67 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2535
Figure imgf000070_0001
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Figure imgf000071_0001
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Figure imgf000072_0001
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Figure imgf000073_0001
- 71 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2598
Figure imgf000074_0001
- 72 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2606
Figure imgf000075_0001
- 73 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2612
Figure imgf000076_0001
- 74 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2651
Figure imgf000077_0001
- 75 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-2678
Figure imgf000078_0001
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Figure imgf000079_0001
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Figure imgf000080_0001
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Figure imgf000081_0001
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Figure imgf000082_0001
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Figure imgf000083_0001
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Figure imgf000084_0001
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Figure imgf000085_0001
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Figure imgf000086_0001
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Figure imgf000087_0001
- 85 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3094
Figure imgf000088_0001
- 86 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3153
Figure imgf000089_0001
- 87 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3197
Figure imgf000090_0001
- 88 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3235
Figure imgf000091_0001
- 89 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3353
Figure imgf000092_0001
- 90 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3460
Figure imgf000093_0001
- 91 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3465
Figure imgf000094_0001
- 92 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3470
Figure imgf000095_0001
- 93 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3586
Figure imgf000096_0001
- 94 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3812
Figure imgf000097_0001
- 95 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3882
Figure imgf000098_0001
- 96 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CDD-3887
Figure imgf000099_0001
stereocenter can exist independently in either the (R) or (S) configuration. In certain embodiments, compounds described herein are present in optically active or racemic forms. It 5 is to be understood that the compounds described herein encompass racemic, optically-active, regioisomeric and stereoisomeric forms, or combinations thereof that possess the therapeutically useful properties described herein. Preparation of optically active forms is achieved in any suitable manner, including by way of non-limiting example, by resolution of the racemic form with recrystallization techniques, synthesis from optically-active starting 10 materials, chiral synthesis, or chromatographic separation using a chiral stationary phase. In certain embodiments, a mixture of one or more isomer is utilized as the therapeutic compound described herein. In other embodiments, compounds described herein contain one or more chiral centers. These compounds are prepared by any means, including stereoselective synthesis, enantioselective synthesis and/or separation of a mixture of 15 enantiomers and/ or diastereomers. Resolution of compounds and isomers thereof is achieved by any means including, by way of non-limiting example, chemical processes, enzymatic processes, fractional crystallization, distillation, and chromatography. The methods and formulations described herein include the use of N-oxides (if appropriate), crystalline forms (also known as polymorphs), solvates, amorphous phases, 20 and/or pharmaceutically acceptable salts of compounds having the structure of any compound(s) described herein, as well as metabolites and active metabolites of these compounds having the same type of activity. Solvates include water, ether (e.g., tetrahydrofuran, methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetates and the like. In certain embodiments, the compounds described herein exist in solvated forms with 25 pharmaceutically acceptable solvents such as water, and ethanol. In other embodiments, the compounds described herein exist in unsolvated form. In certain embodiments, the compound(s) described herein can exist as tautomers. All tautomers are included within the scope of the compounds presented herein. - 97 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) In certain embodiments, compounds described herein are prepared as prodrugs. A “prodrug” refers to an agent that is converted into the parent drug in vivo. In certain embodiments, upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically or therapeutically active form of the compound. In other 5 embodiments, a prodrug is enzymatically metabolized by one or more steps or processes to the biologically, pharmaceutically or therapeutically active form of the compound. In certain embodiments, sites on, for example, the aromatic ring portion of compound(s) described herein are susceptible to various metabolic reactions. Incorporation of appropriate substituents on the aromatic ring structures may reduce, minimize or eliminate 10 this metabolic pathway. In certain embodiments, the appropriate substituent to decrease or eliminate the susceptibility of the aromatic ring to metabolic reactions is, by way of example only, a deuterium, a halogen, or an alkyl group. Compounds described herein also include isotopically-labeled compounds wherein one or more atoms is replaced by an atom having the same atomic number, but an atomic 15 mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds described herein include and are not limited to 2H, 3H, 11C, 13C, 14C, 36Cl, 18F, 123I, 125I, 13N, 15N, 15O, 17O, 18O, 32P, and 35S. In certain embodiments, isotopically-labeled compounds are useful in drug and/or substrate tissue distribution studies. In other embodiments, substitution with heavier isotopes such as 20 deuterium affords greater metabolic stability (for example, increased in vivo half-life or reduced dosage requirements). In yet other embodiments, substitution with positron emitting isotopes, such as 11C, 18F, 15O, and 13N, is useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds are prepared by any suitable method or by processes using an appropriate isotopically-labeled 25 reagent in place of the non-labeled reagent otherwise employed. In certain embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels. The compounds described herein, and other related compounds having different 30 substituents are synthesized using techniques and materials described herein and as described, for example, in Fieser & Fieser’s Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd’s Chemistry of Carbon Compounds, Volumes 1-5 and Supplementals (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), Larock’s Comprehensive Organic Transformations (VCH Publishers Inc., - 98 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 1989), March, Advanced Organic Chemistry 4th Ed., (Wiley 1992); Carey & Sundberg, Advanced Organic Chemistry 4th Ed., Vols. A and B (Plenum 2000,2001), and Green & Wuts, Protective Groups in Organic Synthesis 3rd Ed., (Wiley 1999) (all of which are incorporated by reference for such disclosure). General methods for the preparation of 5 compound as described herein are modified by the use of appropriate reagents and conditions, for the introduction of the various moieties found in the formula as provided herein. Compounds described herein are synthesized using any suitable procedures starting from compounds that are available from commercial sources, or are prepared using procedures described herein. 10 In certain embodiments, reactive functional groups, such as hydroxyl, amino, imino, thio or carboxy groups, are protected in order to avoid their unwanted participation in reactions. Protecting groups are used to block some or all of the reactive moieties and prevent such groups from participating in chemical reactions until the protective group is removed. In other embodiments, each protective group is removable by a different means. Protective 15 groups that are cleaved under totally disparate reaction conditions fulfill the requirement of differential removal. In certain embodiments, protective groups are removed by acid, base, reducing conditions (such as, for example, hydrogenolysis), and/or oxidative conditions. Groups such as trityl, dimethoxytrityl, acetal and t-butyldimethylsilyl are acid labile and are used to 20 protect carboxy and hydroxy reactive moieties in the presence of amino groups protected with Cbz groups, which are removable by hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic acid and hydroxy reactive moieties are blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl, in the presence of amines that are blocked with acid labile groups, such as t-butyl carbamate, or with carbamates that are both 25 acid and base stable but hydrolytically removable. In certain embodiments, carboxylic acid and hydroxy reactive moieties are blocked with hydrolytically removable protective groups such as the benzyl group, while amine groups capable of hydrogen bonding with acids are blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties are protected by conversion to simple ester 30 compounds as exemplified herein, which include conversion to alkyl esters, or are blocked with oxidatively-removable protective groups such as 2,4-dimethoxybenzyl, while co- existing amino groups are blocked with fluoride labile silyl carbamates. Allyl blocking groups are useful in the presence of acid- and base- protecting groups since the former are stable and are subsequently removed by metal or pi-acid catalysts. For - 99 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) example, an allyl-blocked carboxylic acid is deprotected with a palladium-catalyzed reaction in the presence of acid labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate is attached. As long as the residue is attached to the resin, that functional group is blocked 5 and does not react. Once released from the resin, the functional group is available to react. Typically blocking/protecting groups may be selected from: .
Figure imgf000102_0001
o the creation of protecting groups and their removal are described in Greene & Wuts, Protective 10 Groups in Organic Synthesis, 3rd Ed., John Wiley & Sons, New York, NY, 1999, and Kocienski, Protective Groups, Thieme Verlag, New York, NY, 1994, which are incorporated herein by reference for such disclosure. Compositions 15 In one aspect, the present disclosure provides a pharmaceutical composition comprising at least one compound of the present disclosure and at least one pharmaceutically acceptable carrier. The compositions containing the compound(s) described herein include a pharmaceutical composition comprising at least one compound as described herein and at 20 least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises Kolliphor EL, and aqueous buffer, or a combination thereof. In certain embodiments, the aqueous buffer comprises phosphate buffered saline (PBS). In some embodiments, the aqueous buffer comprises 1x PBS. In certain embodiments, the - 100 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) pharmaceutical composition comprises about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, or about 40% Kolliphor EL. In some embodiments, the pharmaceutical composition comprises about 20% Kolliphor EL in 1x PBS. In certain embodiments, the composition is formulated for an administration route 5 such as oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal, intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. In some embodiments, the composition is formulated 10 as a pill, tablet, gelcap, or capsule for oral administration. Methods of Treatment, Amelioration, and/or Prevention In one aspect, the present disclosure relates to a method of promoting male contraception in a male subject. In another aspect, the present disclosure relates to a method 15 of promoting male sterility in a male subject. In yet another aspect, the present disclosure relates to a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject. In yet another aspect, the present disclosure relates to a method of reducing testes size in a male subject. In yet another aspect, the present disclosure relates to a method of modulating a RET, STK33, and/or CLK kinase function and/or activity in a subject. 20 In certain embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound of the disclosure. In certain embodiments, the method comprises administering to the male subject a therapeutically effective amount of a compound of the disclosure. In certain embodiments, the method comprises administering to the subject a 25 therapeutically effective amount of a compound of Formula (I). In certain embodiments, the method comprises administering to the male subject a therapeutically effective amount of a compound of Table A. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound 30 disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an - 101 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) effective amount of a compound of Formula (II) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some 5 embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (V) and/or the pharmaceutical composition 10 thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting STK33 in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VII) 15 and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound disclosed herein, and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method 20 comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (II), and/or a pharmaceutical composition thereof. In some 25 embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 30 compound of Formula (IV), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (V), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method - 102 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 5 compound of Formula (VII), and/or a pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of 10 promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 15 compound of Formula (II) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception 20 and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (V) and/or the 25 pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of promoting male contraception and/or infertility in a male 30 subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VII) and/or the pharmaceutical composition thereof. In some embodiments, the method provides a contraceptive effect in the male subject. In some embodiments, the compound is administered orally to the male subject. In some embodiments, provided herein is a method of minimizing and/or reducing - 103 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the 5 method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (II) and/or the 10 pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa 15 number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a 20 compound of Formula (V) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of 25 minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of a compound of Formula (VII) and/or the pharmaceutical composition thereof. In some embodiments, the method provides a contraceptive effect in the male subject. In some embodiments, the compound is administered orally to the male subject. 30 In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (I) and/or the - 104 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (II) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method 5 comprising administering the mammal an effective amount of a compound of Formula (III) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, 10 the method comprising administering the mammal an effective amount of a compound of Formula (V) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of inhibiting RET in 15 a mammal, the method comprising administering the mammal an effective amount of a compound of Formula (VII) and/or the pharmaceutical composition thereof. RET inhibitors can be used to treat, ameliorate, and/or prevent cancer, including but limited to medullary thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A). In some embodiments, provided herein is a method of 20 treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression and/or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, the method comprises treating a RET mediated cancer. In some embodiments, the RET mediated cancer comprises medullary 25 thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A). In some embodiments, the disease or disorder comprises Hirschprung Disease, central hypoventilation syndrome, or renal agenesis. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the 30 method comprising administering the mammal a therapeutically effective amount of a compound of Formula (I) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of - 105 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Formula (II) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (III) and/or the 5 pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (IV) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or 10 preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (V) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method 15 comprising administering the mammal a therapeutically effective amount of a compound of Formula (VI) and/or the pharmaceutical composition thereof. In some embodiments, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of a compound of Formula (VII) and/or the 20 pharmaceutical composition thereof. CLKs family comprises four homologous proteins, CLK1, CLK2, CLK3, and CLK4. They contribute to cell cycle progression, cell growth and disease occurrence via the regulation of splicing. In addition, human CLK2 may play a role in fatty liver disease through participating in fatty acid oxidation and ketogenesis. CLKs act as important regulators and 25 exert important functions which are essential in catalyzing splicing and modulating phosphorylation which is key in many diseases like Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and cancers (renal cancer, breast cancer, melanoma). In one aspect, provided herein is a method of modulating at least one CLK family 30 protein such as CLK1, CLK2, CLK3, and/or CLK4. In one aspect, provided herein is a method of inhibiting at least one of CLK1, CLK2, CLK3, or CLK4 in a mammal, the metho comprising administering the mammal an effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof. In another aspect, provided herein is a method of treating, ameliorating, and/or preventing a disease or disorder caused by - 106 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound disclosed herein and/or the pharmaceutical composition thereof. In some embodiments, the disease or disorder comprises Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and 5 renal cancer, breast cancer, or a melanoma. In some embodiments, the compound of formula (I) is a compound of formula (II). Exemplary compounds of formula (II) are described elsewhere herein. In other embodiments, the compound of formula (I) is a compound of formula (III). Exemplary compounds of formula (III) are described elsewhere herein. In yet other embodiments, the compound of 10 formula (I) is a compound of formula (IV). Exemplary compounds of formula (IV) are described elsewhere herein. In yet other embodiments, the compound of formula (I) is a compound of formula (V). Exemplary compounds of formula (V) are described elsewhere herein. In yet other embodiments, the compound of formula (I) is a compound of formula (VI). Exemplary compounds of formula (VI) are described elsewhere herein. In yet other 15 embodiments, the compound of formula (I) is a compound of formula (VII). Exemplary compounds of formula (VII) are described elsewhere herein. The methods described herein include administering to the subject a therapeutically effective amount of at least one compound described herein, which is optionally formulated in a pharmaceutical composition. In various embodiments, a therapeutically effective amount 20 of at least one compound described herein present in a pharmaceutical composition is the only therapeutically active compound in a pharmaceutical composition. In certain embodiments, the method further comprises administering to the subject an additional agent that treats, ameliorates, and/or prevents the contemplated condition. The compound of the disclosure can be administered to the subject using any 25 administration route known to a person of skill in the art. Exemplary routes of administration are described elsewhere herein. In some embodiments, a composition comprising a compound of the disclosure is orally administered to the subject. In some embodiments, a pill, tablet, gelcap, or capsule comprising a compound of the disclosure is orally administered to the subject. 30 The compound of the disclosure can be administered to the subject in any dosage with any timing of dosage administration necessary to inhibit STK, or a combination thereof and to provide a desired therapeutic effect. In some embodiments, the compound of the disclosure is administered to the male subject in order to provide a contraceptive effect in the subject. Although not wishing to be limited by theory, it is believed that inhibition of STK33 function - 107 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) in spermatocytes and spermatids leads to a reduction in spermatozoa number and motility. In certain embodiments, the subject is a mammal. In other embodiments, the mammal is a human. 5 Administration/Dosage/Formulations The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the subject either prior to or after the onset of the disease or disorder. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a 10 bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation. Administration of the compositions described herein to a patient, preferably a mammal, more preferably a human, may be carried out using known procedures, at dosages 15 and for periods of time effective to treat the disease or disorder in the patient. An effective amount of the therapeutic compound necessary to achieve a therapeutic effect may vary according to factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat the disease or disorder in the patient. Dosage regimens may be adjusted to provide the optimum therapeutic 20 response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A non- limiting example of an effective dose range for a therapeutic compound described herein is from about 1 and 5,000 mg/kg of body weight/per day. One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective 25 amount of the therapeutic compound without undue experimentation. Actual dosage levels of the active ingredients in the pharmaceutical compositions described herein may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. 30 In particular, the selected dosage level depends upon a variety of factors including the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in - 108 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) the medical arts. A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds 5 described herein employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as 10 used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the compound(s) described herein are dictated by and directly dependent on (a) the unique characteristics of the therapeutic compound and the particular 15 therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding/formulating such a therapeutic compound. In certain embodiments, the compositions described herein are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions described herein comprise a therapeutically effective amount of 20 a compound described herein and a pharmaceutically acceptable carrier. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, C3-C7 aliphatic or aryl n-hydroxyl compound, wherein n is 1 to 10, e.g. glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by 25 the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it is preferable to include isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as 30 mannitol and sorbitol, in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin. In certain embodiments, the compositions described herein are administered to the patient in dosages that range from one to five times per day or more. In other embodiments, - 109 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) the compositions described herein are administered to the patient in range of dosages that include, but are not limited to, once every day, every two, days, every three days to once a week, and once every two weeks. It is readily apparent to one skilled in the art that the frequency of administration of the various combination compositions described herein varies 5 from individual to individual depending on many factors including, but not limited to, age, disease or disorder to be treated, gender, overall health, and other factors. Thus, administration of the compounds and compositions described herein should not be construed to be limited to any particular dosage regime and the precise dosage and composition to be administered to any patient is determined by the attending physician taking all other factors 10 about the patient into account. In certain embodiments, a composition as described herein is a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound described herein, alone or in combination with a second pharmaceutical agent; and instructions for using the compound to treat, or reduce one or more 15 symptoms of a disease or disorder in a patient. Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration, known to the art. The pharmaceutical preparations may be sterilized and if 20 desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other analgesic agents. Routes of administration of any of the compositions described herein include oral, 25 nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical. The compounds for use in the compositions described herein can be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, 30 subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, - 110 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions described herein are not limited to the particular formulations and compositions that are 5 described herein. Oral Administration For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps. The compositions intended for oral use may be prepared according to any method known in the art and such compositions may contain 10 one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients that are suitable for the manufacture of tablets. Such excipients include, for example an inert diluent such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate. The tablets may be uncoated or they may be coated by known techniques for elegance or to 15 delay the release of the active ingredients. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert diluent. For oral administration, the compound(s) described herein can be in the form of tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., polyvinylpyrrolidone, hydroxypropylcellulose or 20 hydroxypropyl methylcellulose); fillers (e.g., cornstarch, lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrates (e.g., sodium starch glycollate); or wetting agents (e.g., sodium lauryl sulphate). If desired, the tablets may be coated using suitable methods and coating materials such as OPADRY™ film coating systems available from Colorcon, West Point, Pa. (e.g., OPADRY™ OY Type, OYC 25 Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and OPADRY™ White, 32K18400). Liquid preparation for oral administration may be in the form of solutions, syrups or suspensions. The liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agent (e.g., 30 lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbic acid). Parenteral Administration For parenteral administration, the compounds as described herein may be formulated for injection or infusion, for example, intravenous, intramuscular or subcutaneous injection or - 111 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) infusion, or for administration in a bolus dose and/or continuous infusion. Suspensions, solutions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing and/or dispersing agents may be used. Sterile injectable forms of the compositions described herein may be aqueous or 5 oleaginous suspension. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1, 3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer’s 10 solution and isotonic sodium chloride solution. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or di-glycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils, such as olive oil or castor oil, especially in their 15 polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv or similar alcohol. Additional Administration Forms Additional dosage forms suitable for use with the compound(s) and compositions described herein include dosage forms as described in U.S. Patents Nos.6,340,475; 20 6,488,962; 6,451,808; 5,972,389; 5,582,837; and 5,007,790. Additional dosage forms suitable for use with the compound(s) and compositions described herein also include dosage forms as described in U.S. Patent Applications Nos.20030147952; 20030104062; 20030104053; 20030044466; 20030039688; and 20020051820. Additional dosage forms suitable for use with the compound(s) and compositions described herein also include dosage forms as 25 described in PCT Applications Nos. WO 03/35041; WO 03/35040; WO 03/35029; WO 03/35177; WO 03/35039; WO 02/96404; WO 02/32416; WO 01/97783; WO 01/56544; WO 01/32217; WO 98/55107; WO 98/11879; WO 97/47285; WO 93/18755; and WO 90/11757. Dosing 30 The therapeutically effective amount or dose of a compound described herein depends on the age, sex and weight of the patient, the current medical condition of the patient and the progression of the disease or disorder in the patient being treated. The skilled artisan is able to determine appropriate dosages depending on these and other factors. A suitable dose of a compound described herein can be in the range of from about - 112 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1,000 mg, for example, from about 1 mg to about 500 mg, such as about 5 mg to about 250 mg per day. The dose may be administered in a single dosage or in multiple dosages, for example from 1 to 4 or more times per day. When multiple dosages are used, the amount of each dosage may be 5 the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses, with about a 12-hour interval between doses. It is understood that the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 5 mg per day dose may 10 be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on. In the case wherein the patient’s status does improve, upon the doctor’s discretion the administration of the compound(s) described herein is optionally given continuously; alternatively, the dose of drug being administered is temporarily reduced or temporarily 15 suspended for a certain length of time (i.e., a “drug holiday”). The length of the drug holiday optionally varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday includes 20 from 10%-100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. Once improvement of the patient’s conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, is reduced to a level at which the improved disease is retained. In certain embodiments, 25 patients require intermittent treatment on a long-term basis upon any recurrence of symptoms and/or infection. The compounds described herein can be formulated in unit dosage form. The term “unit dosage form” refers to physically discrete units suitable as unitary dosage for patients undergoing treatment, with each unit containing a predetermined quantity of active material 30 calculated to produce the desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose or one of multiple daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose. Toxicity and therapeutic efficacy of such therapeutic regimens are optionally - 113 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) determined in cell cultures or experimental animals, including, but not limited to, the determination of the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD50 and 5 ED50. The data obtained from cell culture assays and animal studies are optionally used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage optionally varies within this range depending upon the dosage form employed and the route of administration utilized. 10 Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of this disclosure and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction 15 times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, with art- recognized alternatives and using no more than routine experimentation, are within the scope of the present application. It is to be understood that wherever values and ranges are provided herein, all values 20 and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present disclosure. