Pyrazole derivatives as RET kinase inhibitors

Abstract

This document discloses compounds of formula I or pharmaceutically acceptable salts thereof, wherein variables are as defined herein. These compounds may be used to treat RET-related cancers. Formulations containing compounds of formula I and methods for preparing compounds of formula I are also disclosed.

Description

[0001] background

[0002] The RET kinase rearranged during transfection is a single-transmembrane receptor belonging to the tyrosine kinase superfamily, which is required for the normal development, maturation, and maintenance of several tissue and cell types. The extracellular portion of the RET kinase contains four calcium-dependent cadherin-like repeats involved in ligand binding and a juxtamembranous cysteine-rich region essential for the proper folding of the RET extracellular domain, while the cytoplasmic portion of the receptor includes two tyrosine kinase subdomains.

[0003] RET signaling is mediated by the binding of a group of 35 soluble proteins to the glial cell line-derived neurotrophic factor (GDNF) family ligands (GFL), including nerve growth factor antibody (neurturin (NTRN)), leptospirin (ARTN) and persehin2 (PSPN). Unlike other receptor tyrosine kinases, RET does not directly bind to GFL and requires an additional co-receptor, which can be one of the four GDNF family receptor-α (GFRα) family members linked to the cell surface via a glycosylphosphatidylinositol bond. GFL and the GFRα family member form a binary complex, which then binds to RET and recruits it to the cholesterol-rich membrane subdomain where RET signaling occurs.

[0004] Following binding to the ligand-co-receptor complex, RET dimerization and autophosphorylation at intracellular tyrosine residues recruit adaptor proteins and signal transduction proteins to stimulate a variety of downstream pathways. Binding of adaptor proteins to these docking sites leads to activation of the Ras-MAPK and PI3K-Akt / mTOR signaling pathways, or recruitment of the CBL family of ubiquitin ligases, which plays a role in RET-mediated downregulation of RET function.

[0005] Disruptions to normal RET activity caused by aberrant RET expression, attributed to genetic alterations in RET kinases (including protein-gene fusions and activating point mutations), lead to overactive RET signaling and uncontrolled cell growth, such as in various cancer types and certain gastrointestinal disorders like irritable bowel syndrome (IBS). The ability to inhibit aberrant RET activity in patients with cancer or other disorders associated with overactive RET signaling would be highly beneficial. Furthermore, some genetic alterations in RET kinases are changing their conformational structure so that known RET kinase inhibitors may be less effective (or ineffective). In such cases, novel RET kinase inhibitors effective against modified RET kinases would greatly benefit patients. Summary of the Invention

[0006] This article discloses compounds of formula I or pharmaceutically acceptable salts thereof:

[0007]

[0008] in

[0009] A is C6-C 10 Aryl or C5-C6 heteroaryl;

[0010] Each R1 is independently hydrogen, halogen, C1-C6 alkyl, C1-C6 heteroalkyl, -(C1-C4 alkyl)(C5-C6 heteroalkyl), -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 heteroalkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 heteroalkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C4-C 10 Bicyclol), -(C0-C4 alkyl)(C5-C6 aryl), -(C0-C4 alkyl)(C5-C6 heteroaryl), -(C0-C4 alkyl)(C4-C 10 (heterocyclic), C5-C 12 Spiroalkyl, C5-C 12 Spiroalkyl, dimethylphosphoryl, adamantyl, C1-C6 alkoxy, -(C1-C4 alkyl)-SO2-(C1-C4 alkyl), difluoromethylthioalkyl or pentafluorothioalkyl, wherein each R1 is unsubstituted or, when it can be substituted, is substituted by one or more substituents independently selected from the group consisting of halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, methylamine, N,N-dimethylmethylamine, mono-, di- or tri-halomethoxy, mono-, di- or tri-halomethyl and C0-C3 alkyl-pyrrolidinyl, wherein the pyrrolidinyl group is unsubstituted or substituted by one, two or three independently selected halogen atoms, and wherein two R1 groups may be fused to form a ring structure including a portion of A and optionally aromatic, and n is 1, 2, 3, 4, 5 or 6;

[0011] X1, X2, X3, and X4 are each independently N, CH, C-CH3, C-CH2-OH, C-OCH3, C-CH2-OCH3, or C-halogen; and

[0012] R2 is a C1-C6 alkyl, -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C4-C7 heterocycloalkyl), -(C0-C4 alkyl)(C4-C 10(Bicyclic), each optionally substituted with one or more of halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, cyclopropyl or mono-, di- or tri-halomethyl.

[0013] This document further discloses methods for treating cancer using compounds of Formula I or pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof, and particularly provides methods for treating cancers with aberrant RET expression (e.g., RET-related cancers such as medullary thyroid carcinoma or RET-fused lung cancer). The methods comprise administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof to a patient in need.

[0014] This document also provides compounds of Formula I or pharmaceutically acceptable salts thereof for use in therapies. Furthermore, this document provides compounds of Formula I or pharmaceutically acceptable salts thereof for the treatment of cancers, particularly cancers with aberrant RET expression (e.g., RET-related cancers such as medullary thyroid carcinoma or RET-fusion lung cancer). Use of compounds of Formula I or pharmaceutically acceptable salts thereof in the preparation of medicaments for the treatment of cancers, particularly cancers with aberrant RET expression (e.g., RET-related cancers such as medullary thyroid carcinoma or RET-fusion lung cancer) is also provided.

