CDK9 inhibitors

Abstract

A compound represented by formula (I), or a salt (e.g., a pharma- ceutically acceptable salt) or stereoisomer thereof, wherein X, Y, R 1a , R 1b , R 2 , R 3 , R 4s , R 5 , p, and n are as defined herein. The use of the compounds as components of pharmaceutical compositions and methods for their use are also provided. JPEG2025511975000113.jpg41159

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 329,139, filed April 8, 2022, entitled "CDK9 INHIBITORS," which is incorporated by reference in its entirety herein. [Background technology]

[0002] Technical Field The present disclosure relates generally to compounds that inhibit the activity of protein kinases, such as cyclin-dependent protein kinases (CDKs), and compositions and methods for treating cancers and other conditions associated with CDKs.

[0003] 2. Description of Related Art Cyclin-dependent kinases (CDKs) are serine-threonine kinases that function to regulate multiple cellular functions and phosphorylate substrates essential for progression through the cell cycle. The activity of specific CDKs at specific times is essential for both initiation and coordinated progression through the cell cycle. For example, CDK7, CDK8, and CDK9 play a role in regulating transcription, driving the expression of numerous target genes, thereby further influencing cell proliferation and survival. The relevance of CDKs to cancer proliferation and survival has generated widespread interest in the generation of CDK inhibitors.

[0004] A number of CDK9 inhibitors have been widely used in tumor cells for the rapid induction of apoptosis, however, the complex toxicity profile associated with CDK9-suppressing activity has hindered development.

[0005] Thus, there is a need for the rational design of specific and selective CDK9 inhibitors for the treatment of cancer and other conditions mediated and / or related. The present disclosure fulfills these needs and provides other related advantages. Summary of the Invention

[0006] Briefly, the present disclosure provides CDK9 inhibitor compounds, including stereoisomers or salts thereof (e.g., pharma- ceutically acceptable salts), which may be used alone or in combination with a pharma- ceutically acceptable carrier. Methods for the use of the CDK9 inhibitor compounds for the treatment of various diseases or conditions, such as bladder cancer, prostate cancer, and leukemia, are also provided.

[0007] In one embodiment, a compound having the following structure (I): [ka] or a salt (e.g., a pharma- ceutically acceptable salt) or stereoisomer thereof, wherein X, Y, R 1a , R 1b , R 2 , R 3 , R 4s , R 5 , p, and n are as defined herein. Use of the compounds as components of pharmaceutical compositions and methods for their use are also provided. Pharmaceutical compositions comprising one or more of the compounds of structure (I) above and a therapeutic agent are also provided.

[0008] In other embodiments, the disclosure provides a method for administering a therapeutic agent to a patient in need thereof, comprising preparing a composition comprising a compound of structure (I) and a therapeutic agent, and delivering the composition to the patient.

[0009] These and other aspects of the present disclosure will become evident upon reference to the following detailed description.

[0010] In the drawings, identical reference numbers identify similar elements. The sizes and relative positions of elements in the drawings are not necessarily drawn to scale, and some of these elements have been arbitrarily enlarged and positioned to improve drawing legibility. Furthermore, the particular shapes of the elements as depicted are not intended to convey any information regarding the actual shape of the particular elements and have been selected solely for ease of recognition in the drawings. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 is a graph showing the cellular anti-proliferative efficacy of compound I-2 in six prostate cancer cell lines across a range of concentrations. [Diagram 2] 1 is a graph showing the cellular anti-proliferative efficacy of benchmark compound AZD-4573 in six prostate cancer cell lines across a range of concentrations. [Diagram 3] 1 is a bar graph showing the cellular anti-proliferative efficacy of compound I-2 in 22RV-1 colonies. [Figure 4] 1 is a bar graph showing the cellular anti-proliferative efficacy of compound I-2 in LASCPC-1 colonies. [Diagram 5] A series of images from LASCPC-01 cells. [Figure 6] 1 is a set of graphs showing Annexin V / 7-AAD staining in LASCPC-01. [Figure 7] 1 is a set of graphs showing Annexin V / 7-AAD staining in 22Rev1. [Figure 8] This is an image of a Western blot analysis of N-MYC. [Figure 9] Images of Western blot analysis of N-MYC and C-MYC. [Figure 10] 1 is a bar graph showing RT-PCR analysis in 22Rv1 and LASCPC-01. [Figure 11] 13 is a series of images of staining in LASCPC-01 cells. [Figure 12]1 is an image of a Western blot from 22Rv1 with compounds I-2 and AZD-4573. [Figure 13] 1 is an image of a Western blot from LASCPC-01 with compounds I-2 and AZD-4573. [Figure 14] 22Rv1 mean tumor volume comparison. [Figure 15] 1 is a graph showing LASCPC-01 mean tumor volume comparison. [Figure 16] The relative body weights of treated and untreated mouse groups are shown. [Figure 17] 2 shows the mean tumor volumes of 22Rv1 over a 21 day study with different doses of compound I-2. [Figure 18] Tumor growth inhibition plotted against dose and tumor weight is shown. [Figure 19] The relative body weights of treated and untreated mouse groups are shown. [Figure 20] 1 shows the mean tumor volumes of LASCPC-01 in a 21-day study with different doses of compound I-2. [Figure 21] Tumor growth inhibition plotted against dose and tumor weight is shown. [Figure 22] Figure 1 shows the effect of changing from (50 mg / kg, orally, daily) to (30 mg / kg, orally, biweekly) compound I-2 on tumor volume in C4-2 xenografts in male NOD.SCID mice. Statistical analysis was performed using GraphPad Prism software version 5.0, two-way analysis of variance with Bonferroni post-hoc test. All values ​​are expressed as mean ± standard error, 7-8 mice per group, *P<0.05, ***P<0.001 vs. vehicle control. [Figure 23] FIG. 1 shows the effect of changing from compound I-2 (50 mg / kg, po, daily) to (30 mg / kg, po, biweekly) on tumor volume in C4-2 xenografts in male NOD.SCID mice. [Figure 24]Figure 1 shows the effect of compound I-2 (50mg / kg, orally, daily) on tumor volume in C4-2 xenografts in male NOD.SCID mice (30mg / kg, orally, biweekly). Statistical analysis was performed using GraphPad Prism software version 5.0, t-test. All values ​​are expressed as mean ± standard error, 8 mice per group, *P<0.05 vs. vehicle control. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] In the following description, certain specific details are set forth to provide a thorough understanding of various embodiments of the present disclosure. However, those skilled in the art will understand that embodiments of the present disclosure may be practiced without such details. As used herein, the following terms have the meanings ascribed to them unless otherwise specified.

[0013] Unless the context otherwise requires, throughout this specification and claims, the word "comprise" and variations thereof, such as "comprises" and "comprising," are to be interpreted in an open and inclusive sense, i.e., "including but not limited to."

[0014] References throughout this specification to "one embodiment" or "an embodiment" mean that a particular feature, structure, or characteristic described in connection with that embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0015] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. As used in this specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0016] "Hydroxy" or "hydroxyl" refers to the --OH radical.

[0017] "Amino" refers to the -NH2 radical.

[0018] "Cyano" refers to the -CN radical.

[0019] "Alkyl" refers to a saturated straight or branched chain hydrocarbon radical consisting solely of carbon and hydrogen atoms having from one to six carbon atoms (C1-C6 alkyl) and attached to the remainder of the molecule by a single bond. Examples of hydrocarbon chain radicals include methyl, ethyl, n-propyl, 1 methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1 dimethylethyl (t-butyl), iso-pentyl, n-hexyl, and the like. Unless otherwise stated specifically in the specification, alkyl groups are optionally substituted.

[0020] "Cycloalkyl" refers to a saturated cyclic hydrocarbon radical having three to eight carbon atoms (C3-C8 cycloalkyl) and attached to the remainder of the molecule by a single bond. Examples of saturated cyclic hydrocarbon radicals include cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. Unless otherwise stated specifically in the specification, cycloalkyl groups are optionally substituted.

[0021] "Bridged cycloalkyl" refers to a cycloalkyl as defined above that also includes an alkylene bridge between at least two carbon atoms of the cycle. Bridged cycloalkyl radicals include, for example, bicyclo[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl, bicyclo[3.1.1]heptanyl, (1R,5S)-bicyclo[3.2.1]octanyl, and bicyclo[2.2.2]octanyl. Unless otherwise specifically stated in the specification, bridged cycloalkyl groups are optionally substituted.

[0022] "Halo" refers to fluoro, chloro, bromo, or iodo. Halo is in Group 17 of the periodic table.

[0023] "Alkoxy" means a group of the formula -OR a A radical of formula (wherein R a is an alkyl radical having 1 to 12 carbon atoms (C1-C 12 "C1-C6 alkoxy" refers to an alkyl radical as defined above containing one to eight carbon atoms (C1-C8 alkoxy), one to six carbon atoms (C1-C6 alkoxy), or any value within these ranges. Unless stated otherwise specifically in the specification, an alkoxy group is optionally substituted.

[0024] "Haloalkyl" refers to an alkyl radical, as defined above, substituted by one or more halo radicals, such as trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. Unless stated otherwise specifically in the specification, a haloalkyl group is optionally substituted.

[0025] "Haloalkoxy" refers to a radical having the formula: -O-haloalkyl, where haloalkyl is as defined above. Unless stated otherwise specifically in the specification, a haloalkoxy group is optionally substituted.

[0026] "Hydroxylalkyl" or "hydroxyalkyl" refers to an alkyl radical, as defined above, substituted with one or more hydroxyl radicals. The hydroxyalkyl radical is attached to the backbone via an alkyl carbon atom. Unless otherwise specifically stated in the specification, the hydroxyalkyl group is optionally substituted.

[0027] "Aryl" refers to an all-carbon monocyclic or fused-ring polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group of 6 to 12 carbon atoms having a completely conjugated pi-electron system. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and anthracenyl. Aryl groups can be substituted or unsubstituted. When substituted, aryl groups are substituted with one or more substituents, more preferably one, two, or three, and even more preferably one or two, independently selected from the group consisting of alkyl (wherein alkyl may be optionally substituted with one or two substituents), haloalkyl, halo, hydroxy, alkoxy, mercapto, alkylthio, cyano, acyl, nitro, phenoxy, heteroaryl, heteroaryloxy, haloalkyl, haloalkoxy, carboxy, alkoxycarbonyl, amino, alkylaminodialkylamino, aryl, heteroaryl, carbocycle, or heterocycle (wherein aryl, heteroaryl, carbocycle, or heterocycle may be optionally substituted), as this term is defined below.

[0028] "Heterocyclyl" or "heterocycle" refers to a stable 3- to 18-membered non-aromatic ring radical, which consists of 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless stated otherwise specifically in the specification, a heterocyclyl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems, the nitrogen, carbon or sulfur atoms in the heterocyclyl radical can be optionally oxidized, the nitrogen atom can be optionally quaternized, and the heterocyclyl radical can be partially or fully saturated. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in the specification, heterocyclyl groups may be optionally substituted.

[0029] "N-heterocyclyl" refers to a heterocyclyl radical as defined above, wherein at least one heteroatom or heteroatoms is nitrogen. Examples of N-heterocyclyl radicals include azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, and the like. Unless stated otherwise specifically in the specification, N-heterocyclyl groups can be optionally substituted.

[0030] "Heteroaryl" refers to a 5-18 membered, e.g., 5-6 membered, ring system radical containing 1 to 13 ring carbon atoms, 1 to 6 ring heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. The heteroaryl radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which can include fused or bridged ring systems, and the nitrogen, carbon or sulfur atoms in the heteroaryl radical can be optionally oxidized and the nitrogen atom can be optionally quaternized. Examples include azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindoleyl. Examples of heteroaryl include, but are not limited to, aryl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl). Unless stated otherwise specifically in the specification, heteroaryl groups are optionally substituted.

[0031] As used herein, the term "substituted" refers to any of the above groups (e.g., alkyl, cycloalkyl, or heterocyclyl) in which at least one hydrogen atom is replaced with a halogen atom, such as F, Cl, Br, and I, an oxo group (=O), a hydroxyl group (-OH), an alkoxy group (-OR), or an aryl group (-OR). a , where R a is C1-C 12 alkyl or cycloalkyl), carboxyl group (-OC(=O)R a Or -C(=O)OR a , where R a , H, C1-C 12 alkyl or cycloalkyl), an amine group (-NR a R b , where R a and R b are each independently H, C1-C 12 alkyl or cycloalkyl), C1-C 12 In some embodiments, the substituent is a C-C alkyl group, such as, but not limited to, a cycloalkyl group, or a cycloalkyl group. 12 In other embodiments, the substituent is an alkyl group. In other embodiments, the substituent is a cycloalkyl group. In other embodiments, the substituent is a halo group, such as fluoro. In other embodiments, the substituent is an oxo group. In other embodiments, the substituent is a hydroxyl group. In other embodiments, the substituent is an alkoxy group. In other embodiments, the substituent is a carboxyl group. In other embodiments, the substituent is an amine group.

[0032] "Optional" or "optionally" (e.g., optionally substituted) means that the subsequently described event or circumstance may or may not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted alkyl" means that the alkyl radical may be substituted or unsubstituted, and the description includes both substituted and unsubstituted alkyl radicals.

[0033] "Prodrug" is meant to indicate a compound that can be converted under physiological conditions or by solvent degradation to a biologically active compound of the present disclosure. Thus, the term "prodrug" refers to a metabolic precursor of a compound of the present disclosure that is pharma- ceutically acceptable. A prodrug may be inactive when administered to a subject in need thereof, but is converted in vivo to an active compound of the present disclosure. Prodrugs are typically rapidly transformed in vivo to obtain the parent compound of the present disclosure, for example, by hydrolysis in blood. Prodrug compounds often offer advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). A discussion of prodrugs is provided in Higuchi, T., et al., ACS Symposium Series, Vol. 14, and Bioreversible Carriers in Drug Design, Ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987.

[0034] The term "prodrug" is also meant to include any covalently bonded carrier that releases the active compound of the present disclosure in vivo when such prodrug is administered to a mammalian subject. Prodrugs of the compounds of the present disclosure can be prepared by modifying functional groups present in the compounds of the present disclosure in such a way that the modifications are cleaved to the parent compounds of the present disclosure, either by routine manipulation or in vivo. Prodrugs include compounds of the present disclosure in which a hydroxy group, an amino group, or a mercapto group is bonded to any group that cleaves to form a free hydroxy group, a free amino group, or a free mercapto group, respectively, when a prodrug of the compound of the present disclosure is administered to a mammalian subject. Examples of prodrugs include, but are not limited to, acetate, formate, and benzoate derivatives of alcohols, or amide derivatives of amine functional groups in the compounds of the present disclosure, etc.

[0035] The embodiments disclosed herein are also meant to encompass all pharma- ceutically acceptable compounds of the compounds of structure (I) that are isotopically labeled by having one or more atoms replaced by an atom having a different atomic mass or mass number. Examples of isotopes that may be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, e.g., 2 H, 3 H, 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 I. These radiolabeled compounds may be useful, for example, to aid in the determination or measurement of the effectiveness of a compound by characterizing the site or mode of action, or by affinity binding to a pharmacologically important site of action. Certain isotopically labeled compounds of structure (I), for example those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e. 3 H, and carbon-14, i.e. 14 C are particularly useful for this purpose given their ease of incorporation and ready means of detection.

[0036] Deuterium, i.e. 2 Substitution with heavier isotopes, such as H, can offer certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances.

[0037] 11 C. 18 F, 15 O, and 13Substitution with positron emitting isotopes, such as N, can be useful in Positron Emission Topography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the preparations and examples below, substituting the appropriate isotopically labeled reagent for the non-labeled reagent used above.

[0038] The embodiments disclosed herein are also intended to encompass in vivo metabolic products of the disclosed compounds. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound, primarily through enzymatic processes. Thus, the present disclosure includes compounds resulting from a process comprising administering a compound of the present disclosure to a mammal for a period of time sufficient to yield a metabolic product thereof. Such products are typically identified by administering a detectable dose of a radiolabeled compound of the present disclosure to an animal, such as a rat, mouse, guinea pig, monkey, or human, allowing a period of time sufficient for metabolism to occur, and isolating the conversion product from urine, blood, or other biological sample.

[0039] "Stable compound" and "stable structure" are meant to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0040] "Mammal" includes both humans and non-domestic animals such as domestic animals, eg, laboratory animals and pets (eg, cats, dogs, pigs, cows, sheep, goats, horses, rabbits), and wild animals.

[0041] A "pharmaceutically acceptable carrier, diluent, or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonicity agent, solvent, or emulsifier approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary medicine.

[0042] "Pharmaceutically acceptable salt" includes both acid and base addition salts.

[0043] "Pharmaceutically acceptable acid addition salts" refers to salts that retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and include inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, as well as acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentic acid, glucoheptonic acid, gluconic acid. , glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid (TFA), undecylenic acid, and the like.

[0044] "Pharmaceutically acceptable base addition salts" refer to salts that retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from the addition of an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Preferred inorganic salts are the ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzetine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0045] "Pharmaceutical composition" refers to a formulation of a compound of the present disclosure and a medium generally accepted in the art for delivering the biologically active compound to a mammal, e.g., a human. Such a medium includes any pharma- ceutically acceptable carrier, diluent, or excipient therefor.

[0046] "Effective amount" or "therapeutically effective amount" refers to an amount of a compound of the present disclosure that is sufficient to affect treatment in a mammal, preferably a human, when administered to the mammal, preferably a human. The amount of lipid nanoparticles of the present disclosure that constitutes a "therapeutically effective amount" varies depending on the compound, the condition and its severity, the method of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art, taking into account their own knowledge and this disclosure.