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application. The following examples further illustrate aspects of the present disclosure. However, 25 they are in no way a limitation of the teachings or disclosure of the present disclosure as set forth herein. EXPERIMENTAL EXAMPLES The disclosure is further described in detail by reference to the following 30 experimental examples. These examples are provided for purposes of illustration only, and are not intended to be limiting unless so specified. Thus, the disclosure should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein. - 114 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the compounds of the present disclosure and practice the claimed methods. The following working examples therefore, specifically point out the preferred embodiments of the present 5 disclosure, and are not to be construed as limiting in any way the remainder of the disclosure. Materials and Methods Reagents and solvents purchased from commercial supplies were used as received. All reactions involving air-sensitive reagents were carried out in anhydrous solvents under an 10 atmosphere of nitrogen. Reactions were monitored by thin-layer chromatography (TLC) on Baker-flex® silica gel plates (IB2-F) using UV-light (254 and 365 nm) detection or liquid chromatography/mass spectrometry (LC-MS). Column chromatography was carried out using Teledyne ISCO CombiFlash system equipped with either a silica or C-18 column. NMR spectra were recorded at room temperature using a Bruker Avance III HD 600 MHz 15 spectrometer (1H NMR at 600 MHz and 13C NMR at 150 MHz) or Bruker Avance 800 MHz spectrometer (1H NMR at 800 MHz and 13C NMR at 200 MHz). Chemical shifts (δ) are reported in parts per million (ppm) with reference to solvent signals [1H-NMR: CDCl3 (7.26 ppm), CD3OD (3.31 ppm), and DMSO-d6 (2.50 ppm); 13C-NMR: CDCl3 (77.00 ppm), CD3OD (49.15 ppm), and DMSO-d6 (39.51 ppm)]. Signal patterns are reported as s (singlet), 20 d (doublet), t (triplet), q (quartet), h (heptet), m (multiplet), and br (broad). Coupling constants (J) are given in Hz. High-resolution mass spectra (HRMS) were recorded on a ThermoFisher Scientific Q Exactive hybrid quadrupole-Orbitrap mass spectrometer using ESI (electrospray ionization). Abbreviation Definition AcOH acetic acid Boc t-butyloxycarbonyl CH2Cl2 dichloromethane CH3OH methanol Cs2CO3 cesium carbonate CuCl copper(I) chloride CuI copper(I) iodide DHP 3,4-dihydro-2H-pyran - 115 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) DIEA N,N-diisopropylethylamine DME 1,2-dimethoxyethane DMF N,N-dimethylformamide EtOAc ethyl acetate O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium HATU hexafluorophosphate HCl hydrochloric acid KHF2 potassium bifluoride K2CO3 potassium carbonate KOH potassium hydroxide LiOH lithium hydroxide NaHCO3 sodium bicarbonate Na2SO4 sodium sulfate Pd(OAc)2 palladium(II) acetate PdCl2(PPh3)2 bis(triphenylphosphine)palladium(II) dichloride Pd(PPh3)4 tetrakis(triphenylphosphine)palladium(0) RuPhos 2-dicyclohexylphosphino-2’,6’-diisopropoxybiphenyl TEA trimethylamine THF tetrahydrofuran THP tetrahydropyranyl p-TsOH p-toluenesulfonic acid Xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene General synthetic procedures (a) General procedure for THP protection: A mixture of indazole (5.0 mmol, 1.0 equiv.), DHP (7.5 mmol, 1.5 equiv.), and p-TsOH 5 monohydrate (1.0 mmol, 0.2 equiv.) in anhydrous CH2Cl2 (10 mL) was stirred under nitrogen at room temperature for 16 h. The mixture was quenched by the addition of saturated aqueous NaHCO3 and extracted twice with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography on silica (EtOAc/hexanes, 0:100 to 40:60) to afford the desired 10 product. (b) General procedure for Sonogashira coupling: - 116 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) A oven dried microwave vial equipped with magnetic stir bar was charged with halide (0.5 mmol, 1.0 equiv.), substituted alkyne (0.6 mmol, 1.2 equiv.), CuI (0.05 mmol, 0.1 equiv.), and PdCl2(PPh3)2 (0.05 mmol, 0.1 equiv.). The vial was sealed with a microwave cap, evacuated and backfilled with nitrogen, followed by addition of anhydrous DMF (1 mL) and 5 TEA (1 mL) via syringe. The mixture was vacuum-purged and refilled with nitrogen for three cycles and then heated by microwave reactor at 80 oC for 1 h (or at indicated temperature for indicated period of time). After completion of the reaction (monitored by TLC and LC-MS), the reaction mixture was filtered through a pad of celite, and the filtrate was extracted between EtOAc and 1N HCl aqueous solution. The organic layer was washed with saturated 10 aqueous NaHCO3 and brine, dried over anhydrous Na2SO4, filtered, and concentred under reduced pressure. The residue was purified by flash chromatography on silica with EtOAc/hexanes or CH3OH/CH2Cl2 gradient elution to afford the desired product. (c) General procedure for hydrolysis without THP-protected amine: To a solution of methyl ester (0.5 mmol, 1.0 equiv.) in THF/water (2 mL, 1:1, v/v) was added 15 KOH (5.0 mmol, 10.0 equiv.), and the mixture was vigorously stirred at 40 oC for 48 h or until the disappearance of the starting material (monitored by TLC and LC-MS). The reaction mixture was neutralized to pH 7 with 4N HCl aqueous solution and concentrated in vacuo. The crude carboxylic acid was directly used in the next step. (d) General procedure for hydrolysis with THP-protected amine: 20 To a solution of methyl ester (0.5 mmol, 1.0 equiv.) in THF/water (2 mL, 1:1, v/v) was added LiOH monohydrate (1 mmol, 2.0 equiv.), and the mixture was vigorously stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC and LC-MS), the reaction mixture was neutralized to pH 7 with 4N HCl aqueous solution and concentrated in vacuo. The residue was purified by reverse-phase column chromatography on C-18 25 (CH3OH/water, 5:95 to 100:0) to afford the desired product. (e) General procedure for amide coupling: To a solution of carboxylic acid (0.1 mmol, 1.0 equiv.), substituted amine (0.12 mmol, 1.2 equiv.), and HATU (0.12 mmol, 1.2 equiv.) in anhydrous DMF (0.5 mL) was added DIEA (0.15 mmol, 1.5 equiv.) under nitrogen. The reaction mixture was stirred at room temperature 30 for 16 h. The mixture was quenched by the addition of water and extracted twice with EtOAc. The combined organic layers were washed with saturated aqueous NaHCO3 and brine, dried over anhydrous Na2SO4, filtered, and evaporated under reduced pressure. The residue was purified by flash chromatography on silica (CH3OH/CH2Cl2, 0:100 to 10:90) or reverse-phase column chromatography on C-18 (CH3OH/water, 5:95 to 100:0) to afford the desired product. - 117 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (f) General procedure for removal of THP and/or Boc-protecting group: A mixture of THP- and/or Boc-protected amine (0.1 mmol) and 4N HCl solution in 1,4- dioxane (0.5 mL) was stirred at room temperature for 1 h. After completion of the reaction (monitored by TLC and LC-MS), the resulting mixture was neutralized to pH 7 with 4M 5 NaOH aqueous solution and concentrated in vacuo. The residue was purified by reverse- phase column chromatography on C-18 (CH3OH/water, 5:95 to 100:0) to afford the desired product. Biological and characterization data of synthesized compounds 10 All compounds were summarized into three series A, B, and C based on modification of the scaffold. Preparation of compounds were achieved via synthetic route 1, route 2, or as described in synthesis. IC50 values and corresponding Kd values of biochemical activity against STK33 were calculated from Lanthascreen binding assay, and IC50 values of cellular activity against STK33 were calculated from NanoBRET Target Engagement intracellular 15 kinase assay. ND means not determined either for compounds not passing kinase assay screening against STK33 at 500 nM or for compounds with IC50 >300 nM in the NanoBRET assay against STK33. Example 1: Discovery of potent STK33 inhibitors from DNA-encoded chemical library 20 screening and hit optimization To identify small-molecule binders to the kinase domain (KD) of STK33, DNA- Encoded Chemistry Technology (DEC-Tec) was used to uncover potent and selective kinase inhibitors. Using a full length His-tagged STK33 protein at either 0.1 μM or 0.5 μM (with or 25 without staurosporine, a broad spectrum ATP-competitive kinase inhibitor), selections of 36 or 45 unique libraries were performed, respectively, each containing 3.9 billion unique DNA- encoded molecules. Calculation of dissociation constant (Kd) values for kinase inhibitors that bind to the ATP pocked of a kinase from LanthaScreen binding data. 30 Dynafit was used to fit complex equilibria where a tracer is displaced by a dilution series of a kinase inhibitors. This method relies on the concentration of inhibitor, as well as the concentration of kinase and tracer, and Kd of the tracer. The latter three parameters are tabulated by ThermoFisher for each kinase in the LanthaScreen assay panel. Calculation of inhibition constant (Ki) values for kinase inhibitors from Z’- - 118 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) LYTE and Adapta inhibition data. The Morrison equation with [ATP] = Km app for competitive inhibition (Graphpad Prism7 software) was used to get Ki values. Since Kd and Ki values are defined the same way, i.e., Kd = [E] [I] / [EI] = Ki, Kd and Ki values can be compared. 5 NanoBRET™ Target Engagement intracellular kinase assay HEK293T cells were cultured in DMEM (Gibco11965-092) with 10% FBS and 1% Pen-Strep (Gibco 15140-122) in 5% CO2, 37°C. The cell line underwent authentication by short tandem repeat (STR) profiling at Cytogenetics and Cell Authentication Core (CCAC) MD Anderson Cancer Center. NanoBRET™ TE intracellular kinase assay, K-10 assay (Cat. 10 # N2641), K-9 assay (Cat. # N2631), transfection reagent FuGENE® HD (Cat. # E2311), transfection carrier DNA (Cat. # E4881), NanoLuc®-STK33 fusion vector (Cat. # NV2111), NanoLuc®- RET fusion vector (Cat. # NV1951), NanoLuc®-CLK1 fusion vector (Cat. # NV1131), NanoLuc®-CLK2 fusion vector (Cat. # NV1141) and NanoLuc®-CLK4 fusion vector (Cat. # NV1151) were purchased from Promega. The measurement of test compound 15 engagement with cellular target protein was performed in a 384-well format according to manufacturer’s instructions. HEK293 cells were transiently transfected with NanoLuc® (STK33, RET, CLK1, CLK2, and CLK4) fusion vectors. After 36 hours of transfection, cells were treated with fixed concentration of tracer (recommended by Promega) followed by compounds (inhibitors) for 2 hours of incubation. To determine the test compound affinity, 20 cells were titrated with varying concentrations of test compounds ranging from 40 µM to 0.4 nM at a fixed tracer concentration. Staurosporine was measured in parallel as a positive control. Freshly prepared NanoBRET Nano-Glo substrate plus extracellular NanoLuc inhibitor was then added to initiate the subsequent bioluminescence resonance energy transfer (BRET) measurements using a CLARIOstar Plus BMG LABTECH plate reader. Data 25 analysis was done by measuring the ratio of acceptor emission to donor emission (BRET ratio) and normalized by subtracting no-tracer-control-background. NanoBRET Target Engagement K192 assay. HEK293 cells (ATCC) were cultured in DMEM (Gibco) + 10% FBS (Seradigm), and incubated in a humidified 37°C/5% CO2 incubator. The NanoBRET Target Engagement K192 Kinase Selectivity System (Promega) 30 contains 192 unique transfection-ready NanoLuc/Kinase fusions, pre-diluted in either transfection carrier DNA or relevant cyclin DNA. These NanoLuc/Kinase fusion constructs were transfected into HEK293 cells using Fugene HD (Promega) according to the manufacturer’s protocol. Briefly, 10 µL of a 20 µg/mL DNA solution was added to white, TC-treated 96 well assay plates (Corning), followed by 30 µL of a diluted Fugene HD - 119 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) solution (20 µL Fugene/1 mL Opti-MEM). After transfection complex formation, HEK293 cells were resuspended at 3.3 × 105 c/mL in Opti-MEM + 1% FBS and added at a volume of 60 µL per well, to result in approximately 20000 cells/well. Cells were incubated in a humidified 37°C/5% CO2 incubator for 20 hours. Tracer K-10 (Promega) was prepared at a 5 100× concentration in DMSO (Sigma) and diluted to a working concentration of 10× in tracer dilution buffer (12.5 mM HEPES, 31.25% PEG400, pH 7.5) (Promega). Test compounds were prepared as concentrated stock solutions in DMSO and diluted in Opti-MEM to prepare 10× working stocks. Cells were equilibrated with test compounds and Tracer K-10 for 2 hours prior to BRET measurements. For target engagement analysis, Tracer K-10 was added 10 to cells at one of 4 different concentrations, ranging from 25 nM to 1 µM, based on target affinity, according to manufacturer’s protocol. To measure BRET, NanoBRET NanoGlo Substrate and Extracellular NanoLuc Inhibitor (Promega) were added according to the manufacturer’s protocol, and filtered luminescence was measured on a GloMax Discover luminometer equipped with 450 nm BP filter (donor) and 600 nm LP filter (acceptor), using 15 0.5s integration time. Milli-BRET units are calculated by multiplying the raw BRET units by 1000. For Fractional Occupancy determination, the following equation was used: % Occupancy = [1-(X-Z)/(Y-Z)]×100 Where X = BRET in the presence of the test compound and Tracer K10, Y = BRET in the presence of Tracer K10 only, and Z= Full occupancy, or BRET in the absence of the test 20 compound and Tracer K10. In this case, an untagged NanoLuc construct (Promega) was used for the full occupancy control. DEC-Tec affinity selections For these studies, two independent DEC-Tec selections of our in-house libraries were performed using full length recombinant human STK33 protein with an N-terminal His6 tag 25 (Eurofins #14-671). In Experiment 1, selection of 45 libraries (3.940 billion compounds) was performed with STK33 protein at 0.5 μM in the absence or presence of staurosporine (10 μM). In Experiment 2, selection of 36 libraries (3.936 billion compounds) was performed with STK33 protein at 0.1 μM. Selection methods and informatics were performed as described previously. 30 STK33 KD protein expression and purification for crystallography Human STK33 KD (GenBank accession number CAC29064.1, residues 99−383) with tobacco etch virus (TEV) cleavage site and cGFP-Strep-tag®II was expressed in insect SF9 cells (Invitrogen). SF9 cells were cultured in SF900III medium (Gibco) and infected with STK33 KD baculovirus in an optimal virus/ insect cells ratio. Cells were harvested 48 h after - 120 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) infection; pellets were kept at −80°C. For purification, frozen cell pellet was resuspended in lysis buffer (25 mM Tris, pH 8.0, 150 mM NaCl, and 1mM TCEP) and lysed using homogenizer (Avestin). After centrifugation at 48,000g for 1 h, the soluble fraction was loaded onto pre-equilibrated Strep-Tactin Superflow resin (IBA Lifesciences GmbH) and 5 eluted with elution buffer (25 mM Tris, pH 8.0, 150 mM NaCl, 1 mM TCEP, and 2 mM desthibiotin). Fractions containing the protein were treated with TEV protease (1:60 weight ratio, overnight) to remove the GFP tag and the mixture was subjected to Superdex™ 75 Increase 10/300 GL gel filtration column (Cytiva) equilibrated with the abovementioned lysis buffer. The peak corresponding to the estimated molecular mass of approximately 70 kDa 10 was pooled and used for co-crystallization. From library qDOS28_1 in both selection experiments, several enriched hits were discovered that shared the identical building block 2 (BB2) and building block 3 (BB3) and similar/identical building block 1 (BB1) that only differed in their linker (FIG.1 and FIGs. 5A-5B). These hits were absent in the selection performed in the presence of staurosporine, 15 indicating competitive binding of the hits. The hit with a short linker, CDD-2110 (FIG.2A), along with the hit featuring a long linker, CDD-3348 (Table B), were synthesized off-DNA and confirmed to be potent binders of STK33 in a LanthaScreen binding assay performed at ThermoFisher (Kd = 0.1 and 0.06 nM, respectively; FIG.2C and Table B) and a Promega cell-based NanoBRET assay performed in-house (IC50 = 38 and 169 nM, respectively; FIG. 20 2C and Table B). Compared to two STK33 inhibitors, namely compound 1 (Kd = 1.7 nM and IC50 = 748 nM; Table B) and ML281 (Kd = 39.6 nM and IC50 = 7707 nM; Table B), both CDD-2110 and CDD-3348 demonstrated prominent potency toward STK33 in both biochemical and cellular assays. 25 Table B. Chemical properties, biochemical activity, cellular activity, and metabolism data of the hit with a long linker, CDD-3348, and two STK33 inhibitors, namely compound 1 and ML281. Kd and Ki values were calculated from LanthaScreen binding assay and Z’-LYTE assay, respectively, as described in the method section; IC50 values were calculated from NanoBRET assay; t1/2 was measured using either mouse liver microsomal or human liver 30 microsomal stability assay. Reported IC50 values of compound 1 and ML281 are 7 and 14 nM, respectively. ND means not determined. - 121 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159)
Figure imgf000124_0001
. . . LanthaScreen STK33 0.06 ± 0.01 1.7 ± 0.2 39.6 ± 1.5 Kd (nM) CLK4 26.2 ± 1.5 1.2 ± 0.1 >1000 Biochemical a CLK2 50.4 ± 3.1 8.9 ± 0.9 >1000 activity Z’LYTE RET 5.7 ± 0.5 2.6 ± 0.2 >1000 Ki (nM) CLK1 480 ± 209 21.0 ± 4.0 >1000 STK33 169 748 7707 CLK4 3846 (23X) 41 ND Cellular NanoBRET CLK2 2531 (15X) 202 ND activity IC50 (nM) RET 6411 (38X) 956 ND CLK1 6732 (40X) 406 ND Metabolic MLM, t1/2 (min) 14 ND ND stability HLM, t1/2 (min) 13 ND ND Although CDD-2110 and CDD-3348 demonstrate potent activity, both molecules possess racemic formats, have large molecular weights (>500 Da), and are metabolically 5 labile when incubated in mouse liver microsomes (MLM) and human liver microsomes (HLM), respectively (FIG.2C and Table B). To potentially overcome these drawbacks, two truncated enantiomers, CDD-2211 (R-isomer) and CDD-2212 (S-isomer) (FIG.2B), were subsequently synthesized and assayed, with CDD-2211 revealing outstanding inhibitory activity against STK33 in LanthaScreen (Kd = 0.02 nM) and NanoBRET (IC50 = 5 nM) 10 assays when compared to the antipode CDD-2212 (Kd = 1.9 nM and IC50 = 999 nM), as well as the original hits CDD-2110 and CDD-3348 (FIG.2C). Despite its potent binding to STK33, the half-life of CDD-2211 remained short (t1/2 ≤10 min; FIG.2C) in MLM and HLM metabolic stability assays relevant to contraceptive in mice or men. - 122 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Example 2: Chemical synthesis of STK33 inhibitors Further medicinal chemistry efforts were invested to improve metabolic stability of 5 this hit series. A summary of the data obtained is shown in Table C, Table D, Table E, and Table F showing the structure activity relationship (SAR). All synthesized compounds were summarized into three series based on modification of the scaffold. Preparation of compounds were achieved via synthetic route 1, route 2, or as described in the synthesis. The synthetic route and characterization is shown further below. 10 (1) Synthetic route 1:
Figure imgf000125_0002
(2) Synthetic route 2: 15
Figure imgf000125_0003
1-(1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide (A1, CDD-2110) 20
Figure imgf000125_0001
. , methyl 3-bromo-1H-indazole-5-carboxylate and 2-ethynyl-1,1’-biphenyl were utilized in the general procedure for Sonogashira coupling - 123 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) to give methyl 3-([1,1’-biphenyl]-2-ylethynyl)-1H-indole-5-carboxylate (52%) as a pale yellow solid; 1H NMR (600 MHz, CD3OD) δ 8.19 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.74 (d, J = 7.7 Hz, 1H), 7.69 (d, J = 7.6 Hz, 2H), 7.58–7.38 (m, 7H), 3.99 (s, 3H). In step 2, the cross- coupling intermediate was subjected to the general procedure for hydrolysis without THP- 5 protected amine to yield 3-([1,1’-biphenyl]-2-ylethynyl)-1H-indazole-5-carboxylic acid. In step 3, the resulting acid and N-methyl-1-(pyrrolidin-3-yl)piperidine-4-carboxamide were utilized in the general procedure for amide coupling to afford the title compound (49% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, mixture of diastereomers and rotamers) δ 7.78 (d, J = 7.6 Hz, 1H), 7.74–7.58 (m, 4H), 7.56–7.45 (m, 6H), 7.45–7.34 (m,10 2H), 3.89–3.77 (m, 0.5H), 3.75–3.66 (m, 0.5H), 3.56–3.46 (m, 1H), 3.34–3.14 (m, 3H), 3.01– 2.81 (m, 2H), 2.79–2.68 (m, 0.5H), 2.54 (dd, J = 27.7, 4.4 Hz, 3H), 2.23–2.14 (m, 0.5H), 2.12–1.72 (m, 4H), 1.71–1.44 (m, 4H); 13C NMR (150 MHz, DMSO-d6, mixture of diastereomers and rotamers) δ 174.8, 174.7, 168.4, 168.3, 143.4, 140.2, 140.0, 132.8, 130.4, 130.0, 129.7, 129.4, 129.1 (2 ×), 128.6, 128.3 (2 ×), 127.8, 127.7, 126.2, 126.1, 123.5, 120.2, 15 118.7, 110.8, 92.7, 84.0, 64.2.62.9, 52.8, 51.7, 51.6, 50.8, 50.7, 49.9, 48.1, 44.9, 42.0, 41.9, 40.5, 29.9, 28.5, 28.1, 25.4; HRMS (ESI) m/z calcd for C33H34N5O2 [M + H]+ 532.2713, found 532.2700. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(pyrrolidin-1-yl)methanone (A2, 20 CDD-2134)
Figure imgf000126_0001
pplied. The same procedure for the synthesis of A1 (CDD- 2110) was followed using pyrrolidine in the amide coupling reaction to afford the title compound (58% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79 25 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.5 Hz, 2H), 7.60 (d, J = 8.6 Hz, 1H), 7.57–7.45 (m, 6H), 7.42–7.36 (m, 2H), 3.58–3.53 (m, 2H), 3.28–3.22 (m, 2H), 1.97–1.90 (m, 2H), 1.85–1.77 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 168.2, 143.3, 140.1, 140.0, 132.8, 130.6, 129.6, 129.4, 129.1 (2 ×), 128.6, 128.2 (2 ×), 127.6, 127.6, 126.2, 123.4, 120.1, 118.5, 110.7, 92.7, 83.9, 49.1, 46.0, 26.0, 24.0; HRMS (ESI) m/z calcd for C26H22N3O [M + H]+ 392.1763, - 124 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) found 392.1753. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (A3, CDD-2211) 5 d. The same procedure for the synthesis of A1 (CDD-
Figure imgf000127_0001
2110) was followed using (R)-N,N-dimethyl-3-pyrrolidinamine in the amide coupling reaction to afford the title compound (77% over two steps) as a colorless oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.78 (dd, J = 7.7, 1.4 Hz, 1H), 7.68 (d, J = 7.6 Hz, 2H),10 7.60 (d, J = 8.6 Hz, 1H), 7.55–7.44 (m, 6H), 7.43–7.37 (m, 2H), 3.81–3.77 (m, 0.5H), 3.73– 3.67 (m, 0.5H), 3.56–3.50 (m, 0.5H), 3.43–3.28 (m, 2H), 3.21–3.12 (m, 0.5H), 2.81–2.68 (m, 0.5H), 2.68–2.58 (m, 0.5H), 2.22 (s, 3H), 2.14–1.98 (m, 4H), 1.85–1.75 (m, 0.5H), 1.75–1.63 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.4, 143.2, 140.6, 140.0, 132.7, 130.1, 129.6, 129.3, 129.0 (2 ×), 128.4, 128.2 (2 ×), 127.6 (2 ×), 125.9, 125.7, 123.6, 15 120.3, 118.6, 111.0, 92.6, 84.3, 65.1, 63.9, 53.0, 50.2, 48.2, 45.0, 43.9, 43.6, 30.5, 28.4; HRMS (ESI) m/z calcd for C28H27N4O [M + H]+ 435.2185, found 435.2174. (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (A4, CDD-2212) 20
Figure imgf000127_0002
d. The same procedure for the synthesis of A1 (CDD- 2110) was followed using (S)-N,N-dimethyl-3-pyrrolidinamine in the amide coupling reaction to afford the title compound (19% over two steps) as a colorless oil; 1H NMR (600 MHz, CD3OD, two rotamers) δ 7.73 (dd, J = 7.7, 1.4 Hz, 1H), 7.66 (d, J = 7.0 Hz, 2H), 7.59 (dd, J = - 125 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 8.6, 6.0 Hz, 1H), 7.54 (dd, J = 8.6, 1.5 Hz, 1H), 7.51–7.44 (m, 4H), 7.44–7.38 (m, 2H), 7.30 (d, J = 14.4 Hz, 1H), 3.98 (dd, J = 12.1, 7.3 Hz, 0.5H), 3.94–3.85 (m, 0.5H), 3.76–3.64 (m, 0.5H), 3.56–3.36 (m, 2H), 3.26–3.20 (m, 0.5H), 2.99–2.92 (m, 0.5H), 2.84–2.77 (m, 0.5H), 2.38 (s, 3H), 2.35–2.12 (m, 4H), 2.00–1.91 (m, 0.5H), 1.87–1.77 (m, 0.5H); 13C NMR (150 5 MHz, DMSO-d6, two rotamers) δ 168.3, 143.3, 140.3, 140.0, 132.8, 130.3, 129.9, 129.6, 129.4, 129.1 (2 ×), 128.6, 128.2 (2 ×), 127.7, 127.7, 126.2, 126.0, 123.5, 120.2, 118.7, 110.9, 92.7, 84.0, 65.2, 64.0, 53.0, 50.3, 48.2, 45.0, 43.9, 43.7, 30.5, 28.5; HRMS (ESI) m/z calcd for C28H27N4O [M + H]+ 435.2185, found 435.2172. 10 3-([1,1’-Biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-N-methyl-1H- indazole-5-carboxamide (A5, CDD-2277) The same procedure for the synthesis of A1 (CDD-
Figure imgf000128_0001
2110) was followed using N1,N1,N2-trimethylethane-1,2-diamine in the amide coupling 15 reaction to afford the title compound (28% over two steps) as a colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.80–7.75 (m, 1H), 7.75–7.70 (m, 1H), 7.67 (d, J = 7.5 Hz, 1H), 7.64– 7.25 (m, 9H), 3.23 (d, J = 13.0 Hz, 3H), 3.10–2.81 (m, 2H), 2.67–2.50 (m, 2H), 2.37–2.14 (m, 3H), 2.15–1.58 (m, 3H); HRMS (ESI) m/z calcd for C27H27N4O [M + H]+ 423.2185, found 423.2172. 20 3-([1,1’-Biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-1H-indazole-5- carboxamide (A6, CDD-2278)
Figure imgf000128_0002
. The same procedure for the synthesis of A1 (CDD- - 126 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 2110) was followed using N1,N1-dimethylethane-1,2-diamine in the amide coupling reaction to afford the title compound (63% over two steps) as a colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 8.46 (t, J = 5.7 Hz, 1H), 8.21–8.14 (m, 1H), 7.87 (dd, J = 8.8, 1.6 Hz, 1H), 7.81 (dd, J = 7.7, 1.3 Hz, 1H), 7.77–7.71 (m, 2H), 7.62 (d, J = 8.7 Hz, 1H), 7.59–7.51 (m, 4H), 5 7.48 (td, J = 7.4, 1.6 Hz, 1H), 7.43–7.36 (m, 1H), 3.45–3.37 (m, 2H), 2.45 (t, J = 6.9 Hz, 2H), 2.20 (s, 6H); 13C NMR (150 MHz, DMSO-d6) δ 166.2, 143.2, 140.9, 139.6, 133.2, 129.7, 129.5, 129.0 (2 ×), 128.9, 128.4, 128.3 (2 ×), 127.8, 127.6, 126.0, 123.7, 120.0, 119.4, 110.6, 92.4, 83.9, 58.3, 45.3 (2 ×), 37.6; HRMS (ESI) m/z calcd for C26H25N4O [M + H]+ 409.2028, found 409.2015. 10 3-([1,1’-Biphenyl]-2-ylethynyl)-N,N-dimethyl-1H-indazole-5-carboxamide (A7, CDD-2481)
Figure imgf000129_0001
applied. The same procedure for the synthesis of A1 (CDD- 15 2110) was followed using dimethylamine in the amide coupling reaction to afford the title compound (19% over two steps) as a pale yellow solid; 1H NMR (600 MHz, DMSO-d6) δ 7.79–7.21 (m, 12H), 3.07 (s, 3H), 2.87 (s, 3H); 13C NMR (150 MHz, DMSO-d6) δ 170.1, 143.4, 140.0, 139.9, 132.7, 129.6, 129.3, 129.1, 128.9, 128.3, 128.2, 127.6, 127.6, 126.1, 123.5, 120.2, 118.4, 110.9, 92.7, 83.9; HRMS (ESI) m/z calcd for C24H20N3O [M + H]+ 20 366.1606, found 366.1595. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin- 1-yl)methanone (A8, CDD-2535)
Figure imgf000129_0002
- 127 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Synthetic route 2 was applied. In step 1, 3-iodo-1H-indazole-5-carboxylic acid and tert-butyl (R)-methyl(pyrrolidin-3-yl)carbamate were utilized in the general procedure for amide coupling. In step 2, the general procedure for Sonogashira coupling was employed using 2-ethynyl-1,1'-biphenyl. In step 3, the cross-coupling intermediate was subjected to the 5 general procedure for removal of Boc-protecting group to afford the title compound (41% over three steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.78 (d, J = 7.6 Hz, 1H), 7.67 (d, J = 7.6 Hz, 2H), 7.64–7.34 (m, 9H), 3.70–3.48 (m, 1H), 3.46– 3.34 (m, 2H), 3.29–3.20 (m, 1H), 3.10–3.02 (m, 1H), 2.33 (s, 1.5H), 2.15 (s, 1.5H), 2.09–2.00 (m, 0.5H), 1.97–1.86 (m, 0.5H), 1.85–1.74 (m, 0.5H), 1.74–1.64 (m, 0.5H); 13C NMR (150 10 MHz, DMSO-d6, two rotamers) δ 168.5, 143.4, 140.5, 140.0, 132.7, 130.4, 130.3, 129.6, 129.3, 129.1 (2 ×), 128.5, 128.3 (2 ×), 127.7, 127.7, 126.0, 123.6, 120.3, 118.6, 118.5, 111.0, 92.7, 92.6, 84.2, 59.4, 57.9, 54.5, 51.7, 47.6, 44.6, 34.4, 34.2, 31.5, 29.7; HRMS (ESI) m/z calcd for C27H25N4O [M + H]+ 421.2028, found 421.2016. 15 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(piperazin-1-yl)methanone (A9, CDD-2577)
Figure imgf000130_0001
ied. The same procedure for the synthesis of A8 (CDD- 2535) was followed using tert-butyl piperazine-1-carboxylate in the amide coupling reaction 20 to afford the title compound (56%) as a white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.99 (s, 1H), 7.79 (dd, J = 7.6, 1.3 Hz, 1H), 7.70–7.67 (m, 2H), 7.66 (d, J = 8.6 Hz, 1H), 7.56 (td, J = 7.5, 1.4 Hz, 1H), 7.52–7.47 (m, 5H), 7.44–7.39 (m, 2H), 3.6 (br s, 4H), 3.16 (br s, 4H); 13C NMR (150 MHz, DMSO-d6) δ 169.5, 143.5, 140.1, 133.0, 129.8, 129.6, 129.2, 128.4, 128.2, 127.8, 126.2, 123.7, 120.2, 119.1, 111.3, 92.9, 83.9, 42.9; HRMS (ESI) m/z calcd for 25 C26H23N4O [M + H]+ 407.1872, found 407.1860. 4-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperazin-2-one (A10, CDD-2578) - 128 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ied. The same procedure for the synthesis of A8 (CDD- 2535) w
Figure imgf000131_0001
as followed using piperazin-2-one in the amide coupling reaction to afford the title compound (55%) as a white foam; 1H NMR (600 MHz, DMSO-d6) 13.68 (s, 1H), 8.20 (s, 5 1H), 7.79 (dd, J = 7.6, 1.3 Hz, 1H), 7.66 (t, J = 8.6 Hz, 3H), 7.55 (td, J = 7.5, 1.4 Hz, 1H), 7.51–7.44 (m, 5H), 7.38 (t, J = 7.5 Hz, 1H), 7.33 (s, 1H), 4.48–3.55 (m, 4H), 3.33–3.06 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 169.7, 143.9, 140.5, 140.4, 133.2, 130.1, 129.9, 129.5, 129.1, 128.7, 128.1, 128.1, 126.4, 124.0, 120.5, 119.2, 111.6, 93.3, 84.2, 39.4; HRMS (ESI) m/z calcd for C26H21N4O [M + H]+ 421.1665, found 421.1653. 10 (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1- yl)methanone (A11, CDD-2579)
Figure imgf000131_0002
15 Synthetic route 2 was applied. The same procedure for the synthesis of A8 (CDD- 2535) was followed using tert-butyl (R)-pyrrolidin-3-ylcarbamate in the amide coupling reaction to afford the title compound (52%) as a white foam; 1H NMR (600 MHz, DMSO-d6, two rotamers) 8.27 (br s, 1H), 8.11 (br s, 1H), 7.78 (d, J = 7.
Figure imgf000131_0003
7 Hz, 1H), 7.67 (d, J = 7.5 Hz, 2H), 7.64 (d, J = 8.6 Hz, 1H), 7.57–7.53 (m, 2H), 7.51–7.38 (m, 6H), 3.97–3.75 (m, 2H), 20 3.72–3.60 (m, 2H), 3.42-3.28 (m, 1H), 2.38–2.15 (m, 1H), 2.13–1.91 (m, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.7, 158.5, 143.5, 140.3, 140.0, 133.0, 129.8, 129.7, 129.2 (2 ×), 128.8, 128.4 (2 ×), 127.9, 127.8, 126.3, 123.5, 120.2, 119.0, 111.1, 92.8, 83.9, 52.4, 50.0, 49.7, 48.7, 46.8, 44.0, 30.2, 28.2; HRMS (ESI) m/z calcd for C26H23N4O [M + H]+ 407.1872, found 407.1861. - 129 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1- yl)methanone (A12, CDD-2594) 5 d. The same procedure for the synthesis of A8 (CDD-
Figure imgf000132_0001
2535) was followed using (R)-pyrrolidin-3-ol in the amide coupling reaction to afford the title compound (43%) as a white foam; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.79– 7.75 (m, 1H), 7.65 (t, J = 7.3 Hz, 2H), 7.62–7.59 (m, 2H), 7.56–7.52 (m, 2H), 7.50–7.45 (m, 4H), 7.44–7.38 (m, 1H), 7.36 (d, J = 3.7 Hz, 1H), 4.40 (br s, 0.5H), 4.23 (br s, 0.5H), 3.68– 10 3.62 (m, 2H), 3.43–3.41 (m, 1H), 3.29–3.23 (m, 0.5H), 3.11–3.07 (m, 0.5H), 2.04–1.98 (m, 0.5H), 1.94–1.83 (m, 1H), 1.84–1.75 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 169.0, 143.9, 143.9, 140.5, 140.4, 133.2, 133.1, 132.5, 131.9, 131.9, 130.9, 130.1, 129.9, 129.5, 129.4, 129.2, 129.2, 128.7, 128.2, 128.2, 128.1, 126.7, 123.9, 120.5, 119.1, 118.9, 111.2, 93.3, 84.3, 69.8, 68.6, 57.7, 54.9, 47.6, 44.6, 34.8, 32.7; HRMS (ESI) m/z calcd 15 for C26H22N3O2 [M + H]+ 408.1712, found 408.1700. (S)-1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N- methylpyrrolidine-2-carboxamide (A13, CDD-2595) 20
Figure imgf000132_0002
ed. The same procedure for the synthesis of A8 (CDD- 2535) was followed using (S)-N-methylpyrrolidine-2-carboxamide in the amide coupling reaction to afford the title compound (48%) as a white foam; 1H NMR (600 MHz, CD3OD) δ 7.75 (d, J = 7.6 Hz, 1H), 7.70–7.63 (m, 5H), 7.58 (d, J = 23.8 Hz, 2H), 7.52–7.46 (m, 5H), 7.43 (t, J = 7.4 Hz, 3H), 4.59 (t, J = 7.1 Hz, 1H), 3.85 (t, J = 7.0 Hz, 1H), 3.62 (q, J = 7.5 Hz, - 130 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 1H), 3.47 (d, J = 10.9 Hz, 1H), 2.83 (s, 3H), 2.37 (d, J = 8.8 Hz, 2H), 2.05 (s, 2H); HRMS (ESI) m/z calcd for C28H25N4O2 [M + H]+ 449.1978, found 449.1965. 1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-one (A14, 5 CDD-2596) ied. The same procedure for the synthesis of A8 (CDD-
Figure imgf000133_0001
2535) was followed using pyrrolidin-3-one in the amide coupling reaction to afford the title compound (55%) as a white solid; 1H NMR (600 MHz, CDCl3) δ 7.74 (d, J = 7.0 Hz, 1H), 10 7.67 (d, J = 7.5 Hz, 2H), 7.61 (s, 1H), 7.52–7.38 (m, 6H), 7.37–7.34 (m, 1H), 7.31 (t, J = 7.4 Hz, 1H), 4.33–4.00 (m, 2H), 3.74 (s, 2H), 2.63 (s, 2H); 13C NMR (150 MHz, CDCl3) δ 207.8, 168.3, 142.2, 138.8, 138.7, 131.2, 128.8, 127.8, 127.6, 127.6, 127.4, 126.2, 125.5, 125.4, 125.2, 122.1, 118.6, 118.4, 114.4, 109.1, 92.8, 80.7, 44.0, 42.0; HRMS (ESI) m/z calcd for C26H20N3O2 [M + H]+ 406.1556, found 406.1542. 15 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-oxa-6-azaspiro[3.4]octan-6- yl)methanone (A15, CDD-2597)
Figure imgf000133_0002
d. The same procedure for the synthesis of A8 (CDD- 20 2535) was followed using 2-oxa-6-azaspiro[3.4]octane in the amide coupling reaction to afford the title compound (52%) as a white foam; 1H NMR (600 MHz, CDCl3) δ 7.74 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.5 Hz, 2H), 7.56 (d, J = 8.2 Hz, 1H), 7.49–7.41 (m, 4H), 7.40 (t, J = 7.5 Hz, 2H), 7.38–7.34 (m, 1H), 7.31 (t, J = 7.4 Hz, 1H), 4.84 (d, J = 5.9 Hz, 1H), 4.66 (d, J = 6.1 Hz, 1H), 4.59 (d, J = 6.3 Hz, 1H), 4.55–4.45 (m, 1H), 3.96 (s, 1H), 3.76 (d, J = 7.5 - 131 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Hz, 1H), 3.53 (s, 1H), 3.35 (s, 1H), 2.31 (d, J = 7.5 Hz, 1H), 2.20 (d, J = 6.9 Hz, 1H); 13C NMR (150 MHz, CDCl3) δ 169.8, 144.0, 133.1, 129.6, 129.4, 129.1, 128.1, 127.4, 127.2, 120.7, 120.1, 80.7, 80.0, 58.3, 57.0, 54.9, 48.4, 45.0, 44.3, 36.2, 34.1; HRMS (ESI) m/z calcd for C28H24N3O2 [M + H]+ 434.1869, found 434.1856. 5 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,4-diazepan-1-yl)methanone (A16, CDD-2598) ied. The same procedure for the synthesis of A8 (CDD-
Figure imgf000134_0001
10 2535) was followed using tert-butyl 1,4-diazepane-1-carboxylate in the amide coupling reaction to afford the title compound (42%) as a white foam; 1H NMR (600 MHz, CD3OD) δ 7.74–7.70 (m, 1H), 7.67 – 7.63 (m, 2H), 7.62 (d, J = 8.7 Hz, 1H), 7.53 (dd, J = 8.7, 1.5 Hz, 1H), 7.50–7.42 (m, 4H), 7.40 (td, J = 7.5, 1.5 Hz, 2H), 7.35 (s, 1H), 4.07 (s, 1H), 3.97 (s, 1H), 3.63–3.47 (m, 3H), 3.34 (d, J = 6.1 Hz, 2H), 2.25 (s, 1H), 2.05 (s, 2H); 13C NMR (150 15 MHz, CD3OD) δ 172.6, 144.2, 140.7, 140.5, 132.5, 129.8, 129.3, 129.0, 129.0, 128.4, 127.9, 127.3, 127.1, 125.9, 123.7, 120.5, 119.4, 110.8, 93.3, 82.7, 53.4, 48.3, 48.2, 45.5, 44.5, 42.0, 25.7; HRMS (ESI) m/z calcd for C27H25N4O [M + H]+ 421.2028, found 421.2018. 3-([1,1’-Biphenyl]-2-ylethynyl)-N-(4-methoxyphenyl)-1H-indazole-5- 20 carboxamide (A17, CDD-2606)
Figure imgf000134_0002
The same procedure for the synthesis of A8 (CDD- 2535) was followed using 4-methoxyaniline in the amide coupling reaction to afford the title compound (42%) as a white solid; 1H NMR (600 MHz, CDCl3) δ 7.99 (s, 1H), 7.75–7.55 (m, - 132 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 7H), 7.46–7.38 (m, 6H), 7.20 (d, J = 7.5 Hz, 1H), 6.98 (s, 2H), 3.86 (s, 3H); 13C NMR (150 MHz, CDCl3) δ 129.5, 128.1, 127.7, 122.6, 114.3, 93.2, 82.5, 55.5; HRMS (ESI) m/z calcd for C29H22N3O2 [M + H]+ 444.1712, found 444.1700. 5 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-4- yl)methanone (A18, CDD-2607) lied. In step 1, methyl 3-iodo-1-(tetrahydro-2H-pyran-2-yl)-
Figure imgf000135_0001
1H-indazole-5-carboxylate, derived from methyl 3-iodo-1H-indazole-5-carboxylate via the 10 general procedure for THP protection, and 2-ethynyl-1,1'-biphenyl were utilized in the general procedure for Sonogashira coupling. In step 2, the cross-coupling intermediate was subjected to the general procedure for hydrolysis with THP-protected amine to yield 3-([1,1'- biphenyl]-2-ylethynyl)-1-(tetrahydro-2H-pyran-3-yl)-1H-indazole-5-carboxylic acid (75% over two steps) as a pale yellow solid; 1H NMR (600 MHz, DMSO-d6) δ 13.06 (s, 1H), 8.15 15 (s, 1H), 8.01 (dd, J = 8.8, 1.5 Hz, 1H), 7.87 (d, J = 8.8 Hz, 1H), 7.81 (dd, J = 7.7, 1.3 Hz, 1H), 7.76–7.65 (m, 2H), 7.60–7.51 (m, 4H), 7.48 (td, J = 7.5, 1.5 Hz, 1H), 7.46–7.40 (m, 1H), 5.93 (dd, J = 9.5, 2.5 Hz, 1H), 3.91–3.83 (m, 1H), 3.80–3.70 (m, 1H), 2.40–2.27 (m, 1H), 2.08–1.89 (m, 2H), 1.80–1.66 (m, 1H), 1.65–1.52 (m, 2H). In step 3, the acid and tert- butyl 4,7-diazaspiro[2.5]octane-7-carboxylate were utilized in the general procedure for 20 amide coupling. In step 4, the general procedure for removal of THP and Boc-protecting group was employed to afford the title compound (35% over two steps) as a yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.6 Hz, 2H), 7.59 (d, J = 8.6 Hz, 1H), 7.56–7.35 (m, 8H), 3.37 (s, 2H), 2.92–2.53 (m, 4H), 1.17–0.22 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 143.2, 140.7, 139.9, 132.9, 129.7, 129.4, 129.1, 128.4, 128.2, 25 127.7, 125.9, 123.7, 120.3, 118.7, 110.9, 92.5, 84.3, 13.8; HRMS (ESI) m/z calcd for C28H25N4O [M + H]+ 433.2028, found 433.2016. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-7- yl)methanone (A19, CDD-2608) - 133 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ied. The same procedure for the synthesis of A18 (CDD- 2607) w
Figure imgf000136_0001
as followed using tert-butyl 4,7-diazaspiro[2.5]octane-4-carboxylate in the amide coupling reaction to afford the title compound (78% over two steps) as a pale yellow solid; 1H 5 NMR (600 MHz, DMSO-d6) δ 7.77 (d, J = 7.6 Hz, 1H), 7.67 (d, J = 7.5 Hz, 2H), 7.60 (d, J = 8.6 Hz, 1H), 7.56–7.17 (m, 8H), 3.12 (s, 2H), 2.98–2.54 (m, 4H), 0.77–0.01 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 143.4, 140.1, 140.0, 132.7, 129.7, 129.4, 129.1, 128.4, 128.3, 127.7, 127.7, 125.9, 123.7, 120.2, 118.4, 111.1, 92.7, 84.1, 11.7; HRMS (ESI) m/z calcd for C28H25N4O [M + H]+ 433.2028, found 433.2015. 10 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-8- yl)methanone (A20, CDD-2609)
Figure imgf000136_0002
ied. The same procedure for the synthesis of A18 (CDD- 15 2607) was followed using tert-butyl 5,8-diazaspiro[3.5]nonane-5-carboxylate in the amide coupling reaction to afford the title compound (72% over two steps) as a yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76 (s, 1H), 7.68 (d, J = 6.9 Hz, 2H), 7.62 (d, J = 7.4 Hz, 1H), 7.5– 7.42 (m, 5H), 7.38 (t, J = 7.6 Hz, 1H), 7.35–7.26 (m, 2H), 3.21 (s, 4H), 2.88–2.52 (m, 2H), 2.16–0.94 (m, 6H); 13C NMR (150 MHz, DMSO-d6) δ 169.6, 143.2, 141.1, 140.1, 132.7, 20 129.6, 129.2, 129.1, 128.2, 128.1, 127.7, 127.6, 125.2, 123.9, 120.5, 118.4, 111.6, 92.5, 84.8, 56.5, 31.1, 12.9; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2171.