[0015] describe

[0016] This article describes novel RET kinase inhibitor compounds. These new compounds can meet the need for potent and effective treatment of disorders or cancers associated with abnormal RET activity, such as IBS or cancers, particularly cancers caused by overactive RET signaling (i.e., RET-related cancers). More specifically, these new compounds can meet the need for potent and effective treatment of RET-related cancers such as lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), thyroid cancer (e.g., papillary thyroid carcinoma, medullary thyroid carcinoma, differentiated thyroid carcinoma, recurrent thyroid carcinoma, or refractory differentiated thyroid carcinoma), thyroid adenoma, endocrine gland neoplasm, lung adenocarcinoma, bronchioloalveolar cell carcinoma, type 2A or 2B multiple endocrine neoplasm (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer, mammary cancer, mammary carcinoma, mammary neoplasm, colorectal cancer (e.g., metastatic colorectal cancer), papillary renal cell carcinoma, ganglioneuroma of the gastrointestinal mucosa, inflammatory myofibroblastoma, or cervical cancer.

[0017] This article provides compounds of formula I or pharmaceutically acceptable salts thereof:

[0018]

[0019] in

[0020] A is C6-C 10 Aryl or C5-C6 heteroaryl;

[0021] Each R1 is independently hydrogen, halogen, C1-C6 alkyl, C1-C6 heteroalkyl, -(C1-C4 alkyl)(C5-C6 heteroalkyl), -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 heteroalkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 heteroalkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C4-C 10 Bicyclol), -(C0-C4 alkyl)(C5-C6 aryl), -(C0-C4 alkyl)(C5-C6 heteroaryl), -(C0-C4 alkyl)(C4-C 10 (heterocyclic), C5-C 12 Spiroalkyl, C5-C 12 Spiroalkyl, dimethylphosphoryl, adamantyl, C1-C6 alkoxy, -(C1-C4 alkyl)-SO2-(C1-C4 alkyl), difluoromethylthioalkyl or pentafluorothioalkyl, wherein each R1 is unsubstituted or, when it can be substituted, is substituted by one or more substituents independently selected from the group consisting of halogens, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, methylamine, N,N-dimethylmethylamine, mono-, di- or tri-halomethoxy, mono-, di- or tri-halomethyl and C0-C3 alkyl-pyrrolidinyl, wherein the pyrrolidinyl group is unsubstituted or substituted by one, two or three independently selected halogen atoms, and wherein two R1 groups may be fused to form a ring structure including a portion of A and optionally aromatic, and n is 1, 2, 3, 4, 5 or 6;

[0022] X1, X2, X3, and X4 are each independently N, CH, C-CH3, C-CH2-OH, C-OCH3, C-CH2-OCH3, or C-halogen; and

[0023] R2 is a C1-C6 alkyl, -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C4-C7 heterocycloalkyl), -(C0-C4 alkyl)(C4-C 10 (Bicyclic), each optionally substituted with one or more of halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, cyclopropyl or mono-, di- or tri-halomethyl.

[0024] In some respects, in the compounds of Formula I and pharmaceutically acceptable salts, at least one of X1, X2, X3, and X4 is CH. Sometimes, X1, X2, X3, and X4 are each CH. Alternatively, at least one of X1, X2, X3, and X4 is N. In one embodiment, X1 is N and each of X2, X3, and X4 is CH. In another embodiment, X2 is N and each of X1, X3, and X4 is CH. In some compounds, at least one of X1 and X2 is C-Cl or CF. In other compounds, X1 and X2 are independently selected from the group consisting of C-Cl and CF. In still other compounds, X1 and X2 are independently selected from the group consisting of C-Cl and CF, and X3 and X4 are both CH. Alternatively, X1 and X3 are independently selected from the group consisting of C-Cl and CF. In some compounds, X2 is C-OCH3 or C-CH2-OH.

[0025] In certain compounds of Formula I and pharmaceutically acceptable salts, X2 and X3 are both N. In other compounds, X2 and X3 are both N and X1 and X4 are independently selected from CH, C-Cl, CF, C-OCH3, and C-CH2-OH. In still other compounds, X2 and X3 are both N and X1 and X4 are both CH. In still other compounds, X2 and X3 are both N and X1 and X4 are independently CH, C-Cl, or CF. In some compounds, X1 and X4 are both N. In other compounds, X1 and X4 are both N and X2 and X3 are independently selected from CH, C-Cl, CF, C-OCH3, and C-CH2-OH. In still other compounds, X1 and X4 are both N and X2 and X3 are both CH. In still other compounds, X1 and X4 are both N and X2 and X3 are independently CH, C-Cl, or CF.

[0026] In certain compounds of Formula I and pharmaceutically acceptable salts, X1 and X3 are both N, and X2 and X4 are independently selected from CH, C-Cl, CF, C-OCH3, and C-CH2-OH. In other compounds, X1 and X3 are both N, and X2 and X4 are both CH. In other compounds, X1 and X3 are both N, and X2 and X4 are independently CH, C-Cl, or CF.

[0027] In compounds of formula I and pharmaceutically acceptable salts, or pharmaceutically acceptable salts thereof, A-(R1) n yes

[0028]

[0029] The wavy line indicates the connection to the main chain, and the maximum value of n depends on the number of substituted positions on the A-ring.

[0030] In one respect, in compounds and salts of formula I, A-(R1) n yes

[0031]

[0032] And the maximum value of n depends on the number of substituted positions on the A-ring.

[0033] In one respect, among compounds of formula I and pharmaceutically acceptable salts, A-(R1) n yes:

[0034]

[0035] And the maximum value of n depends on the number of substituted positions on the A-ring.