[0047] "Protein kinase-mediated condition" or "disease" as used herein refers to any disease or other deleterious condition in which a protein kinase is known to play a role and which is alleviated by treatment with a protein kinase inhibitor. In certain embodiments, the cancer is colon cancer, breast cancer, gastric cancer, prostate cancer, pancreatic cancer, or ovarian tissue cancer.

[0048] As used herein, "treating" or "treatment" includes the treatment of a disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest; (i) preventing a disease or condition from occurring in a mammal, particularly when such a mammal is susceptible to the condition but has not yet been diagnosed as having it; (ii) inhibiting a disease or condition, i.e., arresting its development; (iii) alleviating the disease or condition, i.e., causing regression of the disease or condition; or (iv) Relieving symptoms caused by a disease or condition, i.e., relieving pain without treating the underlying disease or condition. As used herein, the terms "disease" and "condition" may be used interchangeably or may differ in that a particular malady or condition may not have a known causative agent (and thus the etiology is yet to be elucidated) and thus is not yet recognized as a disease, but only as an undesirable state or syndrome, with a more or less specific set of symptoms identified by clinicians.

[0049] The compounds of the present disclosure or their salts (e.g., pharma- ceutically acceptable salts) may contain one or more asymmetric centers and thus may give rise to enantiomers, diastereomers, and other stereoisomeric forms that may be defined in terms of absolute stereochemistry as (R)- or (S)- or (D)- or (L)- in amino acids. The present disclosure is intended to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents or resolved using prior art techniques such as chromatography and fractional crystallization. Prior art techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers, likewise, all tautomeric forms are intended to be included.

[0050] "Stereoisomer" refers to a compound that consists of the same atoms linked by the same bonds, but has different three-dimensional structures that are not interchangeable. The present disclosure contemplates various stereoisomers, and mixtures thereof, including "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another. The present disclosure also contemplates "diastereomers," which refer to non-mirror images of non-identical stereoisomers. Diastereomers occur when two or more stereoisomers of a compound have different configurations at one or more equivalent stereocenters and are not mirror images of each other.

[0051] "Tautomer" refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any subject compounds.

[0052] compound In some embodiments, the present disclosure provides CDK9 inhibitor compounds, including stereoisomers or salts thereof (e.g., pharma- ceutically acceptable salts), which may be used alone or in combination with a pharma- ceutically acceptable carrier. Methods for the use of CDK9 inhibitor compounds for the treatment of various diseases or conditions, such as bladder cancer, prostate cancer, and leukemia, are also provided.

[0053] One embodiment is a compound having the following structure (I): [ka] As a stereoisomer or salt thereof, [ka] represents a double or single bond such that all valences are satisfied, forming a heteroaryl ring; X is N or CR 4b and Y is N or CR 4c and R 1a is absent or is C1-C6 alkyl; R 1b is hydrogen, halo, hydroxy, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 hydroxyalkyl; R 2 is hydrogen, or optionally substituted C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, or C1-C6 hydroxyalkyl; R 3 is C3-C5 cycloalkyl, C6-C8 bridged cycloalkyl, or hydroxyl, amino, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3a or R 3is hydroxyl, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3b is a C3-C8 cycloalkyl optionally substituted with R 3a is an optionally substituted 3- to 10-membered heterocyclyl or an optionally substituted 3- to 10-membered heteroaryl; R 3b is an optionally substituted 3- to 10-membered N-heterocyclyl or an optionally substituted 3- to 10-membered heteroaryl; R 3c is hydrogen or optionally substituted C1-C6 alkyl; R 4a , R 4b , and R 4c are each independently hydrogen, halo, or optionally substituted C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy; R 5 each occurrence is independently halo or optionally substituted C1-C6 haloalkyl; n is 0, 1, 2, 3, or 4; Compounds are provided wherein p is 0, 1, or 2.

[0054] One embodiment is a compound having the structure (I'): [ka] As a stereoisomer or salt thereof (e.g., a pharma- ceutically acceptable salt), X is N or CH; R 1 is hydrogen, halo, hydroxy, or optionally substituted C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy; R 2is hydrogen, or optionally substituted C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, or C1-C6 hydroxyalkyl; R 3 is C3-C5 cycloalkyl, or hydroxyl, amino, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3a or R 3 is hydroxyl, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3b is a C3-C8 cycloalkyl optionally substituted with R 3a is an optionally substituted 3- to 10-membered heterocyclyl or an optionally substituted 3- to 10-membered heteroaryl; R 3b is an optionally substituted 3- to 10-membered N-heterocyclyl or an optionally substituted 3- to 10-membered heteroaryl; R 3c is hydrogen or optionally substituted C1-C6 alkyl; R 4 is hydrogen, or optionally substituted C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy; m is 0, 1, or 2; wherein n is 0, 1, 2, 3, or 4.

[0055] In some embodiments, the compound has one of the following structures (Ia) or (Ib): [ka]

[0056] In certain embodiments, R 1a is absent. In some embodiments, R 1a is hydrogen. In some embodiments, R 1a is C1-C6 alkyl. In certain embodiments, R 1a is methyl.

[0057] In some embodiments, R 1 is hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl. In certain embodiments, R 1 is hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, methyl, or ethyl. 1 is hydrogen, cyclopropyl, cyclobutyl, isopropyl, or methyl. In certain embodiments, R 1 is non-substituted.

[0058] In some embodiments, R 1b is hydrogen, C1-C6 alkyl, or C3-C8 cycloalkyl. In certain embodiments, R 1b is hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, methyl, or ethyl. 1b is hydrogen, cyclopropyl, cyclobutyl, isopropyl, or methyl. In certain embodiments, R 1b is non-substituted.

[0059] In some embodiments, R 2 is hydrogen or C1-C6 alkyl. In more specific embodiments, R 2 is C1-C6 alkyl. In some embodiments, R 2 is -CH3. In certain embodiments, R 2 is methyl, ethyl, iso-propyl, or n-propyl. 2is hydrogen. In certain embodiments, R 2 is non-substituted.

[0060] In some embodiments, R 1 or R 2 At least one of R 1 and R 2 are both hydrogen.

[0061] In some embodiments, R 1b or R 2 At least one of R 1b and R 2 are both hydrogen.

[0062] In some embodiments, n is 0 or 1. In certain embodiments, n is 0. In some embodiments, n is 2, 3, or 4.

[0063] In some embodiments, R 3 is C3-C5 cycloalkyl, or hydroxyl, amino, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3a is a 3- to 10-membered heterocyclyl optionally substituted with

[0064] In some embodiments, R 3 is C3-C5 cycloalkyl, C6-C7 bridged cycloalkyl, or hydroxyl, amino, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3a is a 3- to 10-membered heterocyclyl optionally substituted with

[0065] In certain embodiments, R 3is hydroxyl, amino, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3a In some embodiments, R 3 is a C cycloalkyl optionally substituted with hydroxyl or amino. In some embodiments, R 3 is a C cycloalkyl optionally substituted with amino. In some embodiments, R 3 is hydroxyl, cyano, halo, C1-C6 alkyl, C3-C8 cycloalkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, or -N(R 3c )R 3b In some embodiments, R 3 is 5-6 membered heterocyclyl. In some embodiments, R 3 is substituted. In a more specific embodiment, R 3 is non-substituted.

[0066] In certain embodiments, R 3 has one of the following structures: [ka]

[0067] In certain embodiments, R 3 has one of the following structures: [ka]

[0068] In some embodiments, R 3 has one of the following structures: [ka]

[0069] In some embodiments, R 3 has one of the following structures: [ka]

[0070] In some embodiments, R 3 has the following structure: [ka]

[0071] In certain embodiments, R 3 has the following structure: [ka]

[0072] In some embodiments, R 3 has the following structure: [ka]

[0073] In certain embodiments, R 3 has the following structure: [ka]

[0074] In some embodiments, R 3 has the following structure: [ka]

[0075] In certain embodiments, R 3 has the following structure: [ka]

[0076] In some embodiments, R 3 has the following structure: [ka]

[0077] In some embodiments, m is 1 or 2. In certain embodiments, R 4 Each occurrence of is independently halo. In some embodiments, R 4 Each occurrence of is independently fluoro, chloro, or bromo. In some more specific embodiments, R 4 In some embodiments, m is 0.

[0078] In some embodiments, R 4a is hydrogen. In some embodiments, R 4a is halo (e.g., chloro, fluoro, or bromo).

[0079] In some embodiments, X is N. In some embodiments, X is CR 4b and R 4b is hydrogen or halo. In some embodiments, X is CR 4b and R 4b is hydrogen or fluoro. In certain embodiments, X is CF. In some embodiments, X is CH.

[0080] In some embodiments, Y is N. In some embodiments, Y is CR 4b and R 4b is hydrogen or halo. In some embodiments, Y is CR 4b and R 4bis hydrogen or fluoro. In certain embodiments, Y is CF. In some embodiments, Y is CH.

[0081] In some embodiments, p is 1. In some embodiments, p is 2. In certain embodiments, R 5 Each occurrence of is halo (e.g., chloro, fluoro, or bromo). In some embodiments, p is 0. In some embodiments, p is 1 and R 5 is chloro.

[0082] In some embodiments, the compound is in free base form. In certain embodiments, the compound is a pharma- ceutically acceptable salt. In some embodiments, the compound is a trifluoroacetate salt. In some embodiments, the compound is a trifluoroacetate salt, a hydrochloride salt, or a formate salt. In certain embodiments, the compound is a tautomer.

[0083] In various different embodiments, the compound has one of the structures (or a stereoisomer or salt thereof) set forth in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] † Also available as trifluoroacetate ‡ Also available as the hydrochloride salt # Also available as formate salt I C 50 value: ++++ represents values ​​less than 0.05 μM +++ represents values ​​between 0.05 and 0.10 μM ++ indicates values ​​between 0.10 and 1.00 μM + indicates values ​​greater than 1 μM

[0084] It is understood that any embodiment of the compound of structure (I) above, and any particular substituent and / or variable in the compound of structure (I) above, may be independently combined with the substituents and / or variables of other embodiments and / or compounds of structure (I) to form embodiments of the disclosure not specifically set forth above. Furthermore, when a list of substituents and / or variables is recited for any particular R group or variable n or m in a particular embodiment and / or claim, it is understood that each individual substituent and / or variable may be deleted from the particular embodiment and / or claim, and the remaining list of substituents and / or variables is considered to be within the scope of the disclosure. It is understood that combinations of substituents and / or variables of the depicted formulae are permissible herein only if such contributions result in stable compounds.

[0085] Pharmaceutical Compositions Other embodiments are directed to pharmaceutical compositions. The pharmaceutical compositions include any (or more) of the compounds described above and a pharma- ceutically acceptable carrier. In some embodiments, the pharmaceutical compositions are formulated for oral administration. In other embodiments, the pharmaceutical compositions are formulated for injection. In still other embodiments, the pharmaceutical compositions include a compound disclosed herein and an additional therapeutic agent (e.g., an anti-cancer agent). Non-limiting examples of such therapeutic agents are described herein below.

[0086] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, otic, nasal, and topical administration.Further, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injection, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injection.

[0087] In certain embodiments, the compounds described herein are administered in a local rather than systemic manner, for example, via injection of the compound directly into an organ, often in a depot preparation or sustained release formulation. In certain embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. In yet other embodiments, the compounds are delivered in targeted drug delivery systems, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to and selectively taken up by the organ. In yet other embodiments, the compounds described herein are provided in the form of a rapid release formulation, a sustained release formulation, or an intermediate release formulation. In yet other embodiments, the compounds described herein are administered locally.

[0088] In the methods of treatment according to embodiments of the present disclosure, an effective amount of at least one compound of structure (I) is administered to a subject suffering from or diagnosed with such a disease, disorder, or condition. Effective amounts or doses can be ascertained by methods such as modeling, dose escalation studies, or clinical trials, for example, by consideration of the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease, disorder, or condition, the subject's previous or ongoing treatments, the subject's health status and response to the drug, and the judgment of the treating physician.

[0089] The compounds according to the present disclosure are effective over a wide dosage range. For example, in the treatment of adult humans, dosages of 10-5000 mg, 100-5000 mg, 1000 mg-4000 mg, and 1000-3000 mg per day are exemplary dosages used in some embodiments. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the weight of the subject being treated, and the preference and experience of the attending physician.

[0090] In some embodiments, the compound of the present disclosure is administered in a single dose.Typically, such administration is carried out by injection, for example, intravenous injection, in order to rapidly introduce the drug.However, other routes can be used as necessary.A single dose of the compound of the present disclosure can also be used to treat acute conditions.

[0091] In some embodiments, the compound of the present disclosure is administered multiple times. In some embodiments, administration is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, administration is about once a month, once every two weeks, once a week, or once every other day. In another embodiment, the compound of the present disclosure and another agent (e.g., an anticancer agent) are administered together about once a day to about six times a day. In another embodiment, administration of the compound of the present disclosure and the agent continues for less than about 7 days. In yet another embodiment, administration continues for more than about 6, 10, 14, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous dosing is performed and maintained as long as necessary.

[0092] Administration of the disclosed compound may continue as long as necessary. In some embodiments, the disclosed compound is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the disclosed compound is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the disclosed compound is administered continuously, chronically, for example, to treat chronic effects.

[0093] In some embodiments, the compounds of the present disclosure are administered in individual dosage forms. It is known in the art that optimal treatment requires individualization of dosing regimens due to intra-subject variability in compound pharmacokinetics.

[0094] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions.In certain embodiments, the pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and auxiliary agents that facilitate the processing of the disclosed compounds into preparations that can be used pharmaceutically.The appropriate formulation depends on the route of administration selected. Any pharma- ceutically acceptable technique, carrier, and excipient may be used as suitable for formulating the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0095] Provided herein are pharmaceutical compositions comprising one or more compounds of structure (I) and a pharma- ceutically acceptable carrier.

[0096] Provided herein is a pharmaceutical composition comprising one or more compounds selected from the compound of structure (I) and pharma- ceutically acceptable diluent(s), excipient(s), and carrier(s).In certain embodiments, the described compounds are administered as pharmaceutical compositions, in which one or more compounds selected from the compound of structure (I) are mixed with other active ingredients, as in combination therapy.All combinations of active agents described in the combination therapy section below and throughout this disclosure are encompassed herein.In certain embodiments, the pharmaceutical composition comprises one or more compounds of structure (I).

[0097] As used herein, a pharmaceutical composition refers to a mixture of one or more compounds selected from the compounds of structure (I) with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, a therapeutically effective amount of one or more compounds selected from the compounds of structure (I) provided herein is administered in a pharmaceutical composition to a mammal having a disease, disorder, or medical condition to be treated. In certain embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount varies depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of a mixture.

[0098] In one embodiment, one or more compounds selected from the compounds of structure (I) are formulated in an aqueous solution. In certain embodiments, the aqueous solution is selected from physiologically compatible buffers, such as, by way of example only, Hanks' solution, Ringer's solution, or physiological saline buffer. In other embodiments, one or more compounds selected from the compounds of structure (I) are formulated for transmucosal administration. In certain embodiments, the transmucosal formulation includes a penetrant appropriate for the barrier to be permeated. In yet other embodiments, where the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In certain embodiments, such solutions include physiologically compatible buffers and / or excipients.

[0099] In another embodiment, the compounds described herein are formulated for oral administration.The compounds described herein are formulated by combining the active compounds with, for example, a pharma-ceutically acceptable carrier or excipient.In various embodiments, the compounds described herein are formulated in oral dosage forms, including, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, and the like.

[0100] In certain embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid excipients with one or more of the compounds described herein, optionally grinding the resulting mixture, and optionally adding suitable auxiliary agents to obtain tablets or dragee cores, followed by processing the mixture of granules. Suitable excipients are in particular fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol, cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or others, such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In certain embodiments, disintegrants are optionally added. Disintegrants include, by way of example only, cross-linked sodium croscarmellose, polyvinylpyrrolidone, agar, or alginic acid, or its salts, such as sodium alginate.

[0101] In one embodiment, dosage forms such as dragee cores and tablets are provided with one or more suitable coatings. In certain embodiments, concentrated sugar solutions are used to coat dosage forms. The sugar solutions optionally contain additional ingredients such as, for example only, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyes and / or pigments are also optionally added to the coating for identification purposes. Furthermore, dyes and / or pigments are optionally utilized to characterize different combinations of active compound doses.

[0102] In certain embodiments, a therapeutically effective amount of at least one of the compounds described herein is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin and soft sealed capsules made of gelatin and plasticizers (such as glycerol or sorbitol). In certain embodiments, the push-fit capsules contain the active ingredient mixed with one or more fillers. Fillers include, by way of example only, binders such as lactose, starch, and / or lubricants such as talc or magnesium stearate, and optionally, stabilizers. In other embodiments, the soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. Additionally, stabilizers are optionally added.

[0103] In yet other embodiments, the compounds described herein are formulated for parenteral injection, including formulations suitable for bolus injection or continuous infusion. In certain embodiments, the formulations for injection are presented in unit dosage form (e.g., ampoules) or in multi-dose containers. Preservatives are optionally added to the injection formulation. In still other embodiments, the pharmaceutical compositions are formulated in a form suitable for parenteral injection as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle. Parenteral injection formulations optionally contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents. In certain embodiments, pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. In additional embodiments, a suspension of one or more compounds selected from the compounds of structure (I) is prepared as a suitable oily injection suspension. Lipophilic solvents or vehicles suitable for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. In certain embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension contains suitable stabilizers or agents that increase the solubility of the compounds, allowing for the preparation of highly concentrated solutions. Alternatively, in other embodiments, the active ingredient is in powder form for constitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0104] The pharmaceutical composition comprises at least one pharma- ceutically acceptable carrier, diluent or excipient and one or more compounds selected from the compounds of structure (I) described herein as an active ingredient. The active ingredient is in the form of a free acid or free base, or in the form of a pharma- ceutically acceptable salt. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. Additionally, the compounds described herein include unsolvated and solvated forms with pharma- ceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds presented herein are also considered to be disclosed herein. In addition, the pharmaceutical composition optionally comprises other medicinal or pharmaceutical agents, carriers, adjuvants, such as preservatives, stabilizers, wetting agents, or emulsifiers, solubility enhancers, salts for adjusting osmotic pressure, buffers, and / or other therapeutically valuable substances.