Figure imgf000136_0003
(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,6-diazabicyclo[3.1.1]heptan- 3-yl)methanone (A21, CDD-2610) - 134 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) lied; yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.82– 7.74 (m
Figure imgf000137_0001
, 1H), 7.71–7.58 (m, 3H), 7.58–7.32 (m, 8H), 4.02–3.77 (m, 2H), 3.73–3.62 (m, 4H), 2.13–1.67 (m, 1H), 1.56–1.39 (m, 1H); 13C NMR (150 MHz, DMSO-d6) δ 171.4, 143.4, 5 140.0, 139.9, 132.8, 131.5, 131.5, 130.6, 129.7, 129.4, 129.1, 129.1, 128.8, 128.7, 128.4, 128.3, 127.7, 127.7, 125.7, 123.6, 120.2, 118.0, 111.0, 92.6, 84.1, 54.8, 54.3, 52.6, 48.7, 30.2; HRMS (ESI) m/z calcd for C27H23N4O [M + H]+ 419.1872, found 419.1861. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- 10 yl)methanone (A22, CDD-2611)
Figure imgf000137_0002
lied; yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J = 7.5 Hz, 1H), 7.71–7.57 (m, 3H), 7.57–7.25 (m, 8H), 3.27–2.79 (m, 6H), 2.10–1.46 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 169.1, 143.3, 140.3, 140.0, 132.8, 132.1, 131.5, 131.5, 15 130.0, 129.7, 129.4, 129.1, 129.0, 128.8, 128.7, 128.4, 128.3, 128.3, 127.8, 127.7, 127.7, 126.0, 125.1, 123.9, 120.3, 117.9, 111.3, 111.1, 92.7, 84.2, 50.8, 47.9, 43.9, 26.2, 25.9, 24.7; HRMS (ESI) m/z calcd for C28H25N4O [M + H]+ 433.2028, found 433.2015. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- 20 yl)methanone (A23, CDD-2612) - 135 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) lied; yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J = 7.6 H
Figure imgf000138_0001
z, 1H), 7.67 (d, J = 7.6 Hz, 2H), 7.60 (d, J = 8.5 Hz, 1H), 7.55–7.43 (m, 5H), 7.40 (t, J = 7.4 Hz, 1H), 7.32 (d, J = 8.5 Hz, 1H), 7.24 (s, 1H), 4.41–4.27 (m, 1H), 3.31–3.12 (m, 5 4H), 3.02–2.90 (m, 1H), 1.79–1.31 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 170.4, 143.3, 140.5, 140.1, 132.7, 129.6, 129.6, 129.3, 129.1, 128.2, 127.7, 125.4, 123.8, 120.4, 118.3, 111.3, 92.6, 84.4, 53.6, 28.0; HRMS (ESI) m/z calcd for C28H25N4O [M + H]+ 433.2028, found 433.2016. 10 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-3-azabicyclo[3.2.1]octan- 3-yl)methanone (A24, CDD-2613)
Figure imgf000138_0002
lied; yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.5 Hz, 2H), 7.61 (d, J = 8.5 Hz, 1H), 7.55–7.43 (m, 5H), 7.43– 15 7.34 (m, 2H), 7.31 (s, 1H), 4.62–4.08 (m, 4H), 3.41–3.00 (m, 2H), 2.06–1.31 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 170.5, 143.2, 140.7, 140.1, 132.7, 129.7, 129.3, 129.1, 128.9, 128.3, 128.2, 127.7, 127.7, 125.5, 123.9, 120.3, 118.6, 111.4, 92.6, 84.4, 73.2, 27.1; HRMS (ESI) m/z calcd for C28H24N3O2 [M + H]+ 434.1869, found 434.1854. 20 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1- yl)methanone (A25, CDD-2614) - 136 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79 (d, J = 6.5 H
Figure imgf000139_0001
z, 1H), 7.72–7.58 (m, 3H), 7.58–7.07 (m, 13H), 4.52 (s, 1H), 3.19–2.63 (m, 6H); 13C NMR (150 MHz, DMSO-d6) 169.0, 143.4, 140.1, 140.0, 132.8, 129.7, 129.4, 129.3, 129.1, 5 128.5, 128.3, 128.3, 127.8, 127.7, 127.5, 126.9, 126.0, 123.7, 120.2, 118.5, 111.2, 92.8, 84.0; HRMS (ESI) m/z calcd for C32H27N4O [M + H]+ 483.2185, found 483.2171. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-2- yl)methanone (A26, CDD-2674) 10
Figure imgf000139_0002
colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.81– 7.76 (m, 1H), 7.71–7.66 (m, 2H), 7.63–7.57 (m, 1H), 7.57–7.44 (m, 7H), 7.43–7.32 (m, 2H), 3.65–3.59 (m, 2H), 3.09 (s, 2H), 2.79–2.65 (m, 2H), 2.61 (s, 1H), 2.45 (s, 1H), 1.87–1.78 (m, 1H), 1.78–1.65 (m, 1H), 1.60–1.42 (m, 2H), 1.39–1.22 (m, 2H); HRMS (ESI) m/z calcd for 15 C30H29N4O [M + H]+ 461.2341, found 461.2327. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,9-diazaspiro[5.5]undecan-2- yl)methanone (A27, CDD-2675) 20
Figure imgf000139_0003
colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.78 - 137 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.5 Hz, 2H), 7.62 (d, J = 8.6 Hz, 1H), 7.58–7.52 (m, 1H), 7.52–7.45 (m, 4H), 7.42–7.35 (m, 2H), 7.29 (s, 1H), 3.70–3.59 (m, 2H), 2.92 (s, 4H), 2.14 (s, 1H), 1.67–1.33 (m, 7H), 1.30–1.03 (m, 2H); HRMS (ESI) m/z calcd for C31H31N4O [M + H]+ 475.2498, found 475.2484. 5 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-5- yl)methanone (A28, CDD-2676) lied; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77
Figure imgf000140_0001
10 (d, J = 7.6 Hz, 1H), 7.71 (d, J = 7.7 Hz, 2H), 7.59 (d, J = 8.5 Hz, 1H), 7.56–7.33 (m, 8H), 3.16 (s, 2H), 2.92 (s, 2H), 2.49–2.35 (m, 2H), 2.27–2.09 (m, 4H), 1.84–1.60 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 170.7, 143.0, 141.5, 140.0, 132.7, 130.0, 129.6, 129.2, 129.1, 128.2, 128.1, 127.7, 127.6, 125.6, 123.9, 120.5, 119.0, 111.5, 92.3, 84.9, 60.0, 50.3, 46.6, 45.9, 30.7, 14.5; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2171. 15 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6,9-diazaspiro[4.5]decan-6- yl)methanone (A29, CDD-2677)
Figure imgf000140_0002
lied; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 20 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.7 Hz, 2H), 7.59 (d, J = 8.6 Hz, 1H), 7.56–7.43 (m, 5H), 7.43–7.29 (m, 3H), 3.19–3.14 (m, 2H), 2.69 (s, 2H), 2.68–2.61 (m, 2H), 2.20–2.06 (m, 2H), 2.00–1.84 (m, 4H), 1.62–1.50 (m, 2H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. - 138 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone (A30, CDD-2678) lied; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.74
Figure imgf000141_0001
5 (d, J = 7.6 Hz, 1H), 7.68 (d, J = 7.6 Hz, 2H), 7.62 (d, J = 8.8 Hz, 1H), 7.56–7.23 (m, 13H), 3.15–2.59 (m, 7H); 13C NMR (150 MHz, DMSO-d6) δ 143.1, 140.7, 140.0, 139.6, 132.8, 129.6, 129.3, 129.0, 128.5, 128.2, 128.1, 127.7, 127.7, 127.2, 126.6, 124.9, 123.8, 120.4, 118.2, 111.5, 92.5, 84.5, 45.8; HRMS (ESI) m/z calcd for C32H27N4O [M + H]+ 483.2185, found 483.2171. 10 (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4- chlorophenyl)piperazin-1-yl)methanone (A31, CDD-2679) l 1
Figure imgf000141_0002
ied; colorless oil; H NMR (600 MHz, DMSO-d6) δ 7.75 15 (d, J = 7.6 Hz, 1H), 7.67 (d, J = 7.5 Hz, 2H), 7.62 (d, J = 8.6 Hz, 1H), 7.58–7.23 (m, 12H), 3.15–2.58 (m, 7H); 13C NMR (150 MHz, DMSO-d6) δ 143.2, 140.5, 140.0, 138.8, 132.8, 131.4, 129.6, 129.4, 129.3, 129.2, 129.1, 128.3, 128.2, 128.2, 127.7, 125.1, 123.8, 120.3, 118.3, 111.4, 92.5, 84.3, 45.8; HRMS (ESI) m/z calcd for C32H26ClN4O [M + H]+ 517.1795, found 517.1784. 20 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[4.5]decan-2- yl)methanone (A32, CDD-2765) - 139 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76 (d, J =
Figure imgf000142_0001
7.6 Hz, 1H), 7.68 (d, J = 7.5 Hz, 2H), 7.57 (t, J = 7.0 Hz, 1H), 7.54–7.36 (m, 8H), 3.67–3.51 (m, 2H), 3.22–3.13 (m, 2H), 2.75 (s, 1H), 2.63 (s, 1H), 2.01–1.85 (m, 1H), 1.82– 5 1.67 (m, 1H), 1.62–1.55 (m, 1H), 1.53–1.47 (m, 1H), 1.45–1.17 (m, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[4.5]decan-2- yl)methanone (A33, CDD-2766) 10 ; light yellow oil; 1H NMR (600 MHz, DMSO-d6
Figure imgf000142_0002
) δ 7.84–7.64 (m, 3H), 7.57 (d, J = 8.5 Hz, 1H), 7.53–7.33 (m, 8H), 3.63–3.55 (m, 2H), 3.11– 2.96 (m, 2H), 2.46–2.33 (m, 2H), 2.04–1.85 (m, 1H), 1.83–1.68 (m, 1H), 1.59 (s, 1H), 1.52– 1.29 (m, 4H), 1.15 (s, 1H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 15 461.2329. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,8-diazaspiro[4.5]decan-1- yl)methanone (A34, CDD-2767) 20
Figure imgf000142_0003
plied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.78 - 140 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.5 Hz, 2H), 7.60 (d, J = 8.6 Hz, 1H), 7.57–7.39 (m, 7H), 7.34 (s, 1H), 3.30 (t, J = 6.7 Hz, 5H), 3.26–3.17 (m, 2H), 3.08–2.97 (m, 2H), 2.11 (t, J = 6.8 Hz, 2H), 1.82–1.73 (m, 2H), 1.73–1.65 (m, 2H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2327. 5 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-1-azaspiro[4.5]decan-1- yl)methanone (A35, CDD-2768) lied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.78
Figure imgf000143_0001
10 (d, J = 7.6 Hz, 1H), 7.69 (d, J = 7.5 Hz, 2H), 7.58 (d, J = 8.6 Hz, 1H), 7.56–7.45 (m, 5H), 7.43–7.37 (m, 2H), 7.32 (s, 1H), 3.90 (dd, J = 11.4, 5.0 Hz, 2H), 3.46 (t, J = 12.2 Hz, 2H), 3.27 (t, J = 6.6 Hz, 3H), 3.15–3.03 (m, 2H), 2.09 (t, J = 6.8 Hz, 2H), 1.78–1.69 (m, 2H), 1.39 (d, J = 12.4 Hz, 2H); HRMS (ESI) m/z calcd for C30H28N3O2 [M + H]+ 462.2182, found 462.2365. 15 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-8- yl)methanone (A36, CDD-2769)
Figure imgf000143_0002
light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76 20 (d, J = 7.7 Hz, 1H), 7.69 (d, J = 7.5 Hz, 2H), 7.58 (t, J = 9.2 Hz, 1H), 7.53–7.25 (m, 8H), 3.63–3.56 (m, 2H), 3.17–3.05 (m, 2H), 2.48–2.36 (m, 2H), 1.78–1.32 (m, 6H), 1.23 (s, 2H); (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. 3-([1,1’-Biphenyl]-2-ylethynyl)-N-(7-azaspiro[3.5]nonan-2-yl)-1H-indazole-5- - 141 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) carboxamide (A37, CDD-2770) ght yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76
Figure imgf000144_0001
(d, J = 7.7 Hz, 1H), 7.69 (d, J = 7.5 Hz, 2H), 7.58 (t, J = 9.2 Hz, 1H), 7.53–7.25 (m, 8H), 5 3.63–3.56 (m, 2H), 3.17–3.05 (m, 3H), 2.48–2.36 (m, 2H), 1.78–1.32 (m, 6H), 1.23 (s, 1H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2327. (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1- yl)methanone (A38, CDD-2771) 10 lied; light yellow oil; 1H NMR (600 MHz, DMSO-d6
Figure imgf000144_0002
) δ 7.79 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.5 Hz, 2H), 7.65 (d, J = 8.6 Hz, 1H), 7.58–7.35 (m, 8H), 4.71–4.26 (m, 1H), 3.55 (s, 2H), 3.27–2.89 (m, 4H), 2.35 (s, 1H), 1.16–0.66 (m, 6H); HRMS (ESI) m/z calcd for C29H29N4O [M + H]+ 449.2341, found 449.2326. 15 (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1- yl)methanone (A39, CDD-2772)
Figure imgf000144_0003
plied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79 20 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.5 Hz, 2H), 7.65 (d, J = 8.5 Hz, 1H), 7.58–7.44 (m, 7H), - 142 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 7.41 (t, J = 7.4 Hz, 1H), 4.48 (s, 1H), 3.56 (s, 2H), 3.25–2.89 (m, 4H), 2.35 (s, 1H), 1.13–0.59 (m, 6H); HRMS (ESI) m/z calcd for C29H29N4O [M + H]+ 449.2341, found 449.2328. (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- 5 yl)methanone (A40, CDD-2773) d; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.81
Figure imgf000145_0001
(d, J = 7.9 Hz, 1H), 7.73–7.67 (m, 2H), 7.60–7.44 (m, 6H), 7.41 (t, J = 7.5 Hz, 1H), 7.14 (br s, 7H), 3.32–2.99 (m, 9H); HRMS (ESI) m/z calcd for C33H29N4O [M + H]+ 497.2341, found 10 497.2326. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- yl)methanone (A41, CDD-2774) 15
Figure imgf000145_0002
d; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.82 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.6 Hz, 2H), 7.60–7.43 (m, 6H), 7.41 (t, J = 7.4 Hz, 1H), 7.15 (br s, 7H), 3.35–2.94 (m, 9H); HRMS (ESI) m/z calcd for C33H29N4O [M + H]+ 497.2341, found 497.2329. 20 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2S,6S)-2,6-dimethylpiperazin- 1-yl)methanone (A42, CDD-2775) - 143 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) lied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79 (d, J =
Figure imgf000146_0001
7.6 Hz, 1H), 7.71 (d, J = 7.5 Hz, 2H), 7.65 (d, J = 8.6 Hz, 1H), 7.58–7.46 (m, 7H), 7.41 (t, J = 7.4 Hz, 1H), 4.12–4.02 (m, 2H), 3.40–3.36 (m, 2H), 3.19–3.10 (m, 2H), 1.31 (s, 5 3H), 1.30 (s, 3H); HRMS (ESI) m/z calcd for C28H27N4O [M + H]+ 435.2185, found 435.2170. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[3.5]nonan-2- yl)methanone (A43, CDD-2806) 10
Figure imgf000146_0002
; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79 (d, J = 7.7 Hz, 1H), 7.72 (d, J = 7.6 Hz, 2H), 7.69 (s, 1H), 7.66 (d, J = 8.2 Hz, 1H), 7.61 (d, J = 8.7 Hz, 1H), 7.58–7.50 (m, 4H), 7.48 (td, J = 7.4, 1.7 Hz, 1H), 7.43 (t, J = 7.4 Hz, 1H), 3.85 (s, 2H), 3.79 (s, 2H), 2.68–2.52 (m, 4H), 1.62 (t, J = 5.4 Hz, 4H); 13C NMR (150 MHz, 15 DMSO-d6) δ 169.2, 143.2, 140.8, 139.8, 133.0, 129.7, 129.5, 129.0 (2 x), 128.8, 128.3 (2 x), 127.8, 127.7, 126.7, 126.4, 123.6, 120.1, 119.7, 110.9, 92.6, 84.0, 63.4, 59.0, 42.8 (2 x), 36.1 (2 x), 34.3; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2173. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- 20 yl)methanone (A44, CDD-2807) - 144 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) . The same procedure for the synthesis of A18 (CDD- 2607) w
Figure imgf000147_0001
as followed using tert-butyl 2,6-diazaspiro[3.5]nonane-6-carboxylate in the amide coupling reaction to afford the title compound (31% over two steps) as a white solid; 1H 5 NMR (600 MHz, DMSO-d6) δ 7.80 (dd, J = 7.7, 1.3 Hz, 1H), 7.72 (dd, J = 8.2, 1.3 Hz, 2H), 7.69–7.64 (m, 2H), 7.61 (dd, J = 8.6, 0.9 Hz, 1H), 7.58–7.50 (m, 4H), 7.48 (td, J = 7.4, 1.6 Hz, 1H), 7.46–7.42 (m, 1H), 3.84 (d, J = 8.4 Hz, 1H), 3.81–3.74 (m, 2H), 3.70 (d, J = 9.7 Hz, 1H), 2.79–2.65 (m, 2H), 2.59–2.51 (m, 2H), 1.68 (br s, 2H), 1.46–1.23 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 169.4, 143.1, 141.3, 139.8, 132.9, 129.7, 129.4, 129.0 (2 ×), 128.6, 10 128.3 (2 ×), 127.8, 127.6, 126.5, 126.0, 123.7, 120.2, 119.6, 111.2, 92.5, 84.4, 62.2, 57.7, 54.6, 45.3, 34.7, 34.1, 23.2; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2173. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazaspiro[3.5]nonan-2- 15 yl)methanone (A45, CDD-2808)
Figure imgf000147_0002
ied; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.79 (d, J = 7.6 Hz, 1H), 7.74–7.69 (m, 3H), 7.67 (d, J = 8.8 Hz, 1H), 7.62 (d, J = 8.6 Hz, 1H), 7.58– 7.50 (m, 4H), 7.50–7.42 (m, 2H), 3.98–3.92 (m, 1H), 3.92–3.84 (m, 2H), 3.82–3.77 (m, 1H), 20 2.68–2.62 (m, 1H), 2.61–2.54 (m, 1H), 1.67–1.58 (m, 2H), 1.55–1.48 (m, 1H), 1.48–1.40 (m, 1H), 1.38–1.31 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 169.4, 143.3, 140.7, 139.8, 133.1, 129.7, 129.6, 129.0 (2 ×), 128.9, 128.3 (2 ×), 127.9, 127.7, 127.0, 126.5, 123.6, 120.1, 119.8, 110.9, 92.7, 83.9, 64.3, 60.1, 53.3, 42.3, 34.5, 24.9, 21.3; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2171. - 145 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone (A46, CDD-2809) 5 d; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77
Figure imgf000148_0001
(d, J = 7.7 Hz, 1H), 7.72 (d, J = 7.5 Hz, 2H), 7.69 (s, 1H), 7.59 (s, 2H), 7.55–7.49 (m, 4H), 7.49–7.40 (m, 2H), 3.92 (s, 2H), 3.85 (s, 2H), 2.50 (br s, 4H), 1.72 (t, J = 5.3 Hz, 4H); HRMS (ESI) m/z calcd for C29H26N3O2 [M + H]+ 448.2025, found 448.2012. 10 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-2-azaspiro[4.5]decan-2- yl)methanone (A47, CDD-2810) ; light yellow oil; 1
Figure imgf000148_0002
H NMR (600 MHz, DMSO-d6) δ 7.76 (s, 1H), 7.69 (s, 2H), 7.58 (d, J = 8.7 Hz, 1H), 7.54–7.41 (m, 7H), 7.38 (s, 1H), 3.63 (s, 6H), 15 3.18–3.12 (m, 2H), 1.91 (t, J = 7.3 Hz, 1H), 1.78 (s, 1H), 1.68–1.50 (m, 2H), 1.41 (s, 2H); HRMS (ESI) m/z calcd for C30H28N3O2 [M + H]+ 462.2182, found 462.2166. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone (A48, CDD-2811) 20
Figure imgf000148_0003
- 146 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Synthetic route 1 was applied; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76– 7.71 (m, 1H), 7.71–7.65 (m, 2H), 7.63–7.56 (m, 1H), 7.53–7.38 (m, 9H), 7.37–7.18 (m, 4H), 3.17–2.94 (m, 5H), 2.81 (s, 1H), 2.68 (s, 1H); HRMS (ESI) m/z calcd for C32H27N4O [M + H]+ 483.2185, found 483.2172. 5 (R)- or (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[4.5]decan-2-yl)methanone (A49 and A50, CDD-2830 and CDD-2831) DD-2831) are undetermined R- and S-enantiomers
Figure imgf000149_0001
10 isolated from A32 (CDD-2765) by chiral separation. A49: light yellow oil; HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2331. A50: light yellow oil; HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2325. 15 7-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,7- diazaspiro[3.5]nonan-1-one (A51, CDD-2921)
Figure imgf000149_0002
; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76 20 (dd, J = 7.6, 1.4 Hz, 1H), 7.70–7.65 (m, 2H), 7.59 (dd, J = 8.5, 0.9 Hz, 1H), 7.54–7.43 (m, 5H), 7.40–7.35 (m, 1H), 7.34–7.30 (m, 1H), 7.28 (t, J = 1.2 Hz, 1H), 3.69–3.64 (m, 4H), 3.15–3.03 (m, 4H), 2.69 (s, 2H); HRMS (ESI) m/z calcd for C29H22N4O2 [M + H]+ 458.1743, found 458.1726. - 147 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,7- diazaspiro[3.5]nonan-7-yl)methanone (A52, CDD-2922) ; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.73
Figure imgf000150_0001
5 (d, J = 7.4 Hz, 1H), 7.71–7.67 (m, 2H), 7.56 (d, J = 8.5 Hz, 1H), 7.50–7.43 (m, 5H), 7.39– 7.33 (m, 1H), 7.28 (d, J = 1.4 Hz, 1H), 7.19 (dd, J = 8.5, 1.5 Hz, 1H), 3.24–3.12 (m, 4H), 2.96 (s, 4H), 2.22 (s, 3H), 1.80–1.47 (m, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. 10 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (A53, CDD-2923) ; c 1
Figure imgf000150_0002
olorless oil; H NMR (600 MHz, DMSO-d6) δ 7.79 (dd, J = 7.8, 1.3 Hz, 1H), 7.74–7.67 (m, 3H), 7.65 (dd, J = 8.7, 1.6 Hz, 1H), 7.60 (d, J = 8.6 15 Hz, 1H), 7.57–7.49 (m, 4H), 7.48 (td, J = 7.3, 1.7 Hz, 1H), 7.45–7.40 (m, 1H), 3.86 (s, 2H), 3.79 (s, 2H), 2.23 (br s, 4H), 2.10 (s, 3H), 1.71 (t, J = 5.4 Hz, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2329. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7- 20 diazaspiro[3.5]nonan-2-yl)methanone (A54, CDD-2924) - 148 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.78 (d, J = 7
Figure imgf000151_0001
.6 Hz, 1H), 7.72 (d, J = 7.9 Hz, 3H), 7.66–7.58 (m, 2H), 7.56–7.38 (m, 6H), 3.96– 3.91 (m, 2H), 3.86–3.59 (m, 2H), 2.74–2.50 (m, 3H), 2.33–2.12 (m, 1H), 1.88–1.73 (m, 1H), 5 1.62 (s, 1H), 1.56–1.45 (m, 1H), 0.89 (dd, J = 19.0, 6.8 Hz, 3H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2329. (S)-4-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3- phenylpiperazin-2-one (A55, CDD-2925) 10 ied; white solid; 1H NMR (600 MHz, DMSO-d6
Figure imgf000151_0002
) δ 8.37 (d, J = 3.3 Hz, 1H), 7.71 (d, J = 7.4 Hz, 1H), 7.64 (d, J = 7.5 Hz, 2H), 7.61–7.54 (m, 1H), 7.52– 7.10 (m, 13H), 3.22–3.06 (m, 5H); HRMS (ESI) m/z calcd for C32H25N4O2 [M + H]+ 497.1978, found 497.1965. 15 (S)-3-([1,1’-Biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)- 1H-indazole-5-carboxamide (A56, CDD-2926)
Figure imgf000151_0003
d; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.85– 20 7.78 (m, 1H), 7.75 (dt, J = 7.7, 1.7 Hz, 1H), 7.73–7.66 (m, 2H), 7.60–7.25 (m, 13H), 3.68 (s, - 149 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 1H), 2.99 (s, 3H), 2.90 (s, 3H); HRMS (ESI) m/z calcd for C32H27N4O2 [M + H]+ 499.2134, found 499.2120. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-methyl-2,6- 5 diazaspiro[3.5]nonan-2-yl)methanone (A57, CDD-3009) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.76 (d,
Figure imgf000152_0001
J = 7.5 Hz, 1H), 7.72 (d, J = 7.5 Hz, 2H), 7.67 (s, 1H), 7.58–7.48 (m, 6H), 7.48–7.41 (m, 2H), 3.85–3.73 (m, 4H), 2.47–2.17 (m, 4H), 2.16 (s, 3H), 1.64–1.38 (m, 4H; HRMS (ESI) 10 m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,5- diazaspiro[3.5]nonan-2-yl)methanone (A58, CDD-3010) 15 ied; colorles 1
Figure imgf000152_0002
s oil; H NMR (600 MHz, DMSO-d6) δ 7.76 (d, J = 7.6 Hz, 1H), 7.75–7.71 (m, 3H), 7.59–7.50 (m, 6H), 7.49–7.44 (m, 1H), 7.44–7.39 (m, 1H), 4.21 (dd, J = 35.4, 9.7 Hz, 2H), 3.72 (dd, J = 48.7, 9.7 Hz, 2H), 2.45–2.33 (m, 2H), 2.29 (s, 3H), 1.72 (d, J = 10.0 Hz, 2H), 1.53–1.33 (m, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. 20 (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,5- diazaspiro[3.5]nonan-5-yl)methanone (A59, CDD-3011) - 150 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) pplied; colorless oil; 1H NMR (600 MHz, DMSO-d6) δ 7.77 (d, J =
Figure imgf000153_0001
7.5 Hz, 1H), 7.74–7.69 (m, 2H), 7.60 (d, J = 8.5 Hz, 1H), 7.56–7.42 (m, 7H), 7.38 (dd, J = 8.5, 1.5 Hz, 1H), 3.59–3.54 (m, 2H), 3.17–3.13 (m, 2H), 2.90 (d, J = 7.4 Hz, 2H), 2.30 (s, 5 3H), 1.98 (t, J = 6.0 Hz, 2H), 1.76–1.65 (m, 2H), 1.25–1.13 (m, 2H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2326. 1-(1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- N,N-dimethylpiperidine-4-carboxamide (A60, CDD-3347) 10 ow oil; 1H NMR (800 MHz, CD3
Figure imgf000153_0002
OD) δ 7.74 (dd, J = 7.6, 0.6 Hz, 1H), 7.66 (t, J = 8.5 Hz, 2H), 7.60 (dd, J = 11.6, 6.5 Hz, 1H), 7.54 (t, J = 9.2 Hz, 1H), 7.51–7.48 (m, 3H), 7.47 (d, J = 6.9 Hz, 1H), 7.42 (dd, J = 14.9, 7.6 Hz, 2H), 7.31 (t, J = 12.5 Hz, 1H), 4.07–3.84 (m, 1H), 3.70–3.48 (m, 2H), 3.46–3.34 (m, 1H), 3.29– 15 3.11 (m, 3H), 3.11–2.92 (m, 5H), 2.90–2.57 (m, 3H), 2.36–2.25 (m, 1H), 2.25–2.13 (m, 1H), 2.12–2.00 (m, 1H), 1.98–1.58 (m, 4H); HRMS (ESI) m/z calcd for C34H36N5O2 [M + H]+ 546.2869, found 546.2857. 3-(1-(1-(1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- 20 yl)piperidine-4-carbonyl)piperidin-4-yl)-N-methylpropanamide (A61, CDD-3348) - 151 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) ure for the synthesis of A18 (CDD-
Figure imgf000154_0001
2607) was followed using N-methyl-3-(1-(1-(pyrrolidin-3-yl)piperidine-4-carbonyl)piperidin- 4-yl)propanamide, derived from N-methyl-3-(piperidin-4-yl)propenamide and 1-(1-(tert- 5 butoxycarbonyl)pyrrolidin-3-yl)piperidine-4-carboxylic acid via amide coupling and subsequent removal of Boc-protecting group, in the amide coupling reaction to afford the title compound (71% over two steps) as a pale yellow solid; 1H NMR (800 MHz, CD3OD, mixture of diastereomers and rotamers) δ 7.73 (d, J = 7.6 Hz, 1H), 7.65 (d, J = 7.5 Hz, 2H), 7.59 (t, J = 8.2 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.44–7.50 (m, 4H), 7.42 (t, J = 7.3 Hz, 2H), 10 7.30 (d, J = 7.4 Hz, 1H), 4.49 (dd, J = 48.1, 13.4 Hz, 1H), 4.08–3.96 (m, 1H), 3.90 (dd, J = 21.9, 11.1 Hz, 1H), 3.71–3.24 (m, 4H), 3.17–2.86 (m, 4H), 2.77–2.49 (m, 6H), 2.38–2.27 (m, 1H), 2.26–2.14 (m, 3H), 2.07 (dq, J = 49.8, 11.6, 11.0, 11.0 Hz, 1H), 1.98–1.44 (m, 10H), 1.19–0.72 (m, 2H); 13C NMR (200 MHz, CD3OD, mixture of diastereomers and rotamers) δ 176.6, 175.2, 171.9, 171.8, 145.8, 145.7, 145.5, 142.2, 142.2, 142.0 (br), 141.6, 134.3, 133.8, 15 131.3 (br), 131.1, 130.8, 130.7, 130.6, 130.5, 130.4, 130.4, 130.2, 129.5, 129.5, 129.4, 129.4, 129.1, 128.8, 128.8, 128.5, 128.4, 127.4, 127.3, 125.4, 125.1, 125.1, 122.0, 122.0, 120.7, 120.7, 120.5, 112.0 (br), 94.8, 94.7, 84.2, 78.2, 65.8, 64.8, 56.3, 54.3, 54.2, 53.2, 52.9, 52.4, 52.1, 51.1, 49.8, 46.9, 46.8, 46.4, 43.4, 43.3, 39.2, 39.2, 36.9, 36.9, 34.2, 34.1, 34.0, 33.4, 33.3, 32.9, 32.8, 31.1, 29.6, 29.5, 29.3, 29.3, 26.3, 26.3; HRMS (ESI) m/z calcd for 20 C41H47N6O3 [M + H]+ 671.3710, found 671.3694. 3-(4-(2-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)phenyl)-N-methylpropanamide (A62, CDD-3408) - 152 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) R (600 MHz, DMSO-d6) δ 7.79 (d,
Figure imgf000155_0001
J = 7.6 Hz, 1H), 7.76–7.69 (m, 3H), 7.65–7.40 (m, 8H), 7.27 (d, J = 7.9 Hz, 2H), 7.23 (d, J = 7.8 Hz, 2H), 4.04–3.44 (m, 8H), 2.82 (t, J = 7.9 Hz, 2H), 2.54 (d, J = 4.6 Hz, 3H), 2.35 (t, J = 5 7.8 Hz, 2H), 1.85 (s, 2H), 1.50 (s, 2H); HRMS (ESI) m/z calcd for C40H38N5O3 [M + H]+ 636.2975, found 636.2956. 3-(4-(2-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,7- diazaspiro[3.5]nonane-7-carbonyl)phenyl)-N-methylpropanamide (A63, CDD-3410) 10
Figure imgf000155_0002
NMR (600 MHz, DMSO-d6) δ 7.80 (d, J = 7.6 Hz, 1H), 7.78–7.74 (m, 1H), 7.74–7.71 (m, 2H), 7.70–7.65 (m, 2H), 7.62 (d, J = 8.7 Hz, 1H), 7.57–7.50 (m, 4H), 7.50–7.42 (m, 2H), 7.28–7.23 (m, 4H), 4.02–3.79 (m, 4H), 3.70–3.40 (m, 4H), 2.83 (t, J = 7.8 Hz, 2H), 2.55 (d, J = 4.6 Hz, 3H), 2.37 (t, J = 7.8 Hz, 2H), 15 1.91–1.62 (m, 4H); HRMS (ESI) m/z calcd for C40H38N5O3 [M + H]+ 636.2975, found 636.2958. 4-(2-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide (A64, CDD- 20 3459) - 153 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) NMR (600 MHz, DMSO-d6) δ 7.80 (d,