[0036] In another respect, among compounds of formula I and pharmaceutically acceptable salts, A-(R1) n yes

[0037]

[0038] In one embodiment, n does not exceed 5. In another embodiment, n is 1-3 or 1-2. In another embodiment, when n is 3, each R1 group is independently halogen, methyl, or methoxy. In another embodiment, when n is 2, at least one R1 is halogen, while the other R1 is methyl or methoxy. In another embodiment, n is 2 and one R1 is CF3. In yet another embodiment, n is 2, one R1 is CF3, and the other R1 group is -CH2-piperazinyl, wherein the piperazineyl is optionally substituted with a C1-C4 alkyl group such as methyl or ethyl.

[0039] In some respects, A-(R1) is applicable to compounds of formula I and pharmaceutically acceptable salts. n yes

[0040]

[0041] And the maximum value of n depends on the number of substituted positions on the A-ring.

[0042] In some respects, A-(R1) is applicable to compounds of formula I and pharmaceutically acceptable salts. n yes

[0043]

[0044] And the maximum value of n depends on the number of substituted positions on the A-ring.

[0045] Compounds of Formula I and pharmaceutically acceptable salts encompass compounds of Formulas Ia, Ib, Ic, and Id and pharmaceutically acceptable salts:

[0046]

[0047]

[0048] In the compounds and pharmaceutically acceptable salts of formulas Ia, Ib, Ic and Id, X1, X2, X3 and X4 of formula I are CH.

[0049] Compounds of Formula I and pharmaceutically acceptable salts also include compounds of Formulas Ie, Tf, Ig, Ih, Ii, Ij, Ik, and Il, and their pharmaceutically acceptable salts:

[0050]

[0051] Compounds of formulas Ie to Il and pharmaceutically acceptable salts correspond to such formula I, wherein one of X1, X2, X3 and X4 is N, and the others are CH.

[0052] Compounds of Formula I and pharmaceutically acceptable salts also include compounds of Formulas Im, In, Io, Ip, Iq, Ir, Is, and It, and their pharmaceutically acceptable salts:

[0053]

[0054] Compounds of formulas Im, In, Io, Ip, Iq, Ir, Is, and It, and pharmaceutically acceptable salts, correspond to compounds of formula I, wherein X2 and X3 are both N and X1 and X4 are both CH, or X2 and X3 are both CH and X1 and X4 are both N.

[0055] Compounds of formula I also include compounds of formulas Iu, Iv, Iw, and Ix:

[0056]

[0057] Compounds of formulas Iu, Iv, Iw, and Ix, and pharmaceutically acceptable salts, correspond to compounds of formula I, wherein X1 and X3 are both N, and X2 and X4 are both CH.

[0058] In compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is -C1-C6 alkyl, -C1-C4 alkyl-tetrahydrofuranyl, -C1-C6 hydroxyalkyl, -C1-C6 haloalkyl, -C1-C6 alkyl-O-C1-C2 alkyl, -C3-C6 cycloalkyl, tetrahydrofuranyl, pyranyl, -C1-C4 alkyl-tetrahydropyranyl, -C1-C4 alkyl-C3-C6 cycloalkyl, and aziridine, wherein each is unsubstituted or substituted by one or two independently selected groups from the group consisting of: halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, cyclopropyl, or mono-, di-, or tri-halomethyl. In one embodiment, the -C1-C6 alkyl group is an ethyl group.

[0059] In one respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is tert-butyl, -CH(CH3)-cyclopropyl, -2-tetrahydrofuranyl, -3-tetrahydrofuranyl, -3-pyranyl, -4-pyranyl, -CH(CH3)CH2OH, -C(CH3)2CH2OH, -CH2-(2-tetrahydrofuranyl), -CH2-(3-tetrahydrofuranyl), -(CH2)3OH, -C(CH3)2CH2OCH3, -CH(CH3)CF3, -CH2CF3, -C(CH3)2CH2OCH3, cyclopentyl,

[0060]

[0061] In another respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is

[0062]

[0063] In one respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is

[0064]

[0065] In another respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is

[0066]

[0067] In yet another respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is In another respect, among the compounds of formulas I and Ia to Ix and pharmaceutically acceptable salts, R2 is...

[0068] In yet another aspect, in the compounds of formula I and Ia to Ix and pharmaceutically acceptable salts thereof, R2 is

[0069] In one aspect, the compounds of formula I and pharmaceutically acceptable salts thereof encompass the compounds of formula IIa to IIx and IIIa to IIIx and pharmaceutically acceptable salts thereof:

[0070]

[0071] The compounds of formula IIa, IIb, IIc and IId and pharmaceutically acceptable salts thereof correspond to the compounds of formula I wherein X1, X2, X3 and X4 are CH and R2 is isopropyl.

[0072] The compounds of formula I and pharmaceutically acceptable salts thereof also encompass the compounds of formula IIe, IIf, IIg, IIh, IIi, IIj, IIk, Iil and pharmaceutically acceptable salts thereof:

[0073]

[0074]

[0075] The compounds of formula IIe to IIl and pharmaceutically acceptable salts thereof correspond to the compounds of formula I wherein one of X1, X2, X3 and X4 is N and the others are CH and R2 is isopropyl.

[0076] The compounds of formula I and pharmaceutically acceptable salts thereof also encompass the compounds of formula IIm, IIn, IIo, IIp, IIq, IIr, IIs, IIt and pharmaceutically acceptable salts thereof:

[0077]

[0078] The compounds of formula IIm, IIn, IIo, IIp, IIq, IIr, IIs and IIt and pharmaceutically acceptable salts thereof correspond to the compounds of formula I wherein both X2 and X3 are N and both X1 and X4 are CH, or both X2 and X3 are CH and both X1 and X4 are N, and R2 is isopropyl.