[0105] Methods for preparing compositions containing the compounds described herein include formulating the compounds with one or more inert pharma- ceutically acceptable excipients or carriers to form a solid, semi-solid, or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. The pharmaceutical compositions described herein may be in the form of liquid solutions or suspensions, solid forms suitable for dissolving or suspending in liquid prior to use, or as emulsions. These compositions also optionally contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and the like.

[0106] In some embodiments, the pharmaceutical composition comprising one or more compounds selected from the compounds of structure (I) is illustratively in the form of a liquid in which the drug is in solution, in suspension, or both. Typically, when the composition is administered as a suspension, a first portion of the drug is in solution, and a second portion of the drug is in the form of particles suspended in a liquid matrix. In some embodiments, the liquid composition comprises a gel formulation. In other embodiments, the liquid composition is aqueous.

[0107] In certain embodiments, the aqueous suspension contains one or more polymers as suspending agents. Polymers include water-soluble polymers such as cellulosic polymers (e.g., hydroxypropylmethylcellulose) and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Certain pharmaceutical compositions described herein include mucoadhesive polymers, such as carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylic acid), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0108] The pharmaceutical composition also optionally includes a solubilizing agent that aids in the dissolution of one or more compounds selected from the compounds of structure (I). The term "solubilizing agent" generally includes agents that result in the formation of a micellar or true solution of the drug. Certain acceptable non-ionic surfactants, such as polysorbate 80, are useful as solubilizing agents, as are ophthalmically acceptable glycols, polyglycols, such as polyethylene glycol 400, and glycol ethers.

[0109] In addition, the pharmaceutical compositions optionally contain one or more pH adjusting or buffering agents, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid, bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane, and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition, within the acceptable range.

[0110] The compositions also optionally contain one or more salts in an amount necessary to bring the osmolality of the composition into an acceptable range, including those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions, with suitable salts including sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0111] Other pharmaceutical compositions optionally include one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal, stabilized chlorine dioxide, and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0112] The composition may contain one or more surfactants to enhance physical stability or for other purposes. Suitable non-ionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil, and polyoxyethylene alkyl ethers and alkylphenyl ethers, such as Octoxynol 10 and Octoxynol 40.

[0113] The compositions may also include, where necessary, one or more antioxidants to enhance chemical stability. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0114] In certain embodiments, the aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multi-dose reclosable containers are used, in which case a preservative is typically included in the composition.

[0115] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are utilized. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also utilized. In additional embodiments, the compounds described herein are delivered using sustained release systems, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. A variety of sustained release materials are useful herein. In some embodiments, sustained release capsules release the compound for several weeks up to over 100 days. Depending on the chemical nature and biological stability of the therapeutic reagent, additional strategies for protein stabilization are utilized.

[0116] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizing agents. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

[0117] In some embodiments, the concentration of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure is 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25% 19%, 18.75%, 18.50%, 18.25% 18%, 17.75%, 17.50%, 17.25% 17%, 16.75%, 16.50%, 16.25% 16%, 15.75%, 15.50%, 15.25% 15%, 14.75%, 14.50%, 14.25% 14%, 13.75%, 13.50%, 13.25% 13%, 12.75%, 12.50%, 12.25% 12%, 11.75%, 11.50%, 11.25% 11%, 10.75%, 10.50%, 10.25% 10%, 9.75%, 9.50%, 9.25% 9%, 8.75%, 8.50%, 8.25% 8%, 7.75%, 7.50%, 7.25% 7%, 6.75%, 6.50%, 6.25% 6%, 5.75%, 5.50%, 5.25% 5%,4.75%,4.50%,4.25%,4%,3.75%,3.50%,3.25%,3%,2.75%,2.50%,2.25%,2%,1.75%,1.50%,125%,1%,0.5%,0.4%,0.3%,0.2%,0.1%,0.09%,0.08%,0.07%,0.06%,0.05%,0.04%,0.03%,0.02%,0 or greater than 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or 0.0001% w / w, w / v, or v / v.

[0118] In some embodiments, the concentration of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical composition of the present disclosure is from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.07% to about 24%, from about 0.08% to about 23%, from about 0.09% to about 22%, from about 0.1% to about 21%, from about 0.2% to about 20%, from about 0.3% to about 19%, from about 0.4% to about 18%, from about 0.5% to about 17%, from about 0.6% to about 16%, from about 0.7% to about 15%, from about 0.8% to about 14%, from about 0.9% to about 12%, from about 1% to about 10% w / w, w / v or v / v.

[0119] In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical composition of the present disclosure is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, , 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g, or 0.0001g.

[0120] In some embodiments, the amount of one or more compounds selected from the compounds of structure (I) provided in the pharmaceutical compositions of the present disclosure is in the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[0121] Packaging materials for use in packaging the pharmaceutical compositions described herein include, for example, those found in U.S. Patent Nos. 5,323,907, 5,052,558, and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for the selected formulation and intended method of administration and treatment. For example, the container(s) contain one or more compounds described herein, optionally in a composition or in combination with another agent disclosed herein. The container(s) optionally have a sterile access port (e.g., the container is an intravenous solution bag or a vial with a stopper that can be pierced by a hypodermic needle). Such kits optionally contain the compound together with an identifying description or label or instructions for its use in the methods described herein.

[0122] For example, a kit typically includes one or more additional containers, each containing one or more of various materials (optionally in concentrated form, such as reagents and / or devices) that are desirable from a commercial and user standpoint for use of the compounds described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers describing the contents and / or instructions for use, packaging, containers, vials and / or tube labels, and package inserts with instructions for use. A set of instructions for use is also typically included. The label is optionally on or associated with the container. For example, a label is on a container when letters, numbers, or other characters forming the label are affixed, cast, or etched into the container itself, and a label is associated with a container when the label is contained within a receptacle or carrier that also holds the container, for example, as a package insert. Furthermore, the label is used to indicate that the contents are used for a particular therapeutic application. In addition, the label indicates how to use the contents, such as the methods described herein. In certain embodiments, the pharmaceutical composition is present in a pack or dispenser device that contains one or more unit dosage forms containing the compound provided herein. For example, the pack contains metal or plastic foil, such as a blister pack. Otherwise, the pack or dispenser device is accompanied by instructions for administration. Otherwise, the pack or dispenser is accompanied by a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects the approval by the agency of the form of the drug for human or veterinary administration. Such notice is, for example, labeling approved as a prescription drug by the U.S. Food and Drug Administration, or an approved product insert. In some embodiments, a composition containing the compound provided herein, formulated in a compatible pharmaceutical carrier, is prepared, placed in a suitable container, and labeled for the treatment of an indicated condition.

[0123] One embodiment provides a method of treating neuroendocrine prostate cancer (NEPC). Some embodiments provide a method for treating overexpression of N-MYC or MCL-1. Some embodiments provide a method for treating metastatic castration-resistant prostate cancer, or CRPC. In some embodiments, the disorder or disease comprises treating tumors with drug resistance. In some embodiments, NEPC results from prostate adenocarcinoma, androgen deprivation therapy (ADT), or aberrant expression and activation of various kinases. Some embodiments provide a method for treating hematological or solid tumors.

[0124] In some embodiments, the methods include inhibiting or silencing pCDK9, pSer2, P-TEFb, MYC oncogene transcriptional activity, or repression of an active super-enhancer complex.

[0125] In some embodiments, the methods include promoting prostate cancer cell death and overcoming drug resistance (eg, chemotherapy resistance and other current targeted therapies).

[0126] In certain embodiments, the method comprises inhibiting CDK9, N-MYC, C-MYC, and their associated super-enhancer gene expression profiles. In some embodiments, the method comprises increasing median survival to greater than 12, 13, 13.5, 13.6, 13.7, 14, 18, 20, or 36 months. In some embodiments, the method further comprises administering a compound of structure (I) in combination with another chemotherapeutic agent (e.g., docetaxel, abiraterone, enzalutamide).

[0127] In some embodiments, the method includes inhibiting CDK9 as a therapeutic agent for prostate tumors. In some embodiments, the method includes increasing prostate cancer cell death and overcoming resistance caused by chemotherapy and current targeted therapies. In some embodiments, the method includes inhibiting the negative elongation factor (NELF) complex, DRB sensitivity inducer factor (DSIF), and the CDK9-cyclin T complex that phosphorylates Ser2 of the CTD of RNAPII, thereby affecting the removal of the elongation block.

[0128] In some embodiments, the method includes reversibly binding to and inhibiting CDK9. In some embodiments, the method includes treating malignant prostate cancer cells, particularly prostate cancer cells originating from CRPC, mCRPC, NEPC, and treatment-resistant subtypes, and does not exhibit toxicity to normal prostate cells. In some embodiments, the method includes regulating the transcription of N-MYC, C-MYC, and MCL-1 (e.g., in NEPC cells such as 22RV1, LASCPC-01, C4-2, and C4-2B from prostate cancer patients). In certain embodiments, the method includes orally administering a compound of structure (I). In some embodiments, the method includes globally regulating transcription and silencing and inhibiting CDK9. In some embodiments, the method includes dually inhibiting CDK9, MYC, and associated super-enhancer genes. In some embodiments, the method includes inhibiting tumor growth.

[0129] In certain embodiments, the methods include binding or targeting residues within the CDK9 ATP-binding site, such as the gatekeeper Phe103, hinge residues Asp104, Phe105, Cys106, and the DFG loop (residues 167, 168, 169) (including back pocket K48 and sugar-binding pocket residues Glu107, His108, and Asp109).

[0130] As discussed above, the compounds and compositions of the present disclosure find utility in a wide range of diseases and conditions mediated by protein kinases, including, but not limited to, cancers such as lung cancer, NSCLC (non-small cell lung cancer), oat cell cancer, bone cancer, pancreatic cancer, skin cancer, dermatofibrosarcoma protuberans, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, colorectal cancer, anal region cancer, gastric cancer, colon cancer, breast cancer, gynecological tumors (e.g., uterine sarcoma, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, or vulvar cancer), Hodgkin's disease, hepatocellular carcinoma, esophageal cancer, small intestine cancer, endocrine system cancer (e.g., thyroid cancer, pancreatic cancer, parathyroid cancer, or adrenal cancer), soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer (particularly hormonal cancer), and other cancers. The conditions may include cancers, including cancers refractory to leukemia, chronic or acute leukemia, childhood solid tumors, hypereosinophilia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer (e.g., renal cell carcinoma, renal pelvic cancer), childhood malignancies, central nervous system tumors (e.g., primary CNS lymphoma, spinal axis tumor, medulloblastoma, brain stem glioma, or pituitary adenoma), Barrett's esophagus (premalignant syndrome), neoplastic skin diseases, psoriasis, mycosis, and benign prostatic hyperplasia, diabetes-related diseases (such as diabetic retinopathy, retinal ischemia, and retinal neovascularization), liver cirrhosis, angiogenesis, cardiovascular diseases (such as atherosclerosis), immune diseases (such as autoimmune diseases and renal diseases).

[0131] In some embodiments, the pharmaceutical composition comprises the above-mentioned compound approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary medicine, and a pharma- ceutically acceptable carrier, including, for example, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonicity agent, solvent, or emulsifier.

[0132] In some embodiments, a method for treating a disease or disorder, the method comprising administering to a subject in need of treatment an effective amount of a compound or pharmaceutical composition described herein.

[0133] In some embodiments, the disease or disorder is a kinase-expressing cancer. In some particular embodiments, the cancer is bladder cancer. In some other particular embodiments, the cancer is prostate cancer. In some other particular embodiments, the cancer is a hematological malignancy, such as acute myeloid leukemia. In some other particular embodiments, the disease or disorder is an autoimmune disease or an inflammatory disease.

[0134] Preparation of compounds Methods of preparation for the above compounds and compositions are described herein below and / or known in the art. It will be appreciated by those skilled in the art that in the processes described herein, functional groups of intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include hydroxy, amino, mercapto, and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl, or trimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitable protecting groups for amino, amidino, and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C(O)-R" (wherein R" is alkyl, aryl, or arylalkyl), p-methoxybenzyl, trityl, and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl, or arylalkyl esters. Protecting groups can be added or removed according to standard techniques known to those skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T W and P G M Hutz, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley. As will be appreciated by those skilled in the art, the protecting group can also be a polymeric resin, such as a Wang resin, a Rink resin, or a 2-chlorotrityl-chloride resin.

[0135] It will also be understood by those skilled in the art that such protected derivatives of the compounds of the present disclosure may not have such pharmacological activity, but they may be administered to a mammal and then metabolized in the body to form a compound of the present disclosure that is pharmacologically active. Such derivatives may therefore be described as "prodrugs." All prodrugs of the compounds of the present disclosure are included within the scope of the present disclosure.

[0136] Additionally, all compounds of the present disclosure that exist in a free base or acid form can be converted to their pharma- ceutically acceptable salts by treatment with an appropriate inorganic or organic base or acid by methods known to those skilled in the art. Salts of the compounds of the present disclosure can be converted to their free base or acid forms by standard techniques.

[0137] The following reaction scheme illustrates the synthesis of compounds of the present disclosure, i.e., compounds of structure (I): [ka] or a salt (e.g., a pharma- ceutically acceptable salt) or stereoisomer thereof, wherein X, Y, R 1a , R 1b , R 2 , R 3 , R 4s , R 5, p, and n are as defined herein. It is understood that those skilled in the art can make these compounds by similar methods or by combining other methods known to those skilled in the art. It is also understood that those skilled in the art can make other compounds of structure (I) not specifically shown below by using appropriate starting components and modifying the parameters of the synthesis as necessary in a similar manner to that described below. In general, the starting components can be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or can be synthesized according to sources known to those skilled in the art (e.g., Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)), or can be prepared as described in this disclosure.

[0138] The following examples are offered by way of illustration and not by way of limitation.

[0139] Abbreviation: °C (degrees Celsius), 1 H NMR (proton nuclear magnetic resonance), DCM (dichloromethane), DMSO (dimethyl sulfoxide), eq (equivalent), EtOAc (ethyl acetate), g (gram), h (hour), MeOH (methanol), mg (milligram), min (minute), mL (milliliter), mmol (millimole), TFA (trifluoroacetic acid), THF (tetrahydrofuran), TLC (thin layer chromatography), LDA (lithium diisopropylamide), AcOH (acetic acid), mCPBA (3-chloroperbenzoic acid).

[0140] Example 1 Synthesis of Compound I-1 [ka] Synthesis of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (20) [ka] To a stirred solution of 5-bromo-2-methyl-3-(propan-2-yl-2H-indazole) (10) (127 mg, 0.50 mmol, 1.0 equiv.) and bis(pinacolato)diborane (152 mg, 0.60 mmol, 1.2 equiv.) in 1,4-dioxane (8 mL) in a microwave vial was added potassium acetate (119 mg, 1.25 mmol, 2.5 equiv.), Pd(dppf)Cl2.DCM (20 mg, 0.025 mmol, 0.05 equiv.). The vial was then sealed with a cap and degassed with argon for 5 minutes. The reaction mixture was stirred at 90° C. for 16 hours. After completion of the reaction by TLC, the reaction mixture was diluted with water and ethyl acetate, and the resulting slurry was filtered through a pad of Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (20) (150 mg, 100% yield) as a yellow solid.

[0141] Synthesis of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]-1H-indazole (30) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (20) (150 mg, 0.50 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (8 mL) in a microwave vial, 4-chloro-2-(methylthio)pyrimidine (96 mg, 0.60 mmol, 1.2 equiv.), K2CO3 (173 mg, 0.25 mmol, 2.5 equiv.), Pd(dppf)Cl2DCM (20 mg, 0.025 mmol, 0.05 equiv.) were added and degassed with argon for 5 min. The reaction mixture was then stirred at 100° C. for 16 h. The reaction mixture was diluted with water and ethyl acetate and the resulting slurry was filtered through a pad of Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product, which was purified by Combiflash chromatography (4 g column) to give 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]-1H-indazole (30) (130 mg, 87% yield) as a yellow solid. TLC system: Hexane:EtOAc (2:1), R f Value: about 0.2.

[0142] Synthesis of a mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)indazole and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)indazole (40) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]-1H-indazole (30) (98 mg, 0.33 mmol, 1.0 equiv.) in DCM (4 mL) cooled to 0° C. was added 3-chloroperbenzoic acid (mCPBA) (purity, 77%) (96 mg, 0.43 mmol, 1.3 equiv.). The reaction mixture was stirred at 0° C. for 4 h. After completion of the reaction by TLC, the reaction mixture was quenched with saturated aqueous NaHCO3 (10 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine solution, dried over sodium sulfate and concentrated to give a crude mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)indazole and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)indazole (40) as a yellow solid (104 mg, 100% yield). TLC system: EtOAc (100%)R f Value: approximately 0.01 and 0.2.

[0143] Synthesis of tert-butyl (1S,3R)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50a) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)indazole and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)indazole (40) (30 mg, 0.10 mmol, 1 equiv.) in DMSO (5 mL) was added (1S,3S)-3-amino-1-(BOC-amino)cyclopentane (28 mg, 0.14 mmol, 1.5 equiv.) and cesium fluoride (21 mg, 0.14 mmol, 1.5 equiv.) at room temperature. The reaction mixture was then stirred at 95° C. for 16 h. After completion of the reaction by TLC, the reaction mixture was cooled to room temperature and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over anhydrous sodium sulfate, and then concentrated to give the product, which was purified by Combiflash chromatography (4 g column) to give tert-butyl (1S,3R)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50a) (23 mg, 53%) as a yellow solid. TLC system: hexane:EtOAc (1:1), R f Value: about 0.2.