Figure imgf000156_0001
J = 7.7 Hz, 1H), 7.72 (d, J = 7.7 Hz, 2H), 7.68–7.59 (m, 3H), 7.59–7.39 (m, 6H), 4.07–3.44 (m, 8H), 2.84–2.60 (m, 1H), 2.55 (dd, J = 9.9, 4.5 Hz, 3H), 2.29–2.14 (m, 1H), 2.09–1.14 (m, 5 12H); 13C NMR (150 MHz, DMSO-d6) δ 174.6, 140.8, 139.8, 133.0, 129.7, 129.5, 129.0, 128.3, 127.9, 127.7, 126.6, 126.3, 123.6, 120.1, 83.9, 56.7, 43.4, 34.9, 33.4, 26.1, 25.9, 25.6, 25.4; HRMS (ESI) m/z calcd for C38H40N5O3 [M + H]+ 614.3131, found 614.3113. 4-(2-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,8-10 diazaspiro[4.5]decane-8-carbonyl)-N-methylcyclohexane-1-carboxamide (A65, CDD- 3460) 1
Figure imgf000156_0002
id; H NMR (600 MHz, DMSO-d6) δ 7.79 (dd, J = 7.8, 3.5 Hz, 1H), 7.69 (dd, J = 7.8, 3.8 Hz, 2H), 7.60 (d, J = 8.6 Hz, 1H), 7.57–7.45 15 (m, 7H), 7.41–7.37 (m, 1H), 3.68–3.45 (m, 6H), 3.15 (s, 2H), 2.72–2.61 (m, 1H), 2.59–2.52 (m, 3H), 2.32–2.13 (m, 1H), 1.97–1.84 (m, 2H), 1.81–1.25 (m, 12H); HRMS (ESI) m/z calcd for C39H42N5O3 [M + H]+ 628.3288, found 628.3270. 3-(4-(8-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,8- 20 diazaspiro[4.5]decane-2-carbonyl)phenyl)-N-methylpropanamide (A66, CDD-3461) - 154 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) id; 1H NMR (600 MHz, DMSO-d6) δ 7.83–
Figure imgf000157_0001
7.71 (m, 2H), 7.71–7.58 (m, 3H), 7.56–7.23 (m, 11H), 3.81–3.48 (m, 4H), 3.08 (d, J = 6.2 Hz, 1H), 2.83 (dt, J = 25.7, 7.9 Hz, 2H), 2.55 (d, J = 4.7 Hz, 3H), 2.42–2.28 (m, 2H), 2.11– 5 1.18 (m, 7H); 13C NMR (150 MHz, DMSO-d6) δ 171.5, 171.5, 143.3, 140.0, 132.8, 129.7, 129.4, 129.1, 128.2, 128.2, 127.7, 127.2, 126.8, 120.2, 110.8, 92.7, 84.0, 41.3, 36.6, 32.7, 30.8, 25.4; HRMS (ESI) m/z calcd for C41H40N5O3 [M + H]+ 650.3131, found 650.3115. 3-(4-(2-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,8- 10 diazaspiro[4.5]decane-8-carbonyl)phenyl)-N-methylpropanamide (A67, CDD-3462) olid 1
Figure imgf000157_0002
; H NMR (600 MHz, DMSO-d6) δ 7.83– 7.72 (m, 2H), 7.71–7.64 (m, 2H), 7.60 (dd, J = 8.7, 6.0 Hz, 1H), 7.56–7.35 (m, 8H), 7.34– 7.25 (m, 2H), 7.24–7.15 (m, 2H), 3.89–3.46 (m, 4H), 3.17 (s, 2H), 2.82 (dt, J = 26.8, 7.8 Hz, 15 2H), 2.59–2.52 (m, 3H), 2.36 (dt, J = 24.4, 7.8 Hz, 2H), 1.92 (t, J = 7.6 Hz, 1H), 1.80 (s, 1H), 1.71–1.28 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 171.6, 169.0, 168.6, 143.3, 142.9, 142.8, 140.2, 140.0, 133.9, 132.9, 132.8, 130.4, 129.7, 129.5, 129.1, 128.6, 128.3, 128.2, 128.1, 127.7, 126.9, 126.7, 126.1, 123.6, 123.5, 120.1, 118.8, 92.8, 84.0, 59.0, 55.6, 47.3, 44.2, 41.3, 36.6, 33.0, 30.8, 25.4; HRMS (ESI) m/z calcd for C41H40N5O3 [M + H]+ 650.3131, 20 found 650.3111. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[3.5]nonan-7- - 155 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) yl)methanone (A68, CDD-3463) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.78
Figure imgf000158_0001
(dd, J = 7.7, 1.3 Hz, 1H), 7.69–7.64 (m, 2H), 7.62 (d, J = 8.5 Hz, 1H), 7.56–7.50 (m, 1H), 5 7.50–7.42 (m, 4H), 7.40–7.33 (m, 2H), 7.28 (s, 1H), 3.45–3.07 (m, 8H), 1.95–1.46 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 169.1, 143.3, 140.0, 140.0, 132.8, 129.7, 129.6, 129.4, 129.1, 128.4, 128.3, 127.7, 125.8, 123.6, 120.2, 118.2, 111.1, 92.7, 84.0, 55.1; HRMS (ESI) m/z calcd for C29H27N4O [M + H]+ 447.2185, found 447.2173. 10 3-(4-(7-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,7- diazaspiro[3.5]nonane-2-carbonyl)phenyl)-N-methylpropanamide (A69, CDD-3464)
Figure imgf000158_0002
H NMR (600 MHz, DMSO-d6) δ 7.81–7.73 (m, 2H), 7.70–7.65 (m, 2H), 7.61 (d, J = 8.6 Hz, 1H), 7.58–7.55 (m, 2H), 7.53 (dd, 15 J = 7.5, 1.4 Hz, 1H), 7.51–7.43 (m, 4H), 7.40–7.35 (m, 2H), 7.31–7.24 (m, 3H), 4.09 (s, 2H), 3.83 (br s, 6H), 2.84 (t, J = 7.7 Hz, 2H), 2.55 (d, J = 4.6 Hz, 3H), 2.41–2.31 (m, 2H), 2.06– 1.45 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 171.5, 169.2, 169.0, 144.6, 143.3, 140.1, 140.0, 132.8, 130.9, 129.7, 129.5, 129.4, 129.1, 128.3, 128.2, 127.9, 127.7, 125.7, 123.7, 120.2, 118.2, 111.1, 92.7, 84.0, 62.4, 58.1, 36.5, 34.0, 30.8, 25.4; HRMS (ESI) m/z calcd for 20 C40H38N5O3 [M + H]+ 636.2975, found 636.2957. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4-(4-aminophenoxy)piperidin- 1-yl)methanone (A70, CDD-3465) - 156 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) olid; 1H NMR (600 MHz, DMSO-d6) 13C NMR (1
Figure imgf000159_0001
50 MHz, DMSO-d6) δ 7.79 (dd, J = 7.7, 1.3 Hz, 1H), 7.71–7.65 (m, 2H), 7.62 (dd, J = 8.6, 0.8 Hz, 1H), 7.57–7.52 (m, 1H), 7.52–7.45 (m, 4H), 7.45–7.39 (m, 2H), 7.33–7.28 (m, 5 1H), 6.76–6.68 (m, 2H), 6.53–6.48 (m, 2H), 4.65 (s, 2H), 4.46–4.30 (m, 1H), 4.23–3.53 (m, 2H), 2.12–1.38 (m, 4H); HRMS (ESI) m/z calcd for C33H29N4O2 [M + H]+ 513.2291, found 513.2276. N1-(4-((1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperidin-4- 10 yl)oxy)phenyl)-N4-methylcyclohexane-1,4-dicarboxamide (A71, CDD-3466)
Figure imgf000159_0002
600 MHz, DMSO-d6) 9.79– 9.50 (m, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.71–7.38 (m, 13H), 7.31 (s, 1H), 7.00–6.90 (m, 2H), 4.66–4.56 (m, 1H), 4.07 (br s, 2H), 3.26–3.20 (m, 2H), 2.62–2.51 (m, 3H), 2.45–2.35 (m, 15 1H), 2.30–2.19 (m, 1H), 2.14–1.18 (m, 12H); 13C NMR (150 MHz, DMSO-d6) δ 175.3, 174.7, 173.6, 173.5, 169.2, 152.4, 143.4, 140.1, 140.0, 133.1, 133.0, 132.8, 129.7, 129.5, 129.4, 129.1, 128.4, 128.3, 127.7, 125.8, 123.7, 120.8, 120.7, 120.6, 120.2, 118.3, 116.4, 116.2, 116.2, 111.1, 92.7, 84.0, 72.1, 44.1, 43.3, 41.8, 40.3, 28.4, 26.2, 26.2, 25.6, 25.4; HRMS (ESI) m/z calcd for C42H42N5O4 [M + H]+ 680.3237, found 680.3220. 20 N-(4-((1-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperidin-4- yl)oxy)phenyl)-4-(3-(methylamino)-3-oxopropyl)benzamide (A72, CDD-3467) - 157 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) MHz, DMSO-d6) δ 10.06 (s, 1H)
Figure imgf000160_0001
, 7.87–7.82 (m, 2H), 7.79 (dd, J = 7.7, 1.5 Hz, 2H), 7.71–7.64 (m, 4H), 7.64–7.59 (m, 1H), 7.57–7.39 (m, 8H), 7.36–7.28 (m, 3H), 7.06–6.93 (m, 2H), 4.75–4.55 (m, 1H), 4.08 (s, 5 2H), 3.23 (d, J = 6.3 Hz, 2H), 2.88 (t, J = 7.7 Hz, 2H), 2.55 (d, J = 4.6 Hz, 3H), 2.39 (t, J = 7.7 Hz, 2H), 2.23–1.38 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 171.5, 169.2, 165.0, 153.0, 145.2, 143.4, 140.1, 140.1, 132.8, 132.7, 132.6, 129.7, 129.5, 129.4, 129.1, 128.4, 128.3, 128.2, 127.7, 127.7, 127.6, 125.8, 123.7, 122.0, 120.7, 120.2, 118.3, 116.4, 116.1, 111.1, 92.7, 84.1, 72.1, 36.6, 30.9, 25.5; HRMS (ESI) m/z calcd for C44H40N5O4 [M + H]+ 10 702.3080, found 702.3061. N1-(6-(4-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperazin-1- yl)pyridin-3-yl)-N4-methylcyclohexane-1,4-dicarboxamide (A73, CDD-3468) 15
Figure imgf000160_0002
NMR (600 MHz, DMSO-d6) δ 9.79–9.58 (m, 1H), 8.33 (dd, J = 6.4, 2.6 Hz, 1H), 7.82 (dt, J = 9.2, 2.0 Hz, 1H), 7.78 (dd, J = 7.7, 1.3 Hz, 1H), 7.73–7.57 (m, 4H), 7.56–7.28 (m, 8H), 6.85 (d, J = 9.1 Hz, 1H), 3.78 (s, 2H), 2.59–2.52 (m, 3H), 2.45–2.40 (m, 1H), 2.31–2.21 (m, 1H), 2.12–2.04 (m, 1H), 1.93– 1.72 (m, 4H), 1.59–1.32 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 175.3, 174.8, 173.8, 20 173.7, 169.3, 155.3, 155.3, 143.3, 140.3, 140.0, 139.1, 139.0, 132.8, 130.0, 129.7, 129.4, 129.1, 129.1, 128.5, 128.2, 127.7, 127.6, 127.5, 126.0, 123.7, 120.2, 118.6, 111.2, 107.3, 92.7, 84.1, 44.0, 43.3, 41.6, 40.4, 28.4, 26.2, 26.2, 25.6, 25.4; HRMS (ESI) m/z calcd for - 158 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) C40H40N7O3 [M + H]+ 666.3193, found 666.3174. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4-(6-aminopyridin-3- yl)piperazin-1-yl)methanone (A74, CDD-3469) 5 ellow solid; 1H NMR (600 MHz, DMSO-d6) δ
Figure imgf000161_0001
7.78 (dd, J = 7.6, 1.4 Hz, 1H), 7.70–7.59 (m, 4H), 7.57–7.40 (m, 6H), 7.40–7.30 (m, 2H), 7.20 (dd, J = 8.8, 3.0 Hz, 1H), 6.42 (d, J = 8.9 Hz, 1H), 5.46 (s, 2H), 3.81 (br s, 4H), 3.07– 2.80 (m, 4H); 13C NMR (150 MHz, DMSO-d6) δ 169.1, 154.8, 143.3, 140.2, 140.0, 138.6, 10 136.8, 132.8, 129.7, 129.4, 129.2, 129.1, 128.8, 128.5, 128.3, 127.7, 127.7, 125.9, 123.7, 120.2, 118.6, 111.2, 108.4, 92.8, 84.1, 50.7; HRMS (ESI) m/z calcd for C31H27N6O [M + H]+ 499.2246, found 499.2234. N-(5-(4-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperazin-1- 15 yl)pyridin-2-yl)-4-(3-(methylamino)-3-oxopropyl)benzamide (A75, CDD-3470)
Figure imgf000161_0002
R (600 MHz, DMSO-d6) δ 10.51 (s, 1H), 8.13 (d, J = 3.0 Hz, 1H), 8.06 (d, J = 9.0 Hz, 1H), 7.97–7.92 (m, 2H), 7.82– 7.76 (m, 2H), 7.71–7.62 (m, 3H), 7.58–7.43 (m, 7H), 7.40–7.34 (m, 2H), 7.32 (d, J = 8.2 Hz, 20 2H), 3.86 (br s, 4H), 3.24 (br s, 4H), 2.93–2.82 (m, 2H), 2.56 (d, J = 4.6 Hz, 3H), 2.41 (t, J = 7.7 Hz, 2H); 13C NMR (150 MHz, DMSO-d6) δ 171.5, 169.2, 165.2, 145.5, 144.9, 143.6, - 159 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 143.3, 140.2, 140.0, 135.7, 132.8, 131.9, 129.7, 129.4, 129.1, 129.1, 128.3, 128.2, 127.9, 127.7, 127.7, 126.0, 125.4, 123.7, 120.2, 118.6, 115.2, 111.2, 92.8, 84.0, 48.6, 36.5, 30.9, 25.5; HRMS (ESI) m/z calcd for C42H38N7O3 [M + H]+ 688.3036, found 688.3018. 5 (R)-1-(1-(3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)-3-phenylurea (A76, CDD-3525) Synthetic 600 MHz, DMSO-d6, two
Figure imgf000162_0001
rotamers) δ 8.87–8.61 (m, 1H), 7.76 (d, J = 7.6 Hz, 1H), 7.72–7.63 (m, 2H), 7.59 (d, J = 8.610 Hz, 1H), 7.54–7.30 (m, 10H), 7.28–7.10 (m, 2H), 7.00–6.77 (m, 2H), 4.41–4.04 (m, 1H), 3.86– 3.52 (m, 2H), 3.52–3.41 (m, 1H), 3.41–3.34 (m, 0.5H), 3.28–3.11 (m, 0.5H), 2.30–2.05 (m, 1H), 2.03–1.78 (m, 1H); HRMS (ESI) m/z calcd for C33H28N5O2 [M + H]+ 526.2243, found 526.2233. 15 (R)-3-(1-(3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)-3,4-dihydroquinazolin-2(1H)-one (A77, CDD-3526) Synthetic route 1 was
Figure imgf000162_0002
600 MHz, DMSO-d6, two rotamers) δ 9.45–9.25 (m, 1H), 7.77 (d, J = 7.6 Hz, 1H), 7.73–7.64 (m, 2H), 7.63–7.55 (m,20 1H), 7.53–7.40 (m, 8H), 7.21–7.04 (m, 2H), 6.95–6.68 (m, 2H), 5.14–4.71 (m, 1H), 4.47– 4.28 (m, 2H), 3.89–3.69 (m, 1H), 3.67–3.57 (m, 1H), 3.52 – 3.36 (m, 2H), 2.30–1.94 (m, 2H); HRMS (ESI) m/z calcd for C34H28N5O2 [M + H]+ 538.2243, found 538.2235. (R)-(3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin- - 160 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) 2-yl)amino)pyrrolidin-1-yl)methanone (A78, CDD-3527) Synthetic route 1 was 600 MHz, DMSO-d6, two
Figure imgf000163_0001
rotamers) δ 7.77 (dd, J = 10.8, 7.5 Hz, 1H), 7.67 (dd, J = 20.1, 7.6 Hz, 2H), 7.62–7.27 (m, 5 10H), 6.42 (s, 0.5H), 6.28 (s, 0.5H), 4.59–4.49 (m, 0.5H), 4.40–4.29 (m, 0.5H), 3.91–3.74 (m, 1H), 3.67–3.57 (m, 1H), 3.53–3.43 (m, 1H), 3.40–3.33 (m, 0.5H), 3.22–3.16 (m, 0.5H), 2.24 (s, 3H), 2.21–2.18 (m, 0.5H), 2.15–2.11 (m, 0.5H), 2.10 (s, 3H), 2.06–2.00 (m, 0.5H), 1.91– 1.84 (m, 0.5H); HRMS (ESI) m/z calcd for C32H29N6O [M + H]+ 513.2403, found 513.2393. 10 (R)-N-(1-(3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)-2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide (A79, CDD-3585) Synthetic route 1 wa
Figure imgf000163_0002
0 MHz, DMSO-d6, two rotamers) δ 8.51–8.34 (m, 1H), 7.75 (d, J = 7.6 Hz, 1H), 7.72–7.64 (m, 2H), 7.62–7.54 (m, 15 1H), 7.53–7.29 (m, 10H), 6.97–6.81 (m, 1H), 4.61–4.49 (m, 0.5H), 4.37–4.31 (m, 0.5H), 4.31–4.18 (m, 4H), 3.87–3.75 (m, 1H), 3.67–3.62 (m, 1H), 3.58–3.49 (m, 1H), 3.36–3.21 (m, 1H), 2.31–2.19 (m, 0.5H), 2.18–2.01 (m, 1H), 2.00–1.89 (m, 0.5H); HRMS (ESI) m/z calcd for C35H29N4O4 [M + H]+ 569.2189, found 569.2180. 20 (R)-N-(2-((1-(3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin- 3-yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide (A80, CDD-3586) - 161 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) Synthetic route Hz, DMSO-d6, two
Figure imgf000164_0001
rotamers) δ 9.10–8.88 (m, 1H), 8.38–8.08 (m, 3H), 7.96–7.88 (m, 1H), 7.79 (d, J = 7.6 Hz, 1H), 7.77–7.65 (m, 3H), 7.63–7.58 (m, 1H), 7.56–7.38 (m, 8H), 4.47–4.36 (m, 0.5H), 4.25– 5 4.15 (m, 0.5H), 3.98–3.69 (m, 3H), 3.67–3.59 (m, 0.5H), 3.58–3.52 (m, 0.5H), 3.46–3.42 (m, 1H), 3.19–3.07 (m, 1H), 2.26–2.16 (m, 0.5H), 2.12–2.03 (m, 0.5H), 2.00–1.91 (m, 0.5H), 1.87–1.76 (m, 0.5H); HRMS (ESI) m/z calcd for C36H29F3N5O3 [M + H]+ 636.2222, found 636.2214. 10 (3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(vinylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone (A81, CDD-3880) Synthetic route 1 was ap
Figure imgf000164_0002
(600 MHz, DMSO-d6) δ 7.80 (dd, J = 7.7, 1.3 Hz, 1H), 7.71 (d, J = 6.8 Hz, 2H), 7.69–7.65 (m, 2H), 7.65–7.61 (m, 15 1H), 7.59–7.50 (m, 4H), 7.48 (td, J = 7.4, 1.6 Hz, 1H), 7.44 (t, J = 7.4 Hz, 1H), 6.79 (dd, J = 16.5, 10.0 Hz, 1H), 6.16 (d, J = 10.0 Hz, 1H), 6.12 (d, J = 16.5 Hz, 1H), 3.94–3.74 (m, 4H), 3.21–3.08 (m, 2H), 3.02–2.90 (m, 2H), 1.79–1.64 (m, 2H), 1.64–1.48 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 169.4, 143.3, 140.7, 139.8, 133.0, 132.9, 129.7, 129.6, 129.1, 129.1 (2 ×), 129.0, 128.3 (2 ×), 127.8, 127.7, 126.6, 126.5, 123.6, 120.1, 119.8, 110.9, 92.8, 83.8, 20 61.2, 56.9, 52.9, 45.1, 34.4, 32.2, 21.7; HRMS (ESI) m/z calcd for C31H29N4O3S [M + H]+ 537.1960, found 537.1953. (3-([1,1'-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(ethylsulfonyl)-2,6- - 162 - 53366970.3 Attorney Docket. No.046641-7059WO1(00159) diazaspiro[3.5]nonan-2-yl)methanone (A82, CDD-3881) Synthetic route 1 was ap (600 MHz, DMSO-d6) δ
Figure imgf000165_0001
7.80 (dd, J = 7.7, 1.3 Hz, 1H), 7.72–7.65 (m, 4H), 7.63 (dd, J = 8.6, 0.9 Hz, 1H), 7.57–7.50 5 (m, 4H), 7.48 (td, J = 7.4, 1.6 Hz, 1H), 7.44 (t, J = 7.4 Hz, 1H), 3.94–3.73 (m, 4H), 3.31–3.26 (m, 2H), 3.15–3.07 (m, 2H), 3.05 (q, J = 7.4 Hz, 2H), 1.81–1.69 (m, 2H), 1.62–1.45 (m, 2H), 1.19 (t, J = 7.4 Hz, 3H); 13C NMR (150 MHz, DMSO-d6) δ 169.4, 143.3, 140.7, 139.8, 133.1, 129.7, 129.6, 129.1 (2 ×), 129.0, 128.3 (2 ×), 127.9, 127.7, 126.6, 126.5, 123.6, 120.1, 119.8, 110.9, 92.8, 83.8, 61.2, 56.8, 52.9, 45.1, 43.0, 34.5, 32.5, 22.1, 7.6; HRMS (ESI) m/z calcd 10 for C31H31N4O3S [M + H]+ 539.2117, found 539.2108. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1-methyl-1H-indazol-5-yl)(3- dimethylamino)pyrrolidin-1-yl)methanone (B1, CDD-2533) 15
Figure imgf000165_0002
d. The general procedure for amide coupling was employed using 3-iodo-1-methyl-1H-indazole-5-carboxylic acid and (R)-N,N-dimethyl-3- pyrrolidinamine. The resulting amide was partially purified and subjected to the general procedure for Sonogashira coupling using 2-ethynyl-1,1’-biphenyl to afford the title compound (16% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two 20 rotamers) δ 7.77 (dd, J = 7.7, 1.4 Hz, 1H), 7.72 (dd, J = 8.6, 2.9 Hz, 1H), 7.66 (d, J = 7.5 Hz, 2H), 7.59–7.51 (m, 2H), 7.51–7.44 (m, 4H), 7.43–7.37 (m, 2H), 4.07 (s, 3H), 3.84–3.76 (m, 0.5H), 3.75–3.65 (m, 0.5H), 3.59–3.48 (m, 0.5H), 3.36–3.27 (m, 2H), 3.22–3.12 (m, 0.5H), 2.80–2.69 (m, 0.5H), 2.67–2.59 (m, 0.5H), 2.21 (s, 3H), 2.16–1.96 (m, 4H), 1.84–1.78 (m, 0.5H), 1.74–1.64 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.3, 168.2, - 163 - 53366970.3 143.3, 140.0, 139.9, 132.8, 130.5, 130.1, 129.6, 129.4, 129.1, 129.0, 128.3, 127.7, 127.7, 127.4, 126.2, 126.0, 124.2, 124.1, 120.1, 118.8, 118.8, 110.4, 93.1, 83.5, 65.2, 63.9, 53.0, 50.3, 48.2, 45.0, 43.9, 43.7, 36.0, 30.5, 28.5; HRMS (ESI) m/z calcd for C29H29N4O [M + H]+ 449.2341, found 449.2328. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (B2, CDD-2534) d. The same procedure for the synthesis of B1 (CDD-
Figure imgf000166_0001
2533) was followed using methyl 3-bromo-1H-pyrazolo[3,4-b]pyridine-5-carboxylate in the amide coupling reaction; light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.72–8.66 (m, 1H), 7.82 (td, J = 9.6, 1.7 Hz, 2H), 7.67 (d, J = 7.6 Hz, 2H), 7.64–7.44 (m, 6H), 7.44–7.38 (m, 1H), 3.85–3.75 (m, 0.5H), 3.75–3.68 (m, 0.5H), 3.58–3.47 (m, 1H), 3.32– 3.21 (m, 2H), 2.80–2.73 (m, 0.5H), 2.70–2.61 (m, 0.5H), 2.22 (s, 3H), 2.17–2.12 (m, 0.5H), 2.08 (s, 3H), 2.05–1.98 (m, 0.5H), 1.88–1.78 (m, 0.5H), 1.78–1.66 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 166.4, 166.3, 151.3, 148.9, 148.8, 143.6, 139.9, 132.9, 132.0, 132.0, 131.5, 131.4, 129.7, 129.7, 129.1, 129.1, 128.8, 128.7, 128.3, 128.2, 127.9, 127.8, 127.7, 126.9, 126.5, 119.8, 115.0, 114.9, 93.0, 83.3, 65.2, 63.9, 52.9, 50.4, 48.1, 45.2, 43.9, 43.7, 30.5, 28.5; HRMS (ESI) m/z calcd for C27H26N5O [M + H]+ 436.2137, found 436.2126. 3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazole (B3, CDD-2536)
Figure imgf000166_0002
re for Sonogashira coupling was employed using 3-iodo-1H- indazole and 2-ethynyl-1,1'-biphenyl to afford the title compound (78%) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 13.44 (s, 1H), 7.77 (dd, J = 7.7, 1.4 Hz, 1H), 7.71–7.66 (m, 2H), 7.59–7.43 (m, 7H), 7.40 (ddd, J = 8.2, 6.8, 1.1 Hz, 1H), 7.36 (d, J = 8.1 Hz, 1H), 7.16 (ddd, J = 7.9, 6.8, 0.9 Hz, 1H); 13C NMR (150 MHz, DMSO-d6) δ 143.3, 140.0, 139.9, 132.8, 129.6, 129.3, 129.1, 128.3, 127.8, 127.7, 127.6, 126.8, 124.2, 121.4, 120.3, 119.4, 110.8, 92.1, 84.5; HRMS (ESI) m/z calcd for C21H14N2 [M + H]+ 295.1235, found 295.1224. (R)-1-((3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-5-yl)methyl)-N,N- dimethylpyrrolidin-3-amine (B4, CDD-2761) ed except the amide coupling reaction was replaced with
Figure imgf000167_0001
reductive amination using 3-iodo-1H-indazole-5-carboxaldehyde and (R)-N,N- dimethylpyrrolidin-3-amine; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.76 (d, J = 7.6 Hz, 1H), 7.70 (d, J = 7.6 Hz, 2H), 7.58–7.42 (m, 7H), 7.39–7.32 (m, 1H), 7.21 (s, 1H), 3.66 (d, J = 12.6 Hz, 1H), 3.56 (d, J = 12.7 Hz, 1H), 2.75–2.63 (m, 2H), 2.62–2.54 (m, 1H), 2.47–2.39 (m, 1H), 2.30–2.21 (m, 1H), 2.07 (s, 6H), 1.91–1.79 (m, 1H), 1.67–1.56 (m, 1H); HRMS (ESI) m/z calcd for C28H29N4 [M + H]+ 421.2392, found 421.2379. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(2,8- diazaspiro[4.5]decan-2-yl)methanone (B5, CDD-2803)
Figure imgf000167_0002
The same procedure for the synthesis of A26 (CDD- 2674) was followed using methyl 3-bromo-1H-pyrazolo[3,4-b]pyridine-5-carboxylate in the Sonogashira coupling reaction; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.71–8.61 (m, 1H), 7.95–7.74 (m, 2H), 7.70–7.66 (m, 2H), 7.58–7.53 (m, 1H), 7.51–7.46 (m, 4H), 7.42– 7.35 (m, 1H), 3.62 (t, J = 7.2 Hz, 2H), 3.19 (s, 2H), 2.83–2.51 (m, 4H), 1.84 (t, J = 7.3 Hz, 1H), 1.74 (t, J = 6.8 Hz, 1H), 1.60–1.44 (m, 2H), 1.43–1.31 (m, 2H); HRMS (ESI) m/z calcd for C29H28N5O [M + H]+ 462.2294, found 462.2280. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(2,6- diazaspiro[4.5]decan-2-yl)methanone (B6, CDD-2805) . The same procedure for the synthesis of B5 (CDD-
Figure imgf000168_0001
2803) was followed using tert-butyl 2,6-diazaspiro[4.5]decane-6-carboxylate in the amide coupling reaction; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.71–8.63 (m, 1H), 7.91–7.73 (m, 2H), 7.67 (d, J = 7.6 Hz, 2H), 7.56 (t, J = 7.6 Hz, 1H), 7.52–7.39 (m, 5H), 3.71–3.65 (m, 1H), 3.61–3.56 (m, 2H), 3.26–3.23 (m, 1H), 2.80–2.71 (m, 1H), 2.68–2.51 (m, 1H), 2.04–1.87 (m, 1H), 1.85–1.66 (m, 1H), 1.64–1.19 (m, 6H); HRMS (ESI) m/z calcd for C29H28N5O [M + H]+ 462.2294, found 462.2281. (S)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(2- phenylpiperazin-1-yl)methanone (B7, CDD-2846)
Figure imgf000168_0002
lied. The same procedure for the synthesis of B5 (CDD- 2803) was followed using tert-butyl (S)-3-phenylpiperazine-1-carboxylate in the amide coupling reaction; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.40–8.26 (m, 1H), 7.72–7.63 (m, 4H), 7.48–7.38 (m, 9H), 7.28 (t, J = 7.3 Hz, 2H), 3.59–3.55 (m, 1H), 3.11–2.97 (m, 3H), 2.89–2.65 (m, 3H); HRMS (ESI) m/z calcd for C31H26N5O [M + H]+ 484.2137, (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(7-oxa-2- azaspiro[3.5]nonan-2-yl)methanone (B8, CDD-2870) d. The same procedure for the synthesis of B5 (CDD-
Figure imgf000169_0001
2803) was followed using 7-oxa-2-azaspiro[3.5]nonane in the amide coupling reaction; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.43 (d, J = 2.2 Hz, 1H), 7.84 (d, J = 2.2 Hz, 1H), 7.77–7.72 (m, 2H), 7.70–7.63 (m, 1H), 7.51 (t, J = 7.7 Hz, 2H), 7.47–7.37 (m, 4H), 3.96 (s, 2H), 3.90–3.73 (m, 2H), 3.55–3.48 (m, 4H), 1.73 (t, J = 5.3 Hz, 4H); HRMS (ESI) m/z calcd for C28H25N4O2 [M + H]+ 449.1978, found 449.1965. (R)-(3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (B9, CDD-2933)
Figure imgf000169_0002
d; light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.07 (dd, J = 8.7, 4.1 Hz, 1H), 7.80–7.63 (m, 4H), 7.56–7.49 (m, 2H), 7.48–7.40 (m, 3H), 7.40–7.33 (m, 1H), 4.01 (dd, J = 11.3, 6.7 Hz, 0.5H), 3.87–3.60 (m, 2.5H), 3.26 (dd, J = 11.9, 8.4 Hz, 1H), 2.73–2.64 (m, 1H), 2.20 (s, 3H), 2.08 (s, 3H), 2.07–2.01 (m, 1H), 1.77–1.66 (m, 1H); HRMS (ESI) m/z calcd for C27H26N5O [M + H]+ 436.2137, found 436.2126. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (B10, CDD-2934) ; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.95 (d, J =
Figure imgf000170_0001
8.6 Hz, 1H), 7.79–7.74 (m, 2H), 7.69 (dd, J = 8.2, 1.7 Hz, 2H), 7.52–7.44 (m, 4H), 7.42 (td, J = 7.4, 1.7 Hz, 1H), 7.40–7.36 (m, 1H), 4.42 (s, 2H), 3.75 (s, 2H), 2.46–1.93 (m, 7H), 1.80–1.65 (m, 4H); HRMS (ESI) m/z calcd for C29H28N5O [M + H]+ 462.2294, found 462.2280. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-4-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (B11, CDD-3006) lied; white foam; 1H NMR (600 MHz, DMSO-d6
Figure imgf000170_0002
) δ 7.75– 7.68 (m, 3H), 7.63 (d, J = 8.3 Hz, 1H), 7.51–7.44 (m, 5H), 7.42–7.35 (m, 1H), 7.27–7.20 (m, 1H), 7.02 (d, J = 6.8 Hz, 1H), 3.64 (s, 2H), 2.41–1.64 (m, 7H), 1.54–1.42 (m, 4H); (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2328. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-6-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (B12, CDD-3007)
Figure imgf000170_0003
; white foam; 1H NMR (600 MHz, DMSO-d6) δ 7.81 (s, 1H), 7.73 (d, J = 7.6 Hz, 1H), 7.69 (d, J = 7.3 Hz, 2H), 7.53–7.42 (m, 6H), 7.33 (d, J = 8.2 Hz, 1H), 7.30–7.26 (m, 1H), 4.00 (s, 2H), 3.73 (s, 2H), 2.49–1.99 (m, 6H), 1.70 (s, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2329. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-indazol-7-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (B13, CDD-3008) lied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.79
Figure imgf000171_0001
(dd, J = 7.6, 1.3 Hz, 1H), 7.71–7.66 (m, 2H), 7.64 (d, J = 7.2 Hz, 1H), 7.57–7.43 (m, 7H), 7.21 (t, J = 7.6 Hz, 1H), 4.05 (s, 2H), 3.81 (s, 2H), 2.45–1.83 (m, 7H), 1.76–1.67 (m, 4H); HRMS (ESI) m/z calcd for C30H29N4O [M + H]+ 461.2341, found 461.2327. (3-([1,1’-Biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(2,7- diazaspiro[3.5]nonan-2-yl)methanone (B14, CDD-3034) ; yellow sol 1
Figure imgf000171_0002
id; H NMR (600 MHz, DMSO-d6) δ 8.15 (d, J = 8.8 Hz, 1H), 7.99 (d, J = 8.8 Hz, 1H), 7.77–7.75 (m, 1H), 7.75–7.71 (m, 2H), 7.59– 7.52 (m, 2H), 7.51–7.44 (m, 3H), 7.43–7.36 (m, 1H), 4.39 (s, 2H), 3.78 (s, 2H), 2.75–2.55 (m, 4H), 1.75–1.53 (m, 4H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2126. (3-([1,1'-Biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (B15, CDD-3882) Synthetic route 1 was appl 00 MHz, DMSO-d6) δ 7.86 (dd, J = 8.8, 2.3 Hz, 1H), 7.81–7.7
Figure imgf000172_0001
7 (m, 2H), 7.68 (d, J = 7.5 Hz, 1H), 7.56–7.52 (m, 1H), 7.48–7.36 (m, 6H), 4.40–4.32 (m, 2H), 3.74 (d, J = 9.7 Hz, 1H), 3.66 (d, J = 9.7 Hz, 1H), 2.78–2.51 (m, 4H), 1.73–1.60 (m, 2H), 1.45–1.32 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2130. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5- yl)methanone (C1, CDD-2530)
Figure imgf000172_0002