[0079] The compounds of formula I also encompass the compounds of formula IIu, IIv, IIw and IIx:

[0080] [[ID=3,9]]

[0081] Compounds of formulas IIu, IIv, IIw, and IIx, and their pharmaceutically acceptable salts, correspond to compounds of formula I, wherein X1 and X3 are both N, X2 and X4 are both CH, and R2 is isopropyl. Compounds of formula I and their pharmaceutically acceptable salts encompass compounds of formulas IIIa, IIIb, IIIc, and IIId, and their pharmaceutically acceptable salts:

[0082]

[0083] Compounds of formulas IIIa, IIIb, IIIc, and IIId, and pharmaceutically acceptable salts, correspond to compounds of formula I, wherein X1, X2, X3, and X4 are CH, and R2 is

[0084] Compounds of Formula I and pharmaceutically acceptable salts also include compounds of Formulas IIIe, IIIf, IIIg, IIIh, IIIi, IIIj, IIIk, and IIIl, and their pharmaceutically acceptable salts:

[0085]

[0086] Compounds of formulas IIIe to IIIl and pharmaceutically acceptable salts correspond to compounds of formula I, wherein one of X1, X2, X3, and X4 is N, while the others are CH, and R2 is

[0087] Compounds of Formula I and pharmaceutically acceptable salts also encompass compounds of Formulas IIIm, IIIn, IIIo, IIIp, IIIq, IIIr, IIIs, and IIIt, and their pharmaceutically acceptable salts:

[0088]

[0089]

[0090] Compounds of formulas IIIm, IIIn, IIIo, IIIp, IIIq, IIIr, IIIs, and IIIt, and pharmaceutically acceptable salts thereof, correspond to compounds of formula I, wherein X2 and X3 are both N, and X1 and X4 are both CH, or X2 and X3 are both CH, and X1 and X4 are both N, and R2 is

[0091] Compounds of Formula I also include compounds of Formulas IIIu, IIIv, IIIw, and IIIx:

[0092]

[0093] Compounds of formulas IIIu, IIIv, IIIw, and IIIx, and pharmaceutically acceptable salts thereof, correspond to compounds of formula I, wherein X1 and X3 are both N, and X2 and X4 are both CH, or X2 and X4 are both N, and X1 and X3 are both CH, and R2 is

[0094] In one aspect, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently hydrogen, C1-C6 alkyl, C4-C8 bridged bicycloalkyl, C5-C6 alkyl, C6-C6 alkyl, C7-C8 alkyl, C8-C9 alkyl, C1 ... 12 Spiroane, wherein each R1 is optionally substituted with one or more of a halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, methylamine, N,N-dimethylmethylamine, or mono-, di-, or tri-halomethyl.

[0095] In another aspect, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently 2-chloro-4-methyl-phenyl, 2,4-dichloro-3-fluoro-phenyl, 2-methyl-4-chloro-phenyl, 2-chloro-4-methoxy-phenyl, 2-chloro-4-trifluoromethyl-phenyl, 2-methoxy-4-chloro-phenyl, 2,2-dimethylpropyl; 2-chloro-4-fluoro-phenyl; 2,4-dichloro-phenyl Phenyl; 1,1-dimethyl-2,2,2-trifluoroethyl; 1,1-dimethylethyl; 1,1-dimethylpropyl; trifluoromethyl; 1,1-dimethyl-2,2-difluoropropyl; 1,1-dimethyl-3,3,3-trifluoropropyl; 1-methylcyclopropyl; (1-methylcyclopropyl)methyl; 3-methylbicyclo[1.1.1]pentan-1-yl; 3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl; or (3,3-dimethylcyclobutyl)methyl.

[0096] In another respect, in the compounds of formula I, Ia to Ix, IIa to IIx and IIIa to IIIx and pharmaceutically acceptable salts, each R1 is independently a C4-C8 bridged bicycloalkyl group that is unsubstituted or substituted by one or two groups independently selected from the group consisting of C1-C4 alkyl, CF3, halogen and CN.

[0097] In another respect, in the compounds of formula I, Ia to Ix, IIa to IIx and IIIa to IIIx and pharmaceutically acceptable salts, each R1 is independently a C4-C8 bridged bicycloalkyl group that is unsubstituted or substituted by one or two groups independently selected from the group consisting of methyl, ethyl, CF3, halogen and CN.

[0098] In some respects, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently a bridged C4-C8 bicycloalkyl group.

[0099]

[0100] In other respects, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently C5-C6. 12 Spiroane is an unsubstituted or substituted group, or a group selected independently from the group consisting of: =O, -OH, C1-C4 alkyl, -CF3, and halogen, each R1 being independently C5-C6. 12 Hexaspiranes are unsubstituted or substituted groups selected independently from the group consisting of: =O, -OH, C1-C4 alkyl, -CF3, and halogens. (C5-C) 12 Spiroane or C5-C 12 Examples of heterospiranes include

[0101]

[0102]

[0103] In one respect, among the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, n is 1 and R1 is C5-C6. 12 Spiroane is an unsubstituted or substituted group selected independently from the group consisting of: =O, -OH, methyl, ethyl, CF3 and halogen.

[0104] In yet another aspect, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently a halogen, C1-C6 alkyl, -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(phenyl), -(C1-C4 alkyl)(C5-C6 heteroalkyl), -(C0-C4 heteroalkyl)(C3-C7 cycloalkyl) or -(C0-C4 alkyl)(C5-C6 heteroaryl), wherein each R1 is optionally substituted by one or more substituents independently selected from the group consisting of halogens, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, methylamine, N,N-dimethylmethylamine, and mono-, di- or tri-halomethyl.