[0144] Synthesis of (1S,3R)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate (compound I-1) [ka] To a stirred solution of tert-butyl (1S,3R)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50a) (22 mg, 0.05 mmol, 1 equiv.) in DCM (4 mL) was added trifluoroacetic acid (TFA) (2 mL) and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction by TLC, the solvent was removed in vacuo. Diethyl ether was then added to the reaction, during which a yellow solid appeared in large amounts. The mixture was centrifuged, washed with diethyl ether, and dried at room temperature to give the product (1S,3R)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate salt (compound I-1) (25 mg, 100%) as a yellow solid. 1 HNMR(400MHz,MeOD-d4) δ 8.82(s,1H),8.29(d,J=6.0Hz,1H),8.14(d,J=9.0Hz,1H),7.64(d,J=9.0Hz,1H),754(d,J=6.0Hz,1H),4.59(wide,1H),4.19(s ,3H),3.76-3.74(m,2H),2.79-2.73(m,1H),2.30-2.20(m,2H),2.00-1.87(m,2H),1.80-1.72(m,1H),1.60(d,J=6.8Hz,6H); HRMS(ESI) m / z:[M+H] + C 20 H 26 Calculated value for N6: 351.2292, observed value: 351.2270.

[0145] Example 2 Synthesis of Compound I-2 The precursors 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)indazole and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)indazole (40) were prepared according to the synthesis described in steps 1-3 of Example 1.

[0146] Synthesis of tert-butyl (1S,3S)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50b) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)indazole and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)indazole (40) (30 mg, 0.10 mmol, 1 equiv.) in DMSO (5 mL) was added (1S,3S)-3-amino-1-(BOC-amino)cyclopentane (28 mg, 0.14 mmol, 1.5 equiv.) and cesium fluoride (21 mg, 0.14 mmol, 1.5 equiv.) at room temperature. The reaction mixture was then stirred at 95° C. for 16 h. After completion of the reaction by TLC, the reaction mixture was cooled to room temperature and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over anhydrous sodium sulfate, and then concentrated to give the product, which was purified by Combiflash chromatography (4 g column) to give tert-butyl (1S,3S)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50b) (25 mg, 58%) as a yellow solid. TLC system: hexane:EtOAc (1:1), R f Value: about 0.2.

[0147] Synthesis of (1S,3S)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate (compound I-2) [ka] To a stirred solution of tert-butyl (1S,3S)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (50b) (25 mg, 0.056 mmol, 1 equiv.) in DCM (4 mL) was added trifluoroacetic acid (TFA) (2 mL) and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction by TLC, the solvent was removed in vacuo. Diethyl ether was then added to the reaction, during which a lot of yellow solid appeared. The mixture was centrifuged, washed with diethyl ether, and dried at room temperature to give the product (1S,3S)-[3-[4-(3-isopropyl-2-methylindazol-5-yl)pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate salt (compound I-2) (22 mg, 69%) as a yellow solid. 1 HNMR(400MHz,MeOD-d4) δ 8.79(s,1H),8.28(d,J=6.2Hz,1H),8.15(d,J=9.1Hz,1H),7.64(d,J=9.1Hz,1H),7.52(s,wide,1H),4.71(s,wide,1H),4.19(s,3H),3.89-3.8 2(m,1H),3.70-3.62(m,1H),2.44-2.34(m,2H),2.30-2.18(m,2H),1.92-1.84(m,1H),1.79-1.70(m,1H),1.60(d,J=7.0Hz,6H);HRMS(ESI) m / z:[M+H] + C 20 H 26 Calculated value for N6: 351.2292, observed value: 351.2265.

[0148] Example 3 Synthesis of compound I-3 [ka] [ka] Synthesis of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine (21) [ka] To a stirred solution of 5-bromo-2-methyl-3-propylpyrazolo[4,3-b]pyridine (11) (45 mg, 0.18 mmol, 1.0 equiv.) and bis(pinacolato)diborane (56 mg, 0.22 mmol, 1.2 equiv.) in 1,4-dioxane (8 mL) in a microwave vial, potassium acetate (44 mg, 0.45 mmol, 2.5 equiv.), Pd(dppf)Cl2.DCM (8 mg, 0.01 mmol, 0.05 equiv.) were added. The vial was then sealed with a cap and degassed with argon for 5 minutes. The reaction mixture was stirred at 90° C. for 16 hours. After completion of the reaction by TLC, the reaction mixture was diluted with water and ethyl acetate, and the resulting slurry was filtered through a pad of Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine (21) (54 mg, 100% yield) as a brown oil.

[0149] Synthesis of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine (31) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine (21) (54 mg, 0.18 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (8 mL) in a microwave vial, 4-chloro-2-(methylthio)pyrimidine (43 mg, 0.27 mmol, 1.5 equiv.), K2CO3 (62 mg, 0.45 mmol, 2.5 equiv.), Pd(dppf)Cl2DCM (8 mg, 0.01 mmol, 0.05 equiv.) were added and degassed with argon for 5 min. The reaction mixture was then stirred at 100° C. for 16 h. The reaction mixture was diluted with water and ethyl acetate, and the resulting slurry was filtered through a pad of Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product, which was purified by Combiflash chromatography (4 g column) to give 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine (31) (10 mg, 18% yield) as a yellow solid. TLC system: Hexane:EtOAc (2:1), R f Value: about 0.3.

[0150] Synthesis of a mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine (41) [ka] To a stirred solution of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine (31) (10 mg, 0.033 mmol, 1.0 equiv.) in DCM (4 mL) cooled to 0° C. was added 3-chloroperbenzoic acid (mCPBA) (purity, 77%) (10 mg, 0.043 mmol, 1.3 equiv.). The reaction mixture was stirred at 0° C. for 4 h. After completion of the reaction by TLC, the reaction mixture was quenched with saturated NaHCO3 solution (10 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine solution, dried over sodium sulfate and concentrated to give a crude mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine (41) (10 mg, 100% yield) as a yellow solid. TLC system: EtOAc (100%)R f Value: approximately 0.01 and 0.2.

[0151] Synthesis of tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (51) [ka] To a stirred solution of a mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine (41) (10 mg, 0.033 mmol, 1 equiv.) in DMSO (5 mL) was added (1S,3S)-3-amino-1-(BOC-amino)cyclopentane (10 mg, 0.05 mmol, 1.5 equiv.) and cesium fluoride (8 mg, 0.05 mmol, 1.5 equiv.) at room temperature. The reaction mixture was then stirred at 95° C. for 16 h. After completion of the reaction by TLC, the reaction mixture was cooled to room temperature and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over anhydrous sodium sulfate, and then concentrated to give the product, which was purified by Combiflash chromatography (4 g column) to give tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (51) (10 mg, 67%) as a yellow solid. TLC system: hexane:EtOAc (1:1), R f Value: about 0.2.

[0152] Synthesis of (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate (compound I-17) [ka] To a stirred solution of tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (51) (10 mg, 0.022 mmol, 1 equiv.) in DCM (4 mL) was added trifluoroacetic acid (TFA) (2 mL) and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction by TLC, the solvent was removed in vacuo. Diethyl ether was then added to the reaction, during which a lot of yellow solid appeared. The mixture was centrifuged, washed with diethyl ether, and dried at room temperature to give the product (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate (compound I-3) (5 mg, 38%) as a yellow solid. 1 HNMR(400MHz,MeOD-d4) δ 8.53(d,J=8.9Hz,1H),8.40(d,J=5.6Hz,1H),8.05(d,J=8.9Hz,1H),7.86(J=5.6Hz,1H),4.68(s,wide,1H),4.07(s,3H),3.87-3.8 1(m,1H),3.64-3.57(m,1H),2.41-2.33(m,2H),2.23-2.19(m,2H),1.85-1.72(m,2H),1.54(d,J=7.0Hz,6H);HRMS data:HRMS(ESI) m / z[M+H]+ C 19 H 25 Calculated value for N7: 352.2244, observed value: 352.2217.

[0153] Example 4 Synthesis of compound I-4 Compound I-4 was prepared from the fluorine-substituted derivative of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (20) followed by the same synthesis described in Example 2.

[0154] Example 5 Synthesis of compound I-5 Compound I-21 was prepared from the fluorine-substituted derivative of 5-bromo-2-methyl-3-propylpyrazolo[4,3-b]pyridine (11) followed by the same synthesis described in Example 3.

[0155] Example 6 Synthesis of compound I-6 [ka] Overall reaction scheme: [ka] Synthesis of 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole [ka] A solution of 5-bromo-2-methyl-2H-indazole (500 mg, 2.36 mmol, 1.0 equiv), bis(pinacolato)diborane (722 mg, 2.84 mmol, 1.2 equiv), and potassium acetate (582 mg, 5.92 mmol, 2.5 equiv) in 1,4-dioxane (10 mL) was degassed for 5 min and Pd(dppf)Cl2.DCM (97 mg, 0.118 mmol, 0.05 equiv) was added. The mixture was degassed with nitrogen gas for an additional 5 min and stirred at 80 °C for 16 h. After completion of the reaction by TLC, it was diluted with water (50 mL) and extracted with ethyl acetate (2 x 50 mL). The combined organic layers were washed with brine solution (20 mL), dried over sodium sulfate and concentrated to give 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (630 mg, crude) as a black semi-solid. TLC system: EtOAc / petroleum ether (40:60), R f Value: approx. 0.45 (single spot, expected for boronic ester). Cr: LC / MS: 177.1 [M+H], 74.5% as boronic acid, and 259.2 [M+H], 18% as boronic ester.

[0156] Synthesis of 5-(2-chloropyrimidin-4-yl)-2-methyl-2H-indazole [ka] To a stirred solution of 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (550 mg, 2.13 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (11 mL) in a sealed tube, 2,4-dichloropyrimidine (381 mg, 2.58 mmol, 1.2 equiv.) and K2CO3 (735 mg, 5.32 mmol, 2.5 equiv.) were added at room temperature. Degassed for 5 min and later Pd(dppf)Cl2.DCM (87 mg, 0.106 mmol, 0.05 equiv.) was added. The reaction mixture was stirred at 80° C. for 16 h. After completion of the reaction by TLC, it was diluted with water and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over sodium sulfate, concentrated and purified by silica gel column (60-120 mesh, eluent: 50% ethyl acetate / petroleum ether) to give 5-(2-chloropyrimidin-4-yl)-2-methyl-2H-indazole (450 mg, 88%) as a pale yellow solid. TLC system: EtOAc (100%), R f Value: about 0.2. LC / MS retention time=2.92 minutes, 245.0[M+H] + .

[0157] 1 H NMR (400 MHz, CDCl3) δ 8.6 (d, J = 5.2 Hz, 1H), 8.60-8.59 (m, 1H), 8.06 (s, 1H), 7.94 (dd, J = 9.2 and 1.6 Hz, 1H), 7.79 (dd, J = 8.8 and 0.8 Hz, 1H), 7.67 (d, J = 5.2 Hz, 1H), 4.27 (s, 3H).

[0158] Synthesis of tert-butyl ((1S,3S)-3-((4-(2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-2-methyl-2H-indazole (450 mg, 1.84 mmol, 1 equiv) in IPA (5 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (443 mg, 2.21 mmol, 1.2 equiv) at room temperature and stirred at 110° C. for 60 h. TLC showed a polar spot and the reaction mixture was allowed to reach room temperature. The reaction mixture was evaporated, diluted with water (30 mL) and extracted with EtOAc (2×30 mL). The organic layer was dried over Na2SO4, filtered, evaporated and purified by reverse phase column (eluted with 25%-35% acetonitrile in water and 0.01% FA) to give tert-butyl ((1S,3S)-3-((4-(2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (235 mg, 32%) as an off-white solid. TLC system: EtOAc / petroleum ether (70:30), R f Value: approx. 0.40; LC / MS retention time = 3.0 minutes, 409.2[M+H] + .

[0159] 1 H NMR(400MHz,CDCl3):δ 8.39(s,1H),8.31(d,J=5.2Hz,1H),8.01(s,1H),7.95(d,J=8.0Hz,1H),7.75(d,J=9.2Hz,1H),7.01(d,J=5.6Hz,1H),5.21(d,J=6.8Hz,1H) ,4.59(brs,1H),4.53-4.48(m,1H),4.25(s,3H),4.18-4.11(m,1H),2.34-2.20(m,2H),2.04-1.98(m,2H),1.77-1.48(m,2H),1.45(s,9H).

[0160] Synthesis of (1S,3S)-N1-(4-(2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt [ka] To a stirred solution of tert-butyl ((1S,3S)-3-((4-(2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (230 mg, 0.56 mmol, 1 equiv.) in 1,4-dioxane (2 mL) was added 4M HCl in 1,4-dioxane (0.4 mL) at 0° C. and stirred at room temperature for 2 h. After completion of the starting material, the reaction mixture was evaporated and triturated with diethyl ether (2×3 mL) followed by lyophilization to give (1S,3S)-N1-(4-(2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt (120 mg, 56%) as a yellow solid. TLC system: MeOH / DCM (20:80), R f Value: approx. 0.1; LC / MS retention time = 1.88 minutes, 309.3[M+H] + .

[0161] 1 H NMR(400MHz,CD3OD) δ 8.90(brs,1H),8.62(s,1H),8.31-8.28(m,2H),7.77(d,J=9.2Hz,1H),7.67(d,J=6.8Hz,1H),4.93-4 .89(m,1H),4.31(s,3H),3.92-3.87(m,1H),2.47-2.36(m,2H),2.32-2.28(m,2H),1.94-1.86(m,2H).

[0162] Example 7 Synthesis of compound I-7 [ka] Overall reaction scheme: [ka] Synthesis of 5-bromo-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole [ka] To a stirred solution of 5-bromo-3-isopropyl-2H-indazole (200 mg, 0.84 mmol, 1.0 equiv) in DCM (4 mL) was added dihydropyran (0.2 mL, 1.67 mmol, 2.0 equiv), p-TSA (8 mg, 0.04 mmol, 0.05 equiv) at room temperature and stirred for 16 h. After completion of the reaction by TLC, it was diluted with cold water (30 mL) and DCM (2×30 mL). The organic layer was dried over sodium sulfate, filtered, concentrated and purified by silica gel column (60-120 mesh, eluent: 15% EtOAc / petroleum ether) to give 5-bromo-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (260 mg, 96% yield) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: approx. 0.8. LC / MS retention time=4.48 minutes, 323.0[M+H] + .

[0163] 1 H NMR(400MHz,CDCl3) δ 7.86(t,J=1.2Hz,1H),7.42-7.41(m,2H),5.61(dd,J=9.6 and 2.4Hz,1H),4.06-4.03(m,1H),3.75-3.69(m,1H),3.37-3.30(m ,1H),2.53-2.49(m,1H),2.14-2.12(m,1H),2.04-2.00(m,1H),1.77-1.71(m,2H),1.65-1.62(m,1H),1.44(d,J=7.6Hz,6H).

[0164] Synthesis of 3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole [ka] A stirred solution of 5-bromo-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (260 mg, 0.87 mmol, 1.0 equiv), bis(pinacolato)diborane (224 mg, 1.04 mmol, 1.2 equiv) and potassium acetate (213 mg, 2.17 mmol, 2.5 equiv) in 1,4-dioxane (6 mL) was degassed for 5 min and Pd(dppf)Cl2.DCM (36 mg, 0.043 mmol, 0.05 equiv) was added. The reaction mixture was stirred at 90° C. for 16 h. After completion of the reaction by TLC, it was diluted with water (50 mL) and extracted with ethyl acetate (2×50 mL). The organic layer was dried over sodium sulfate and concentrated to give 3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (325 mg, crude) as a black semi-solid. TLC system: EtOAc / petroleum ether (30:70), R f Value: about 0.5. LC / MS retention time=4.68 min, 371.2[M+H] + Cr: LC / MS: 62.1% boronate ester m / z and 10% boronic acid m / z observed, material carried onto next step without purification.

[0165] Synthesis of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole [ka] To a degassed stirred solution of 3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (320 mg, 0.86 mmol, 1.0 equiv) in 1,4-dioxane and water (3:1) (6 mL) was added 2,4-dichloropyrimidine (154 mg, 1.04 mmol, 1.2 equiv), K2CO3 (298 mg, 2.16 mmol, 2.5 equiv), followed by Pd(dppf)Cl2DCM (35 mg, 0.043 mmol, 0.05 equiv) and stirred for 16 h at 90° C. After completion of the reaction by TLC, it was diluted with water and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over sodium sulfate, concentrated and purified by silica gel column (60-120 mesh, eluent: 25% EtOAc / petroleum ether) to give 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (280 mg, 70% wrt purity) as a white solid. TLC system: EtOAc / petroleum ether (30:70), R f Value: approx. 0.2; LC / MS retention time = 4.77 minutes, 357.2[M+H] + ;purity 76%

[0166] Synthesis of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (280 mg, 0.79 mmol, 1 equiv.) in IPA (4.5 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (190 mg, 0.94 mmol, 1.2 equiv.) and stirred at 110° C. for 36 h. TLC showed a polar spot and the reaction mixture was allowed to reach room temperature. The reaction mixture was evaporated, diluted with water (30 mL) and extracted with EtOAc (3×20 mL). The organic layer was dried over Na2SO4, filtered, evaporated and purified by silica gel column (60-120 mesh, eluent: 15%-70% EtOAc / petroleum ether) to give tert-butyl ((1S,3S)-3-((4-(3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (150 mg, 37% yield) as a yellow sticky liquid. TLC system: EtOAc / petroleum ether (70:30), R f Value: approx. 0.20; LC / MS retention time = 4.67 minutes, 521.3[M+H] + .