d. The general procedure for amide coupling was employed using 3-iodo-1H-indazole-5-carboxylic acid and (R)-N,N-dimethyl-3- pyrrolidinamine. The resulting amide was partially purified and subjected to the general procedure for Sonogashira coupling using phenylacetylene to afford the title compound (8% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.99 (s, 1H), 7.72–7.67 (m, 2H), 7.65 (d, J = 8.6 Hz, 1H), 7.58 (t, J = 8.5 Hz, 1H), 7.50–7.44 (m, 3H), 3.76–3.71 (m, 0.5H), 3.67–3.62 (m, 0.5H), 3.59–3.52 (m, 1H), 3.52–3.45 (m, 1H), 3.27–3.22 (m, 1H), 2.77–2.69 (m, 0.5H), 2.68–2.60 (m, 0.5H), 2.18 (s, 3H), 2.13–1.97 (m, 4H), 1.82– 1.73 (m, 0.5H), 1.73–1.62 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.3, 140.3, 131.5, 129.1, 128.8, 128.6, 126.1, 121.8, 118.9, 110.8, 92.8, 81.1, 65.2, 63.9, 53.0, 50.2, 48.2, 45.0, 43.9, 43.6, 30.4, 28.3; HRMS (ESI) m/z calcd for C22H23N4O [M + H]+ 359.1872, found 359.1860. (R)-(3-((2,6-Dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (C2, CDD-2531) The same procedure for the synthesis of C1 (CDD-
Figure imgf000173_0001
2530) was followed using 1,3-dichloro-2-ethynylbenzene in the Sonogashira coupling reaction to afford the title compound (20% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.96 (d, J = 10.0 Hz, 1H), 7.71–7.47 (m, 5H), 3.76– 3.51 (m, 3H), 3.28–3.25 (m, 1H), 2.83–2.57 (m, 1H), 2.18 (s, 3H), 2.12–1.95 (m, 4H), 1.83– 1.62 (m, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.0, 140.4, 135.8, 131.0, 129.0, 128.3, 128.3, 127.8, 126.7, 123.8, 121.5, 118.7, 111.1, 91.5, 86.8, 65.2, 63.8, 53.2, 50.3, 48.3, 45.2, 43.8, 43.7, 30.4, 28.3; HRMS (ESI) m/z calcd for C22H21Cl2N4O [M + H]+ 427.1092, found 427.1080. (R)-(3-([1,1’-Biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (C3, CDD-2532)
Figure imgf000173_0002
d. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 3-ethynyl-1,1'-biphenyl in the Sonogashira coupling reaction to afford the title compound (26% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.06 (d, J = 4.8 Hz, 1H), 7.97 (s, 1H), 7.77–7.72 (m, 3H), 7.71– 7.64 (m, 2H), 7.61–7.54 (m, 2H), 7.49 (t, J = 7.7 Hz, 2H), 7.40 (t, J = 7.2 Hz, 1H), 3.76–3.71 (m, 0.5H), 3.66–3.62 (m, 0.5H), 3.58–3.51 (m, 0.5H), 3.51–3.44 (m, 0.5H), 3.36–3.34 (m, 1H), 3.27–3.22 (m, 1H), 2.75–2.66 (m, 0.5H), 2.66–2.58 (m, 0.5H), 2.17 (s, 3H), 2.10–1.95 (m, 4H), 1.81–1.72 (m, 0.5H), 1.72–1.62 (m, 0.5H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.4, 140.8, 140.3, 139.1, 130.5, 130.2, 129.6, 129.5, 129.0 (2 ×), 128.6, 128.3, 127.9, 127.5, 126.9 (2 ×), 126.3, 126.2, 123.5, 122.5, 119.0, 110.8, 92.8, 81.4, 65.1, 63.9, 53.0, 50.3, 48.2, 45.1, 43.8, 43.6, 30.4, 28.3; HRMS (ESI) m/z calcd for C28H27N4O [M + H]+ 435.2185, found 435.2172. (R)-(3-([1,1’-Biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (C4, CDD-2563) The same procedure for the synthesis of C1 (CDD-
Figure imgf000174_0001
2530) was followed using 4-ethynyl-1,1'-biphenyl in the Sonogashira coupling reaction to afford the title compound (8% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.03 (s, 1H), 7.78 (s, 4H), 7.74 (d, J = 7.2 Hz, 2H), 7.66 (d, J = 8.6 Hz, 1H), 7.59 (t, J = 8.8 Hz, 1H), 7.50 (t, J = 7.6 Hz, 2H), 7.41 (t, J = 7.3 Hz, 1H), 3.80– 3.71 (m, 0.5H), 3.69–3.61 (m, 0.5H), 3.60–3.46 (m, 2H), 3.29–3.22 (m, 1H), 2.79–2.69 (m, 0.5H), 2.69–2.60 (m, 0.5H), 2.19 (s, 3H), 2.13–1.96 (m, 4H), 1.83–1.63 (m, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.3, 140.6, 140.3, 139.1, 132.1 (2 ×), 130.5, 130.1, 129.0 (2 ×), 128.6, 128.0, 126.9 (2 ×), 126.7 (2 ×), 126.3, 123.5, 120.8, 118.9, 110.8, 92.8, 81.8, 65.2, 63.9, 53.1, 50.3, 48.2, 45.0, 43.9, 43.6, 30.4, 28.3; HRMS (ESI) m/z calcd for C28H27N4O [M + H]+ 435.2185, found 435.2173. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol- 5-yl)methanone (C5, CDD-2564) e same procedure for the synthesis of C1 (CDD- 2530) w
Figure imgf000175_0001
as followed using 5-ethynylisoquinoline in the Sonogashira coupling reaction to afford the title compound (18% over two steps) as a white solid; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 13.90 (s, 1H), 9.46 (s, 1H), 8.73 (s, 1H), 8.27 (d, J = 8.2 Hz, 1H), 8.25–8.17 (m, 2H), 8.11 (d, J = 7.0 Hz, 1H), 7.79 (t, J = 7.7 Hz, 1H), 7.71 (dd, J = 8.6, 2.7 Hz, 1H), 7.64 (dd, J = 13.3, 8.5 Hz, 1H), 3.81–3.46 (m, 3H), 3.42–3.39 (m, 1H),2.80–2.61 (m, 1H), 2.20 (s, 3H), 2.09 (s, 3H), 2.02 (q, J = 6.2 Hz, 1H), 1.75 (dt, J = 43.4, 10.4 Hz, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.2, 153.0, 144.4, 140.4, 134.8, 134.8, 129.0, 128.3, 127.3, 126.4, 123.4, 119.0, 118.4, 117.9, 111.0, 89.3, 87.0, 65.2, 63.8, 53.2, 50.2, 48.2, 45.1, 43.8, 43.7, 30.4, 28.3; HRMS (ESI) m/z calcd for C25H24N5O [M + H]+ 410.1980, found 410.1971. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-(pyrimidin-5-ylethynyl)-1H-indazol-5- yl)methanone (C6, CDD-2568)
Figure imgf000175_0002
. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 5-ethynylpyrimidine in the Sonogashira coupling reaction to afford the title compound (29% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 12.47 (s, 1H), 9.20 (s, 1H), 8.94 (s, 2H), 8.06 (d, J = 12.1 Hz, 1H), 7.57 (d, J = 8.6 Hz, 1H), 7.47 (d, J = 8.6 Hz, 1H), 4.06–3.89 (m, 1H), 3.76–3.42 (m, 3H), 2.91–2.70 (m, 1H), 2.36 (s, 3H), 2.25 (s, 3H), 2.13 (dt, J = 13.7, 6.6 Hz, 1H), 1.92 (q, J = 11.2 Hz, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 169.9, 158.8, 157.0, 140.7, 130.3, 129.4, 126.4, 124.0, 119.8, 119.3, 110.8, 87.7, 86.4, 65.7, 64.5, 54.2, 50.7, 48.9, 45.7, 44.2, 31.1, 29.1; HRMS (ESI) m/z calcd for C20H21N6O [M + H]+ 361.1776, found 361.1765. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol- 5-yl)methanone (C7, CDD-2569) d. The same procedure for the synthesis of C1 (CDD-
Figure imgf000176_0001
2530) was followed using 1-ethynyl-2-fluorobenzene in the Sonogashira coupling reaction to afford the title compound (27% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 12.69 (s, 1H), 8.10 (d, J = 17.6 Hz, 1H), 7.59 (dd, J = 20.3, 8.2 Hz, 3H), 7.37 (tdd, J = 7.4, 5.2, 1.7 Hz, 1H), 7.20–7.12 (m, 2H), 3.97 (ddd, J = 68.6, 12.7, 8.4 Hz, 1H), 3.81–3.42 (m, 3H), 2.90–2.69 (m, 1H), 2.35 (s, 3H), 2.25 (s, 4H), 1.91 (dt, J = 26.6, 10.7 Hz, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 169.9, 163.6, 161.9, 140.8, 133.6, 130.6, 130.6, 130.2, 129.9, 126.6, 124.1, 124.1, 120.0, 119.9, 115.7, 115.6, 111.3, 111.2, 110.8, 87.3, 85.6, 65.8, 64.5, 54.1, 50.7, 48.9, 45.7, 44.2, 44.2, 31.1, 29.2; HRMS (ESI) m/z calcd for C22H22FN4O [M + H]+ 377.1777, found 377.1767. (R)-5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile (C8, CDD-2572)
Figure imgf000176_0002
pplied. To a solution of (R)-(3-(dimethylamino)pyrrolidin-1- yl)(3-iodo-1H-indazol-5-yl)methanone (0.1 mmol) in anhydrous DMA (0.5 mL) were added Zn(CN)2 (0.17 mmol), Zn (0.06 mmol), CuI (0.1 mmol), and Pd(dppf)Cl2 (0.015 mmol). The reaction was heated at 120 oC under microwave for 1 h. Then the solution was filtrated with a pad of celite. The filtrate was concentrated and purified by flash chromatography to afford the title compound (55%); 1H NMR (600 MHz, CDCl3, two rotamers) δ 8.00 (d, J = 8.3 Hz, 1H), 7.61 (dd, J = 8.8, 5.7 Hz, 1H), 7.50 (t, J = 9.3 Hz, 1H), 4.04–3.98 (m, 0.5H), 3.94 (t, J = 10.8 Hz, 0.5H), 3.75–3.67 (m, 0.5H), 3.67–3.54 (m, 2H), 3.43 (t, J = 9.5 Hz, 0.5H), 2.92– 2.76 (m, 1H), 2.37 (s, 3H), 2.26 (s, 3H), 2.24–2.23 (m, 0.5H), 2.17–2.11 (m, 0.5H), 1.98–1.87 (m, 1H); HRMS (ESI) m/z calcd for C15H18N5O [M + H]+ 284.1511, found 284.1503. (R)-(3-([1,1’-Biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone (C9, CDD-2573) he general procedure for amide coupling was
Figure imgf000177_0001
employed using 3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylic acid and (R)- N,N-dimethyl-3-pyrrolidinamine. The resulting amide was partially purified by flash chromatography and used in the next step. A oven dried microwave vial equipped with magnetic stir bar was charged with the amide (0.05 mmol, 1.0 equiv.), [1,1'-biphenyl]-2- ylboronic acid (0.1 mmol, 2.0 equiv.), Pd(PPh3)4 (0.005 mmol, 0.1 equiv.), and K2CO3 (0.1 mmol, 2.0 equiv.). The vial was sealed with a microwave cap, evacuated and backfilled with nitrogen, followed by addition of anhydrous DMF (0.3 mL) and water (0.1 mL, purged with nitrogen) via syringe. The mixture was vacuum-purged and refilled with nitrogen for three cycles and then heated by microwave reactor at 100 oC for 1 h. After completion of the reaction (monitored by TLC and LC-MS), the reaction mixture was filtered through a pad of celite. The filtrate was concentrated and partially purified by flash chromatography to yield (3-([1,1'-biphenyl]-2-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)((R)-3- (dimethylamino)pyrrolidin-1-yl)methanone. The general procedure for removal of THP was applied to afford the title compound (45% over three steps) as a yellow oil; 1H NMR (600 MHz, CDCl3, two rotamers) δ 7.65 (d, J = 7.5 Hz, 1H), 7.54 (d, J = 6.1 Hz, 2H), 7.50–7.32 (m, 4H), 7.21 (br s, 2H), 7.12 (d, J = 7.4 Hz, 3H), 3.94–3.84 (m, 0.5H), 3.84–3.72 (m, 0.5H), 3.69–3.54 (m, 0.5H), 3.46–3.34 (m, 1H), 3.26–3.11 (m, 1.5H), 2.85–2.58 (m, 1H), 2.33 (s, 3H), 2.25–1.99 (m, 4H), 1.88–1.54 (m, 1H); HRMS (ESI) m/z calcd for C26H27N4O [M + H]+ 411.2185, found 411.2176. (R)-2-((5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)- N-phenylbenzamide (C10, CDD-2580) d. The same procedure for the synthesis of C1 (CDD- 2530) w
Figure imgf000178_0001
as followed using 2-ethynyl-N-phenylbenzamide in the Sonogashira coupling reaction to afford the title compound (8% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.94–7.49 (m, 9H), 7.36–6.91 (m, 3H), 3.81–3.62 (m, 1H), 3.55–3.42 (m, 2H), 3.30–3.25 (m, 1H), 2.78–2.58 (m, 1H), 2.28–2.15 (m, 3H), 2.11–1.84 (m, 4H), 1.84–1.52 (m, 1H); HRMS (ESI) m/z calcd for C29H28N5O2 [M + H]+ 478.2243, found 478.2228. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone (C11, CDD-2581) N
Figure imgf000178_0002
d. The same procedure for the synthesis of C1 (CDD- 2530) was followed using (2-ethynylphenyl)(4-methylpiperazin-1-yl)methanone in the Sonogashira coupling reaction to afford the title compound (14% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 7.93 (s, 1H), 7.78–7.71 (m, 1H), 7.66 (d, J = 8.6 Hz, 1H), 7.59 (d, J = 8.7 Hz, 1H), 7.55–7.48 (m, 2H), 7.43–7.36 (m, 1H), 3.78–3.72 (m, 1H), 3.66–3.55 (m, 2H), 3.54–3.46 (m, 2H), 3.30–3.26 (m, 1H), 3.23–3.14 (m, 2H), 2.84–2.58 (m, 1H), 2.42–2.23 (m, 4H), 2.20 (s, 3H), 2.08 (s, 3H), 2.06–1.98 (m, 4H), 1.86–1.61 (m, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 167.3, 140.2, 139.4, 131.8, 129.4, 129.0, 128.4, 126.5, 126.4, 123.7, 118.9, 118.7, 110.9, 90.9, 84.3, 65.2, 63.9, 54.5, 54.0, 52.9, 50.3, 48.2, 46.4, 45.5, 45.0, 43.8, 43.6, 41.0, 30.4, 28.2; HRMS (ESI) m/z calcd for C28H33N6O2 [M + H]+ 485.2665, found 485.2654. (R)-(3-(Cyclopropylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone (C12, CDD-2589) ed. The same procedure for the synthesis of C1 (CDD-
Figure imgf000179_0001
2530) was followed using ethynylcyclopropane in the Sonogashira coupling reaction to afford the title compound (32% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 11.68 (s, 1H), 7.98 (d, J = 8.7 Hz, 1H), 7.58–7.45 (m, 2H), 4.04–3.82 (m, 1H), 3.75–3.39 (m, 3H), 2.74 (ddt, J = 74.9, 16.0, 7.5 Hz, 1H), 2.33 (s, 3H), 2.23 (s, 3H), 2.10 (dd, J = 12.4, 6.3 Hz, 1H), 1.88 (dq, J = 20.3, 10.4 Hz, 1H), 1.56 (tt, J = 8.1, 5.2 Hz, 1H), 0.97–0.89 (m, 4H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 170.0, 140.6, 131.3, 129.8, 126.4, 124.1, 120.2, 110.4, 98.4, 66.7, 65.8, 64.6, 54.3, 50.6, 48.8, 45.6, 44.3, 31.2, 29.3, 8.9, 0.3; HRMS (ESI) m/z calcd for C19H23N4O [M + H]+ 323.1871, found 323.1862. (R)-6-((5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)pyridin-2(1H)-one (C13, CDD-2590)
Figure imgf000179_0002
d. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 6-ethynylpyridin-2(1H)-one in the Sonogashira coupling reaction to afford the title compound (20% over two steps) as a white solid; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 13.87 (s, 1H), 12.14 (s, 1H), 8.20 (d, J = 4.0 Hz, 1H), 7.68 (dd, J = 8.7, 2.0 Hz, 1H), 7.60 (t, J = 8.0 Hz, 1H), 7.50 (dd, J = 9.2, 6.8 Hz, 1H), 6.70 (d, J = 6.8 Hz, 1H), 6.48 (d, J = 9.1 Hz, 1H), 3.82–3.63 (m, 1H), 3.59–3.46 (m, 2H), 3.27 (dd, J = 11.8, 8.4 Hz, 1H), 2.69 (dq, J = 52.1, 7.4 Hz, 1H), 2.20 (s, 3H), 2.08 (s, 4H), 1.84–1.67 (m, 1H); 13C NMR (150 MHz, DMSO-d6, two rotamers) δ 168.4, 162.4, 140.3, 140.2, 130.8, 130.5, 127.4, 126.4, 126.3, 123.9, 119.2, 111.4, 110.9, 87.0, 84.6, 65.1, 63.9, 53.1, 50.2, 48.2, 45.0, 43.9, 43.7, 30.4, 28.4; HRMS (ESI) m/z calcd for C21H25N5O2 [M + H]+ 376.1773, found 376.1764. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)- 1H-indazol-5-yl)methanone (C14, CDD-2591) d. The same procedure for the synthesis of C1 (CDD-
Figure imgf000180_0001
2530) was followed using 1-ethynyl-2-(trifluoromethoxy)benzene in the Sonogashira coupling reaction to afford the title compound (24% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 12.44 (s, 1H), 8.06 (d, J = 9.6 Hz, 1H), 7.68 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 8.2 Hz, 1H), 7.51 (d, J = 8.7 Hz, 1H), 7.42 (t, J = 7.9 Hz, 1H), 7.33 (q, J = 6.3 Hz, 2H), 4.08–3.88 (m, 1H), 3.75–3.41 (m, 3H), 2.80 (dp, J = 59.1, 7.9 Hz, 1H), 2.36 (s, 3H), 2.24 (s, 3H), 2.13 (dd, J = 12.0, 6.3 Hz, 1H), 1.91 (dt, J = 22.4, 11.0 Hz, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 169.9, 149.3, 140.8, 133.8, 130.2, 130.0, 126.9, 126.6, 124.2, 124.1, 121.5, 121.2, 119.8, 119.7, 118.1, 117.3, 110.8, 87.8, 85.8, 65.7, 64.5, 53.9, 50.7, 48.8, 45.6, 44.2, 44.1, 31.1, 29.1; HRMS (ESI) m/z calcd for C23H22F3N4O2 [M + H]+ 443.1694, found 443.1682. (R)-(3-((2-Chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1-yl)methanone (C15, CDD-2593)
Figure imgf000180_0002
y pp ed. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 1-chloro-2-ethynylbenzene in the Sonogashira coupling reaction to afford the title compound (24% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 12.36 (s, 1H), 8.10 (d, J = 14.2 Hz, 1H), 7.62 (d, J = 7.5 Hz, 1H), 7.56 (t, J = 7.9 Hz, 1H), 7.50 (dd, J = 8.7, 4.8 Hz, 1H), 7.44 (dd, J = 8.1, 4.2 Hz, 1H), 7.32–7.23 (m, 2H), 3.95 (ddd, J = 67.3, 12.7, 8.5 Hz, 1H), 3.76–3.40 (m, 3H), 2.88–2.66 (m, 1H), 2.22 (s, 3H), 2.20–2.06 (m, 1H), 1.88 (dp, J = 30.6, 10.1 Hz, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 169.86, 140.81, 135.94, 133.41, 130.39, 130.16, 129.90, 129.81, 129.38, 126.78, 126.72, 126.62, 124.30, 124.26, 122.55, 120.11, 120.03, 110.77, 90.54, 85.60, 65.76, 64.50, 54.11, 50.70, 48.91, 45.67, 44.25, 44.21, 31.16, 29.17; HRMS (ESI) m/z calcd for C22H22ClN4O [M + H]+ 393.1482, found 393.1473. (R)-2-((5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)- N-methylbenzamide (C16, CDD-2599)
Figure imgf000181_0001
d. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 2-ethynyl-N-methylbenzamide in the Sonogashira coupling reaction to afford the title compound (36% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 8.61 (s, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 8.4 Hz, 1H), 7.59–7.53 (m, 4H), 6.64 (s, 1H), 3.98–3.87 (m, 1H), 3.63 (br s, 2H), 3.47 (s, 3H), 3.41 (br s, 1H), 2.80–2.69 (m, 1H), 2.33 (s, 3H), 2.21 (s, 3H), 2.08 (br s, 1H), 1.88 (br s, 1H); HRMS (ESI) m/z calcd for C24H26N5O2 [M + H]+ 416.2087, found 416.2077. (R)-7-((5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)indolin-2-one methylbenzamide (C17, CDD-2603) d. The same procedure for the synthesis of C1 (CDD- 2530) w
Figure imgf000182_0001
as followed using 7-ethynylindolin-2-one in the Sonogashira coupling reaction to afford the title compound (39% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 8.00 (br, 1H), 7.66 (br, 1H), 7.57 (br, 2H), 7.47 (br, 1H), 7.07 (br, 1H), 3.93– 3.80 (m, 1H), 3.64–3.57 (m, 2H), 3.37 (br s, 1H), 2.74 (br s, 1H), 2.65 (br s, 1H), 2.26 (s, 3H), 2.21 (br s, 2H), 2.16 (s, 3H), 2.02 (br s, 1H), 1.82 (br s, 1H); HRMS (ESI) m/z calcd for C24H24N5O2 [M + H]+ 414.1930, found 414.1923. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-(pyridazin-3-ylethynyl)-1H-indazol-5- yl)methanone (C18, CDD-2604)
Figure imgf000182_0002
d. The same procedure for the synthesis of C1 (CDD- 2530) was followed using 3- ethynylpyridazine in the Sonogashira coupling reaction to afford the title compound (22% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 9.15 (t, J = 3.7 Hz, 1H), 8.07 (d, J = 14.2 Hz, 1H), 7.73 (d, J = 8.4 Hz, 1H), 7.63– 7.48 (m, 3H), 4.04–3.83 (m, 1H), 3.73–3.38 (m, 3H), 2.85–2.67 (m, 1H), 2.31 (s, 3H), 2.21 (s, 3H), 2.08 (dt, J = 13.0, 6.2 Hz, 1H), 1.87 (dp, J = 30.7, 10.3 Hz, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 170.0, 149.5, 148.0, 140.9, 130.6, 130.2, 130.0, 128.8, 126.7, 126.4, 126.0, 124.3, 124.2, 119.7, 119.6, 111.2, 89.5, 85.7, 65.7, 64.5, 54.0, 50.7, 48.9, 45.6, 44.3, 44.1, 31.1, 29.1; HRMS (ESI) m/z calcd for C20H21N6O [M + H]+ 361.1776, found 361.1768. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-(piperidin-4-ylethynyl)-1H-indazol-5- yl)methanone (C19, CDD-2605) d. The same procedure for the synthesis of C1 (CDD-
Figure imgf000183_0001
2530) was followed using tert-butyl 4-ethynylpiperidine-1-carboxylate in the Sonogashira coupling reaction to afford the title compound (18% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 7.98 (d, J = 16.6 Hz, 1H), 7.53 (t, J = 7.8 Hz, 1H), 7.47 (t, J = 7.9 Hz, 1H), 4.04–3.80 (m, 1H), 3.74–3.38 (m, 3H), 3.17 (dt, J = 10.9, 4.5 Hz, 2H), 2.91–2.64 (m, 4H), 2.34 (s, 3H), 2.23 (s, 3H), 2.10 (dt, J = 12.8, 6.5 Hz, 1H), 2.03–1.95 (m, 2H), 1.92–1.83 (m, 1H); 13C NMR (150 MHz, CDCl3, two rotamers) δ 170.0, 140.7, 130.9, 129.6, 129.4, 126.2, 124.0, 120.2, 120.0, 110.4, 97.6, 77.2, 65.7, 64.5, 54.2, 50.6, 48.8, 45.6, 44.9, 44.2, 32.5, 31.1, 29.2, 28.2; HRMS (ESI) m/z calcd for C21H28N5O [M + H]+ 366.2294, found 366.2285. (R)-(3-([1,1’-Biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (C20, CDD-2647)
Figure imgf000183_0002
Preparation of potassium ([1,1’-biphenyl]-2-ylmethyl)trifluoroborate: To a solution of 2-(bromomethyl)-1,1’-biphenyl (1.0 mmol, 1.0 equiv.) and bis(pinacolato)diborane (1.25 mmol, 1.25 equiv.) in anhydrous THF (2 mL) were added Xantphos (0.05 mmol, 0.05 equiv.) and CuCl (0.05 mmol, 0.05 equiv.) under nitrogen. The reaction mixture was stirred at room temperature for 16 h and filtered through a pad of celite. The filtrate was concentrated and purified by flash chromatography to afford 2-([1,1’- biphenyl]-2-ylmethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (70%) as a white solid; 1H NMR (600 MHz, CDCl3) δ 7.42 (m, 4H), 7.35 (m, 1H), 7.31 (m, 2H), 7.26 (m, 2H), 2.34 (s, 2H), 1.20 (s, 12H). To a solution of 2-([1,1’-Biphenyl]-2-ylmethyl)-4,4,5,5-tetramethyl-1,3,2- dioxaborolane (0.3 mmol) in CH3OH (2 mL) was added KHF2 (0.2 mL, 4.5 M in water) dropwise. The solution was stirred at room temperature for 1 h and concentrated to yield potassium ([1,1’-biphenyl]-2-ylmethyl)trifluoroborate. This product was subsequently used in the next step without undergoing any purification process. Synthetic route 2 was applied. The general procedure for amide coupling was employed using 3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-5-carboxylic acid and (R)- N,N-dimethyl-3-pyrrolidinamine. The resulting amide was partially purified by flash chromatography and used in the next step. A oven dried microwave vial equipped with magnetic stir bar was charged with the amide (0.1 mmol, 1.0 equiv.), potassium ([1,1’- biphenyl]-2-ylmethyl)trifluoroborate (0.25 mmol, 2.5 equiv.), Pd(OAc)2 (0.01 mmol, 0.1 equiv.), RuPhos (0.02 mmol, 0.2 equiv.), and Cs2CO3 (0.3 mmol, 3.0 equiv.). The vial was sealed with a microwave cap, evacuated and backfilled with nitrogen, followed by addition of anhydrous toluene (3 mL) and water (0.3 mL, purged with nitrogen) via syringe. The mixture was vacuum-purged and refilled with nitrogen for three cycles and then heated by microwave reactor at 120 oC for 1 h. After completion of the reaction (monitored by TLC and LC-MS), the reaction mixture was filtered through a pad of celite. The filtrate was concentrated and partially purified by flash chromatography to yield (3-([1,1’-biphenyl]-2-ylmethyl)-1- (tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)((R)-3-(dimethylamino)pyrrolidin-1- yl)methanone. The general procedure for removal of THP was applied to afford the title compound (29% over three steps) as a yellow oil; 1H NMR (600 MHz, CDCl3, two rotamers) δ 7.52 (dd, J = 8.6, 1.5 Hz, 1H), 7.38–7.26 (m, 10H), 7.21–7.13 (m, 1H), 4.37–4.26 (m, 2H), 4.00–3.90 (m, 0.5H), 3.87–3.76 (m, 0.5H), 3.70–3.59 (m, 0.5H), 3.53–3.37 (m, 1.5H), 3.34– 3.14 (m, 1H), 2.85–2.72 (m, 0.5H), 2.71–2.56 (m, 0.5H), 2.34 (s, 3H), 2.26–1.98 (m, 4H), 1.90–1.64 (m, 1H); HRMS (ESI) m/z calcd for C27H29N4O [M + H]+ 425.2341, found 425.2331. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H- indazol-5-yl)methanone (C21, CDD-2648) d. The same procedure for the synthesis of C1 (CDD- 2530) w
Figure imgf000185_0001
as followed using 4-(2-ethynylphenyl)morpholine in the Sonogashira coupling reaction to afford the title compound (41% over two steps) as a white solid; 1H NMR (600 MHz, CDCl3, two rotamers) δ 8.06 (d, J = 9.6 Hz, 1H), 7.61 (d, J = 7.2 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.34 (m, 1H), 7.16 (m, 1H), 7.02 (t, J = 6.6 Hz, 1H), 6.98 (d, J = 8.4 Hz, 1H), 4.00 (m, 0.5H), 3.89 (t, 4.8 Hz, 4.5H), 3.72–3.62 (m, 1.5H), 3.58–3.48 (m, 1H), 3.42 (m, 0.5H), 3.27 (t, J = 4.2Hz, 4H), 2.81–2.72 (m, 1H), 2.33 (s, 3H), 2.22 (s, 3H), 2.09 (br s, 0.5H), 1.91 (m, 1.5H); HRMS (ESI) m/z calcd for C26H30N5O2 [M + H]+ 444.2400, found 444.2389. (R,E)-(3-(2-([1,1’-Biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone (C22, CDD-2649)
Figure imgf000185_0002
lied. To a solution of (R)-(3-(dimethylamino)pyrrolidin-1- yl)(3-iodo-1H-indazol-5-yl)methanone (0.1 mmol) in anhydrous THF (0.5 mL) were added Pd(OAc)2 (0.01 mmol), PPh3 (0.02 mmol), and 2-vinyl-1,1'-biphenyl (0.15 mmol). The reaction solution was heated at 80 oC under microwave. After 2 h, the solution was filtrated with a pad of celite, and the filtrate was concentrated. The crude product was purified to afford the title compound (65%) as a yellow oil; 1H NMR (600 MHz, CDCl3, two rotamers) δ 7.85 (d, J = 7.8 Hz, 1H), 7.81 (d, J = 9.0 Hz, 1H), 7.56 (m, 2H), 7.46–7.37 (m, 9H), 7.16 (m, 1H), 3.99–3.88 (m, 1H), 3.68 (m, 0.5H), 3.50–3.42 (m, 1.5H), 3.20 (br s, 1H), 2.81–2.68 (m, 1H), 2.37 (s, 3H), 2.20 (s, 3H), 2.04–1.82 (m, 2H); HRMS (ESI) m/z calcd for C28H29N4O [M + H]+ 437.2341, found 437.2331. (R)-N-(2-((5-(3-(Dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide (C23, CDD-2651) d. The same procedure for the synthesis of C1 (CDD-
Figure imgf000186_0001
2530) was followed using N-(2-ethynylphenyl)acetamide in the Sonogashira coupling reaction to afford the title compound (14% over two steps) as a light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.37 (dd, J = 8.6, 2.7 Hz, 1H), 8.16 (s, 1H), 7.83 (t, J = 9.6 Hz, 1H), 7.66–7.53 (m, 1H), 7.42 (d, J = 7.7 Hz, 1H), 7.22–7.15 (m, 1H), 6.84–6.74 (m, 1H), 6.65–6.54 (m, 1H), 3.79–3.70 (m, 1H), 3.70–3.62 (m, 1H), 3.52–3.46 (m, 2H), 3.28– 3.23 (m, 1H), 2.76 (s, 3H), 2.73–2.62 (m, 1H), 2.19 (s, 3H), 2.09–2.07 (m, 3H), 2.07–1.98 (m, 1H), 1.81–1.66 (m, 1H); HRMS (ESI) m/z calcd for C24H26N5O2 [M + H]+ 416.2087, found 416.2075. (3,6-Diazabicyclo[3.1.1]heptan-3-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C24, CDD-2762)
Figure imgf000186_0002
plied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.90–7.85 (m, 2H), 7.75 (d, J = 8.6 Hz, 1H), 7.62 (t, J = 7.8 Hz, 1H), 7.58–7.49 (m, 3H), 4.27 (s, 1H), 4.14–3.96 (m, 2H), 3.95–3.69 (m, 3H), 2.82–2.69 (m, 1H), 1.81–1.72 (m, 1H); HRMS (ESI) m/z calcd for C22H18F3N4O2 [M + H]+ 427.1382, found 427.1371. (2,8-Diazaspiro[4.5]decan-2-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C25, CDD-2763) light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 7.98–7.