[0105] In one aspect, in the compounds of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx and pharmaceutically acceptable salts, each R1 is unsubstituted. In another aspect, each R1 is substituted by no more than four groups, or no more than three groups, or no more than two groups, or only one group.

[0106] In another respect, among the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently -CH2C(CH3)3, -CH(CH2CH3)2, -CF2CH3, -CH2CH2CF3, -C(CH3)2F, -C(CH3)2CF3, -CH2-cyclopentyl, -CH2-cyclohexyl, -CH2-cyclopropyl, -CH2-cyclobutyl, -CH2-morpholinyl, -CH2-pyrrolidinyl, phenyl, naphthyl, pyran-4-yl, 4-Methylphenyl, 4-methoxyphenyl, 2-fluorophenyl, 2-fluoro-4-chlorophenyl, 3-trifluoromethylphenyl, 3-fluoro-5-trifluoromethylphenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 2,4-dimethylphenyl, 4-chlorophenyl, 2-methylphenyl, 3-fluorophenyl, 3-chlorophenyl, 2-methoxyphenyl, 4-difluoromethoxyphenyl, -C(CH3)2SO2CH3, 4-fluorophenyl, 2,4-difluorophenyl, 2,4-dichlorophenyl, 3-chlorophenyl, methyl, ethyl, tert-butyl, isopropyl, trifluoromethyl, 3- Methoxyphenyl, 3-bromophenyl, 4-bromophenyl, 2,4,6-trifluoromethylphenyl, 3-tetrahydrofuranyl, -C(CH3)2CH2CH3, -C(CH3)2OH, 3,3-dimethylcyclobut-1-yl, 2,3-dichlorobenzyl, 3,3-difluoromethylcyclobutyl, 2,2-dimethylcyclopropyl-1-yl, 2,2-difluorocyclopropyl, N-methylimidazolyl, 2-methyl-4-chlorophenyl, 2,4-dichloro-3-fluorophenyl, -CH2OCH3, -CH2CF3, -CF2CH3, -CF2C(CH3) 3. -CH(CH3)cyclopropyl, 2,6-difluorophenyl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 3-chloropyridin-2-yl, 2-methylpyridin-3-yl, 4-methylpyridin-3-yl, 3,5-dichloropyridin-2-yl, 3,3-difluorocyclopentyl, fluorine, chlorine, bromine, 2-methoxy-4-chlorophenyl, 2-fluoro-4-chlorophenyl, 3-fluoro-4-chlorophenyl, benzyl, piperidinyl, 4-methyl-piperidinyl, 4-methoxy-piperidinyl, N-methylpyrazol-3-yl, -C(O)-piperidinyl, -OCHF2,

[0107]

[0108] In one respect, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, R1 is 2,2-dimethylpropyl or

[0109] In another respect, in the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, R1 is 2,2-dimethylpropyl.

[0110] In another aspect, among the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx, and IIIa to IIIx, R1 is...

[0111] In another respect, among the compounds and pharmaceutically acceptable salts of formulas I, Ia to Ix, IIa to IIx and IIIa to IIIx, each R1 is independently...

[0112]

[0113] In another respect, in the compounds of formula I, Ia to Ix, IIa to IIx and IIIa to IIIx and pharmaceutically acceptable salts, R1 is a C1-C6 alkyl group that is unsubstituted or substituted by one or more substituents independently selected from the group consisting of halogen, cyano, hydroxy, oxo, methyl, methoxy, hydroxymethyl, ethyl, ethoxy, hydroxyethyl, methylamine, N,N-dimethylmethylamine, and mono-, di- and tri-halomethyl.

[0114] In another respect, the compounds of formula I and pharmaceutically acceptable salts are:

[0115]

[0116]

[0117] In yet another respect, the compounds of formula I and pharmaceutically acceptable salts are:

[0118]

[0119]

[0120] definition

[0121] The specific chemical nomenclature rules used in this article are intended to be familiar to those skilled in the art of chemistry. Some terms have been defined specifically for clarity.

[0122] As used herein, the term alkyl refers to a saturated straight-chain or branched monovalent hydrocarbon group having one to six atoms, e.g., “C1-C6 alkyl”. In cases indicating zero, e.g., C0-C4 or C0-C3 alkyl, the component with substituents may be absent; thus, if the C5 heterocyclic alkyl substituent is at position R2 in formula (I), the C5 heterocyclic alkyl substituent may be described by a -(C0-C4 alkyl)(C3-C7 heterocyclic alkyl) substituent as described with respect to R2 (i.e., the substituent would be -(C0)(C5 heterocyclic alkyl)). Examples include, but are not limited to, methyl, ethyl, propyl, 1-propyl, isopropyl, and butyl. Similarly, as used herein, the term heteroalkyl refers to a saturated straight-chain or branched monovalent alkyl molecule as defined herein, containing one or more heteroatoms replacing carbon atoms in the alkyl chain. Typically, no more than four heteroatoms or no more than three heteroatoms are present in the heteroalkyl group.

[0123] The term aryl, as used herein, denotes a group containing six to ten carbon atoms (e.g., C6-C). 10 A heteroaryl group is a functional group or substituent of an aromatic ring having five to six carbon atoms (e.g., C5-C6 aryl) and no heteroatoms. Examples of aryl groups include phenyl and naphthyl. The term heteroaryl as used herein refers to a functional group or substituent derived from an aromatic ring containing a carbon atom and one or more heteroatoms (e.g., nitrogen, oxygen, or sulfur) as part of the aromatic ring. Examples include rings having five or six atoms, wherein at least one atom is a heteroatom. Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, furanyl, pyrroleyl, thiopheneyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, and thiazolyl.