[0167] Synthesis of (1S,3S)-N1-(4-(3-isopropyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt [ka] To a stirred solution of ((1S,3S)-3-((4-(3-isopropyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (150 mg, 0.29 mmol, 1 equiv.) in MeOH (0.7 mL) was added concentrated HCl (0.7 mL) at 0° C. and stirred at room temperature for 18 h. After completion of the starting material, the volatiles were removed, triturated with diethyl ether (2×5 mL) and purified by reverse phase column (eluent: 8% ACN and 0.01% FA in water) to give (1S,3S)-N1-(4-(3-isopropyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt (28 mg, 29% yield) as a yellow sticky solid. TLC system: MeOH / DCM (30:70), R f Value: approx. 0.1; LC / MS retention time = 2.32 minutes, 337.4[M+H] + .

[0168] 1 H NMR(400MHz,CD3OD) δ 8.84(s,1H),8.38(brs,1H),8.29(d,J=6.8Hz,1H),7.73(d,J=6.4Hz,1H),7.65(d,J=9.2Hz,1H),3.91-3.87(m,1H),3 .59-3.53(m,1H),3.35-3.33(m,1H),2.48-2.36(m,2H),2.32-2.27(m,2H),1.93-1.80(m,2H),1.49(d,J=6.8Hz,6H).

[0169] Example 8 Synthesis of compound I-3 [ka] Overall reaction scheme: [ka] Synthesis of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine [ka] To a stirred solution of 5-bromo-2-methyl-3-propylpyrazolo[4,3-b]pyridine (45 mg, 0.18 mmol, 1.0 equiv.) and bis(pinacolato)diborane (56 mg, 0.22 mmol, 1.2 equiv.) in 1,4-dioxane (8 mL) in a microwave vial, potassium acetate (44 mg, 0.45 mmol, 2.5 equiv.), Pd(dppf)Cl2.DCM (8 mg, 0.01 mmol, 0.05 equiv.) were added. The vial was then sealed with a cap and degassed with argon for 5 minutes. The reaction mixture was stirred at 90° C. for 16 hours. After completion of the reaction by TLC, the reaction mixture was diluted with water and ethyl acetate, and the resulting slurry was filtered through Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine (54 mg, 100% yield) as a brown oil.

[0170] Synthesis of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazolo[4,3-b]pyridine (54 mg, 0.18 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (8 mL) in a microwave vial, 4-chloro-2-(methylthio)pyrimidine (43 mg, 0.27 mmol, 1.5 equiv.), K2CO3 (62 mg, 0.45 mmol, 2.5 equiv.), Pd(dppf)Cl2DCM (8 mg, 0.01 mmol, 0.05 equiv.) were added and degassed with argon for 5 minutes. The reaction mixture was then stirred at 100° C. for 16 hours. The reaction mixture was diluted with water and ethyl acetate and the resulting slurry was filtered through Celite® (i.e., diatomaceous earth). The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to give the crude product, which was purified by Combiflash chromatography (4 g column) to give 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine (10 mg, 18% yield) as a yellow solid. TLC system: Hexane:EtOAc (2:1), R f Value: about 0.3;

[0171] Synthesis of a mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine [ka] To a stirred solution of 3-isopropyl-2-methyl-5-[2-(methylthio)pyrimidin-4-yl]pyrazolo[4,3-b]pyridine (10 mg, 0.033 mmol, 1.0 equiv.) in DCM (4 ml) cooled to 0° C. was added 3-chloroperbenzoic acid (mCPBA) (purity, 77%) (10 mg, 0.043 mmol, 1.3 equiv.). The reaction mixture was stirred at 0° C. for 4 h. After completion of the reaction by TLC, the reaction mixture was quenched with saturated NaHCO3 solution (10 mL) and extracted with ethyl acetate. The combined organic layers were washed with brine solution, dried over sodium sulfate and concentrated to give a crude mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine as a yellow solid (10 mg, 100% yield). TLC system: EtOAc (100%)R f Value: approximately 0.01 and 0.2.

[0172] Synthesis of tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate [ka] To a stirred solution of a mixture of 3-isopropyl-2-methyl-5-(2-methylsulfinylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine and 3-isopropyl-2-methyl-5-(2-methylsulfonylpyrimidin-4-yl)pyrazolo[4,3-b]pyridine (10 mg, 0.033 mmol, 1 equiv.) in DMSO (5 mL) was added (1S,3S)-3-amino-1-(BOC-amino)cyclopentane (10 mg, 0.05 mmol, 1.5 equiv.) and cesium fluoride (8 mg, 0.05 mmol, 1.5 equiv.) at room temperature. The reaction mixture was then stirred at 95° C. for 16 hours. After completion of the reaction by TLC, the reaction mixture was cooled to room temperature and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over anhydrous sodium sulfate, then concentrated to give the product, which was purified by CombiFlash chromatography (4 g column) to give tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (10 mg, 67%) as a yellow solid. TLC system: hexane:EtOAc (1:1), Rf value: approx. 0.2.

[0173] Synthesis of (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate [ka] To a stirred solution of tert-butyl (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]aminocarboxylate (10 mg, 0.022 mmol, 1 equiv.) in DCM (4 mL) was added trifluoroacetic acid (TFA) (2 ml) and the reaction mixture was stirred at room temperature for 24 h. After completion of the reaction by TLC, the solvent was removed in vacuo. Diethyl ether was then added to the reaction, during which a yellow solid appeared in large amounts. The mixture was centrifuged, washed with diethyl ether, and dried at room temperature to give the product (1S,3S)-[3-[4-[3-isopropyl-2-methylpyrazolo[4,3-b]pyridin-5-yl]pyrimidin-2-yl]aminocyclopentan-1-yl]-1,3-diamine trifluoroacetate (5 mg, 38%) as a yellow solid. 1 HNMR(400MHz,MeOD-d4) δ 8.53(d,J=8.9Hz,1H),8.40(d,J=5.6Hz,1H),8.05(d,J=8.9Hz,1H),7.86(J=5.6Hz,1H),4.68(s,wide,1H),4.07(s,3H),3.87-3.8 1(m,1H),3.64-3.57(m,1H),2.41-2.33(m,2H),2.23-2.19(m,2H),1.85-1.72(m,2H),1.54(d,J=7.0Hz,6H);HRMS data:HRMS(ESI) m / z[M+H]+ C 19 H 25 Calculated value for N7 is 352.2244, measured value is 352.2217

[0174] Example 9 Synthesis of compound I-9 [ka] Overall reaction scheme [ka] Synthesis of 5-bromo-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole [ka] To a stirred solution of 5-bromo-3-methyl-2H-indazole (500 mg, 2.36 mmol, 1.0 equiv) in DCM (5 mL), dihydropyran (0.43 mL, 4.73 mmol, 2.0 equiv), p-TSA (22 mg, 0.04 mmol, 0.05 equiv) were added at room temperature and stirred for 16 h. After completion of the reaction by TLC, it was diluted with cold water (30 mL) and DCM (2×30 mL). The organic layer was dried over sodium sulfate, filtered, concentrated and purified by silica gel column (60-120 mesh, eluent: 10% EtOAc / petroleum ether) to give 5-bromo-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (550 mg, 79% yield) as a yellow liquid. TLC system: EtOAc / petroleum ether (40:60), R f Value: about 0.8.

[0175] 1 H NMR(400MHz,CDCl3) δ 7.78(dd,J=0.8 and 2Hz,1H),7.46-7.39(m,2H),5.59(dd,J=10 and 2.8Hz,1H),4.07-4.04(m,1H),3.75-3.6 9(m,1H),2.56-2.50(m,4H),2.15-2.12(m,1H),2.05-2.00(m,1H),1.77-1.71(m,2H),1.65-1.62(m,1H).

[0176] Synthesis of 3-methyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole [ka] A stirred solution of 5-bromo-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (550 mg, 1.86 mmol, 1.0 equiv), bis(pinacolato)diborane (570 mg, 2.23 mmol, 1.2 equiv), and potassium acetate (460 mg, 4.66 mmol, 2.5 equiv) in 1,4-dioxane (11 mL) was degassed for 5 min and Pd(dppf)Cl2.DCM (76 mg, 0.093 mmol, 0.05 equiv) was added. The reaction mixture was stirred at 90° C. for 16 h. After completion of the reaction by TLC, it was diluted with water (50 mL) and extracted with ethyl acetate (2×50 mL). The organic layer was dried over sodium sulfate and concentrated to give 3-methyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (630 mg, crude) as a black semi-solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: approx. 0.3. LC / MS retention time=4.19 min, 343.2[M+H] + The material was carried forward to the next step.

[0177] Synthesis of 5-(2-chloropyrimidin-4-yl)-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole [ka] To a degassed stirred solution of 3-methyl-2-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (630 mg, 1.84 mmol, 1.0 equiv) in 1,4-dioxane and water (3:1) (12 mL) was added 2,4-dichloropyrimidine (329 mg, 2.21 mmol, 1.2 equiv) and K2CO3 (635 mg, 4.60 mmol, 2.5 equiv). The reaction mixture was degassed for 5 min and Pd(dppf)Cl2DCM (75 mg, 0.092 mmol, 0.05 equiv) was added. The reaction mixture was stirred at 90° C. for 16 h. After completion of the reaction by TLC, it was diluted with water and extracted with ethyl acetate (2×50 mL). The combined organic layers were dried over sodium sulfate, concentrated and purified by silica gel column (60-120 mesh, eluent: 50% EtOAc / petroleum ether) to give 5-(2-chloropyrimidin-4-yl)-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (550 mg, semi-pure) as an off-white solid. TLC system: EtOAc / petroleum ether (30:70), R f Value: approx. 0.3; LC / MS retention time = 3.76 minutes, 329.1[M+H] + 69% purity. The material was carried on to the next step.

[0178] Synthesis of tert-butyl ((1S,3S)-3-((4-(3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazole (550 mg, 1.67 mmol, 1 equiv) in IPA (8 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (335 mg, 2.01 mmol, 1.2 equiv) and stirred at 110° C. for 24 h. TLC showed a polar spot and the RM was allowed to reach room temperature. The reaction mixture was evaporated, diluted with water (30 mL) and extracted with EtOAc (3×20 mL). The organic layer was dried over Na2SO4, filtered, evaporated and purified by silica gel column (60-120 mesh, eluent: 15%-60% EtOAc / petroleum ether) to give tert-butyl ((1S,3S)-3-((4-(3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (240 mg, semi-pure) as an off-white solid. TLC system: EtOAc / petroleum ether (70:30), R f Value: approx. 0.20; LC / MS retention time = 4.06 minutes, 493.3[M+H] + purity 58%. The material was carried on to the next step.

[0179] Synthesis of (1S,3S)-N1-(4-(3-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt [ka] To a stirred solution of tert-butyl ((1S,3S)-3-((4-(3-methyl-2-(tetrahydro-2H-pyran-2-yl)-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (170 mg, 0.29 mmol, 1 equiv.) in MeOH (1 mL) was added concentrated HCl (1 mL) at 0° C. and stirred at room temperature for 12 h. After completion of the starting material, it was removed of volatiles, triturated with diethyl ether (2×5 mL) and purified by preparative HPLC (method ABC). Ammonium salts were avoided by working up and fractions were lyophilized by adding aqueous HCl to give (1S,3S)-N1-(4-(3-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt (32 mg, 5% yield for four steps) as a yellow solid. TLC system: MeOH / DCM (30:70), R f Value: approx. 0.1; LC / MS retention time = 1.78 minutes, 309.2[M+H] + .

[0180] 1 H NMR(400MHz,CD3OD) δ 8.83(s,1H),8.42(d,J=6.4Hz,1H),8.31(d,J=6.8Hz,1H),7.73(d,J=6.8Hz,1H),7.67(d,J=8.8Hz,1H),3.9 1-3.88(m,1H),3.29-3.28(m,1H),2.69(s,3H),2.48-2.38(m,2H),2.30(t,J=6.8Hz,2H),1.93-1.82(m,2H).

[0181] Example 10 Synthesis of compound I-4 [ka] Overall reaction scheme [ka] [ka] Synthesis of 5-bromo-2,3-difluorobenzaldehyde [ka] To a stirred solution of 2,3-difluorobenzaldehyde (5 g, 35.2 mmol, 1.0 equiv) in sulfuric acid (38 mL) was added NBS (31.3 mL, 52.78 mmol, 1.5 equiv) at room temperature and stirred at 60 °C for 16 h. After completion of the reaction by TLC, it was diluted with cold water (50 mL) and extracted with EtOAc (2 x 100 mL). The organic layer was washed with NaHCO3 solution, dried over sodium sulfate and concentrated to give 7 g of material. The crude compound was purified by silica gel column (100-200 mesh, eluent: 2% EtOAc / Hexane) to give 5-bromo-2,3-difluorobenzaldehyde (3.5 g, 45%) as a yellow liquid. TLC system: EtOAc / petroleum ether (05:95), R f Value: about 0.8.

[0182] 1 H NMR (400MHz, CDCl3) δ 10.30 (s, 1H), 7.79-7.77 (m, 1H), 7.63-7.58 (m, 1H).

[0183] Synthesis of 5-bromo-7-fluoro-2H-indazole [ka] To a stirred solution of 5-bromo-2,3-difluorobenzaldehyde (2) (3.4 g, 15.5 mmol, 1.0 equiv) in EtOH (34 mL, 10 vol) was added 85% hydrazine hydrate (17 mL, 5 vol) and stirred at 100 °C for 16 h. After completion of the reaction by TLC, the RM was evaporated, diluted with water (100 mL) and extracted with ethyl acetate (3 x 50 mL). The organic layer was separated, dried over sodium sulfate and concentrated to give 4.1 g of material. The crude compound was purified by silica gel column (60-120 mesh, eluent: 25%-35% EtOAc / Hexane) to give 5-bromo-7-fluoro-2H-indazole (1.7 g, 51%) as an off-white solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.2. LC / MS retention time=3.10 minutes, 214.9[M+H] + .

[0184] Synthesis of 5-bromo-7-fluoro-2-methyl-2H-indazole [ka] To a stirred solution of 5-bromo-7-fluoro-2H-indazole (1.7 g, 7.94 mmol, 1.0 equiv) in EtOAc (54 mL, 30 vol), trimethyloxonium tetrafluoroborate (7.05 g, 47.7 mmol, 6.0 equiv) was added at room temperature and stirring was continued for 16 h. After completion of the reaction by TLC, the reaction mixture was evaporated and purified by silica gel column (60-120 mesh, eluent: 15%-18% EtOAc / Hexane) to give 5-bromo-7-fluoro-2-methyl-2H-indazole (750 mg, 42%) as an off-white solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.5.

[0185] 1H NMR (400 MHz, CDCl3) δ 7.91 (d, J = 2.8 Hz, 1H), 7.61 (d, J = 1.6 Hz, 1H), 7.08 (dd, J = 10.4 and 1.6 Hz, 1H), 4.26 (s, 3H). Confirmed by nOe.

[0186] Synthesis of 5-bromo-7-fluoro-3-iodo-2-methyl-2H-indazole [ka] To a stirred solution of 5-bromo-7-fluoro-2-methyl-2H-indazole (0.75 g, 3.28 mmol, 1.0 equiv) in DCM (15 mL, 20 vol) was added pyridine (1.7 g, 4.93 mmol, 1.5 equiv) and PIFA (1.7 g, 3.95 mmol, 1.2 equiv) at 0° C. The reaction mixture was stirred at 0° C. for 30 min, after which iodine (0.5 g, 3.95 mmol, 1.2 equiv) was added and stirred at room temperature for 18 h. After completion of the reaction by TLC, the reaction mixture was diluted with saturated NaHCO3 (50 mL) and extracted with DCM (2×20 mL). The organic layer was separated, dried over sodium sulfate, and concentrated to give the crude product. Purification by silica gel column (60-120 mesh, eluent: 5% EtOAc / hexane) gave 5-bromo-7-fluoro-3-iodo-2-methyl-2H-indazole (700 mg, 60%) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.8.

[0187] 1 H NMR (400MHz, CDCl3) δ 7.38 (d, J = 1.6 Hz, 1H), 7.10 (dd, J = 10.4 and 1.6 Hz, 1H), 4.27 (s, 3H).

[0188] Synthesis of 5-bromo-7-fluoro-2-methyl-3-(prop-1-en-2-yl)-2H-indazole [ka] To a stirred solution of 5-bromo-7-fluoro-3-iodo-2-methyl-2H-indazole (700 mg, 1.98 mmol, 1.0 equiv) in 1,4-dioxane (14 mL) was added 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane (398 mg, 2.37 mmol, 1.2 equiv) and potassium carbonate (682 mg, 4.94 mmol, 2.5 equiv) and degassed for 5 min. Then Pd(dppf)Cl2.DCM (80 mg, 0.098 mmol, 0.05 equiv) was added. The reaction mixture was stirred at 90° C. for 16 h. After completion of the reaction by TLC. The volatiles were evaporated and the residue was diluted with water (80 mL) and extracted with ethyl acetate (2×50 mL). The organic layer was separated, washed with brine solution (30 mL), dried over sodium sulfate and concentrated. The crude compound was purified by silica gel column (60-120 mesh, eluent: 10%-12% EtOAc / Hexane) to give 5-bromo-7-fluoro-2-methyl-3-(prop-1-en-2-yl)-2H-indazole (450 mg, 85%) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: approx. 0.50. LC / MS retention time=3.99 minutes, 269.0[M+H] + .

[0189] 1 H NMR (400MHz, CDCl3) δ 7.54(s,1H),7.04(d,J=10Hz,1H),5.63(s,1H),5.28(s,1H),4.16(s,3H),2.22(s,3H).