Figure imgf000187_0001
84 (m, 2H), 7.72–7.66 (m, 1H), 7.66–7.49 (m, 4H), 3.64–3.58 (m, 2H), 3.56–3.53 (m, 2H), 3.13 (s, 2H), 3.07–2.95 (m, 2H), 1.92–1.85 (m, 1H), 1.84–1.76 (m, 2H), 1.74–1.68 (m, 1H), 1.65–1.54 (m, 2H); HRMS (ESI) m/z calcd for C25H24F3N4O2 [M + H]+ 469.1851, found 469.1838. (S)-(2-Phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C26, CDD-2764) lied; l 1
Figure imgf000187_0002
ight yellow oil; H NMR (600 MHz, DMSO-d6) δ 7.83 (d, J = 7.8 Hz, 1H), 7.77 (s, 1H), 7.70 (d, J = 8.6 Hz, 1H), 7.64–7.50 (m, 3H), 7.50–7.33 (m, 5H), 7.26 (t, J = 7.3 Hz, 1H), 3.62–3.52 (m, 2H), 3.10–2.92 (m, 3H), 2.85–2.75 (m, 1H), 2.66 (dt, J = 11.5, 6.3 Hz, 1H); HRMS (ESI) m/z calcd for C27H22F3N4O2 [M + H]+ 491.1695, found 491.2679. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C27, CDD-2871)
Figure imgf000187_0003
Synthetic route 1 was applied; light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.70–8.66 (m, 2H), 7.82 (dt, J = 7.1, 1.3 Hz, 1H), 7.74–7.70 (m, 2H), 7.61–7.50 (m, 5H), 7.45 (d, J = 8.6 Hz, 1H), 3.78–3.75 (m, 1H), 3.70–3.64 (m, 1H), 3.23–3.20 (m, 1H), 2.78 (s, 1H), 2.66 (s, 1H), 2.21 (s, 3H), 2.17–1.97 (m, 4H), 1.90–1.62 (m, 1H); HRMS (ESI) m/z calcd for C27H26N5O [M + H]+ 436.2137, found 436.2126. (3-((2-(Pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[3.5]nonan- 2-yl)methanone (C28, CDD-2872) ; light yellow oil; 1H NMR (600 MHz, DMSO-d6) 8.72
Figure imgf000188_0001
(d, J = 5.7 Hz, 2H), 7.82 (d, J = 7.3 Hz, 1H), 7.76 (d, J = 6.1 Hz, 3H), 7.63–7.50 (m, 5H), 3.87 (s, 2H), 3.76 (s, 2H), 2.68–2.53 (m, 4H), 1.72–1.53 (m, 4H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2126. (3-((2-(Pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(7-oxa-2- azaspiro[3.5]nonan-2-yl)methanone (C29, CDD-2873)
Figure imgf000188_0002
d; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.74–8.69 (m, 2H), 7.84 (d, J = 7.4 Hz, 1H), 7.79–7.73 (m, 3H), 7.70–7.53 (m, 5H), 3.95 (s, 2H), 3.84 (s, 2H), 3.55–3.52 (m, 2H), 3.50–3.48 (m, 2H), 1.80–1.67 (m, 4H); HRMS (ESI) m/z calcd for C28H25N4O2 [M + H]+ 449.1978, found 449.1966. (S)-(2-Phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone (C30, CDD-2884) ied; light yellow oil; 1H NMR (600 MHz, DMSO-d6) δ 8.71–8.
Figure imgf000189_0001
64 (m, 2H), 7.82 (d, J = 7.5 Hz, 1H), 7.74–7.68 (m, 2H), 7.66–7.38 (m, 10H), 7.26 (t, J = 7.3 Hz, 1H), 3.61–3.49 (m, 2H), 3.15–2.94 (m, 3H), 2.84–2.66 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 149.6, 147.2, 140.4, 140.1, 139.4, 133.2, 129.8, 129.7, 129.5, 128.9, 128.5, 128.1, 127.1, 126.6, 125.5, 123.9, 123.7, 120.0, 118.1, 111.3, 91.7, 84.7, 45.7; HRMS (ESI) m/z calcd for C31H26N5O [M + H]+ 484.2137, found 484.2125. (R)-(3-(Dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C31, CDD-2885) d; light yellow oi 1
Figure imgf000189_0002
l; H NMR (600 MHz, DMSO-d6, two rotamers) δ 8.90 (d, J = 2.3 Hz, 1H), 8.61 (dt, J = 4.6, 2.5 Hz, 1H), 8.11 (dt, J = 7.8, 2.0 Hz, 1H), 7.86–7.81 (m, 1H), 7.63–7.51 (m, 6H), 7.45 (d, J = 13.9 Hz, 1H), 3.80 (dd, J = 11.8, 7.1 Hz, 0.5H), 3.70 (t, J = 11.1 Hz, 0.5H), 3.57–3.51 (m, 0.5H), 3.48–3.43 (m, 2H), 3.27–3.25 (m, 0.5H), 2.84–2.74 (m, 0.5H), 2.70–2.60 (m, 0.5H), 2.22 (s, 3H), 2.17–2.11 (m, 0.5H), 2.07 (s, 3H), 2.05–1.98 (m, 0.5H), 1.89–1.65 (m, 1H); HRMS (ESI) m/z calcd for C27H26N5O [M + H]+ 436.2137, found 436.2126. (R)-(3-(Cyclohexylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone (C32, CDD-2886) d; light yellow oil; 1H NMR (600 MHz, DMSO-d6, two rotamer
Figure imgf000190_0001
s) δ 7.78 (d, J = 3.6 Hz, 1H), 7.57 (d, J = 8.6 Hz, 1H), 7.51 (t, J = 9.8 Hz, 1H), 3.75– 3.67 (m, 2H), 3.65–3.59 (m, 2H), 2.84–2.57 (m, 3H), 2.18 (s, 3H), 2.08 (s, 4H), 1.93–1.84 (m, 2H), 1.79–1.65 (m, 3H), 1.61–1.45 (m, 3H), 1.43–1.32 (m, 3H); HRMS (ESI) m/z calcd for C22H29N4O [M + H]+ 365.2341, found 365.2329. (E)-(3-(2-([1,1’-Biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone (C33, CDD-2942)
Figure imgf000190_0002
d. To a solution of methyl 3-iodo-1H-indazole-5- carboxylate (0.1 mmol) in anhydrous THF (0.5 mL) were added Pd(OAc)2 (0.01 mmol), PPh3 (0.02 mmol), and 2-vinyl-1,1'-biphenyl (0.15 mmol). The reaction solution was heated at 80 oC under microwave. After 2 h, the solution was filtrated with a pad of celite, and the filtrate was concentrated. The crude product was dissolved in THF/MeOH/H2O (4:1:1, 0.1 M), and the solution was added 2M LiOH (5 equiv.). After overnight, the solution was acidified with 1N HCl to pH = 2. The solution was extracted with CH2Cl2, and the combined organic layers were concentrated and purified by flash chromatography to yield (E)-3-(2-([1,1'-biphenyl]-2- yl)vinyl)-1H-indazole-5-carboxylic acid; 1H NMR (600 MHz, CDCl3) δ 7.69 (d, J = 7.2 Hz, 1H), 7.64 (d, J = 7.2 Hz, 1H), 7.49–7.44 (m, 3H), 7.40–7.32 (m, 6H), 6.75 (q, 1H), 5.75 (d, J = 17.4 Hz, 1H), 5.23 (d, J = 17.4 Hz, 1H). The general procedure for amide coupling was employed using the resulting acid and 7-methyl-2,7-diazaspiro[3.5]nonane to afford the title compound (80%) as a white solid; 1H NMR (600 MHz, CDCl3) δ 7.97 (s, 1H), 7.87 (d, J = 7.8 Hz, 1H), 7.77 (d, J = 9.0 Hz, 1H), 7.58 (d, J = 16.8 Hz, 1H), 7.50–7.40 (m, 10H), 3.94 (s, 2H), 3.74 (s, 2H), 2.35 (s, 5H), 1.88 (s, 6H); HRMS (ESI) m/z calcd for C30H31N4O [M + H]+ 463.2498, found 463.2488. (E)-(3-(2-([1,1’-Biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2- azaspiro[3.5]nonan-2-yl)methanone (C34, CDD-2953) lied. The same procedure for the synthesis of C33 (CDD-
Figure imgf000191_0001
2942) was followed using 7-oxa-2-azaspiro[3.5]nonane in the amide coupling reaction to afford the title compound (78%) as a white solid; 1H NMR (600 MHz, CDCl3) δ 7.99 (s, 1H), 7.87 (d, J = 7.2 Hz, 1H), 7.61 (d, J = 6.0 Hz, 1H), 7.61 (d, J = 16.8 Hz, 1H), 7.52–7.40 (m, 10H), 4.00 (s, 2H), 3.79 (s, 2H), 3.63 (s, 4H), 1.80 (s, 4H); HRMS (ESI) m/z calcd for C29H28N3O2 [M + H]+ 450.2182, found 450.2170. (7-Methyl-2,7-diazaspiro[3.5]nonan-2-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone (C35, CDD-2986)
Figure imgf000191_0002
; light yellow oil; 1H NMR (600 MHz, CD3OD) δ 8.92– 8.88 (m, 1H), 8.64 (dd, J = 4.9, 1.6 Hz, 1H), 8.21 (dt, J = 7.9, 1.9 Hz, 1H), 7.86–7.76 (m, 1H), 7.75–7.71 (m, 2H), 7.65–7.51 (m, 5H), 4.05 (s, 2H), 3.97 (s, 2H), 2.51–2.38 (m, 4H), 2.27 (s, 3H), 1.91 (br s, 4H); HRMS (ESI) m/z calcd for C29H28N5O [M + H]+ 462.2294, found 462.2285. (7-Methyl-2,7-diazaspiro[3.5]nonan-2-yl)(3-((3-phenylpyridin-2-yl)ethynyl)-1H- indazol-5-yl)methanone (C36, CDD-2987) ; light yellow oil; 1H NMR (600 MHz, CD3OD) δ 8.63 (dd, J =
Figure imgf000192_0001
4.8, 1.6 Hz, 1H), 7.99 (dd, J = 7.9, 1.7 Hz, 1H), 7.79–7.74 (m, 3H), 7.73 (dd, J = 8.7, 1.6 Hz, 1H), 7.64 (dd, J = 8.7, 0.9 Hz, 1H), 7.61–7.55 (m, 3H), 7.55–7.49 (m, 1H), 4.00 (s, 2H), 3.97 (s, 2H), 2.53 (br s, 4H), 2.28 (s, 3H), 1.90 (br s, 4H); HRMS (ESI) m/z calcd for C29H28N5O [M + H]+ 462.2294, found 462.2285. (3-((2-(Pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan- 2-yl)methanone (C37, CDD-3094)
Figure imgf000192_0002
; pale yellow solid; 1H NMR (600 MHz, DMSO-d6) δ 8.91 (d, J = 2.3 Hz, 1H), 8.65 (dd, J = 4.8, 1.6 Hz, 1H), 8.15 (dt, J = 7.9, 2.0 Hz, 1H), 7.83 (d, J = 7.6 Hz, 1H), 7.65–7.59 (m, 4H), 7.59–7.50 (m, 4H), 3.90 (d, J = 8.5 Hz, 1H), 3.85 (d, J = 8.4 Hz, 1H), 3.77 (d, J = 9.7 Hz, 1H), 3.70 (d, J = 9.7 Hz, 1H), 2.85–2.68 (m, 2H), 2.62–2.52 (m, 2H), 1.77–1.60 (m, 2H), 1.45–1.27 (m, 2H); 13C NMR (150 MHz, DMSO-d6) δ 169.0, 148.9, 148.3, 141.7, 139.1, 136.1, 135.1, 132.5, 129.3, 129.0, 128.0, 127.7, 125.8, 125.2, 123.3, 122.9, 120.4, 118.9, 111.2, 91.3, 84.9, 61.8, 57.2, 54.2, 44.9, 34.2, 33.7, 22.8; HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2133. (2,6-Diazaspiro[3.5]nonan-2-yl)(3-((4-(trifluoromethoxy)-[1,1’-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone (C38, CDD-3134) llow solid; 1H NMR (600 MHz, DMSO-d6) δ 7.83 (d
Figure imgf000193_0001
, J = 2.6 Hz, 1H), 7.75–7.69 (m, 3H), 7.67 (dd, J = 8.7, 1.5 Hz, 1H), 7.66–7.61 (m, 2H), 7.59–7.52 (m, 3H), 7.49–7.45 (m, 1H), 3.85 (d, J = 8.5 Hz, 1H), 3.83–3.76 (m, 2H), 3.71 (d, J = 9.7 Hz, 1H), 2.73 (d, J = 5.1 Hz, 2H), 2.57–2.54 (m, 1H), 2.49–2.44 (m, 1H), 1.68 (s, 2H), 1.45–1.31 (m, 2H); HRMS (ESI) m/z calcd for C30H26F3N4O2 [M + H]+ 531.2008, found 531.1990. (3-((2-(Naphthalen-2-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C39, CDD-3135)
Figure imgf000193_0002
; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.09 (d, J = 8.7 Hz, 1H), 8.02–7.93 (m, 3H), 7.81 (dd, J = 7.6, 1.4 Hz, 1H), 7.69–7.61 (m, 2H), 7.59–7.46 (m, 6H), 3.73–3.69 (m, 2H), 3.66–3.59 (m, 2H), 2.64 (s, 2H), 2.55–2.51 (m, 2H), 1.61–1.56 (m, 2H), 1.40–1.23 (m, 2H); HRMS (ESI) m/z calcd for C33H29N4O [M + H]+ 497.2341, found 497.2327. (2,6-Diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1’-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone (C40, CDD-3136)
le yellow solid; 1H NMR (600 MHz, DMSO-d6) δ
Figure imgf000194_0001
8.03 (d, J = 7.9 Hz, 1H), 7.86–7.82 (m, 2H), 7.82–7.75 (m, 2H), 7.70–7.65 (m, 2H), 7.64 (dd, J = 8.5, 1.0 Hz, 1H), 7.57 (dd, J = 8.4, 6.9 Hz, 2H), 7.53–7.47 (m, 1H), 3.84 (d, J = 8.5 Hz, 1H), 3.81–3.76 (m, 2H), 3.71 (d, J = 9.7 Hz, 1H), 2.73 (d, J = 6.2 Hz, 2H), 2.57–2.52 (m, 2H), 1.68 (s, 2H), 1.47–1.28 (m, 2H); HRMS (ESI) m/z calcd for C30H26F3N4O [M + H]+ 515.2059, found 515.2042. (3-((4’-Methyl-5-(trifluoromethyl)-[1,1’-biphenyl]-2-yl)ethynyl)-1H-indazol-5- yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone (C41, CDD-3137) hite 1
Figure imgf000194_0002
solid; H NMR (600 MHz, DMSO-d6) δ 7.95 (d, J = 8.0 Hz, 1H), 7.78 (dd, J = 8.0, 1.8 Hz, 1H), 7.76 (s, 1H), 7.67 (d, J = 7.8 Hz, 2H), 7.63 (s, 1H), 7.59 (d, J = 8.6 Hz, 1H), 7.52 (dd, J = 8.6, 1.5 Hz, 1H), 7.37 (d, J = 7.8 Hz, 2H), 3.83 (d, J = 8.4 Hz, 1H), 3.80–3.73 (m, 2H), 3.68 (d, J = 9.8 Hz, 1H), 2.78–2.65 (m, 2H), 2.56–2.51 (m, 2H), 2.39 (s, 3H), 1.67 (s, 2H), 1.42–1.28 (m, 2H); HRMS (ESI) m/z calcd for C31H28F3N4O [M + H]+ 529.2215, found 529.2199. (2,6-Diazaspiro[3.5]nonan-2-yl)(3-((3’-(trifluoromethoxy)-[1,1’-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone (C42, CDD-3153) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.84 (d, J = 7.6
Figure imgf000195_0001
Hz, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.70–7.51 (m, 8H), 7.42 (dd, J = 8.4, 2.3 Hz, 1H), 3.86 (d, J = 8.4 Hz, 1H), 3.80 (d, J = 8.4 Hz, 1H), 3.76 (d, J = 9.8 Hz, 1H), 3.68 (d, J = 9.7 Hz, 1H), 2.72 (s, 2H), 2.54 (t, J = 5.1 Hz, 2H), 1.72–1.57 (m, 2H), 1.46–1.28 (m, 2H); HRMS (ESI) m/z calcd for C30H26F3N4O2 [M + H]+ 531.2008, found 531.1992. (2,6-Diazaspiro[3.5]nonan-2-yl)(3-((2’-(trifluoromethoxy)-[1,1’-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone (C43, CDD-3154)
Figure imgf000195_0002
; white solid; 1H NMR (600 MHz, DMSO-d6) δ 7.80 (dd, J = 7.5, 1.6 Hz, 1H), 7.66–7.50 (m, 8H), 7.46 (s, 1H), 7.44 (dd, J = 7.4, 1.5 Hz, 1H), 3.84 (d, J = 8.5 Hz, 1H), 3.79 (d, J = 9.8 Hz, 2H), 3.72 (d, J = 9.8 Hz, 1H), 2.74 (s, 2H), 2.58–2.51 (m, 2H), 1.68 (s, 2H), 1.46–1.32 (m, 2H); HRMS (ESI) m/z calcd for C30H26F3N4O2 [M + H]+ 531.2008, found 531.1993. 2’-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methyl-[1,1’-biphenyl]-2-carboxamide (C44, CDD-3155) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.05–
Figure imgf000196_0001
7.99 (m, 1H), 7.72 (dd, J = 7.1, 1.9 Hz, 1H), 7.63 (dd, J = 8.7, 1.5 Hz, 1H), 7.58 (d, J = 8.7 Hz, 1H), 7.57–7.53 (m, 2H), 7.51 (td, J = 7.5, 1.4 Hz, 1H), 7.48 (s, 1H), 7.46–7.40 (m, 3H), 7.33 (dd, J = 7.2, 1.8 Hz, 1H), 3.86–3.76 (m, 3H), 3.70 (d, J = 9.7 Hz, 1H), 2.72 (s, 2H), 2.57 (d, J = 4.6 Hz, 3H), 2.57–2.51 (m, 2H), 1.75–1.61 (m, 2H), 1.42–1.32 (m, 2H); HRMS (ESI) m/z calcd for C31H30N5O2 [M + H]+ 504.2400, found 504.2388. 2’-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methyl-[1,1’-biphenyl]-3-carboxamide (C45, CDD-3156) ; white solid 1
Figure imgf000196_0002
; H NMR (600 MHz, DMSO-d6) δ 8.54– 8.49 (m, 1H), 8.12 (s, 1H), 7.91 (dd, J = 8.0, 4.3 Hz, 2H), 7.83 (d, J = 7.7 Hz, 1H), 7.67–7.64 (m, 2H), 7.64–7.56 (m, 4H), 7.52 (s, 1H), 3.89 (d, J = 8.5 Hz, 1H), 3.82 (t, J = 9.6 Hz, 2H), 3.72 (d, J = 9.8 Hz, 1H), 2.85 (s, 2H), 2.74 (d, J = 4.5 Hz, 3H), 2.64 (t, J = 5.5 Hz, 2H), 1.77– 1.64 (m, 2H), 1.51–1.38 (m, 2H); HRMS (ESI) m/z calcd for C31H30N5O2 [M + H]+ 504.2400, found 504.2388. 2’-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methyl-[1,1’-biphenyl]-4-carboxamide (C46, CDD-3157) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.57– 8.52 (m
Figure imgf000197_0001
, 1H), 8.02–7.98 (m, 2H), 7.85–7.76 (m, 4H), 7.69 (dd, J = 8.8, 1.5 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), 7.59–7.55 (m, 2H), 7.55–7.48 (m, 1H), 3.89 (d, J = 8.4 Hz, 1H), 3.83 (d, J = 8.4 Hz, 1H), 3.79 (d, J = 9.7 Hz, 1H), 3.70 (d, J = 9.7 Hz, 1H), 2.82 (d, J = 4.5 Hz, 3H), 2.80–2.70 (m, 2H), 2.59–2.52 (m, 2H), 1.78–1.57 (m, 2H), 1.44–1.31 (m, 2H); HRMS (ESI) m/z calcd for C31H30N5O2 [M + H]+ 504.2400, found 504.2388. (3-((2-(6-Amino-4-methylpyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C47, CDD-3197) ; white soli 1
Figure imgf000197_0002
d; H NMR (600 MHz, DMSO-d6) δ 7.81 (s, 1H), 7.75–7.67 (m, 2H), 7.62–7.55 (m, 2H), 7.50–7.40 (m, 2H), 7.34 (d, J = 7.5 Hz, 1H), 6.42 (s, 1H), 5.80 (s, 2H), 3.98–3.87 (m, 2H), 3.73–3.69 (m, 2H), 2.85–2.67 (m, 2H), 2.63– 2.50 (m, 2H), 2.10 (s, 3H), 1.78–1.59 (m, 2H), 1.36 (s, 2H); HRMS (ESI) m/z calcd for C29H29N6O [M + H]+ 477.2403, found 477.2391. (3-((2-(2-Aminopyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C48, CDD-3198) ; white solid; 1H NMR (600 MHz, DMSO-d6) 8.05 (d, J = 5.2 H
Figure imgf000198_0001
z, 1H), 7.87 (s, 1H), 7.80 (d, J = 7.5 Hz, 1H), 7.69 (dd, J = 8.8, 1.4 Hz, 1H), 7.63 (d, J = 8.7 Hz, 1H), 7.59–7.45 (m, 3H), 6.85 (dd, J = 5.2, 1.5 Hz, 1H), 6.70 (s, 1H), 5.99 (s, 2H), 3.89 (d, J = 8.4 Hz, 1H), 3.84 (d, J = 8.4 Hz, 1H), 3.76 (d, J = 9.8 Hz, 1H), 3.70 (d, J = 9.7 Hz, 1H), 2.78 (d, J = 11.7 Hz, 1H), 2.72 (d, J = 11.9 Hz, 1H), 2.57–2.51 (m, 2H), 1.72 (s, 1H), 1.66 (s, 1H), 1.42–1.36 (m, 1H), 1.36–1.31 (m, 1H); 13C NMR (150 MHz, DMSO-d6) δ 169.1, 159.9, 148.2, 147.7, 141.6, 140.8, 133.3, 129.6, 129.1, 128.8, 128.4, 126.9, 126.6, 123.5, 119.8, 119.7, 112.4, 110.9, 107.8, 92.0, 84.5, 62.2, 57.7, 54.5, 45.3, 34.7, 34.0, 23.2; HRMS (ESI) m/z calcd for C28H27N6O [M + H]+ 463.2246, found 463.2236. (3-((2-(2-Aminopyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C49, CDD-3232)
Figure imgf000198_0002
; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.01 (d, J = 5.0 Hz, 1H), 7.74 (dd, J = 7.3, 1.7 Hz, 1H), 7.55 (d, J = 8.7 Hz, 1H), 7.51–7.34 (m, 6H), 6.76–6.51 (m, 1H), 5.69 (br s, 1H), 5.44 (s, 1H), 3.97–3.79 (m, 2H), 3.76–3.67 (m, 2H), 2.92–2.68 (m, 2H), 2.66–2.51 (m, 2H), 1.82–1.51 (m, 2H), 1.45–1.30 (m, 2H); HRMS (ESI) m/z calcd for C28H27N6O [M + H]+ 463.2246, found 463.2236. (3-((2-(2-(Pyrrolidin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C50, CDD-3233) N ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.16 (d,
Figure imgf000199_0001
J = 4.9 Hz, 1H), 7.68 (d, J = 7.3 Hz, 1H), 7.56 (d, J = 8.7 Hz, 1H), 7.53–7.47 (m, 2H), 7.47– 7.34 (m, 4H), 6.80–6.73 (m, 1H), 3.92–3.73 (m, 2H), 3.73–3.63 (m, 2H), 3.11–2.93 (m, 4H), 2.74 (s, 2H), 2.62–2.51 (m, 2H), 1.73–1.60 (m, 6H), 1.44–1.29 (m, 2H); HRMS (ESI) m/z calcd for C32H33N6O [M + H]+ 517.2716, found 517.2704. (3-((2-(2-(Piperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C51, CDD-3234)
Figure imgf000199_0002
; white solid; 1H NMR (600 MHz, DMSO-d6) δ 8.27 (dd, J = 4.8, 1.9 Hz, 1H), 7.75 (d, J = 7.6 Hz, 1H), 7.68 (dd, J = 7.3, 1.9 Hz, 1H), 7.62–7.38 (m, 6H), 7.12–6.95 (m, 1H), 3.88–3.74 (m, 2H), 3.74–3.65 (m, 2H), 3.24 (s, 4H), 2.90 (s, 4H), 2.75 (s, 2H), 2.60–2.52 (m, 2H), 1.80–1.62 (m, 2H), 1.45–1.30 (m, 2H); HRMS (ESI) m/z calcd for C32H34N7O [M + H]+ 532.2825, found 532.2814. N-(2’-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1’- biphenyl]-2-yl)acetamide (C52, CDD-3235) ; white solid; 1H NMR (600 MHz, DMSO-d6) δ 9.13 (s,
Figure imgf000200_0001
1H), 7.78–7.70 (m, 1H), 7.66 – 7.59 (m, 1H), 7.53 (d, J = 8.6 Hz, 1H), 7.51–7.41 (m, 4H), 7.40–7.33 (m, 2H), 7.32–7.21 (m, 2H), 3.81–3.71 (m, 2H), 3.71–3.64 (m, 2H), 2.72 (s, 2H), 2.61–2.51 (m, 2H), 1.85 (s, 3H), 1.68 (s, 2H), 1.45–1.29 (m, 2H); HRMS (ESI) m/z calcd for C31H30N5O2 [M + H]+ 504.2400, found 504.2387. N-(2’-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1’- biphenyl]-2-yl)pivalamide (C53, CDD-3236) 1
Figure imgf000200_0002
; white solid; H NMR (600 MHz, DMSO-d6) δ 8.58– 8.45 (m, 1H), 7.77–7.70 (m, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.58–7.48 (m, 2H), 7.48–7.37 (m, 5H), 7.37–7.27 (m, 2H), 3.84–3.73 (m, 2H), 3.72–3.64 (m, 2H), 2.72 (s, 2H), 2.63–2.52 (m, 2H), 1.68 (s, 2H), 1.48–1.27 (m, 2H), 0.98 (s, 9H); HRMS (ESI) m/z calcd for C34H36N5O2 [M + H]+ 546.2869, found 546.2856. (3-((2-(2-(4-Methylpiperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5- yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone (C54, CDD-3353)
; pale yellow solid; 1H NMR (600 MHz, DMSO-d6) δ
Figure imgf000201_0001
8.31–8.24 (m, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.69 (dd, J = 7.4, 1.9 Hz, 1H), 7.62–7.55 (m, 1H), 7.55–7.47 (m, 3H), 7.47–7.40 (m, 2H), 7.13–6.99 (m, 1H), 3.86–3.74 (m, 4H), 3.73– 3.66 (m, 2H), 3.01–2.94 (m, 4H), 2.74 (s, 1H), 2.58–2.52 (m, 1H), 2.18–2.08 (m, 4H), 2.06 (s, 3H), 1.70 (s, 2H), 1.45–1.26 (m, 2H); HRMS (ESI) m/z calcd for C33H36N7O [M + H]+ 546.2981, found 546.2968. (3-([1,1'-Biphenyl]-3-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C55, CDD-3808) Synthetic route 2 was appl
Figure imgf000201_0002
R (600 MHz, DMSO-d6) δ 7.50 (s, 1H), 7.35 (s, 1H), 7.14 (d, J = 7.7 Hz, 1H), 6.94–6.90 (m, 2H), 6.89–6.85 (m, 2H), 6.83–6.78 (m, 2H), 6.68 (t, J = 7.7 Hz, 2H), 6.58 (t, J = 7.3 Hz, 1H), 3.21 (d, J = 8.5 Hz, 1H), 3.17 (d, J = 8.1 Hz, 1H), 2.90 (d, J = 9.7 Hz, 1H), 2.83 (d, J = 9.7 Hz, 1H), 1.96–1.81 (m, 2H), 1.74–1.67 (m, 2H), 0.90–0.74 (m, 2H), 0.60–0.44 (m, 2H); HRMS (ESI) m/z calcd for C27H27N4O [M + H]+ 423.2185, found 423.2176. (3-([1,1'-Biphenyl]-4-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C56, CDD-3809) Synthetic route 2 was ap R (600 MHz, DMSO-d6) δ
Figure imgf000202_0001
8.35 (s, 1H), 8.09 (d, J = 8.3 Hz, 2H), 7.86 (d, J = 8.3 Hz, 2H), 7.79–7.74 (m, 2H), 7.70 (dd, J = 8.7, 1.4 Hz, 1H), 7.65 (d, J = 8.7 Hz, 1H), 7.51 (t, J = 7.7 Hz, 2H), 7.40 (t, J = 7.4 Hz, 1H), 4.03 (d, J = 8.4 Hz, 1H), 3.98 (d, J = 8.4 Hz, 1H), 3.75 (d, J = 9.7 Hz, 1H), 3.68 (d, J = 9.8 Hz, 1H), 2.78–2.67 (m, 2H), 2.58–2.51 (m, 2H), 1.74–1.60 (m, 2H), 1.42–1.31 (m, 2H); HRMS (ESI) m/z calcd for C27H27N4O [M + H]+ 423.2185, found 423.2175. (3-((2'-Morpholino-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C57, CDD-3810) Synthetic route 2 was app
Figure imgf000202_0002
0 MHz, DMSO-d6) δ 7.73 (dd, J = 7.6, 1.3 Hz, 1H), 7.59–7.53 (m, 1H), 7.53–7.44 (m, 3H), 7.43–7.31 (m, 4H), 7.21– 7.09 (m, 2H), 3.82–3.74 (m, 2H), 3.74–3.66 (m, 2H), 3.39–3.33 (m, 4H), 2.80–2.65 (m, 6H), 2.60–2.51 (m, 2H), 1.73–1.63 (m, 2H), 1.47–1.27 (m, 2H); HRMS (ESI) m/z calcd for C33H34N5O2 [M + H]+ 532.2713, found 532.2710. 7-(2-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)isoindolin-1-one (C58, CDD-3811) Synthetic route 2 was appl R (600 MHz, DMSO-d6) δ
Figure imgf000203_0001
7.74–7.34 (m, 10H), 7.27 (s, 1H), 4.37 (s, 2H), 3.86–3.75 (m, 2H), 3.74–3.69 (m, 2H), 2.81– 2.64 (m, 2H), 2.61–2.51 (m, 2H), 1.75–1.56 (m, 2H), 1.47–1.25 (m, 2H); HRMS (ESI) m/z calcd for C31H28N5O2 [M + H]+ 502.2243, found 502.2235. 2'-((5-(2,6-Diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N,N- dimethyl-[1,1'-biphenyl]-2-carboxamide (C59, CDD-3812) Synthetic route 2 was app
Figure imgf000203_0002
0 MHz, DMSO-d6) δ 7.79– 7.71 (m, 1H), 7.66–7.40 (m, 9H), 7.31–7.25 (m, 1H), 3.79–3.73 (m, 2H), 3.72–3.67 (m, 2H), 2.80–2.50 (m, 10H), 1.75–1.59 (m, 2H), 1.47–1.27 (m, 2H); HRMS (ESI) m/z calcd for C32H32N5O2 [M + H]+ 518.2556, found 518.2549. (3-((2'-(Pyrrolidin-1-ylmethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C60, CDD-3813) - 201 -
Synthetic route 2 was appl R (600 MHz, DMSO-d6) δ
Figure imgf000204_0001
7.73–7.67 (m, 1H), 7.59 (d, J = 7.6 Hz, 1H), 7.52 (d, J = 8.7 Hz, 1H), 7.48–7.37 (m, 4H), 7.36–7.30 (m, 2H), 7.27 (dd, J = 7.5, 1.5 Hz, 1H), 7.21 (s, 1H), 3.89–3.60 (m, 6H), 2.79–2.67 (m, 2H), 2.60–2.52 (m, 2H), 2.26 (s, 4H), 1.75–1.65 (m, 2H), 1.54 (s, 4H), 1.44–1.30 (m, 2H); HRMS (ESI) m/z calcd for C34H36N5O [M + H]+ 530.2920, found 530.2919. (3-((2'-(Morpholinomethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone (C61, CDD-3814) Synthetic route 2 was app
Figure imgf000204_0002
0 MHz, DMSO-d6) δ 7.73 (dd, J = 7.2, 1.8 Hz, 1H), 7.58–7.34 (m, 8H), 7.30 (dd, J = 7.3, 1.6 Hz, 1H), 7.27–7.22 (m, 1H), 3.84–3.64 (m, 4H), 3.42–3.27 (m, 6H), 2.79–2.67 (m, 2H), 2.61–2.53 (m, 2H), 2.14 (s, 4H), 1.76–1.64 (m, 2H), 1.45–1.33 (m, 2H); HRMS (ESI) m/z calcd for C34H36N5O2 [M + H]+ 546.2869, found 546.2869. (3-((2-(Pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan- 2-yl)methanone (C62, CDD-3883) Synthetic route 2 was appl 00 MHz, DMSO-d6) δ 8.76– 8.67 (m, 2H), 7.84 (d, J = 7.2 Hz,
Figure imgf000205_0001
1H), 7.79–7.69 (m, 3H), 7.64–7.53 (m, 5H), 3.86 (d, J = 8.4 Hz, 1H), 3.81 (d, J = 8.4 Hz, 1H), 3.76 (d, J = 9.8 Hz, 1H), 3.68 (d, J = 9.8 Hz, 1H), 2.81–2.68 (m, 2H), 2.58–2.51 (m, 2H), 1.76–1.62 (m, 2H), 1.42–1.31 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2133. (3-((2-Phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C63, CDD-3884) Synthetic route 2 was appl
Figure imgf000205_0002
00 MHz, DMSO-d6) δ 8.64 (dd, J = 4.7, 1.7 Hz, 1H), 8.15–8.06 (m, 3H), 7.77 (d, J = 3.9 Hz, 1H), 7.60–7.48 (m, 4H), 7.45 (dd, J = 7.8, 4.7 Hz, 1H), 7.40 (dd, J = 8.6, 1.6 Hz, 1H), 3.87–3.83 (m, 1H), 3.80–3.77 (m, 1H), 3.69–3.61 (m, 2H), 2.80–2.62 (m, 2H), 2.57–2.50 (m, 2H), 1.71–1.63 (m, 2H), 1.43– 1.26 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2135. (3-((3-Phenylpyridin-4-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C64, CDD-3885)
Figure imgf000205_0003
Synthetic route 2 was applied; yellow solid; 1H NMR (600 MHz, DMSO-d6) δ 8.69 (s, 1H), 8.60 (d, J = 5.1 Hz, 1H), 7.79 (d, J = 7.5 Hz, 2H), 7.71–7.65 (m, 2H), 7.63–7.56 (m, 3H), 7.55–7.50 (m, 1H), 7.48 (d, J = 8.8 Hz, 1H), 3.85–3.74 (m, 2H), 3.71–3.66 (m, 2H), 2.82–2.63 (m, 2H), 2.61–2.50 (m, 2H), 1.74–1.63 (m, 2H), 1.43–1.29 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2134. (3-((4-Phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C65, CDD-3886) Synthetic route 2 was app 00 MHz, DMSO-d6) δ 8.92 (s,
Figure imgf000206_0001
1H), 8.63 (d, J = 5.1 Hz, 1H), 7.87–7.81 (m, 2H), 7.78–7.72 (m, 1H), 7.65–7.49 (m, 6H), 3.88–3.83 (m, 1H), 3.81–3.79 (m, 1H), 3.71–3.66 (m, 2H), 2.81–2.64 (m, 2H), 2.61–2.50 (m, 2H), 1.72–1.63 (m, 2H), 1.42–1.30 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2134. (3-((3-Phenylpyridin-2-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone (C66, CDD-3887) Synthetic route 2 was appl
Figure imgf000206_0002
00 MHz, DMSO-d6) δ 8.61 (dd, J = 4.8, 1.6 Hz, 1H), 7.87 (dd, J = 7.8, 1.6 Hz, 1H), 7.79–7.75 (m, 2H), 7.59–7.47 (m, 5H), 7.44 (dd, J = 7.8, 4.7 Hz, 1H), 7.34–7.28 (m, 1H), 3.86–3.60 (m, 4H), 2.81–2.63 (m, 2H), 2.59–2.51 (m, 2H), 1.71–1.63 (m, 2H), 1.41–1.28 (m, 2H); HRMS (ESI) m/z calcd for C28H26N5O [M + H]+ 448.2137, found 448.2134. Table C. Validation of STK33/CDD-2211 interactions by structure-activity-relationship (SAR) study of CDD-2211 analogs. The hinge binding of the indazole (CDD-2211 versus CDD-2533), the ortho-orientation of the biphenyl (CDD-2211 versus CDD-2532 and CDD- 2563), and the two-carbon distance between the indazole and the biphenyl (CDD-2211 versus CDD-2573 and CDD-2647) contributed to the potency toward STK33. ND means not determined for compounds not passing kinase assay screening against STK33 at 500 µM. 7
Figure imgf000207_0001
S 33, d N 9.9 ± .3 7.7 ± 0.7 N N 0.002 (nM) NanoBRET STK33, IC50 16 ND 289 8707 ND ND (nM) Table D. Structure activity relationship of substituted amines on indazole molecular scaffold. T
Figure imgf000207_0002
53366970.3 A1 2110 0.1 38 3.1 746 A2 2134 ND ND ND ND A3 2211 0.02 5 0.5 78 A4 2212 1.9 999 2.0 1093 A5 2277 ND 8226 ND ND A6 2278 ND 2171 ND ND A7 2481 ND 1924 ND ND A8 2535 0.01 17 <1 82 A9 2577 0.04 35 0.47 129 A10 2578 ND 803 ND ND A11 2579 0.08 46 0.12 251 A12 2594 ND 647 ND ND A13 2595 ND ND ND ND A14 2596 ND 387 ND 4086 A15 2597 1.2 391 18 194 A16 2598 0.18 124 0.46 575 A17 2606 ND ND ND ND A18 2607 0.35 328 3.8 559 A19 2608 3.2 1033 4.8 ND A20 2609 2.1 864 6.0 ND A21 2610 0.03 28 0.17 222 A22 2611 0.08 95 0.5 318 A23 2612 0.1 47 0.06 100 A24 2613 9.0 1029 54 ND A25 2614 3.6 232 7.4 1035 A26 2674 1.0 1294 20 2804 A27 2675 ND >10000 ND 7759 A28 2676 2.2 640 2.7 573 A29 2677 5.4 1001 5.2 1209 A30 2678 2.9 252 4.3 809 A31 2679 ND 3019 ND 6329 A32 2765 0.07 52 0.2 260 A33 2766 0.66 517 8.2 1680 A34 2767 35 >10000 7.1 4019 A35 2768 11 6130 62 7559 A36 2769 0.84 719 15 2370 A37 2770 24 3909 889 >10000 A38 2771 5.9 1963 8.8 1603 A39 2772 48 7811 43 5045 A40 2773 ND ND ND ND A41 2774 ND ND ND ND A42 2775 2.4 914 8.6 1911 A43 2806 0.08 168 9.3 4292 A44 2807 0.02 9.2 0.32 363 A45 2808 0.15 236 2.6 2345 A46 2809 2.1 925 513 >10000 A47 2810 2.6 868 226 >10000 A48 2811 23 7484 147 6404 A49 2830 0.01 38 0.15 227 A50 2831 1.7 1048 5.0 2661 A51 2921 2.1 1222 8.9 1262 A52 2922 0.37 111 12 2143 A53 2923 0.06 62 31 4007 A54 2924 0.14 126 23 2566 A55 2925 101 6231 26 1371 A56 2926 ND ND ND ND A57 3009 1.5 1383 77 >10000 A58 3010 6.5 1438 73 5311 A59 3011 20 >10000 71 9471 A60 3347 0.08 61 5.4 2282 A61 3348 0.06 169 5.6 6411 A62 3408 11.7 4600 250 >10000 A63 3410 4.0 5268 222 >10000 A64 3459 6.1 7856 178 >10000 A65 3460 0.4 633 10 6783 A66 3461 1.9 2572 25 8330 A67 3462 0.7 769 9.8 5304 A68 3463 0.14 700 1.3 4950 A69 3464 0.5 619 26 6003 A70 3465 2.0 2335 8.3 6266 A71 3466 4.1 2461 31 7108 A72 3467 9.9 ND 66 ND A73 3468 1.5 2361 63 >10000 A74 3469 0.4 231 1.3 7987 A75 3470 4.6 3492 118 >10000 A76 3525 1.0 921 3.6 3701 A77 3526 13 7868 6.1 6820 A78 3527 13 4395 24 >10000 A79 3585 5.2 ND 12 ND A80 3586 32 ND 382 ND A81 3880 1648 >10000 A82 3881 2459 >10000
Figure imgf000210_0001
abe . Structure actvty reatons p o N-susttute an metyene amne susttuted compounds. T
Figure imgf000210_0002
B4 2761 0.7 270 8.4 2122 B5 2803 5.7 7285 127 >10000 B6 2805 0.19 709 0.44 1429 B7 2846 3.4 2436 <1 1729 B8 2870 19 4123 435 >10000 B9 2933 0.13 62 1.9 461 B10 2934 0.51 321 156 >10000 B11 3006 ND ND ND ND B12 3007 ND ND ND ND B13 3008 ND ND ND ND B14 3034 0.64 508 60 >10000 B15 3882 204 1424
Figure imgf000211_0001
. , , , d b d nd methylene amine substituted compounds.