[0124] As used herein, the term C3-C7 cycloalkyl refers to a cycloalkyl molecule containing three to seven carbon atoms. Examples of C3-C7 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, and cyclopentyl. Similarly, as used herein, the terms heterocycloalkyl and cyclohexaalkyl refer to cycloalkyl molecules as defined herein, containing three to seven atoms in total, wherein at least one atom is a heteroatom. Heterocycloalkyl or cyclohexaalkyl groups can be fused to other groups, such as phenyl groups. For example, piperidine fused to a phenyl group can form tetrahydroisoquinoline or tetrahydroquinoline.

[0125] As used herein, the terms bicyclic, bicyclol, and bicycloalkyl refer to groups having two or more fused or bridged rings. (C4-C) 10 Second Ring Road, C4-C 10 Dicyclic and C4-C 10Bicyclic alkyl groups contain four to ten carbon atoms. When the bicyclic groups are fused, the two rings share two adjacent atoms. When the bicyclic groups are bridged (also called bridged bicyclic alkyl groups), the two rings share three or more atoms. Bridged bicyclic alkyl groups can have four to ten carbon atoms or four to eight carbon atoms. Such groups can be referred to as "C4-C" respectively. 10 "Bridged bicycloalkyl" or "C4-C8 bridged bicycloalkyl". The bicyclic molecule can be entirely aliphatic, entirely aromatic, or a mixture of aromatic and aliphatic. The term C4-C8 is also used. 10 Heterocyclic bicycles refer to C4-C bicycles as defined, which also include one or more heteroatoms. 10 Bicyclic groups. Examples of bridging bicyclic alkyl groups that can be used in compounds of formula (I) include, but are not limited to:

[0126]

[0127] C4-C 10 Examples of heterocyclic bicycles include, but are not limited to:

[0128]

[0129] The term C5-C used in this article 12 Spiroalkyl or spiroalkyl denotes a spirocyclic ring system. (C5-C) 12 Spiroalkyl, C5-C 12 Spiroane and C5-C 12 Spiroane refers to a spirocyclic ring system having 5 to 12 carbon rings. The spirocyclic ring system contains at least two and at most four rings, wherein at least two rings must be linked by a helical link. As described herein, C5-C 12 Spiroane can be unsubstituted or substituted.

[0130] C5-C 12 Heterospirolyl refers to a spirocyclic ring system having 5 to 12 ring atoms, wherein at least one ring atom is a heteroatom, i.e., not carbon. The heterospirilyl ring system contains two to four rings, wherein at least two rings must be linked by a spiro link. As described herein, C5-C 12 Heterospinanes can be unsubstituted or substituted. Substituted and unsubstituted C5-C... 12 Spiroane and C5-C 12 Examples of heterospiranes include, but are not limited to:

[0131]

[0132] As used in this article, the term halogen (or halo) refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0133] The term oxo, as used in this article, refers to oxygen bonded to a carbon atom by a double bond, which can be represented as =O.

[0134] Compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx may be deuterated or triturated at one or more sites, and such deuterated or triturated forms are part of this invention. Deuteration involves replacing one or more hydrogen atoms of a drug with a deuterium atom in an attempt to slow down CYP-mediated drug metabolism or reduce the formation of unwanted metabolites. Trituration makes the molecule radioactive and allows researchers to determine the deposition sites and metabolic pathways of drugs in vivo.

[0135] The compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx described herein can form pharmaceutically acceptable salts. It is intended to include such pharmaceutically acceptable salts of compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx. Pharmaceutically acceptable salts and common methods for preparing them are well known in the art (see, e.g., P. Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection and Use, 2nd revised edition (Wiley-VCH, 2011); SMBerge et al., “Pharmaceutical Salts,” Journal of Pharmaceutical Sciences, Vol. 66, No. 1, January 1977).

[0136] Compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIX as described herein are generally effective over a wide dose range. For example, daily doses range from about 0.01 mg / kg to about 200 mg / kg. Furthermore, compounds of formula I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein may be administered, for example, in amounts of about 0.05 mg / kg to about 150, about 0.1 mg / kg to about 150, about 0.5 mg / kg to about 150 mg / kg, about 1 mg / kg to about 150 mg / kg, about 5 mg / kg to about 100 mg / kg, about 20 mg / kg to about 40 mg / kg, about 25 mg / kg to about 35 mg / kg, about 50 mg / kg to about 70 mg / kg, about 55 mg / kg to about 65 mg / kg, about 90 mg / kg to about 110 mg / kg, about 95 mg / kg to about 105 mg / kg, or about 50 mg / kg to about 150 mg / kg.