[0190] Synthesis of 5-bromo-7-fluoro-3-isopropyl-2-methyl-2H-indazole [ka] To a stirred solution of 5-bromo-7-fluoro-2-methyl-3-(prop-1-en-2-yl)-2H-indazole (400 mg, 1.49 mmol, 1.0 equiv) in EtOAc (8 mL, 20 vol), PtO2 (11.3 mg, 0.036 mmol, 0.05 equiv) was added and stirred at room temperature under H2 atmosphere for 7 h. After completion of the reaction by TLC, the reaction mixture was filtered through Celite® (i.e., diatomaceous earth) and washed with EtOAc. The organic layer was evaporated and purified by silica gel column (100-200 mesh, eluent: 50% EtOAc / hexane) to give 5-bromo-7-fluoro-3-isopropyl-2-methyl-2H-indazole (360 mg, 70%) as a brown solid. TLC system: EtOAc / petroleum ether (50:50), R f Value: approx. 0.25; LC / MS retention time = 3.87 minutes, 271.0[M+H] + .

[0191] 1 H NMR (400MHz, CDCl3) δ 7.70(d,J=1.6Hz,1H),7.02(dd,J=10.4 and 1.6Hz,1H),4.16(s,3H),3.45-3.38(m,1H),1.51(d,J=6.8Hz,6H).

[0192] Synthesis of 7-fluoro-3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole [ka] To a stirred solution of 5-bromo-7-fluoro-3-isopropyl-2-methyl-2H-indazole (300 mg, 1.11 mmol, 1.0 equiv) in 1,4-dioxane (6 mL) at room temperature, bis(pinacolato)diborane (335 mg, 1.33 mmol, 1.2 equiv) and potassium acetate (272 mg, 2.77 mmol, 2.5 equiv) were added and degassed for 5 min, followed by the addition of Pd(dppf)Cl2.DCM (45 mg, 0.055 mmol, 0.05 equiv). The reaction mixture was stirred at 100° C. for 16 h. After completion of the reaction by TLC. The volatiles were removed under vacuum, diluted with water (50 mL) and extracted with ethyl acetate (3×30 mL). The organic layer was separated, washed with brine solution (30 mL), dried over sodium sulfate and concentrated to give 7-fluoro-3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (350 mg, crude) as a black semi-solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.45. Cr: LC / MS shows 44% boronic ester and 16.3% boronic acid m / z. LC / MS retention time = 5.39 min, 319.2 [M+H] + Therefore, the crude material was used in the next step.

[0193] Synthesis of 5-(2-chloropyrimidin-4-yl)-7-fluoro-3-isopropyl-2-methyl-2H-indazole [ka] To a stirred solution of 7-fluoro-3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazole (350 mg, 1.10 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (6 mL) was added 4-chloro-2-(methylthio)pyrimidine (208 mg, 1.43 mmol, 1.3 equiv.) and K2CO3 (380 mg, 2.75 mmol, 2.5 equiv.). The reaction mixture was degassed for 5 min and Pd(dppf)Cl2DCM (45 mg, 0.055 mmol, 0.05 equiv.) was added. The reaction mixture was stirred at 100° C. for 16 h. After completion of the reaction by TLC, it was diluted with water and extracted with ethyl acetate (3×20 mL). The organic layer was dried over Na2SO4, evaporated and purified by silica gel column (100-200 mesh, eluent: 25%-30% EtOAc / Hexane) to give 5-(2-chloropyrimidin-4-yl)-7-fluoro-3-isopropyl-2-methyl-2H-indazole (180 mg, 54%) as a yellow solid. TLC system: EtOAc / Hexane (50:50), R f Value: approx. 0.25. LC / MS retention time=3.53 minutes, 305.0[M+H] + .

[0194] 1 H NMR (400MHz, CDCl3) δ 8.61(d,J=5.6Hz,1H),8.41(s,1H),7.61-7.57(m,2H),4.20(s,3H),3.56-3.48(m,1H),1.58(d,J=7.2Hz,6H).

[0195] Synthesis of tert-butyl ((1S,3S)-3-((4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-7-fluoro-3-isopropyl-2-methyl-2H-indazole (150 mg, 0.493 mmol, 1 equiv.) in isopropyl alcohol (2.5 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (120 mg, 0.64 mmol, 1.3 equiv.) and heated at 120° C. for 16 h. TLC showed a polar spot and the reaction mixture was allowed to reach room temperature. The reaction mixture was evaporated, diluted with water (20 mL) and extracted with EtOAc (2×20 mL). The organic layer was dried over Na2SO4, filtered, evaporated and purified by silica gel column (eluent in 40% EtOAc / hexanes) to give tert-butyl tert-butyl ((1S,3S)-3-((4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (130 mg, 56%) as a yellow solid. TLC system: MeOH / DCM (10:90), R f Value: approx. 0.30; LC / MS retention time = 3.50 minutes, 469.2[M+H] + .

[0196] Synthesis of (1S,3S)-N1-(4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine TFA salt [ka] To a solution of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (130 mg, 0.277 mmol, 1 equiv.) in 1,4-dioxane (1.3 mL) was added 4M HCl in 1,4-dioxane (0.3 mL) at 0° C. and stirred at room temperature for 16 h. After completion of the starting material, the reaction mixture was concentrated and purified by preparative HPLC (elution in 32% acetonitrile and 0.1% TFA in water) to give (1S,3S)-N1-(4-(7-fluoro-3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine TFA salt (40 mg, 39%) as a yellow solid. TLC system: MeOH / DCM (10:90), R f Value: approx. 0.1; LC / MS retention time = 2.67 minutes, 369.3[M+H] + .

[0197] 1 H NMR(400MHz,CD3OD) δ 8.55(s,1H),8.29(d,J=6.4Hz,1H),7.85(d,J=13.2Hz,1H),7.47(d,J=5.6Hz,1H),4.71-4.68(m,1H),4.20(s,3H),3.86-3.83 (m,1H),3.69-3.61(m,1H),2.41-2.35(m,2H),2.25-2.19(m,2H),1.88-1.82(m,1H),1.80-1.75(m,1H),1.59(d,J=7.2Hz,6H).

[0198] Example 11 Synthesis of compound I-10 [ka] Overall reaction scheme: [ka] Synthesis of 5-bromo-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 5-bromo-3-isopropyl-2H-pyrazolo[3,4-b]pyridine (0.8 g, 3.33 mmol, 1.0 equiv) in THF (24 mL, 30 vol), 2M NaHMDS in THF (2.4 mL, 4.99 mmol, 1.5 equiv) was added at 0° C., followed by MeI (0.8 mL, 9.99 mmol, 3 equiv) and stirred at room temperature for 4 h. After completion of the reaction by TLC, it was diluted with cold water (100 mL) and extracted with EtOAc (2×80 mL). The organic layer was separated, dried over sodium sulfate, and concentrated to give 5-bromo-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine (600 mg, 71% yield) as a yellow solid. TLC system: EtOAc / petroleum ether (1:1), R f Value: about 0.4;

[0199] 1 H NMR (400MHz, CDCl3) δ 8.61(d,J=2.4Hz,1H),8.25(d,J=2.0Hz,1H),4.13(s,3H),3.43-3.39(m,1H),1.49(d,J=7.2Hz,6H).

[0200] Synthesis of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 5-bromo-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine (600 mg, 2.36 mmol, 1.0 equiv) in 1,4-dioxane (12 mL), bis(pinacolato)diborane (722 mg, 2.83 mmol, 1.2 equiv), potassium acetate (581 mg, 5.90 mmol, 2.5 equiv) were added and degassed for 5 min. Then Pd(dppf)Cl2.DCM (96 mg, 0.12 mmol, 0.05 equiv) was added and stirred at 90 °C for 16 h. After completion of the reaction by TLC. The volatiles were removed under vacuum, diluted with water (70 mL) and extracted with ethyl acetate (3 x 30 mL). The organic layer was separated, washed with brine solution (30 mL), dried over sodium sulfate and concentrated to give 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-pyrazolo[3,4-b]pyridine (700 mg, crude) as a sticky mass. TLC system: EtOAc (100%), R f Value: approx. 0.1; Cr: LC / MS: 55% boronic acid m / z, 220.1 [M+H] + .

[0201] Synthesis of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 3-isopropyl-2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-pyrazolo[3,4-b]pyridine (700 mg, 2.32 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (14 mL), 4-chloro-2-(methylthio)pyrimidine (572 mg, 2.78 mmol, 1.2 equiv.), K2CO3 (1.1 g, 5.80 mmol, 2.5 equiv.) were added and degassed for 5 min. Then, Pd(dppf)Cl2DCM (131 mg, 0.12 mmol, 0.05 equiv.) was added and stirred at 80° C. for 16 h. After completion of the reaction by TLC, the reaction mixture was evaporated, diluted with water (80 mL) and extracted with ethyl acetate (2×70 mL). The organic layer was separated, dried over sodium sulfate and concentrated to give 2.6 g of crude, which was purified by silica gel column (100-200 mesh, eluent: 80% EtOAc / petroleum ether) to give 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine (180 mg, 26% yield) as a viscous liquid. TLC system: EtOAc (100%), R f Value: about 0.2;

[0202] 1 H NMR(400MHz,CDCl3) δ 9.28(d,J=2.4Hz,1H),9.04(d,J=2.4Hz,1H),8.69(d,J=5.2Hz,1H),7.75(d,J=5.2Hz,1H),4.26(s,3H),3.58-3.53(m,1H),1.63(d,J=6.8Hz,6H).

[0203] Synthesis of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine (180 mg, 0.63 mmol, 1 equiv) in IPA (1.8 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (163 mg, 0.82 mmol, 1.3 equiv) and stirred at 110° C. for 48 h. TLC showed a new polar spot and traces of starting material. The volatiles were removed in vacuo, diluted with water (20 mL) and extracted with EtOAc (2×20 mL). The organic layer was dried over Na2SO4, filtered, concentrated and purified by silica gel column (100-200 mesh, eluent: 70-80% EtOAc / petroleum ether) to give tert-butyl ((1S,3S)-3-((4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (200 mg, 70%) as a yellow sticky solid. TLC system: MeOH / DCM (5:95), R f Value: approx. 0.4; LC / MS: retention time = 3.49 minutes, 452.2[M+H] + .

[0204] 1 H NMR(400MHz,CDCl3):δ 9.31(brs,1H),8.83(s,1H),8.38(d,J=5.2Hz,1H),7.06(d,J=5.2Hz,1H),5.26(d,J=6.8Hz,1H),4.62(brs,1H),4.55-4.50(m,1H), 4.24-4.18(m,4H),3.57-3.50(m,1H),2.38-2.25(m,2H),2.08-1.99(m,2H),1.61(d,J=7.2Hz,6H),1.58-1.52(m,2H),1.49(s,9H).

[0205] Synthesis of (1S,3S)-N1-(4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt [ka] To a stirred solution of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (6) (200 mg, 0.44 mmol, 1 equiv) in DCM (2 mL) was added 2 M HCl in EtO (1 mL) at 0° C. and stirred at room temperature for 16 h. After completion of the starting material, the volatiles were removed under vacuum, triturated with diethyl ether (2×3 mL) and lyophilized to give ((1S,3S)-N1-(4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt (94 mg, 50%) as a brown solid. TLC system: MeOH / DCM (10:90), R f Value: approx. 0.1; LC / MS retention time = 2.29 minutes, 352.2[M+H] + .

[0206] 1 H NMR(400MHz,CD3OD) δ 9.79-9.74(m,2H),8.48(d,J=6.8Hz,1H),7.77(d,J=6.4Hz,1H),5.11-5.05(br,1H),4.34(s,3H),3.94-3.8 7(m,1H),3.84-3.77(m,1H),2.49-2.36(m,2H),2.33-2.29(m,2H),1.94-1.81(m,2H),1.66(d,J=6.8Hz,6H).

[0207] Example 12 Synthesis of compound I-11 [ka] Overall reaction scheme: [ka] Synthesis of 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-ol [ka] To a stirred solution of 5-bromo-2-fluoronicotinaldehyde (8 g, 39.2 mmol, 1.0 equiv.) in dry THF (80 mL) was added 1.5 M isopropylmagnesium bromide in THF (31.3 mL, 47.1 mmol, 1.2 equiv.) at 0 °C and stirred at room temperature for 3 h. After completion of the reaction by TLC, the reaction mixture was diluted with saturated NH4Cl (50 mL) and extracted with EtOAc (2 x 100 mL). The organic layer was dried over sodium sulfate, concentrated and purified by silica gel column (60-120 mesh, eluent: 12% EtOAc / petroleum ether) to give 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-ol (2.71 g, 28% yield) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.4. 1 H NMR(400MHz,CDCl3) δ 8.16(dd,J=2.4 and 1.2Hz,1H),8.04-8.01(m,1H),4.72(t,J=4.8Hz,1H),1.99-1.97(m,2H),0.93(d,J=6.8Hz,3H),0.87(d,J=6.8Hz,3H).

[0208] Synthesis of 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-one [ka] To a stirred solution of 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-ol (3) (2.7 g, 10.9 mmol, 1.0 equiv.) in DCM (54 mL) was added DMP (6.95 g, 16.4 mmol, 1.5 equiv.) at 0 °C and stirred at room temperature for 2 h. After completion of the reaction by TLC, it was diluted with saturated NaHCO3 solution (30 mL) and extracted with DCM (2 x 50 mL). The organic layer was separated, dried over sodium sulfate, and concentrated to give 3 g of crude product, which was purified by silica gel column (60-120 mesh, eluent: 4% EtOAc / petroleum ether) to give 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-one (2.3 g, 86% yield) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.85.

[0209] 1 H NMR (400 MHz, CDCl3) δ 8.41 (dd, J = 2.4 and 1.2 Hz, 1H), 8.35 (dd, J = 8.0 and 2.4 Hz, 1H), 3.44-3.40 (m, 1H), 1.20 (d, J = 6.8 Hz, 6H).

[0210] Synthesis of 5-bromo-3-isopropyl-2H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 1-(5-bromo-2-fluoropyridin-3-yl)-2-methylpropan-1-one (2.3 g, 9.38 mmol, 1.0 equiv) in EtOH (23 mL, 10 vol) was added 85% hydrazine hydrate (12 mL, 5 vol) and stirred at 100 °C for 16 h. After completion of the reaction by TLC, the reaction mixture was evaporated, diluted with water (80 mL) and extracted with ethyl acetate (3 x 50 mL). The organic layer was separated, dried over sodium sulfate and concentrated to give 2.6 g of crude, which was purified by silica gel column (60-120 mesh, eluent: 9%-12% EtOAc / petroleum ether) to give 5-bromo-3-isopropyl-2H-pyrazolo[3,4-b]pyridine (1.1 g, 49% yield) as an off-white solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: approx. 0.30. LC / MS retention time=3.25 minutes, 240.0[M+H] + .

[0211] 1 H NMR (400MHz, CDCl3) δ 11.05(brs,1H),8.59(d,J=2.0Hz,1H),8.27(d,J=2.0Hz,1H),3.41-3.36(m,1H),1.48(d,J=6.8Hz,6H).

[0212] Synthesis of 5-bromo-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 5-bromo-3-isopropyl-2H-pyrazolo[3,4-b]pyridine (1.0 g, 4.18 mmol, 1.0 equiv) in EtOAc (40 mL, 40 vol), trimethyloxonium tetrafluoroborate (3.71 g, 16.4 mmol, 4 equiv) was added and stirred at room temperature for 12 h. After completion of the reaction by TLC, it was diluted with cold water (100 mL) and extracted with EtOAc (2×80 mL). The organic layer was separated, dried over sodium sulfate, and concentrated to give 1.5 g of crude, which was purified by silica gel column (60-120 mesh, eluent: 8%-11% EtOAc / petroleum ether) to give 5-bromo-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine (280 mg, 27% yield) as a yellow liquid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.7;

[0213] 1 H NMR (400MHz, CDCl3) δ 8.51(d,J=2.0Hz,1H),8.18(d,J=2.0Hz,1H),4.06(s,3H),3.36-3.29(m,1H),1.43(d,J=7.2Hz,6H).

[0214] Synthesis of 3-isopropyl-1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 5-bromo-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine (280 mg, 1.11 mmol, 1.0 equiv) in 1,4-dioxane (6 mL), bis(pinacolato)diborane (337 mg, 1.32 mmol, 1.2 equiv), potassium acetate (271 mg, 2.77 mmol, 2.5 equiv) were added and degassed for 5 min. Then Pd(dppf)Cl2.DCM (45 mg, 0.055 mmol, 0.05 equiv) was added and stirred at 90 °C for 16 h. After completion of the reaction by TLC. The volatiles were removed under vacuum, diluted with water (50 mL) and extracted with ethyl acetate (3 x 20 mL). The organic layer was separated, washed with brine solution (10 mL), dried over sodium sulfate and concentrated to give 3-isopropyl-1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazolo[3,4-b]pyridine (360 mg, crude) as a black semi-solid. TLC system: EtOAc / petroleum ether (20:80), R f Value: about 0.45. Cr: LC / MS: 56% boronic acid m / z, 220.1 [M+H] + .