Figure imgf000211_0002
STK33 RET T
Figure imgf000211_0003
C4 2563 18 8707 ND ND C5 2564 7.4 3283 45 >10000 C6 2568 ND ND ND ND C7 2569 ND 1939 ND ND C8 2572 ND ND ND ND C9 2573 ND ND ND ND C10 2580 ND 577 ND ND C11 2581 ND ND ND ND C12 2589 ND ND ND ND C13 2590 ND ND ND ND C14 2591 0.1 340 0.23 1381 C15 2593 ND 837 ND ND C16 2599 0.24 209 0.6 575 C17 2603 ND ND ND ND C18 2604 ND ND ND ND C19 2605 ND ND ND ND C20 2647 ND ND ND ND C21 2648 7.1 2093 26 ND C22 2649 0.01 11 0.01 28 C23 2651 ND ND ND ND C24 2762 0.7 145 0.5 145 C25 2763 8.6 3607 37 5821 C26 2764 3.5 766 1.6 178 C27 2871 0.08 45 1.1 380 C28 2872 1.2 3008 108 >10000 C29 2873 9.4 3778 110 >10000 C30 2884 0.35 279 1.8 557 C31 2885 0.13 45 1.8 484 C32 2886 ND ND ND ND C33 2942 0.3 173 5.7 1445 C34 2953 4.5 1190 53 4984 C35 2986 1.0 435 91 >10000 C36 2987 34 >10000 ND >10000 C37 3094 0.09 59 0.7 833 C38 3134 0.37 718 14 >10000 C39 3135 0.64 921 5.8 4228 C40 3136 0.13 177 0.8 2630 C41 3137 11 7155 21 7791 C42 3153 3.7 1255 6.4 3956 C43 3154 24 8511 14 3852 C44 3155 0.86 1825 4.3 >10000 C45 3156 0.22 647 0.7 3191 C46 3157 0.41 3448 4.1 >10000 C47 3197 16 >10000 166 494 C48 3198 0.19 >10000 2.6 6882 C49 3232 0.19 196 3.0 2594 C50 3233 52 >10000 573 >10000 C51 3234 4.7 >10000 494 >10000 C52 3235 ND ND ND ND C53 3236 IC50 = 213 >10000 37.5 5211 C54 3353 5.5 6279 469 >10000 C55 3808 14 2348 5.9 >10000 C56 3809 53 >10000 11 >10000 C57 3810 12 2086 34 >10000 C58 3811 1.3 5418 1.8 >10000 C59 3812 3.7 1713 6.6 >10000 C60 3813 22 >10000 263 >10000 C61 3814 17 >10000 147 >10000 C62 3883 298 2461 C63 3884 186 2539 C64 3885 37 316 C65 3886 45 763 C66 3887 6907 >10000
Figure imgf000213_0001
Example 3: Metabolic, toxicity, and distribution studies of STK33 inhibitors Metabolic stability assay in liver microsomes. The stabilities of CDD-2110, CDD-2211, CDD-2212, CDD-2807, and CDD-3348 in liver microsomes were performed and half-life was calculated as previously reported. Briefly, each CDD compound (2.0 μM) was incubated in the mouse or human liver microsomes (0.5 mg protein/mL) at 37 °C in 1× PBS fortified with NADPH (1.0 mM). The samples were collected at specific time-points 0, 30, and 60 min in duplicate. The reactions were terminated by adding equivalent volume of ice-cold CH3OH and vortexed. After centrifugation at 15000 g for 10 min, 3.0 μL of the supernatant was analyzed by UHPLC-Q Exactive Orbitrap MS equipped with 50 mm × 4.6 mm column (XDB C-18, Agilent Technologies, USA). The column temperature was maintained at 40 °C. Q Exactive MS was operated in positive mode with electrospray ionization. MS data were acquired from 100 to 800 Da in-profile mode. The ion at m/z 371.1012 (positive mode) was used as reference masses during acquisition. Pharmacokinetics of CDD-2807 in mice. The pharmacokinetic studies (50) of CDD-2807 were performed at two doses. CDD- 2807 was dissolved in 10% Captisol containing 5% DMSO to prepare the solutions of 0.5 mg/mL and 2.5 mg/mL. Six male mice (strain: C57BL/6NJ, 8−12 weeks) were divided equally and randomly into 2 groups (n = 3 for each dose). One group of mice were administered with 5 mg/kg CDD-2807 (i.p., 10 µL/g of 0.5 mg/mL solution) and the second group with 25 mg/kg CDD-2807 (i.p., 10 µL/g of 2.5 mg/mL solution). At 0, 15, 30 min, 1, 2, 4, 6, 8, 24 h post-dose, around 20 µL of blood was collected from the tail vein and anti- coagulated by heparin. The blood samples were centrifuged at 2000 g for 3 min at 4 ºC, and the plasma samples were transferred to tubes and stored at −80°C before analysis. All the samples were analyzed by the Thermo TSQ Quantis MS coupled with a Thermo Vanquish UHPLC (San Jose, CA). CDD-2807 was separated on a Luna C18 column (1 mm × 50 mm, 1.6 µm, Phenomenex, Torrance, CA), and eluted by a water-acetonitrile mobile phase system (both containing 0.1% formic acid, v/v) at the flow rate of 0.15 mL/min. CDD-2807 was monitored under the selected reaction monitoring (SRM) mode coupled with a positive electrospray ionization (ESI) source. The SRM ion pairs were 447→321 for CDD-2807. The ion spray voltage was set at 3500 V. High-purity nitrogen was used as the sheath gas (35 arbitrary unit), auxiliary gas (7 arbitrary unit), and high-purity argon was used as the collision gas. The temperatures of the ion transfer tube and the vaporizer were set at 300°C and 275°C, respectively. The concentration of CDD-2807 in plasma was quantified using individual calibration curves. The calibration curve was regressed with a weight of 1/x2 with high linearity (r2 >0.99). PK parameters such as half-time (t1/2), area under the plasma concentration–time curve during the period of observation (AUC0–t), area under the plasma concentration–time curve from zero to infinity (AUC0–∞), clearance normalized by the bioavailability (CL/F), volume of distribution normalized by the bioavailability (Vd/F) and the mean residence time (MRT) were calculated by WinNonlin software (Certara, Princeton, NJ) by noncompartmental analysis. The plasma concentration–time curves were plotted in Prism 9 (GraphPad, San Diego, CA) as mean ± S.D. CDD-2807 concentrations in testis and brain. The testis and brain harvested from day 45 of protocol 1 and day 63 of protocol 2 were weighed and snap-frozen in liquid nitrogen. The samples were stored at −80°C before analysis. Twenty-five mg of testis or brain tissue was homogenized in 150 μL of 50% methanol, and 50 μL of homogenate was added to 200 μL of ice-cold CH3OH containing 0.1 μM of agomelatine (internal standard). The mixture was vortexed and centrifuged at 15000 g for 15 min, and supernatants were transferred into sample vials for analysis. CDD-2807 was analyzed by the same LC-MS/MS method used in the pharmacokinetic study The concentration of CDD-2807 in the testis and brain was determined by using individual calibration curves established with the corresponding tissue homogenates from untreated mice. The calibration curve was regressed with a weight of 1/x2 with high linearity (r2 >0.99). Alanine transaminase (ALT) and aspartate transaminase (AST) assays. On day 45 of protocol 1 and day 63 of protocol 2, three mice from each protocol were euthanized, and blood was collected through cardiac puncture. The serum was isolated by centrifuging the whole blood samples at 1500 g for 10 min and then maintained on wet ice before analysis. ALT and AST levels were in duplicates using assay kits (StanBio, Boerne, TX) according to the manufacturer’s instructions. ALT and AST levels were determined using the rate of NADH oxidation, which was quantified by measuring absorbance at 340 nm using a CLARIOstar plate reader (BMG Labtech in Cary, NC). CDD-2807 (FIG.2B) emerged as a potent STK33 ligand (Kd = 0.02 nM), not only maintaining high potency (IC50 = 9.2 nM) toward STK33 in the NanoBRET cellular assay but also demonstrating excellent metabolic stability with >60 min half-life in both MLM and HLM (FIG.2C). To access cellular kinome-wide selectivity, CDD-2807 was subjected to NanoBRET Target Engagement K192 assay (Promega) at a dose of 1 μM against 192 full- length protein kinases in HEK293 cells. In this assay, STK33 was the most potent binder (95.9% occupancy) although other kinases were engaged by CDD-2807 (FIG.2D). ThermoFisher dose response assays and NanoBRET cellular assays were used to evaluate potency of CDD-2807 against off-target kinases possessing >80% occupancy, including CDC-like kinase 4 (CLK4) (94.7% occupancy), CLK2 (89.7% occupancy), rearranged during transfection (RET) (85.8% occupancy), and CLK1 (83.5% occupancy). Compared to its interaction with CLK4 (Kd = 0.9 nM and IC50 = 85 nM), CLK2 (Ki = 1.3 nM and IC50 = 101 nM), RET (Ki = 0.3 nM and IC50 = 363 nM), and CLK1 (Ki = 3.1 nM and IC50 = 116 nM) in these two assays, CDD-2807 was the most potent ligand of STK33 (FIG.2C). Taken together, CDD-2807 is a potent, stable, and relatively selective STK33 inhibitor, showing >9- fold selectivity versus the other kinases (FIG.2C). Original hits CDD-2110 and CDD-3348, along with truncated enantiomers CDD- 2211 and CDD-2212, were also assayed against off-target kinases CLK1, CLK2, CLK4, and RET (FIG.2C and Table C). All compounds except CDD-2212 exhibited >10-fold selectivity of STK33 versus off-target kinases in cells. The significant difference of biochemical and cellular activities between CDD-2211 (R-isomer) and CDD-2212 (S-isomer) (e.g., 16-fold versus 1.1-fold selectivity of STK33 over RET in NanoBRET assays; FIG.2C) suggested that the chirality played an important role on the selectivity. The increased selectivity of CDD-2110 compared to CDD-3348 (e.g., 96-fold to 437-fold selectivity of STK33 over CLK4 in biochemical assays and 14-fold to 23-fold selectivity of STK33 over CLK4 in the NanoBRET assay; FIG.2C and Table B) also suggested that the selectivity toward STK33 could be improved by modification of the linker at BB1. A summary of the data in relation to other molecular scaffolds is shown in Table B. A summary of the pharmacokinetic (PK) parameters is shown in Table G. Table G. Noncompartmental PK parameters for CDD-2807 (intraperitoneal injection, n = 3, mean ± SD). Parameter Unit 5 mg/kg 25 mg/kg Tmax h 0.33 ± 0.14 0.33 ± 0.14 Cmax µM 2.95 ± 0.64 11.90 ± 0.40 T1/2 h 11.90 ± 4.38 11.30 ± 3.41 AUC0-t h*µM 9.46 ± 1.47 55.48 ± 5.42 AUC0-∞ h*µM 11.17 ± 1.11 67.73 ± 12.41 CL/F mg/(h*µM)/kg 7.77 ± 3.20 5.92 ± 0.79 Vd/F mg/(µM)/kg 0.45 ± 0.04 0.38 ± 0.08 MRT0-t h 5.27 ± 0.48 6.17 ± 0.59 MRT0-∞ h 11.14 ± 4.43 12.24 ± 3.80 Example 4: Production of X-ray co-crystal structure of STK33 One obstacle to a better understanding of the molecular basis of the high affinity of our STK33 inhibitors is the lack of available structures for STK33. To address this conundrum, the crystal structure of the human STK33 kinase domain was determined in complex with CDD-2211 at 2.7 Å resolution (FIG.3 and Table H). STK33 KD protein expression and purification for crystallography. Human STK33 KD (GenBank accession number CAC29064.1, residues 99−383) with tobacco etch virus (TEV) cleavage site and cGFP-Strep-tag®II was expressed in insect SF9 cells (Invitrogen). SF9 cells were cultured in SF900III medium (Gibco) and infected with STK33 KD baculovirus in an optimal virus/ insect cells ratio. Cells were harvested 48 h after infection; pellets were kept at −80°C. For purification, frozen cell pellet was resuspended in lysis buffer (25 mM Tris, pH 8.0, 150 mM NaCl, and 1mM TCEP) and lysed using homogenizer (Avestin). After centrifugation at 48,000g for 1 h, the soluble fraction was loaded onto pre-equilibrated Strep-Tactin Superflow resin (IBA Lifesciences GmbH) and eluted with elution buffer (25 mM Tris, pH 8.0, 150 mM NaCl, 1 mM TCEP, and 2 mM desthibiotin). Fractions containing the protein were treated with TEV protease (1:60 weight ratio, overnight) to remove the GFP tag and the mixture was subjected to Superdex™ 75 Increase 10/300 GL gel filtration column (Cytiva) equilibrated with the abovementioned lysis buffer. The peak corresponding to the estimated molecular mass of approximately 70 kDa was pooled and used for co-crystallization. Crystallization, data collection, and structure solution STK33 KD was co-crystallized with CDD-2211 by hanging drop vapor diffusion method. For crystallization, the purified STK33 at 1.8 mg/mL was mixed with 6 molar excesses of CDD-2211. The protein-inhibitor mixture was concentrated using Amicon Ultra- 15 centrifugal filters (Millipore Sigma) to 14 mg/mL. Mosquito (SPT labtech) was used to dispense equal volumes of protein and reservoir (250 nl each) against 70 μL reservoir buffer in 96 wells crystallization tray (96-Well clear polystyrene microplate from SPT labtech). Crystals were observed after 2 days in drop using JCSGplus -A12 condition (0.2 M potassium nitrate, 20% w/v Polyethylene glycol 3350). The diffraction data were collected at Advanced Light Source (ALS), Beam Line 5.0.2 (UC Berkeley, USA) at wavelength (λ) =1.0000 Å, by using an Dectris Pilatus 6M detector. The data was integrated and scaled by using iMosflm and SCALA, respectively. The crystal structures of the STK33/CDD-2211 complex was determined by molecular replacement using a monomeric model of STK33 KD (residues 99−383) generated by Alpha fold as a search model. CDD-2211 was traced and fitted manually into electron density by using COOT. The final models have gone through several rounds of refinement using phenix.refine (46) followed by manual model building using COOT (47). For all structural analysis and preparation of figures, the visualization program PyMOL was used (48). A summary of data collection is shown herein. Table H. Data collection and refinement statistics. Complex STK33/CDD-2211 Data collection Space group P 212121 Cell Dimensions a, b, c (Å) 84.187.9398.45 α, β, γ (°) 90, 90, 90 Resolution (Å) 43.94–2.70 (2.83–2.70)a Rmerge 0.149 (1.53) I/ σ (I) 7.40 (1.30) CC 1/2 0.994 (0.605) Completeness (%) 96.77 (100) Multiplicity 2.0 (2.0) Refinement No. of unique reflections 19879 (1037) Rwork/Rfreeb 21.46/25.67 (35.01/42.15) No. of atoms Protein 4272 Ligand 66 R.m.s.c deviations Bond length (Å) 0.004 Bond angles (°) 0.77 Ramachandran plot (%) Favored region 96.64 Allowed region 3.17 Outliers 0.19 a Highest resolution shell is shown in parenthesis. b 5% of the observed intensities was excluded from refinement for cross validation purposes. c R.m.s. means root mean square. The STK33:CDD-2211 cocrystal contains one dimer per asymmetric unit, with each monomer showing clear density for bound CDD-2211 (FIG.3B). The two STK33 kinase domain monomers are very similar, showing a root-mean-square deviation (RMSD) values of ~0.5 Å between ∼240 shared CA atoms (FIG.6). Each monomer shows a nearly identical pose for CDD-2211 (FIG.3B) bound in the ATP pocket (FIG.6). All residues in the construct are ordered except for a segment of the activation loop (residues 273-283 in chain A and 274-284 in chain B are disordered). The STK33 dimer is formed by the activation loop segment and the following two-turn helix with the APE motif from one monomer docking onto a surface formed between ^E and ^G helices of the other monomer (FIG.3A). CDD-2211 binds in the active site formed between the small and large lobes (FIGs. 3A-3E and FIG.6). The biphenyl (BB3) binds to the outer lip of the active site near the hinge (FIG.3C). It interacts with L122 and G123 on top and G198, E199, and M245 at the bottom through van der Waals interactions (FIGs.3D and 3E). The indazole (BB2) binds the ribose pocket near the hinge region. It forms hydrogen bonds with backbones of E193 and C195 at the hinge (FIG.3E). The pyrrolidine with dimethyl amine (BB1) contacts the Mg2+ positioning loop, glycine-rich loop, and activation loop. The dimethyl amine hydrogen bonds with D265 that normally interacts with Mg2+. BB1 is shielded by F127 at the Gly rich-loop (top) and E242 and N243 at the catalytic loop (bottom), and it interacts with these residues through van der Waals interactions. This cocrystal structure is consistent with structure-activity-relationship (SAR) studies from a set of CDD-2211 analogs (Table B). Replacement of the -NH group at the indazole (BB2) with the N-methyl group resulted in total loss of STK33 activity at 500 µM, confirming that hydrogen bond interaction at the hinge is critical for STK33 potency. At the biphenyl (BB3), ortho-substitution was proven to be the best orientation since STK33 potency gradually decreased with the movement of the phenyl substituent from the ortho- position to meta- and para-positions. Moreover, no STK33 activity was observed at 500 µM for analogs with shorter linker between the indazole and the biphenyl. Therefore, the interaction at the hinge region involving the indazole fragment, the ortho-orientation of the biphenyl group, and the specific distance hold by the two-carbon acetylene linker between the indazole and biphenyl components within the CDD-2211 scaffold synergistically contributed to its enhanced potency against STK33. A computational model of STK33 bound with CDD-2807 was generated based on the crystal structure of the STK33/CDD-2211 complex (FIG.8). The model suggests that the nitrogen of the piperdine moiety at BB1 would interact with the side chain of E199 through a hydrogen bond while its interactions with the active site at BB2 and BB3 would be similar as in the STK33/CDD-2211 complex. CDD-2211, similar to CDD-2807, demonstrated some (although low) inhibition of off-target kinases CLK1, CLK2, CLK4, and RET, as shown in FIG.2C. To understand its cross activity, crystal structures of RET and CLK4 were aligned as representatives with the STK33/CDD-2211 complex and compared the binding site residues. The structure comparison showed that the majority of the CDD-2211 contacting residues from STK33 are conserved in both RET and CLK4 except for E199 of the linker (immediately following the hinge) and M245 of ^7 (FIG.7). In RET and CLK4, E199 is replaced with a serine (S811 in RET and S247 in CLK4) and M245 is replaced with a leucine (L881 in RET and L295 in CLK4), suggesting the potential of increasing the selectivity toward STK33 by modifying BB1 or BB3 moieties of CDD-2211 near E199. Example 5: Mouse study of contraceptive effects Based on molecular modeling, mouse and human STK33 have identical ATP binding pockets, consistent with evolutionary conservation and function of STK33. The human STK33 amino acids that are in contact with CDD-2211 are identical in mouse STK33 (FIG. 9), suggesting that CDD-2807 would equally inhibit mouse and human STK33. To determine the consequences of STK33 inhibition in vivo, reproductive outcomes in male were evaluated in mice dosed with CDD-2807, a potent and stable compound. Fertility analysis Sexually mature male C57/129 hybrid mice were randomly assigned to either the vehicle control treatment or the CDD-2807 treatment groups. Drug formulation and injections were as follows: mice receiving the 15 mg/kg CDD-2807 twice daily dosing had the compound dissolved in DMSO, which was injected at 5% of the final volume in 10% Captisol and water. For mice receiving a single daily dose of 50 mg/kg CDD-2807 once daily, the compound was dissolved in DMSO and injected at 5% in 20% Captisol and water. All injections were administered as an intraperitoneal (IP) injection, and for the 15 mg/kg CDD-2807 and DMSO control mice, the injections were delivered in the morning and the evening. Mice in the 50 mg/kg dosing received IP injections once in the evening. Weights were collected daily prior to drug administration and monitored throughout the entirety of the injection period. Injections were given for 21 days prior to introduction of female mice. During the fertility assessment, males were housed continuously with two C57/129 sexually mature females for 45 days to generate approximately two months of fertility data. During the fertility assessment, the total number of pups born per litter sired by each male were counted and the total number of litters and pups sired per male over the entire trial was calculated. Mean pup number and SEM were used to determine differences in average number of pups sired using a t-test using Prism 9. Histological analysis Briefly, tissue was fixed in Bouin’s fixative, washed with 70% ethanol, embedded in paraffin, sectioned at 4-µm thickness, and stained by periodic acid-Schiff (PAS)- hematoxylin. Computer-assisted sperm analysis (CASA). Sperm were extracted by mincing both segments of the cauda epididymis 25 times with dissection scissors in 1 mL of Enhance Sperm Wash w/Gentamicin (Vitrolife, Sweden) medium. All media was maintained at 37°C throughout the procedure. After incubation for 15 minutes, the supernatant was diluted, and 6 µL of diluted sample was added into a single chamber of a dual chambered 20 µm-depth Leja semen analysis slide (Spectrum Technologies). Sperm parameters were measured using the Hamilton Thorne CEROS II system. Per sample, a minimum total of 200 sperm were measured, which typically consisted of recording a minimum of five non-overlapping fields. An additional measurement was taken following a 90-minute incubation at 37°C to allow for sperm capacitation. Sperm analysis for scanning electron microscopy (SEM). Sperm were extracted by mincing the cauda of the epididymis 30 times with dissection scissors in 1 mL of Enhance Sperm Wash w/Gentamicin (Vitrolife, Sweden) medium incubated at 37°C. Sperm were allowed to remain in the media at 37°C for a minimum of 15 minutes to allow sperm to disperse from the tissue. Media and sperm were then collected and moved to a separate tube and then sperm were spun down at 300 x g for 5 minutes at room temperature, allowing sperm to collect at the bottom without any epididymis segments being present. Media was removed and sperm cells were resuspended in DPBS to wash. Following the wash, sperm were spun down a second time and then resuspended in fresh 2.5% glutaraldehyde/PBS and fixed for 30 minutes at room temperature on a rocker. After fixation, sperm were dehydrated in increasing concentrations of ethanol form 20%, 30%, 50%, 60%, 70%, 90%, and 100%. During each dehydration step, sperm were incubated at room temperature for 15 minutes. After each increase in percentage of ethanol, the density of the sperm changes and requires higher speeds for each subsequent spin down; when sperm are in 100% ethanol, it requires ≥1200 x g to pellet them. Sperm were incubated overnight in 100% ethanol at 4°C, and the following day the samples were split and then spun down at 1500 x g for 5 minutes at room temperature and then resuspended in 50% tert-butanol/ethanol and incubated for 15 minutes. Samples were then imaged at the Houston Methodist Research Institute Microscopy SEM/AFM core on a FEI Nova NanoSEM 230 instrument. Because CDD-2807 has a plasma t1/2 of over 11 hours with dose-dependent increase of the maximum concentration (Cmax) and area under the curve (AUC) when delivered intraperitoneally (FIG.10 and Table G), adult male mice were subjected to two protocols (FIGs.11A-11B). In protocol 1, adult male mice (n = 6 per cohort) were treated with vehicle control or CDD-2807 at 15 mg/kg twice per day for 21 days, housed continuously with fertile females mice of reproductive age (2 per male), and then evaluated at day 45 for effects of CDD-2807 on the male reproductive tract (after the first litter was born) or continued to be housed with the females until day 66 (FIG.11A). In protocol 2, adult male mice (n = 7 per cohort) were treated with vehicle control or CDD-2807 at 50 mg/kg once per day for 21 days, housed continuously with females (2 per male), and then evaluated at later time points. During the period of CDD-2807 or control delivery, there were no death and no significant weight changes of the two cohorts of mice for protocol 1 (FIG.12A) or protocol 2 (FIG. 12B), suggesting that CDD-2807 is very safe. Whereas the six control treated males in protocol 1 sired an average of 1.83 ± 0.07 litters/male and 6.75 ± 0.19 pups/female/litter in month 1, four of the six CDD-2807-treated males sired litters (either one or two litters; 1.0 ± 0.37 litters/male) and at a lower 0.92 pups/female/litter (FIG.4A). In month 2, litters were born to all females housed with the six control males (2.0 ± 0 litters male; 7.25 ± 0.17 pups/female/litter) (FIG.4A). In contrast, no litters were produced by females housed with males who received CDD-2807 (FIG.4A). For the protocol 2 mice, all seven control males sired litters from both females in month 1 (7.29 ± 0.18 pups/litter/female), whereas only one female became pregnant and delivered a single pup from the seven CDD-2807 treated males (50 mg/kg/day) (0.07 ± 0.07 pups/litter/female) (FIG.4B). In month 2 CDD-2807-treated males did not sire any litters, despite their ability to mate; while litters were born to all females housed with control males (2.71 ± 0.07 litters male; 9.16 ± 0.09 pups/female/litter). Thus, CDD-2807 delivery in both protocols induced a contraceptive effect. For males in protocol 1 evaluated at day 45 of CDD-2807 treatment or day 63 of protocol 2 mice, CDD-2807 easily crossed the blood-testis barrier (89.6 ± 14.1 ng or 114.3 ± 7.9 ng of CDD-2807/mg tissue, respectively), whereas it was extremely low in the brains of the protocol 1 males (0.28 ± 0.09 ng of CDD-2807/mg tissue) and undetectable in the protocol 2 males (FIG.13A). Despite the contraceptive effect of CDD-2807, testis size of CDD-2807-treated mice was not changed versus control males (FIG.4C; FIG.13B). For protocol 1, CDD-2807-treated males showed no difference in body weight versus controls (FIG.12C) or alterations in serum ALT (control: 47.7 ± 5.3 IU/L; CDD-2807: 20.3 ± 5.3 IU/L) or AST (control: 48.2 ± 23.4 IU/L; CDD-2807: 35.8 ± 3.1 IU/L). Likewise, for protocol 2 mice, CDD-2807-treated males showed no statistical difference in body weight versus controls (FIG.12D) or alterations in serum ALT (control: 37.7 ± 13.6 IU/L; CDD- 2807: 46.0 ± 11.0 IU/L) or AST (control: 42.1 ± 14.9 IU/L; CDD-2807: 29.8 ± 6.8 IU/L), confirming the relative safety of CDD-2807. To further understand the cause of the contraceptive effect of CDD-2807, sperm from the cauda epididymides of each mouse were analyzed using Computer Assisted Sperm Analysis (CASA). Although sperm counts were lower in the controls in protocol 1 mice versus CDD-2807-treated mice (FIG.13C), sperm motility (FIG.13D), progressive sperm (FIG.13E), and hyperactivated sperm (FIG.13F) were reduced in the CDD-2807-treated mice. In protocol 2, sperm counts (FIG.4D), sperm motility (FIG.4E), progressive sperm (FIG.4F), and hyperactivated sperm (FIG.4G) were all statistically reduced in the CDD- 2807-treated mice versus control mice. Histologically, the testes and epididymides of the CDD-2807-treated mice in protocol 2 showed no major defects (FIGs.14A-14B), suggesting that CDD-2807 directly caused a functional defect in motility rather than a morphologic defect. By scanning electron microscopy (SEM), sperm analysis of the CDD-2807-treated mice in protocol 2 demonstrated teratozoospermia (FIG.4H). Thus, low dose (CDD-2807 at 15 mg/kg twice per day for 45 days) appears to cause mostly a functional defect in motility, whereas high dose (CDD-2807 at 50 mg/kg per day for 63 days) causes both functional and morphologic defects, consistent with the Stk33 KO and likely a direct role of STK33 in phosphorylation of fibrous sheath proteins A-kinase anchoring protein 3 (AKAP3) and AKAP4. To evaluate the reversibility of the effects of CDD-2807 on male fertility, drug and control treatment of the mice in protocol 1 was halted at day 63, the males were removed from the females for 21 days, and the males were caged with new adult females (FIG.4A). The four control males sired offspring from the females (7.50 ± 0.20 pups/litter/female); likewise, the three previously treated CDD-2807 cohort males sired offspring from the females (7.67 ± 0.61 pups/litter/female) within the first month of breeding. Thus, the effects of CDD-2807 are quickly reversible. In conclusion, examples herein demonstrate that DEC-Tec can identify novel kinase inhibitors that can be used for contraceptive development. In the present example, a hit from DEC-Tec screening was identified that was developed into a potent inhibitor of STK33, showed that our inhibitor (CDD-2807; molecular weight of 447 Da) easily crossed the BTB to cause infertility, and demonstrated that the contraceptive effects were quickly reversible. Because CDD-2807 is more metabolically stable (~10 hours) in HLM versus MLM (~1 hour) (FIG.2C), CDD-2807 could be metabolically more stable in men than mice, allowing us to translate our findings to men. Additionally, a proteolysis-targeting chimera (PROTAC)-based chemical knockdown approach is possible for testis-specific degradation of STK33. Lastly, CDD-2807 represents an excellent chemical probe that can be used to investigate STK33 signaling in various contexts other than male contraception. This work illuminates that kinases can serve as beneficial targets for treating human conditions beyond oncology indications. Sequence Listing STK33 (Human) (SEQ ID NO:1) MADSGLDKKSTKCPDCSSASQKDVLCVCSSKTRVPPVLVVEMSQTSSIGSAESLISLERKKE KNINRDITSRKDLPSRTSNVERKASQQQWGRGNFTEGKVPHIRIENGAAIEEIYTFGRILGK GSFGIVIEATDKETETKWAIKKVNKEKAGSSAVKLLEREVNILKSVKHEHIIHLEQVFETPK KMYLVMELCEDGELKEILDRKGHFSENETRWIIQSLASAIAYLHNNDIVHRDLKLENIMVKS SLIDDNNEINLNIKVTDFGLAVKKQSRSEAMLQATCGTPIYMAPEVISAHDYSQQCDIWSIG VVMYMLLRGEPPFLASSEEKLFELIRKGELHFENAVWNSISDCAKSVLKQLMKVDPAHRITA KELLDNQWLTGNKLSSVRPTNVLEMMKEWKNNPESVEENTTEEKNKPSTEEKLKSYQPWGNV PDANYTSDEEEEKQSTAYEKQFPATSKDNFDMCSSSFTSSKLLPAEIKGEMEKTPVTPSQGT ATKYPAKSGALSRTKKKL RET (SEQ ID NO:2) MAKATSGAAGLRLLLLLLLPLLGKVALGLYFSRDAYWEKLYVDQAAGTPLLYVHALRDAPEE VPSFRLGQHLYGTYRTRLHENNWICIQEDTGLLYLNRSLDHSSWEKLSVRNRGFPLLTVYLK VFLSPTSLREGECQWPGCARVYFSFFNTSFPACSSLKPRELCFPETRPSFRIRENRPPGTFH QFRLLPVQFLCPNISVAYRLLEGEGLPFRCAPDSLEVSTRWALDREQREKYELVAVCTVHAG AREEVVMVPFPVTVYDEDDSAPTFPAGVDTASAVVEFKRKEDTVVATLRVFDADVVPASGEL VRRYTSTLLPGDTWAQQTFRVEHWPNETSVQANGSFVRATVHDYRLVLNRNLSISENRTMQL AVLVNDSDFQGPGAGVLLLHFNVSVLPVSLHLPSTYSLSVSRRARRFAQIGKVCVENCQAFS GINVQYKLHSSGANCSTLGVVTSAEDTSGILFVNDTKALRRPKCAELHYMVVATDQQTSRQA QAQLLVTVEGSYVAEEAGCPLSCAVSKRRLECEECGGLGSPTGRCEWRQGDGKGITRNFSTC SPSTKTCPDGHCDVVETQDINICPQDCLRGSIVGGHEPGEPRGIKAGYGTCNCFPEEEKCFC EPEDIQDPLCDELCRTVIAAAVLFSFIVSVLLSAFCIHCYHKFAHKPPISSAEMTFRRPAQA FPVSYSSSGARRPSLDSMENQVSVDAFKILEDPKWEFPRKNLVLGKTLGEGEFGKVVKATAF HLKGRAGYTTVAVKMLKENASPSELRDLLSEFNVLKQVNHPHVIKLYGACSQDGPLLLIVEY AKYGSLRGFLRESRKVGPGYLGSGGSRNSSSLDHPDERALTMGDLISFAWQISQGMQYLAEM KLVHRDLAARNILVAEGRKMKISDFGLSRDVYEEDSYVKRSQGRIPVKWMAIESLFDHIYTT QSDVWSFGVLLWEIVTLGGNPYPGIPPERLFNLLKTGHRMERPDNCSEEMYRLMLQCWKQEP DKRPVFADISKDLEKMMVKRRDYLDLAASTPSDSLIYDDGLSEEETPLVDCNNAPLPRALPS TWIENKLYGMSDPNWPGESPVPLTRADGTNTGFPRYPNDSVYANWMLSPSAAKLMDTFDS CLK4 (SEQ ID NO:3) MRHSKRTHCPDWDSRESWGHESYRGSHKRKRRSHSSTQENRHCKPHHQFKESDCHYLEARSL NERDYRDRRYVDEYRNDYCEGYVPRHYHRDIESGYRIHCSKSSVRSRRSSPKRKRNRHCSSH QSRSKSHRRKRSRSIEDDEEGHLICQSGDVLRARYEIVDTLGEGAFGKVVECIDHGMDGMHV AVKIVKNVGRYREAARSEIQVLEHLNSTDPNSVFRCVQMLEWFDHHGHVCIVFELLGLSTYD FIKENSFLPFQIDHIRQMAYQICQSINFLHHNKLTHTDLKPENILFVKSDYVVKYNSKMKRD ERTLKNTDIKVVDFGSATYDDEHHSTLVSTRHYRAPEVILALGWSQPCDVWSIGCILIEYYL GFTVFQTHDSKEHLAMMERILGPIPQHMIQKTRKRKYFHHNQLDWDEHSSAGRYVRRRCKPL KEFMLCHDEEHEKLFDLVRRMLEYDPTQRITLDEALQHPFFDLLKKK STK33 (Mouse) (SEQ ID NO:4) MADPSLNDNPTACPHCASSQAGLLCVCPAGKSPVLVVEMSQTSSIGSTEFFASQERKKERNT SRESSLKDLSIRTSNVERKPQAQWSRSNVTVGKIPHIRMDDGAGIEEFYTFGRILGQGSFGM VFEAIDKETGAKWAIKKVNKEKAGSSAMKLLEREVSILKTVNHQHIIHLEQVFESPQKMYLV MELCEDGELKAVMDQRGHFSENETRLIIQSLASAIAYLHNKDIVHRDLKLENIMVKSSFIDD NNEMNLNIKVTDFGLSVQKHGSRSEGMMQTTCGTPIYMAPEVINAHDYSQQCDIWSIGVIMF ILLCGEPPFLANSEEKLYELIKKGELRFENPVWESVSDSAKNTLKQLMKVDPAHRITAKELL DNQWLTGNTLSSARPTNVLEMMKEWKNNPESDEETNTDEETEQSAVYSPSANTAKQPTNAAK KPAAESVGMTSSNSSSSKLLSAESKAEPEKSSETVGHASVAKTTLKSTTLFRGKKRL Enumerated Embodiments The following exemplary embodiments are provided, the numbering of which is not to be construed as designating levels of importance: Embodiment 1 provides a compound of Formula (I): I), wherein: L1 is selected from the g
Figure imgf000226_0001
roup cons st ng o -C C-,-CH=CH- , -CH2-, and a bond; X is selected from the group consisting of -C(=O)-, -C(CH3)2-, -CF2-, -CHF-, - CH(CH3)-, and -CH2; Z1, Z2, or Z3 are independently CH or N, wherein 0 or 1 of Z1, Z2, and Z3 is N; Ra and Rb are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, and optionally substituted 3-to-8 membered heterocyclyl, or Ra and Rb may combine with the nitrogen atom to which they are bound to form optionally substituted 3-to-8 membered heterocyclyl; and Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C2-C10 heteroaryl, or CN; or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof. Embodiment 2 provides the compound of Embodiment 1, wherein the compound of formula (I) is the compound of formula (Ia): a). Embodiment 3 provide
Figure imgf000226_0002
ent 1 or 2, wherein X is -C(=O)-. Embodiment 4 provides the compound of any one of Embodiments 1-3, wherein Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolinyl, optionally substituted quinolinyl, optionally substituted 2-pyridyl, optionally substituted 3-pyridyl, optionally substituted 4-pyridyl, optionally substituted pyrimidinyl, optionally substituted pyrazinyl, or optionally substituted triazinyl. Embodiment 5 provides the compound of any one of Embodiments 1-4, wherein Y is Rc1 Rc5 Rc2 , wherein: c1 c
Figure imgf000227_0001
R , R 2, Rc3, Rc4, and Rc5 are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, - C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3- C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl; Re1 and Re2 are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally substituted 3-to-8 membered heterocyclyl, or optionally substituted C3-C10 heteroaryl. Embodiment 6 provides the compound of any one of Embodiments 1-5, wherein one of Ra or Rb is hydrogen or methyl. Embodiment 7 provides the compound of any one of Embodiments 1-6, wherein Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted 5-, 6-, or-7 membered heterocyclyl. Embodiment 8 provides the compound of any one of Embodiments 1-7, wherein Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted pyrrolidinyl. Embodiment 9 provides the compound of any one of Embodiments 1-3 and 5-8, which is a compound of Formula (II), (III), (IV), or (V):
V), w
Figure imgf000228_0001
Rc1, Rc2, Rc3, Rc4, and Rc5 are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, - C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3- C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally naphthyl, optionally 3-to-8 membered heterocyclyl, and optionally substituted 5-to-8- membered heteroaryl, optionally wherein two of Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, can combine to form an optionally substituted 2-to-7-atom alkylene or heteroalkylene group (e.g., spiro- or fused-bicyclic ring system); X1 is C(Rd3)(Rd4), O, or NRf, wherein Rf is selected from the group consisting of optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl; and X2 is C(Rd5)(Rd6), O, or NRf, wherein Rf is selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl. Embodiment 10 provides the compound of any one of Embodiments 1-9, wherein NRaRb is selected from the group consisting of: , , , ,
Figure imgf000229_0001
, ,
Figure imgf000230_0001
herein the group is selected from the group consisting of:
Figure imgf000230_0002
,
Figure imgf000230_0003
,
Figure imgf000231_0001