[0137] Alternatively, compounds of formula I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein may be administered, for example, in amounts of about 0.1 mg / kg to about 15 mg / kg, or about 0.5 mg / kg to about 10 mg / kg, or 1 mg / kg to about 9 mg / kg, or about 2 mg / kg to about 8 mg / kg, or about 3 mg / kg to 7 mg / kg. Alternatively, dosages of approximately 1 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1.7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 3 mg / kg, and 3.1 mg / kg can be used. kg, 3.2mg / kg, 3.3mg / kg, 3.4mg / kg, 3.5mg / kg, 3.6mg / kg, 3.7mg / kg, 3.8mg / kg, 3.9mg / kg, 4, mg / kg, 4.1mg / kg, 4.2mg / k g, 4.3mg / kg, 4.4mg / kg, 4.5mg / kg, 4.6mg / kg, 4.7mg / kg, 4.8mg / kg, 4.9mg / kg, 5mg / kg, 5.1mg / kg, 5.2mg / kg, 5.3mg / kg, 5 .4mg / kg, 5.5mg / kg, 5.6mg / kg, 5.7mg / kg, 5.8mg / kg, 5.9mg / kg, 6mg / kg, 6.1mg / kg, 6.2mg / kg, 6.3mg / kg, 6.4mg / kg, 6.5 mg / kg, 6.6mg / kg, 6.7mg / kg, 6.8mg / kg, 6.9mg / kg, 7mg / kg, 7.1mg / kg, 7.2mg / kg, 7.3mg / kg, 7.4mg / kg, 7.5mg / kg, 7.6mg / kg, 7.7mg / kg, 7.8mg / kg, 7.9mg / kg, 8mg / kg, 8.1mg / kg, 8.2mg / kg, 8.3mg / kg, 8.4mg / kg, 8.5mg / kg, 8.6mg / kg, 8.7mg / kg , 8.8mg / kg, 8.9mg / kg, 9mg / kg, 9.1mg / kg, 9.2mg / kg, 9.3mg / kg, 9.4mg / kg, 9.5mg / kg, 9.6mg / kg, 9.7mg / kg, 9.8mg / kg, 9.9 mg / kg, approximately 10 mg / kg, approximately 15 mg / kg, approximately 20 mg / kg, approximately 25 mg / kg, approximately 30 mg / kg, approximately 35 mg / kg, approximately 40 mg / kg, approximately 45 mg / kg, approximately 50 mg / kg, approximately 55 mg / kg, approximately 60 mg / kg, approximately 65 mg / kg, approximately 70 mg / kg, approximately 75 mg / kg, approximately 80 mg / kg, approximately 85 mg / kg, approximately 90 mg / kg, approximately 95 mg / kg, approximately 100 mg / kg, approximately 105 mg / kg, approximately 110 mg / kg, approximately 115 mg / kg, approximately Compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein may be administered in amounts of 120 mg / kg, approximately 125 mg / kg, approximately 130 mg / kg, approximately 135 mg / kg, approximately 140 mg / kg, approximately 145 mg / kg, approximately 150 mg / kg, approximately 155 mg / kg, approximately 160 mg / kg, approximately 165 mg / kg, approximately 170 mg / kg, approximately 175 mg / kg, approximately 180 mg / kg, approximately 185 mg / kg, approximately 190 mg / kg, approximately 195 mg / kg, or approximately 200 mg / kg. Daily administration may be once daily or in multiple doses, for example, twice daily (BID). For patients weighing >50 kg, administration may be 160 mg orally twice daily, or for patients weighing <50 kg, administration may be 120 mg orally twice daily. It should be understood that the actual amount of compound administered will be determined by the physician based on relevant circumstances, including the condition to be treated, the chosen route of administration, one or more actual compounds administered, the individual patient's age, weight, and response, and the severity of the patient's symptoms. Possible central nervous system involvement / the need for increased central nervous system exposure is also typically considered.

[0138] Compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein can be formulated into pharmaceutical compositions that can be administered via a variety of routes. Such pharmaceutical compositions and methods of their preparation are well known in the art (see, for example, Remington: The Science and Practice of Pharmacy (A. Gennaro et al., eds., 21st ed., Mack Publishing Co., 2005)). Specifically, compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx, or pharmaceutically acceptable salts thereof, as described herein, can be combined with one or more pharmaceutically acceptable carriers, diluents, or excipients. More specifically, compounds described herein by formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx can be formulated into pharmaceutical compositions. Furthermore, compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx, or pharmaceutically acceptable salts thereof, as described herein, can be combined with one or more other therapeutic agents. For example, compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx, or pharmaceutically acceptable salts thereof, as described herein, may be components in pharmaceutical compositions for the treatment of cancer, in combination with one or more pharmaceutically acceptable carriers, diluents, or excipients, and optionally with one or more additional therapeutic agents. In concomitant therapy, compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx may be administered in combination with a second therapeutic agent when a patient is detected to have multiple driver mutations / alterations—such as when an EGFR mutation and clinically significant RET alterations are detected. Examples of combinations include, but are not limited to, compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx and osimertinib. Other examples of combinations include compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx in combination with one or more platinum-based chemotherapeutic acids and / or non-platinum-based chemotherapeutic agents. Pharmaceutical compositions containing compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein, or pharmaceutically acceptable salts thereof, may be used in the methods described herein.

[0139] The term “treating” (or “treat” or “treatment”) as used in this article means to suppress, slow down, stop or reverse the progression or severity of an existing symptom, condition or disorder.

[0140] As used in this article, the term "irritable bowel syndrome" or "IBS" refers to gastrointestinal disorders, including but not limited to diarrhea-predominant, constipation-predominant, or alternating stool patterns, functional bloating, functional constipation, functional diarrhea, unspecified functional bowel disorder, functional abdominal pain syndrome, chronic idiopathic constipation, functional esophageal disorder, functional gastroduodenal disorder, functional anorectal pain, and inflammatory bowel disease.