[0215] Synthesis of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine [ka] To a stirred solution of 3-isopropyl-1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazolo[3,4-b]pyridine (360 mg, 1.19 mmol, 1 equiv.) in 1,4-dioxane and water (3:1) (7 mL), 4-chloro-2-(methylthio)pyrimidine (213 mg, 1.43 mmol, 1.2 equiv.), K2CO3 (412 mg, 2.99 mmol, 2.5 equiv.) was added and degassed for 5 min. Then, Pd(dppf)Cl2DCM (48 mg, 0.06 mmol, 0.05 equiv.) was added and stirred at 80 °C for 16 h. After completion of the reaction by TLC, the RM was evaporated, diluted with water (30 mL) and extracted with ethyl acetate (2 x 40 mL). The organic layer was separated, dried over sodium sulfate and concentrated to give 2.6 g of crude, which was purified by silica gel column (100-200 mesh, eluent: 18-22% EtOAc / petroleum ether) to give 5-(2-chloropyrimidin-4-yl)-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine (230 mg, semi-pure) as a pale yellow liquid. TLC system: EtOAc / petroleum ether (30:70), R f Value: about 0.2. LC / MS: Retention time = 3.56 minutes, 288.1[M+H] + ;Purity 68%.

[0216] Synthesis of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate [ka] To a stirred solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridine (230 mg, 0.80 mmol, 1 equiv) in IPA (4 mL) was added tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (240 mg, 1.20 mmol, 1.5 equiv) and stirred at 110° C. for 60 h. TLC showed a new polar spot and the absence of starting material. The volatiles were removed in vacuo, diluted with water (20 mL) and extracted with EtOAc (2×20 mL). The organic layer was dried over Na2SO4, filtered, concentrated and purified by reverse phase column (eluent: 32-59% acetonitrile and 0.01% formic acid in water) to give tert-butyl ((1S,3S)-3-((4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (120 mg, 33%) as a brown solid. TLC system: EtOAc / petroleum ether (50:50), R f Value: approx. 0.40; LC / MS retention time = 3.65 minutes, 452.2[M+H] + .

[0217] 1 H NMR(400MHz,CDCl3):δ 9.17(brs,1H),8.70(s,1H),8.34(d,J=4.8Hz,1H),7.02(d,J=5.2Hz,1H),5.32(bs,1H),4.57(brs,1H),4.52- 4.47(m,1H),4.15-4.12(m,4H),3.49-3.40(m,1H),2.34-2.23(m,2H),2.05-1.95(m,2H),1.50-1.45(m,17H).

[0218] Synthesis of (1S,3S)-N1-(4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt [ka] To a stirred solution of tert-butyl ((1S,3S)-3-((4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)amino)cyclopentyl)carbamate (120 mg, 0.27 mmol, 1 equiv.) in 1,4-dioxane (1.2 mL) was added 4M HCl in 1,4-dioxane (0.6 mL) at 0° C. and stirred at room temperature for 2 h. After completion of the starting material, the volatiles were removed under vacuum, triturated with diethyl ether (2×3 mL) and lyophilized. The resulting material was purified by reverse phase column (eluted with 32% acetonitrile and 0.01% formic acid in water) to give (1S,3S)-N1-(4-(3-isopropyl-1-methyl-1H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-yl)cyclopentane-1,3-diamine HCl salt (40 mg, 43%) as a brown sticky solid. TLC system: MeOH / DCM (10:90), R f Value: approx. 0.1; LC / MS retention time = 2.27 minutes, 352.4[M+H] + .

[0219] 1 H NMR(400MHz,CD3OD) δ 9.47(brs,1H),9.15(s,1H),8.36(d,J=6.4Hz,1H),7.77(d,J=6.4Hz,1H),4.55-4.49(br,1H),4.10(s,3H),3.92 -3.88(m,1H),3.54-3.50(m,1H),2.47-2.38(m,2H),2.31-2.28(m,2H),1.94-1.80(m,2H),1.48(d,J=6.8Hz,6H).

[0220] Example 13 Synthesis of compound I-13 [ka] Synthesis of N-((1R,2S,4R)-bicyclo[2.2.1]heptan-2-yl)-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine [ka] In a sealed tube, a solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-indazole (300 mg, 1.04 mmol, 1 equiv.) and (1S,2S,4R)-bicyclo[2.2.1]heptan-2-amine (250 mg, 1.36 mmol, 1.3 equiv.) in isopropyl alcohol (3 mL) was stirred at 110° C. for 16 h. After completion of the reaction by TLC, the reaction mixture was evaporated and purified by reverse phase column [eluted with 56% acetonitrile in 0.1% aqueous formic acid] to give N-((1R,2S,4R)-bicyclo[2.2.1]heptan-2-yl)-4-(3-isopropyl-2-methyl-2H-indazol-5-yl)pyrimidin-2-amine (77 mg, 20% yield) as an off-white solid. TLC system: EtOAc / hexane (1:1), R f Value: approx. 0.2; LC / MS (m / z): 362.3 (M+H) +

[0221] 1 H NMR(400MHz, CDCl3) δ:8.60(brs,1H),8.30(d,J=4.8Hz,1H),7.88(d,J=9.2Hz,1H),7.67(d,J=9 .2Hz,1H),6.99(d,J=5.2Hz,1H),5.08-5.06(m,1H),4.16(s,3H),3.89(brs ,1H),3.53-3.46(m,1H),2.43(brs,1H),2.33(brs,1H),1.95-1.89(m,1H), 1.60-1.57(m,6H),1.54-1.49(m,2H),1.38-1.33(m,2H),1.32-1.28(m,2H).

[0222] Example 14 Synthesis of compound I-14 [ka] Synthesis of N-((1R,2S,4R)-bicyclo[2.2.1]heptan-2-yl)-4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-amine TFA salt (BLX-3051) [ka] A solution of 5-(2-chloropyrimidin-4-yl)-3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridine (300 mg, 1.04 mmol, 1 equiv.) and (1S,2S,4R)-bicyclo[2.2.1]heptan-2-amine (150 mg, 1.35 mmol, 1.3 equiv.) in IPA (5 mL) in a sealed tube was stirred at 110° C. for 36 h. After completion of the reaction by TLC, the reaction mixture was evaporated and purified by reverse phase column [eluted with 45% acetonitrile in 0.1% aqueous formic acid] to give N-((1R,2S,4R)-bicyclo[2.2.1]heptan-2-yl)-4-(3-isopropyl-2-methyl-2H-pyrazolo[3,4-b]pyridin-5-yl)pyrimidin-2-amine TFA salt (89 mg, 24% yield) as a pale yellow solid. TLC system: EtOAc / Hexane (60:40), R f Value: approx. 0.2; LC / MS (m / z): 363.3 (M+H) +

[0223] 1 H NMR(400MHz,DMSO-d6) δ:9.32(s,1H),9.10(brs,1H),8.39(d,J=5.6Hz,1H),7.72(brs,1H),7.48(d,J=4.0Hz,1H),4.17(s,3H),3.79(brs,1H),3.64-3 .57(m,1H),2.48-2.45(m,1H),2.33-2.27(m,2H),1.76-1.71(m,1H),1.56-1.48(m,10H),1.29-1.26(m,1H),1.16-1.12(m,2H).

[0224] Biological Example 1 Cytotoxicity assay Multiplexing of LDH-Glo cytotoxicity assay and CellTiter-Glo 2D cell viability assay. RWPE-1 normal prostate cells were incubated with 10 doses of IC 50 Cells were treated in duplicate with compounds I-2 and AZD-4573 in 3-fold serial dilutions starting at 100 μM for 72 h at 5000 cells / well in a 96-well format in 3x mode. Changes in toxicity were measured using the LDH-Glo cytotoxicity assay by combining 50 μL of diluted sample with 50 μL of LDH detection reagent and recording luminescence after 1 h of incubation. After removing the sample for LDH measurement, an equal volume of CellTiter Glo 2D reagent was added to the remaining cell suspension and luminescence was recorded after a 30 min incubation period.

[0225] The cytotoxicity of compound I-2 was tested in RWPE-1 normal healthy donor prostate cells using a highly sensitive assay that measures the release of lactate dehydrogenase (LDH), a cytoplasmic enzyme that is released into cell culture medium when cell membrane integrity is lost. Data using both the LDH-Glo assay and a cell viability assay using CellTiter-Glo indicate that normal cell cytotoxicity does not occur until compound I-2 reaches more than 15-fold the concentration required to kill most prostate cancer cells (IC5.26 μM in these normal cells). 50 The reference compound AZD-4573 is highly toxic to normal prostate cells. [Table 2]

[0226] Biological Example 2 CellTiter-Glo assay CellTiter-Glo® 2.0 Luminescent Cell Viability Assay Reagent was purchased from Promega, and 22Rv1 and LASCPC-01 cell lines were purchased from ATCC. The test compound, Compound I-2, was administered at 10 doses IC 50All selected cell lines were used in a 96-well format, dosed in duplicate, at 4000 cells / well (adhesive), starting at 100 μM, with 3-fold serial dilutions, and a treatment time of 72 hours. An equal volume of CellTiter-Glo 2D reagent was added to the cell suspension, and luminescence was recorded after a 30-minute incubation period. The results are shown in Figures 1 and 2. Figure 1 shows the cellular anti-proliferative efficacy of compound I-2 in six prostate cancer cell lines across a range of concentrations. Figure 2 shows the cellular anti-proliferative efficacy of the benchmark CDK9 inhibitor AD-4573 in six prostate cancer cell lines across a range of concentrations. The experimental data in Figures 1 and 2 are summarized in Table 3. [Table 3]

[0227] Biological Example 3 Colony formation assay Colony formation assays were performed with two different NEPC cell lines. Cells were seeded at 800 cells per well in 6-well plates and allowed to adhere for 24 hours in complete growth medium. Treatment with compound I-2 ranged from NT to 10.0 μM for 14 days. Cells were fixed with a 3:1 mixture of methanol / acetic acid and stained with methylene blue. SEM from two independent experiments. (*P<0.05, **P<0.01, ***P<0.005 vs. DMSO control). The results showed that compound I-2 inhibited the proliferation and colony formation of 22Rv1 more significantly than that of LASCPC-01, as shown in Figures 3 and 4.

[0228] Biological Example 4 Annexin V / 7-AAD apoptosis assay Apoptosis of two NEPC cells by Annexin V / 7-AAD staining: Apoptosis induced by CDK9 inhibitor Compound I-2 was evaluated using a Guava flow cytometer by Annexin V / 7-AAD double staining and FACS analysis. Treatment with Compound I-2 increased apoptosis in a dose-dependent manner, and real-time measurements of apoptosis induced by treatment with Compound I-2 in prostate cancer cells were determined using an IncuCyte S3 Live-Cell Analysis System. Representative images and bar graphs from LASCPC-01 cells are shown in Figure 5. In addition, apoptosis induced by CDK9 inhibitor Compound I-2 was also evaluated by Annexin V / 7-AAD double staining and FACS analysis. Treatment with Compound I-2 increased apoptosis in a dose-dependent manner, and the results of Annexin V / 7-AAD staining are shown in Figures 6 and 7.

[0229] Treatment with compound I-2 increased apoptosis in a dose-dependent manner, which corresponds to the results obtained by the CellTiter-Glo assay described herein. NEPC cells were treated for 72 hours, and it was shown that LASCPC-01, 22Rv1, and C4-2 cells were highly sensitive to treatment with compound I-2. This treatment resulted in 69, 71, and 84% cell death, respectively, which corresponds to the results of the 2D CellTiter-Glo assay.

[0230] Biological Example 5 Western blot Two NEPC cell lines (22Rv1 and LASCPC-01) were treated with compound I-2 for 6 and 24 hours. Cytoplasmic and nuclear extracts from these cells were separated using the NE-PER Nuclear and Cytoplasmic Extraction Kit (ThermoFisher). Cell extracts were separated on 4-12% Bis-Tris NuPAGE gels and then transferred onto 0.45 nitrocellulose membranes, followed by blocking with Pierce Fast Blocking Buffer for 30 minutes and then incubation with the indicated antibodies at 4°C overnight. Relative protein expression of CDK9, N-Myc, and other downstream signaling were quantified by RT-PCR genes using GraphPad Prism 8 and ImageJ software. The results are shown in Figures 8-10.

[0231] Biological Example 6 Expression Total RNA was extracted using the RNeasy mini kit (Qiagen). cDNA was synthesized using the iScript cDNA synthesis kit (Bio-Rad, Hercules, CA). Quantitative RT-PCR was performed with SsoAdvanced SYBR Green Supermix (Bio-Rad) on a CFX96 Real-Time PCR Detection System (Bio-Rad). Data were normalized to GUS, and percentages of gene expression were determined by the ΔCt method.

[0232] Biological Example 7 ADP-Glo ​​kinase assay ADP-Glo ​​assay kit was purchased from Promega, CDK9 / CycT1 protein and PDKtide substrate were purchased from SignalChem. Compounds were administered at 10 doses IC 50The wells were dosed in duplicate in 3x serial dilutions in 1x assay buffer (50 mM HEPES, 3 mM MgCl2, 3 mM MnCl2, and 1 mM DTT) and started with 3-fold serial dilutions at 100 μM. Reactions were initiated by preincubating 4 μL of protein (20 nM), 2 μL of substrate (100 μM), and 2 μL of serially diluted drug for 30 min in 1x assay buffer (50 mM HEPES, 3 mM MgCl2, 3 mM MnCl2, and 1 mM DTT). After preincubation, 4 μL of ATP (1 μM) was added to each well and incubated for an additional 90 min. ADP-Glo ​​reagent (10 μL) and Kinase Detection Reagent (20 μL) were added to each well and incubated as recommended by Promega. Reactions were quantified by measuring luminescence and IC was calculated using GraphPad Prism software. 50 was calculated.

[0233] Biological Example 8 Immunofluorescence staining LASCPC-01 NEPC cells are treated with compound I-2 starting from NT, 1.1, 3.3 10, and 30 μM for 6 hours. After 6 hours, cells are fixed with 4% formaldehyde and probed with primary N-Myc antibody overnight. Cells are then incubated with secondary antibody conjugated with Alexa Fluor 488 and counterstained with Hoechst 33342. Images are taken using a Nikon automated wide-angle CCD camera and quantified using ImageJ and GraphPad Prism 8 software, which are shown in FIG. 11.

[0234] Biological Example 9 Cell cycle arrest assay NEPC cells were treated with compound I-2 starting at NT, 1, and 3.16 μM for 8 hours. Cells were then harvested in PBS and fixed overnight at -20°C with ice-cold 70% ethanol. Cells were then washed twice with ice-cold PBS and then resuspended in DAPI staining buffer and incubated in the dark for 30 minutes. Cells were analyzed using a BD Fortessa flow cytometer and cell cycle distribution was analyzed using FlowJo and GraphPad Prism 8 software. Western blot analysis of nuclear extracts showed that compound I-2 inhibited CDK9 downstream targets. Representative blots from 22Rv1 and LASCPC-01 are shown in Figures 12-13.

[0235] Biological Example 10 Cell cycle arrest assay A pilot in vivo xenograft 22Rv1 and LASCPC-01 engraftment study is described herein. Animals: Male BALB / c nude mice (8 weeks old). NEPC cell lines: 22Rv1 and LASCPC-01. Number of cells: 22Rv1 (1.2×10 6 ) and LASPC-01(10 6 Sub.Q. Tumor volume was measured twice weekly by caliper and calculated as length × (width). 2 ) / 2. The tumor size was approximately 100-500 mm 3 Upon reaching , mice were randomized and selected for treatment with compound I-2 as a TFA salt at an oral dose of 18 mg / kg. The data from this experiment are shown in Figures 14-15.

[0236] Biological Example 11 Kinase Profiling CDK family kinase profiling assays were performed using a miniaturized radioisotope-based assay platform with Kd KINOMEscan's DiscoveRx matrix (KdELECT) and reaction biology ( 33 P).IC 50Values ​​were estimated based on the best curve fitting available. Compounds I-2 and AZD4573 (CAS number 2057509-72-3) had IC values ​​of less than 4.2 and 5.0 nM, respectively, against CDK9. 50 The values ​​were shown.

[0237] In summary, compound I-2 showed promising 3- to 50-fold selectivity over the closest CDK homologs tested. [Table 4]

[0238] Biological Example 12 Off-target inhibition studies Compound I-2 had an IC of 4.2 nM in cell-free assays. 50 , and in cellular assays, with an IC of 15.58 nM in the NanoBRET target engagement assay 50 and had IC of 132, 58, 28, and 31 nM in inhibiting NEPC-derived 22Rv1, LASCPC-01, C4-2, and C4-2B cells. 50 Compound I-2 has strong activity with value. Compound I-2 reduced pCDK9, pSer2, and its downstream N-MYC, C-MYC, and MCL-1 targets in newly diagnosed resistant prostate cancer patient samples, indicating its response to overcoming resistance and antitumor efficacy. Data suggests that compound I-2 has strong potential for development as a candidate for NEPC therapy.

[0239] Off-target inhibition of other kinases was analyzed using the scanMAX panel, which contains 468 kinases including lipid and atypical kinases. Compound I-2 was assayed against the intended target, CDK9, with an IC 50At 1 μM, activity was found against nine other kinases, including CLK1 (2.9 Kd), DYRKA1 (9 Kd), ERK8 (5.5 Kd), FLT3 / ITD(D835V) (1.2 Kd), Haspin (2.8 Kd), PCTK1 (8.2 Kd), PRKD3 (230 Kd), TAOK1 (19 Kd), and TAOK3 (9.2 Kd), each of which showed 0-1% control activity. These nine off-target kinases were analyzed using their Kd and IC as shown in Table 3 above. 50 The selectivity profiler screen revealed that the major off-target kinase is FLT3. The FDA has approved 48 kinase inhibitors, 11 of which have 2,4-disubstituted pyrimidines like compound I-2 and showed similar off-target profiles. The structure of compound I-2 contains a unique chemotype with high cell permeability, supporting superior overall selectivity and efficacy when compared to the other 11 approved kinase inhibitors.