Rc5 is optionally substituted phenyl. Embodiment 13 provides the compound of any one of Embodiments 5-10 and 12, wherein Rc1, Rc2, Rc3, and Rc4 are each independently hydrogen. Embodiment 14 provides the compound of Embodiment 5, which is a compound of formula (VI): d3 Rd4 R Rd2 d1 H d5 R N R N I). Embodiment 15 p 14, which is a compound of
Figure imgf000232_0001
formula (VII): I). Embodiment 16 prov Embodiments 1-15, which is
Figure imgf000232_0002
selected from the group consisting of: 3-([1,1'-biphenyl]-2-ylethynyl)-N-(4-methoxyphenyl)-1H-indazole-5-carboxamide, (S)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, (R)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(pyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1- yl)methanone, 1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-one, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethylpiperidine-4-carboxamide, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- N,N-dimethylpiperidine-4-carboxamide, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- N,N-dimethylpiperidine-4-carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone, 4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide, (R)-4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide, (S)-4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1’-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-(phenylethynyl)-1H-indazol-5-yl)(pyrrolidin-1-yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-N-methyl-1H-indazole-5- carboxamide, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-1H-indazole-5- carboxamide, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N,N-dimethyl-1H-indazole-5-carboxamide, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5-yl)methanone, (R)-(3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1-yl)methanone, (S)-(3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1-yl)methanone, (3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol- 5-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H- indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H-indazol- 5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, ((3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol- 5-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol- 5-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(piperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,6-diazabicyclo[3.1.1]heptan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-3-azabicyclo[3.2.1]octan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1-yl)methanone, (R)- 3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1- yl)methanone, (S)- 3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1- yl)methanone, 4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperazin-2-one, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,4-diazepan-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-4- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-7- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-8- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,7-diazaspiro[3.5]nonan- 7-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan- 2-yl)methanone, (7-methyl-2,7-diazaspiro[3.5]nonan-2-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H- indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-4-yl)(7-methyl-2,7-diazaspiro[3.5]nonan- 2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7-diazaspiro[3.5]nonan- 2-yl)methanone, (R)- (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone, (S)- (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone, (S)-4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2- one, (R)-4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2- one, 4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2-one, 7-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,7-diazaspiro[3.5]nonan- 1-one, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-6-yl)(7-methyl-2,7-diazaspiro[3.5]nonan- 2-yl)methanone, (2,8-diazaspiro[4.5]decan-2-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol- 5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-5- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6,9-diazaspiro[4.5]decan-6- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (S)-(2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (S)-(2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2S,6S)-2,6-dimethylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2R,6S)-2,6-dimethylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2R,6R)-2,6-dimethylpiperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,9-diazaspiro[5.5]undecan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[4.5]decan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[4.5]decan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,8-diazaspiro[4.5]decan-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-1-azaspiro[4.5]decan-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-8- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-oxa-6-azaspiro[3.4]octan-6- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-2-azaspiro[4.5]decan-2- yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(7-azaspiro[3.5]nonan-2-yl)-1H-indazole-5- carboxamide, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)- 1H-indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-methyl-2,6-diazaspiro[3.5]nonan- 2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,5-diazaspiro[3.5]nonan- 2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,5-diazaspiro[3.5]nonan- 2-yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,5-diazaspiro[3.5]nonan- 5-yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((4-(trifluoromethoxy)-[1,1'-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone, (R)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, (S)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, 2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, (R)-N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, (S)-N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)- 1H-indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[1,1'-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((2'-(trifluoromethoxy)-[1,1'-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone, (3-((2-(naphthalen-2-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan- 2-yl)methanone, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl- [1,1'-biphenyl]-2-carboxamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl- [1,1'-biphenyl]-3-carboxamide, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'- biphenyl]-2-yl)acetamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl- [1,1'-biphenyl]-4-carboxamide, (3-((2-(6-amino-4-methylpyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone, (3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)- 1H-indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[1,1'-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((2'-(trifluoromethoxy)-[1,1'-biphenyl]-2- yl)ethynyl)-1H-indazol-5-yl)methanone, (3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-(pyrrolidin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-(piperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (R)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, (S)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, 2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, (3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(6-amino-4-methylpyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (R)-7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)indolin-2-one, (S)-7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)indolin-2-one, 7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)indolin-2- one, (3-((2-(2-(pyrrolidin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-(piperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'- biphenyl]-2-yl)acetamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl- [1,1'-biphenyl]-3-carboxamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl- [1,1'-biphenyl]-4-carboxamide, (3-((2-(2-(4-methylpiperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (S)-1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine- 2-carboxamide, (R)-1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine- 2-carboxamide, 1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'- biphenyl]-2-yl)pivalamide, 5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (R)-5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (S)-5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1-yl)methanone, (R,E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S,E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin- 1-yl)methanone, (R,Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S,Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone, (E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan- 2-yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan- 2-yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone, (E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7- diazaspiro[3.5]nonan-2-yl)methanone, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea, 3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3,4- dihydroquinazolin-2(1H)-one, (R)-3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- 3,4-dihydroquinazolin-2(1H)-one, (S)-3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- 3,4-dihydroquinazolin-2(1H)-one, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-2,3- dihydrobenzo[b][1,4]dioxine-6-carboxamide, (R)-N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, (S)-N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)- 2,3-dihydrobenzo[b][1,4]dioxine-6-carboxamide, N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (R)-N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (S)-N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (3-([1,1'-biphenyl]-3-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-4-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone, (3-((2'-morpholino-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, 7-(2-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)isoindolin-1-one, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N,N- dimethyl-[1,1'-biphenyl]-2-carboxamide, (3-((2'-(pyrrolidin-1-ylmethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2'-(morpholinomethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(vinylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(ethylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((3-phenylpyridin-4-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((4-phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, and (3-((3-phenylpyridin-2-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone. Embodiment 17 provides a compound selected from Table A, or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof. Embodiment 18 provides a pharmaceutical composition comprising the compound of any one of Embodiments 1-17 and a pharmaceutically acceptable carrier. Embodiment 19 provides a method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17, and/or the pharmaceutical composition of Embodiment 18, optionally wherein the sterilization is temporary. Embodiment 20 provides a method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18, optionally wherein the infertility is temporary. Embodiment 21 provides a method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18. Embodiment 22 provides the method of any one of Embodiments 19-21, wherein the compound provides a contraceptive effect in the male subject. Embodiment 23 provides the method of any one of Embodiments 19-22, wherein the compound is administered orally to the male subject. Embodiment 24 provides a method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of Embodiments 1-16 and/or the pharmaceutical composition of Embodiment 18. Embodiment 25 provides a method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18. Embodiment 26 provides the method of Embodiment 25, wherein the disease or disorder comprises a RET-mediated cancer. Embodiment 27 provides the method of Embodiment 26, wherein the RET-mediated cancer comprises medullary thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A). Embodiment 28 provides the method of Embodiment 25, wherein the disease or disorder comprises Hirschprung Disease, central hypoventilation syndrome, and/or renal agenesis. Embodiment 29 provides a method of inhibiting at least one of CL1, CLK2, CLK3, and CLK4 in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18. Embodiment 30 provides a method of treating, ameliorating, and/or preventing a disease or disorder caused by CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of Embodiments 1-17 and/or the pharmaceutical composition of Embodiment 18. Embodiment 31 provides a method of Embodiment 30, wherein the disease or disorder comprises Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and renal cancer, breast cancer, and/or melanoma. The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this disclosure has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this disclosure may be devised by others skilled in the art without departing from the true spirit and scope of the disclosure. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims
CLAIMS What is claimed is: 1. A compound of Formula (I): I), wherein: 1
Figure imgf000248_0001
L is selected from the group consisting of -C≡C-,-CH=CH- , -CH2-, and a bond; X is selected from the group consisting of -C(=O)-, -C
(CH3)2-, -CF2-, -CHF-, - CH(CH3)-, and -CH2; Z1, Z2, or Z3 are independently CH or N, wherein 0 or 1 of Z1, Z2, and Z3 is N; Ra and Rb are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, and optionally substituted 3-to-8 membered heterocyclyl, or Ra and Rb may combine with the nitrogen atom to which they are bound to form optionally substituted 3-to-8 membered heterocyclyl; and Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C2-C8 heterocycloalkyl, optionally substituted C2-C10 heteroaryl, or CN; or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof. 2. The compound of claim 1, wherein the compound of formula (I) is the compound of formula (Ia): a).
Figure imgf000248_0002
3. The compound of claim 1 or 2, wherein X is -C(=O)-.
4. The compound of any one of claims 1-3, wherein Y is optionally substituted phenyl, optionally substituted naphthyl, optionally substituted isoquinolinyl, optionally substituted quinolinyl, optionally substituted 2-pyridyl, optionally substituted 3-pyridyl, optionally substituted 4-pyridyl, optionally substituted pyrimidinyl, optionally substituted pyrazinyl, or optionally substituted triazinyl. Rc1 Rc5 Rc2
5. The compound of any one of claims 1-4, wherei , wherein c1 c2 c3 c4 c5
Figure imgf000249_0001
R , R , R , R , and R are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, - C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3- C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl; Re1 and Re2 are independently selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally substituted 3-to-8 membered heterocyclyl, or optionally substituted C3-C10 heteroaryl.
6. The compound of any one of claims 1-5, wherein one of Ra or Rb is hydrogen or methyl.
7. The compound of any one of claims 1-6, wherein Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted 5-, 6-, or-7 membered heterocyclyl.
8. The compound of any one of claims 1-7, wherein Ra and Rb combine with the nitrogen atom to which they are bound to form optionally substituted pyrrolidinyl.
9. The compound of any one of claims 1-3 and 5-8, which is a compound of Formula (II), (III), (IV), or (V): V), w
Figure imgf000250_0001
Rc1, Rc2, Rc3, Rc4, and Rc5 are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C1-C8 alkoxy, optionally substituted C3-C8 cycloalkoxy, - C(=O)NRe1Re2, -NRe1C(=O)Re2, optionally substituted phenyl, optionally substituted naphthyl, optionally substituted 3-to-8 membered heterocyclyl and optionally substituted C3- C10 heteroaryl; or two adjacent substituents selected the group consisting of Rc1, Rc2, Rc3, Rc4, and Rc5 may combine with the carbon atoms with which they are bound to form optionally substituted phenyl, optionally substituted C4-C8 heterocyclyl, or optionally substituted C4-C8 heteroaryl Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, are independently selected from the group consisting of hydrogen, halogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted phenyl, optionally naphthyl, optionally 3-to-8 membered heterocyclyl, and optionally substituted 5-to-8- membered heteroaryl, optionally wherein two of Rd1, Rd2, Rd3, Rd4, Rd5, Rd6, Rd7, Rd8, Rd9, Rd10, Rd11, and Rd12, if present, can combine to form an optionally substituted 2-to-7-atom alkylene or heteroalkylene group (e.g., spiro- or fused-bicyclic ring system); X1 is C(Rd3)(Rd4), O, or NRf, wherein Rf is selected from the group consisting of optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl; and X2 is C(Rd5)(Rd6), O, or NRf, wherein Rf is selected from the group consisting of hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C3-C8 cycloalkyl, optionally substituted C6-C10 aryl, optionally substituted C3-C8 heterocycloalkyl, optionally substituted C2-C8 heteroaryl, and optionally substituted C1-C8 acyl.
10. The compound of any one of claims 1-9, wherein NRaRb is selected from the group consisting of: , , , ,
Figure imgf000251_0001
, ,
Figure imgf000252_0001
11. The compound of any one of claims 5-10, wherein t group is selected from the group consisting of:
Figure imgf000252_0002
, ,
Figure imgf000253_0001
12. The compound of any one of claims 5-10, wherein Rc5 is optionally substituted phenyl. 13. The compound of any one of claims 5-10 and 12, wherein Rc1, Rc2, Rc3, and Rc4 are each independently hydrogen. 14. The compound of claim 5, which is a compound of formula (VI): d Rd4 R 3 Rd2 H Rd1 N Rd5 N I).
Figure imgf000254_0001
15. The compound of claim 14, which is a compound of formula (VII): I).
Figure imgf000254_0002
16. The compound of any one of claims 1-15, which is selected from the group consisting of: 3-([1,1'-biphenyl]-2-ylethynyl)-N-(4-methoxyphenyl)-1H-indazole-5-carboxamide, (S)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, (R)-3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H- indazole-5-carboxamide, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)-2-oxo-1-phenylethyl)-1H-indazole-5- carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(pyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-hydroxypyrrolidin-1-yl)methanone, 1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-one, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N- methylpiperidine-4-carboxamide, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethylpiperidine-4-carboxamide, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethylpiperidine-4-carboxamide, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-N,N- dimethylpiperidine-4-carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone, 4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6-diazaspiro[3.5]nonane-6- carbonyl)-N-methylcyclohexane-1-carboxamide, (R)-4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide, (S)-4-(2-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,6- diazaspiro[3.5]nonane-6-carbonyl)-N-methylcyclohexane-1-carboxamide, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1’-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-(phenylethynyl)-1H-indazol-5-yl)(pyrrolidin-1-yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-N-methyl-1H-indazole-5- carboxamide, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(2-(dimethylamino)ethyl)-1H-indazole-5-carboxamide, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[3,4-b]pyridin-5-yl)(3- (dimethylamino)pyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-(methylamino)pyrrolidin-1- yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N,N-dimethyl-1H-indazole-5-carboxamide, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-(phenylethynyl)-1H-indazol-5-yl)methanone, (R)-(3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-((2,6-dichlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-((2-chlorophenyl)ethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol- 5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol- 5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone,
(R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-fluorophenyl)ethynyl)-1H-indazol- 5-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-morpholinophenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, ((3-([1,1'-biphenyl]-3-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-([1,1'-biphenyl]-4-ylethynyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5- yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5- yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-(isoquinolin-5-ylethynyl)-1H-indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(piperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,6-diazabicyclo[3.1.1]heptan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazabicyclo[2.2.2]octan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3,8-diazabicyclo[3.2.1]octan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-3-azabicyclo[3.2.1]octan-3- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1-yl)methanone, (R)- 3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1-yl)methanone, (S)- 3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-phenylpiperazin-1-yl)methanone, 4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)piperazin-2-one, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-aminopyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,4-diazepan-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-4-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(4,7-diazaspiro[2.5]octan-7-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-8-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,7-diazaspiro[3.5]nonan-7- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (7-methyl-2,7-diazaspiro[3.5]nonan-2-yl)(3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-4-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (R)- (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (S)- (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (S)-4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2-one, (R)-4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2-one, 4-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-3-phenylpiperazin-2-one, 7-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-2,7-diazaspiro[3.5]nonan-1-one, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-6-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (2,8-diazaspiro[4.5]decan-2-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5,8-diazaspiro[3.5]nonan-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6,9-diazaspiro[4.5]decan-6-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (S)-(2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (2-phenylpiperazin-1-yl)(3-((2-(trifluoromethoxy)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-(4-chlorophenyl)piperazin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-phenylpiperazin-1-yl)methanone, (S)-(2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (R)-(2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5- yl)methanone, (2-phenylpiperazin-1-yl)(3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2S,6S)-2,6-dimethylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2R,6S)-2,6-dimethylpiperazin-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)((2R,6R)-2,6-dimethylpiperazin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-benzylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-isopropylpiperazin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,9-diazaspiro[5.5]undecan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[4.5]decan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[4.5]decan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(1,8-diazaspiro[4.5]decan-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-1-azaspiro[4.5]decan-1- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,8-diazaspiro[4.5]decan-8-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-oxa-6-azaspiro[3.4]octan-6- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(8-oxa-2-azaspiro[4.5]decan-2- yl)methanone, 3-([1,1'-biphenyl]-2-ylethynyl)-N-(7-azaspiro[3.5]nonan-2-yl)-1H-indazole-5-carboxamide, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,7-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-methyl-2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,5-diazaspiro[3.5]nonan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(5-methyl-2,5-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2-methyl-2,5-diazaspiro[3.5]nonan-5- yl)methanone, (3-((2-(pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((4-(trifluoromethoxy)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (R)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, (S)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, 2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- methylbenzamide, (R)-N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, (S)-N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, N-(2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3- yl)ethynyl)phenyl)acetamide, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(2,5-diazaspiro[3.5]nonan-2-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((2'-(trifluoromethoxy)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (3-((2-(naphthalen-2-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl-[1,1'- biphenyl]-2-carboxamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl-[1,1'- biphenyl]-3-carboxamide, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'-biphenyl]-2- yl)acetamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl-[1,1'- biphenyl]-4-carboxamide, (3-((2-(6-amino-4-methylpyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone, (3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((5-(trifluoromethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((3'-(trifluoromethoxy)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (2,6-diazaspiro[3.5]nonan-2-yl)(3-((2'-(trifluoromethoxy)-[1,1'-biphenyl]-2-yl)ethynyl)-1H- indazol-5-yl)methanone, (3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(2-(pyrrolidin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-(piperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-aminopyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (R)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, (S)-2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, 2-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)-N- phenylbenzamide, (3-((2-(2-aminopyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(6-amino-4-methylpyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (R)-7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)indolin-2-one, (S)-7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)indolin-2-one, 7-((5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazol-3-yl)ethynyl)indolin-2-one, (3-((2-(2-(pyrrolidin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2-(2-(piperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (R)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, (S)-(3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1- carbonyl)phenyl)ethynyl)-1H-indazol-5-yl)methanone, (3-(dimethylamino)pyrrolidin-1-yl)(3-((2-(4-methylpiperazine-1-carbonyl)phenyl)ethynyl)- 1H-indazol-5-yl)methanone, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'-biphenyl]-2- yl)acetamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl-[1,1'- biphenyl]-3-carboxamide, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N-methyl-[1,1'- biphenyl]-4-carboxamide, (3-((2-(2-(4-methylpiperazin-1-yl)pyridin-3-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (S)-1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide,
(R)-1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide, 1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)-N-methylpyrrolidine-2- carboxamide, N-(2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-[1,1'-biphenyl]-2- yl)pivalamide, 5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (R)-5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (S)-5-(3-(dimethylamino)pyrrolidine-1-carbonyl)-1H-indazole-3-carbonitrile, (3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-yl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1-yl)methanone, (3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylmethyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R,E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S,E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (R,Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (S,Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(3-(dimethylamino)pyrrolidin-1- yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone,
(E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-oxa-2-azaspiro[3.5]nonan-2- yl)methanone, (3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (E)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, (Z)-(3-(2-([1,1'-biphenyl]-2-yl)vinyl)-1H-indazol-5-yl)(7-methyl-2,7-diazaspiro[3.5]nonan-2- yl)methanone, 1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3-phenylurea, (R)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea, (S)-1-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3- phenylurea, 3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3,4- dihydroquinazolin-2(1H)-one, (R)-3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3,4- dihydroquinazolin-2(1H)-one, (S)-3-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-3,4- dihydroquinazolin-2(1H)-one, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, (R)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, (S)-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(3-((4,6-dimethylpyrimidin-2- yl)amino)pyrrolidin-1-yl)methanone, N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-2,3- dihydrobenzo[b][1,4]dioxine-6-carboxamide, (R)-N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-2,3- dihydrobenzo[b][1,4]dioxine-6-carboxamide, (S)-N-(1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)-2,3- dihydrobenzo[b][1,4]dioxine-6-carboxamide,
N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3-yl)amino)-2- oxoethyl)-3-(trifluoromethyl)benzamide, (R)-N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (S)-N-(2-((1-(3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazole-5-carbonyl)pyrrolidin-3- yl)amino)-2-oxoethyl)-3-(trifluoromethyl)benzamide, (3-([1,1'-biphenyl]-3-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-4-yl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2-yl)methanone, (3-((2'-morpholino-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan- 2-yl)methanone, 7-(2-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)phenyl)isoindolin- 1-one, 2'-((5-(2,6-diazaspiro[3.5]nonane-2-carbonyl)-1H-indazol-3-yl)ethynyl)-N,N-dimethyl-[1,1'- biphenyl]-2-carboxamide, (3-((2'-(pyrrolidin-1-ylmethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-((2'-(morpholinomethyl)-[1,1'-biphenyl]-2-yl)ethynyl)-1H-indazol-5-yl)(2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(vinylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-indazol-5-yl)(6-(ethylsulfonyl)-2,6- diazaspiro[3.5]nonan-2-yl)methanone, (3-([1,1'-biphenyl]-2-ylethynyl)-1H-pyrazolo[4,3-b]pyridin-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-(pyridin-4-yl)phenyl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((2-phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((3-phenylpyridin-4-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, (3-((4-phenylpyridin-3-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone, and (3-((3-phenylpyridin-2-yl)ethynyl)-1H-indazol-5-yl)(2,6-diazaspiro[3.5]nonan-2- yl)methanone.
17. A compound selected from Table A, or a salt, solvate, stereoisomer, tautomer, or geometric isomer thereof.
18. A pharmaceutical composition comprising the compound of any one of claims 1-17 and a pharmaceutically acceptable carrier.
19. A method of sterilizing a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of claims 1-17, and/or the pharmaceutical composition of claim 18, optionally wherein the sterilization is temporary.
20. A method of promoting male contraception and/or infertility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of claims 1-17 and/or the pharmaceutical composition of claim 18, optionally wherein the infertility is temporary.
21. A method of minimizing and/or reducing spermatozoa number and/or motility in a male subject, the method comprising administering to the male subject a therapeutically effective amount of at least one compound of any one of claims 1-17 and/or the pharmaceutical composition of claim 18.
22. The method of any one of claims 19-21, wherein the compound provides a contraceptive effect in the male subject.
23. The method of any one of claims 19-22, wherein the compound is administered orally to the male subject.
24. A method of inhibiting RET in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of claims 1-16 and/or the pharmaceutical composition of claim 18.
25. A method of treating, ameliorating, and/or preventing a disease or disorder caused by RET overexpression or RET hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of claims 1- 17 and/or the pharmaceutical composition of claim 18.
26. The method of claim 25, wherein the disease or disorder comprises a RET-mediated cancer.
27. The method of claim 26, wherein the RET-mediated cancer comprises medullary thyroid cancer, non-small cell lung cancer, and/or Multiple Endocrine Neoplasia Type IIa (MEN2A).
28. The method of claim 25, wherein the disease or disorder comprises Hirschprung Disease, central hypoventilation syndrome, and/or renal agenesis.
29. A method of inhibiting at least one of CL1, CLK2, CLK3, and CLK4 in a mammal, the method comprising administering the mammal an effective amount of at least one compound of any one of claims 1-17 and/or the pharmaceutical composition of claim 18.
30. A method of treating, ameliorating, and/or preventing a disease or disorder caused by CLK overexpression or CLK hyperactivity, the method comprising administering the mammal a therapeutically effective amount of at least one compound of any one of claims 1- 17 and/or the pharmaceutical composition of claim 18.
31. The method of claim 30, wherein the disease or disorder comprises Duchenne muscular dystrophy, Alzheimer's disease, HIV-1, influenza virus, and renal cancer, breast cancer, and/or melanoma.
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