[0240] Biological Example 13 Target engagement assay in HEK293 cells HEK293 cells were transfected with 1 μg of CDK9 and 9 μg of Cyclin T1 DNA, and the transfected cells were treated with compound I-2 (starting at 100 μM, 10 doses in 3-fold dilutions) and the reference compound dinaciclib (starting at 10 μM, 10 doses in 3-fold dilutions) was used. Curve fitting was performed only if the percentage of NanoBRET signal at the highest concentration of compound was less than 55%. Fresh HEK293 cells were sourced from ATCC, and the compound treatment time was 1 hour. The results were plotted and the IC was calculated as shown in Table 4 below. 50 The data was calculated. [Table 5]

[0241] Biological Example 14 Cell Antiproliferative Efficacy CellTiter-Glo® 2.0 Luminescent Cell Viability Assay Reagent was purchased from Promega, and 22Rv1, LASCPC-01, C4-2, C4-2B, and NCI-H660, DU-145, PC-3, and LNCap cell lines were obtained from ATCC and the Huntsman Cancer Institute Clinical Core Facility. Compound I-2 and reference compound AZD-4573 were administered at 10 doses IC 50 All selected cell lines were tested in duplicate in 96-well format at 2500 cells / well (adhesive), starting at 100 μM with 3-fold serial dilutions and a 72-hour treatment time. Cell viability results were obtained and IC20 ... 50 Values ​​were calculated (see Table 5 below). Compound I-2 was particularly sensitive in all NEPC cells (see, for example, results for 22Rv1, LASCPC-01, NCI-H660, C4-2, C4-2B). [Table 6]

[0242] Biological Example 15 Colony formation assay 2 Prostate cancer cells were seeded at 400 cells per well in 6-well plates and allowed to attach for 24 hours in complete growth medium before being treated with compound I-2 at concentrations ranging from 0.123, 0.37, 1.1, 3.0, and 10 μM for 72 hours. Treatment resulted in complete inhibition of colony formation by all prostate cancer cells.

[0243] Biological Example 16 Fluorescence polarization assay hERG channel biochemical binding studies were performed in the absence of radioligand using the Predictor™ hERG Fluorescence Polarization Assay Kit. Assay performance was validated using a panel of established hERG channel blockers. Results from the Predictor™ assay correlate highly with those obtained from patch clamp techniques. Compound I-2 was found to have no hERG activity, with a measured IC 50 was 57.8 μM.

[0244] Biological Example 17 P450 isoenzyme inhibition assay Since only a single P450 isozyme is expressed on human liver microsomes (HLM), P450 baculosome reagents (i.e., microsomes) were prepared from insect cells infected with recombinant baculovirus containing human P450 isozyme and rabbit NAPDH-P450 reductase. Compound I-2 was incubated with baculosomes + substrate, and the reaction was initiated by adding NAPDH-450. At the end of the incubation period, a stop solution was added to terminate the reaction, resulting in a stable fluorescent signal compared to compound I-2 and solvent control. The inhibitory effect of compound I-2 was evaluated against five P450 isozymes, and the results are provided below. [Table 7]

[0245] Biological Example 17 Pharmacokinetic Studies - Bioavailability Intravenous administration of 1 mg / kg compound I-2 to male beagle dogs showed moderate plasma clearance compared to normal hepatic blood flow (i.e., 40 mL / min / kg). Steady-state volume distribution was found to be higher, which may be one of the reasons for tissue distribution. The mean end of concentration was 24 hours after intravenous bolus injection. Compound I-2 showed a half-life of 4.26 hours in dog plasma. Oral administration of 3 mg / kg compound I-2 to male beagle dogs showed no lag phase in absorption, and the mean time to reach peak concentration was 1 hour. Compound I-2 showed higher systemic availability. Oral bioavailability was found to be 83±22%. Overall, the mean residence time and plasma half-life were comparable in plasma after intravenous and oral administration. [Table 8] C0: Extrapolated concentration at time zero; C0 reported for IV route; C for oral route max was reported a Reported average (min-max) b Nominal IV and oral doses and AUC inf Bioavailability was calculated using F b (%) is the oral bioavailability

[0246] Compound I-2 achieved good systemic exposure after oral administration. The high steady state of distribution volume suggested distribution to tissues. The increased distribution volume helps maintain plasma concentration up to 24 hours after administration, which may be one of the reasons for the moderate to high plasma half-life of 4.26 and 5.81 hours for intravenous and oral administration, respectively. Due to its moderate clearance, compound I-2 showed a very high oral bioavailability of 83% compared to the intravenous route.

[0247] Pharmacokinetic studies in mouse and rat species have been completed, with compound I-2 achieving excellent systemic exposure with weight-dependent exposure of 28% in mice and 38% oral bioavailability in rats.

[0248] Biological Example 18 In vivo tumor efficacy studies Antitumor efficacy of compound I-2 in 22RV1-induced neuroendocrine prostate cancer (NEPC) CDX males Nu / Nu (Balb / c background)

[0249] 22Rv1 cells were cultured in RPMI supplemented with 10% FBS and 1% P / S in a humidified incubator at 37°C and 5% CO2. Nu / nu Balb / c homozygous mice were sourced through Jackson Laboratory (strain 002019). Mice were fed Teklad irradiated (sterile) mouse chow and bedding with Teklad irradiated (sterile) corn cob bedding from Envigo (Indianapolis, IN). Mice were housed in an Optimice rotating sterile room with a filtered air supply in disposable cages from Animal Care Systems, Inc. (Centennial, CO). A 12-hour light / 12-hour dark light cycle was observed and animal handling was only performed during the light cycle.

[0250] 22Rv1 transplantation: On the day of transplantation, cells were trypsinized and removed from the flask. The trypsin was then neutralized with complete medium and the cells were spun at 300×g. The medium was aspirated and the cells were removed at 5×10 7 The cells were resuspended in 50:50 Cultrex RPMI (without supplements) at a concentration of 100 cells / mL. A volume of 100 μL was injected into the right hind flank of each animal (a total of 5 × 10 cells per mouse). 6 Cells were checked for viability using trypan blue exclusion and found to be >98% viable.

[0251] Study arms and treatments: Mean tumor volume ≈225 mm 3Upon reaching this age, mice were stratified and placed into (5) treatment groups as outlined below. [Table 9]

[0252] Treatments were administered by oral gavage (10 μL / gram adjusted volume). Dosing solutions were made up daily with molecular biology grade water as vehicle (a salt correction factor of 1.65 was applied and TFA salts were adjusted).

[0253] Animal Weight: Change in animal weight during the study is an additional indicator of tolerability. Weights of all animals are recorded (see, for example, Figure 16).

[0254] Relative weights of the groups in this study: the 50 mg / kg and 60 mg / kg groups, as well as the single dose of the 65 mg / kg group, had mice taken off the study starting on day 5. Tumor growth inhibition (TGI) data were analyzed versus dose and tumor weight.

[0255] Tumor volume: Tumor volume was measured by digital calipers and the volume was calculated according to the formula [width 2 × length] / 2. All tumor volume measurements are recorded.

[0256] The 22Rv1 prostate cancer model is the first model developed for the growth of human prostate cancer in immunodeficient mice. In this example, tumors were grown subcutaneously for efficacy studies (a four-arm study comparing vehicle with compound I-2 at 30 mg / kg, 50 mg / kg, and 65 mg / kg; an additional group of four mice received a single dose of 60 mg / kg, orally administered once daily). Tolerability issues were noted at dose levels above 50 mg / kg, leading to some mice being removed from the study and some mice having a dosing break to recover. Compound I-2 appears to inhibit tumor growth in a dose-dependent manner, with tumor growth inhibition observed in all treatment groups.

[0257] For PK / PD studies, plasma, tumor, spleen, femur, heart, kidney, liver, lung, and brain were collected and processed 6 hours after the last dose.

[0258] The results for mean body weight compared to day 0, change in mean tumor volume on day 21 of the study, and mean tumor weight are shown in Figures 16-18, respectively.

[0259] Biological Example 19 In vivo Tumor Efficacy Study 2 Antitumor efficacy of compound I-2 in LASCPC-01-induced neuroendocrine prostate cancer (NEPC) CDX male Nu / Nu (Balb / c background): A similar protocol to the 22Rv1 study was performed in the LASCPC-01 study, using doses of 30, 50, 65, and 80 mg / kg.

[0260] Changes in animal weight during the study are an additional indicator of tolerability. Body weights of all animals are recorded. Tumor volumes are measured by digital calipers and volumes are calculated according to the formula [width x width x depth]. 2 All tumor volume measurements are recorded. LASCPC-01 tumors showed robust growth (180 mm in 19 days). 3 From 2700mm 3 Dose-dependent tumor growth inhibition (TGI) was observed.

[0261] The 50 mg / kg and 80 mg / kg groups had mice removed from the study starting on day 5. Vehicle had large tumors removed on day 19, so day 21 does not reflect the average size of all mice in the group.

[0262] Compound I-2 appeared to inhibit tumor growth in a dose-dependent manner, with the 80 mg / kg group having a maximum growth inhibition rate of 117% on day 5. In the mid-dose group (50 mg / kg), tumor growth inhibition of up to 100% was observed early in the study, and remained greater than 80% throughout the course of the study. Plasma, tumors, and bones were collected for PK / PD studies.

[0263] The results for mean body weight compared to day 0, change in mean tumor volume over the 21st day of the study, and mean tumor weight are shown in Figures 19-21, respectively.

[0264] Biological Example 20 In vivo Tumor Efficacy Study 3 This example presents the evaluation and efficacy studies of compound I-2 conducted in the C4-2 human neuroendocrine prostate cancer xenograft model using male NOD.SCID mice.

[0265] Weight loss was observed when compound I-2 (50 mg / kg, orally, daily) was administered from day 1 to day 4. Starting from day 8, compound I-2 was administered at a dose of 30 mg / kg every other week. From day 13 onward, compound I-2 was administered at 30 mg / kg, orally, daily. Compound I-2 (50-30 mg / kg) showed a reduction in tumor volume compared to the vehicle control group from day 8 to day 18. Compound I-2 at 50-30 mg / kg showed a significant reduction in tumor volume from day 18 to day 21 compared to the vehicle control group (see, e.g., FIG. 22). Compound I-2 at doses of 50-30 mg / kg showed an increase in tumor growth inhibition compared to the vehicle control group from day 8 to day 12 (see, e.g., FIG. 23). Compound I-2 at 50-30 mg / kg showed 43.25% tumor growth inhibition from the vehicle control group on the final 21 days. Compound I-2 at 50-30 mg / kg showed a significant reduction in tumor weight of 48.72% compared to the vehicle control group on the 21st day (see, e.g., FIG. 24). Compound I-2 at 50-30 mg / kg showed a significant weight loss compared to the vehicle control group starting on day 4. Death was observed in one mouse in the compound I-2 treatment group, which may be due to severe weight loss (i.e., about 15%). 1.5 hours after the final dose, blood was collected and processed, and plasma and tumor samples were processed for bioanalysis LC-MS / MS, showing promising PK / PD correlation efficacy.

[0266] If necessary, aspects of the embodiments can be modified to provide yet further embodiments by adopting concepts from various patents, applications, and publications. These and other changes can be made to the embodiments in light of the above detailed description. In general, in the following claims, the terms used should not be construed to limit the claims to the specific embodiments disclosed in the specification and claims, but should be construed to include all possible embodiments along with the full scope of equivalents to which such claims are entitled. Thus, the claims are not limited by this disclosure.

Claims

1. A compound having the following structure (I): 【Chemistry 1】 As a stereoisomer or salt thereof, 【Chemistry 2】 represents a double bond or a single bond such that all valences are satisfied, forming a heteroaryl ring; X is N or CR ４ｂ and Y is N or CR ４ｃ and R １ａ does not exist or C １ -C ６ is alkyl, R １ｂ is hydrogen, halo, hydroxy, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy or C １ -C ６ is hydroxyalkyl, R ２ is hydrogen or optionally substituted C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, or C １ -C ６ is hydroxyalkyl, R ３ But, C ３ -C ５ Cycloalkyl, C ６ -C ８ Bridged cycloalkyl, or hydroxyl, amino, cyano, halo, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, or -N(R ３ｃ ) R ３ａ or R ３ Hydroxyl, cyano, halo, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, or -N(R ３ｃ ) R ３ｂ C optionally substituted with ３ -C ８ is cycloalkyl, R ３ａ is optionally substituted 3- to 10-membered heterocyclyl or optionally substituted 3- to 10-membered heteroaryl; R ３ｂ is an optionally substituted 3- to 10-membered N-heterocyclyl or an optionally substituted 3- to 10-membered heteroaryl; R ３ｃ is hydrogen or optionally substituted C １ -C ６ is alkyl, R ４ａ , R ４ｂ , and R ４ｃ are each independently hydrogen, halo, or optionally substituted C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy or C １ -C ６ is haloalkoxy; R ５ Each occurrence of is independently halo or optionally substituted C １ -C ６ is haloalkyl, n is 0, 1, 2, 3, or 4; A compound wherein p is 0, 1, or 2.

2. 2. The compound of claim 1, wherein the compound has one of the following structures (Ia) or (Ib): 【Chemistry 3】

3. R １ａ The compound according to claim 1 or 2, wherein is not present.

4. R １ａ The compound according to claim 1 or 2, wherein is methyl.

5. R １ｂ is hydrogen, C １ -C ６ Alkyl, or C ３ -C ８ The compound according to any one of claims 1 to 4, which is cycloalkyl.

6. R １ｂ The compound according to any one of claims 1 to 5, wherein is hydrogen, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, isopropyl, methyl, or ethyl.

7. R １ｂ The compound according to any one of claims 1 to 6, wherein is hydrogen, cyclopropyl, cyclobutyl, isopropyl, or methyl.

8. R １ｂ The compound of any one of claims 1 to 7, wherein is unsubstituted.

9. R ２ is hydrogen or C １ -C ６ The compound according to any one of claims 1 to 8, which is alkyl.

10. R ２ But, C １ -C ６ The compound according to any one of claims 1 to 9, which is alkyl.

11. R ２ But -CH ３ The compound according to any one of claims 1 to 10,

12. R ２ The compound according to any one of claims 1 to 9, wherein is hydrogen.

13. R ２ The compound of any one of claims 1 to 9, wherein is unsubstituted.

14. R １ｂ Or R ２ The compound according to any one of claims 1 to 13, wherein at least one of is hydrogen.

15. R １ｂ and R ２ The compound according to any one of claims 1 to 9, wherein both of are hydrogen.

16. The compound according to any one of claims 1 to 15, wherein n is 0 or 1.

17. The compound according to any one of claims 1 to 16, wherein n is 0.

18. R ３ But, C ３ -C ５ Cycloalkyl, C ６ -C ７ Bridged cycloalkyl, or hydroxyl, amino, cyano, halo, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, or -N(R ３ｃ ) R ３ａ 18. The compound of any one of claims 1 to 17, which is a 3-10 membered heterocyclyl optionally substituted with

19. R ３ hydroxyl, amino, cyano, halo, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, or -N(R ３ｃ ) R ３ａ C optionally substituted with ５ 20. The compound of claim 18 which is cycloalkyl.

20. R ３ is optionally substituted with hydroxyl or amino; ５ 20. The compound of claim 18 or 19, which is cycloalkyl.

21. R ３ Hydroxyl, cyano, halo, C １ -C ６ Alkyl, C ３ -C ８ Cycloalkyl, C １ -C ６ Haloalkyl, C １ -C ６ Alkoxy, C １ -C ６ Haloalkoxy, or -N(R ３ｃ ) R ３ｂ C optionally substituted with ３ -C ８ The compound of any one of claims 1 to 17, which is cycloalkyl.

22. R ３ The compound according to any one of claims 1 to 17, wherein is 5-6 membered heterocyclyl.

23. R ３ The compound of any one of claims 1 to 22, wherein:

24. R ３ The compound of any one of claims 1 to 22, wherein is unsubstituted.

25. R ３ has the following structure: 【Chemistry 4】 The compound according to any one of claims 1 to 24, having one of the following:

26. R ３ has the following structure: 【Chemistry 5】 The compound according to any one of claims 1 to 25, having one of the following:

27. R ４ａ The compound according to any one of claims 1 to 26, wherein is hydrogen.

28. The compound according to any one of claims 1 to 27, wherein X is N.

29. 28. The compound according to any one of claims 1 to 27, wherein X is CH or CF.

30. The compound according to any one of claims 1 to 29, wherein Y is N.

31. Y is CR ４ｃ The compound according to any one of claims 1 to 29,

32. 30. The compound according to any one of claims 1 to 29, wherein Y is CH or CF.

33. p is 1, R ５ The compound according to any one of claims 1 to 32, wherein is chloro.

34. The compound according to any one of claims 1 to 32, wherein p is 0.

35. The compound has the following structure: 【Chemistry 6-1】 【Chemistry 6-2】 The compound of claim 1 having one of the following formula: Or a stereoisomer or salt thereof.

36. The compound of any one of claims 1 to 35, wherein the compound is in free base form.

37. The compound according to any one of claims 1 to 35, wherein the compound is a pharma- ceutically acceptable salt.

38. The compound according to any one of claims 1 to 35, wherein the compound is a trifluoroacetate, hydrochloride, or formate salt.

39. A pharmaceutical composition comprising a compound according to any one of claims 1 to 38 and a pharma- ceutically acceptable carrier.

40. A method for treating a disease or disorder, comprising administering to a subject in need of treatment an effective amount of a compound or salt according to any one of claims 1 to 38 or a pharmaceutical composition according to claim 39.

41. 41. The method of claim 40, wherein the disease or disorder is a kinase-expressing cancer.

42. 42. The method of claim 41, wherein the cancer is bladder cancer.

43. 42. The method of claim 41, wherein the cancer is prostate cancer.

44. 42. The method of claim 41, wherein the cancer is a hematological malignancy.

45. 45. The method of claim 44, wherein the hematological malignancy is acute myeloid leukemia.

46. 42. The method of claim 41, wherein the disease or disorder is an autoimmune disease or an inflammatory disease.

Images