[0141] The terms “cancer” and “cancerous” as used in this article refer to or describe a physiological condition in a patient that is typically characterized by disordered cell proliferation. Both benign and malignant cancers are included in this definition. “Early-stage cancer” or “early-stage tumor” refers to cancer that is not late-stage or metastatic, or that is classified as stage 0, stage I, or stage II cancer. Examples of cancers include, but are not limited to, lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), thyroid cancer (e.g., papillary thyroid carcinoma, medullary thyroid carcinoma, differentiated thyroid carcinoma, recurrent thyroid carcinoma, or refractory differentiated thyroid carcinoma), thyroid adenoma, endocrine gland vegetations, lung adenocarcinoma, bronchioloalveolar cell carcinoma, type 2A or 2B multiple endocrine neoplasms (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer, mammary cancer, mammary carcinoma, mammary neoplasm, colorectal cancer (e.g., metastatic colorectal cancer), papillary renal cell carcinoma, ganglioneuroma of the gastrointestinal mucosa, inflammatory myofibroblastoma, and cervical cancer.

[0142] As used herein, the term “RET-related disease or disorder” refers to a disease or disorder that is associated with or has the following regulatory abnormalities: abnormal regulation of the expression, activity, or level of the RET gene, RET kinase (also referred to herein as RET kinase protein), or any of them (e.g., one or more) (e.g., any type of regulatory abnormality of the expression, activity, or level of the RET gene, RET kinase, RET kinase domain, or any of them described herein).

[0143] As used herein, the term "RET-related cancer" refers to cancer that is associated with or has abnormal regulation of the expression, activity, or level of the RET gene, RET kinase (also referred to herein as RET kinase protein), or any of these. Non-limiting examples of RET-related cancers are described herein.

[0144] The phrase “abnormal regulation of the expression, activity, or level of the RET gene, RET kinase, or any of them” indicates a gene mutation or alteration. Examples of such gene mutations that cause abnormal regulation of the expression, activity, or level of the RET gene, RET kinase, or any of them include, but are not limited to: RET gene translocations that result in the expression of fusion proteins; deletions in the RET gene that result in the expression of RET proteins with at least one amino acid deletion compared to wild-type RET proteins; mutations in the RET gene that result in the expression of RET proteins with one or more point mutations; or alternative spliced ​​versions of RET mRNA that result in RET proteins with at least one amino acid deletion compared to wild-type RET proteins; or RET gene amplification that results in the overexpression of RET proteins; or autocrine activity resulting from overexpression of RET genes in cells that leads to a pathogenic increase in the activity of the kinase domains of RET proteins in cells (e.g., the constituent active kinase domains of RET proteins). Abnormal regulation of the expression, activity, or level of a RET gene, RET protein, or any of them can be a mutation in a RET gene that encodes a RET protein with constitutive activity or increased activity compared to a protein encoded by a RET gene excluding the mutation. For example, abnormal regulation of the expression, activity, or level of a RET gene, RET protein, or any of them can be the result of a gene or chromosomal translocation leading to the expression of a fusion protein containing a second half / part of a RET gene and a first half of some other non-RET gene. In some embodiments, abnormal regulation of the expression, activity, or level of a RET gene, RET protein, or any of them can be the result of a gene translocation between one RET gene and another non-RET gene.

[0145] Methods are provided for treating RET-related diseases or disorders, particularly IBS or cancers with aberrant RET expression, using compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein. Examples of cancers with aberrant RET expression include cancers caused by abnormal regulation of the expression, activity, or level of RET genes, RET kinases, or any of them. One method of treating RET-related diseases or disorders such as IBS or cancer involves administering a therapeutically effective amount of a compound of formula I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx, or a pharmaceutically acceptable salt thereof, to a patient in need. Another method of treating RET-related diseases or disorders such as IBS or cancer involves: a) detecting abnormal regulation of the expression, activity, or level of RET genes, RET kinases, or any of them in a sample from a patient; and b) administering a therapeutically effective amount of a compound of formula I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx, or a pharmaceutically acceptable salt thereof.

[0146] Abnormal regulation of the expression, activity, or level of RET genes, RET kinases, or any of them can be the result of one or more chromosomal translocations or inversions that result in RET gene fusions (i.e., the gene translocations result in the expression of a fusion protein containing residues from non-RET chaperone proteins and including a minimal amount of a functional RET kinase domain). Non-limiting examples of RET fusion chaperones and their associated cancers include: ACBD5 (papillary thyroid carcinoma); AFAP1 (NSCLC); AFAP1L2 (papillary thyroid carcinoma); AKAP13 (papillary thyroid carcinoma); BCR (chronic myelomonocytic leukemia); C10orf118 (papillary thyroid carcinoma); CCDC6 (also known as PTC1, D10S170, or H4) (NSCLC, colon cancer, papillary thyroid carcinoma, adenocarcinoma, lung adenocarcinoma, metastatic colorectal cancer, adenosquamous carcinoma, breast cancer); CCDC88C (NSCLC); CCDC186-R ET, CEP55 (diffuse gastric cancer); CGNL1 (pancreatic cancer); CLIP1 (adenocarcinoma); CUX1 (lung adenocarcinoma); DLG5 (non-anaplastic thyroid cancer); DOCK1 (NSCLC); EML4 (papillary thyroid carcinoma); ERC1 (also known as ELKS) (papillary thyroid carcinoma, breast cancer); ETV6 (salivary gland carcinoma); FGFR1OP (CMML, primary myelofibrosis with secondary acute myeloid leukemia); FKBP15 (papillary thyroid carcinoma); FOXP4 (lung adenocarcinoma); FRMD4A (NSCLC); GOLGA5 (also known as PTC5) (Papillary thyroid carcinoma, spitzoid growths); H4L (various); HOOK3 (papillary thyroid carcinoma); HRH4-RET (thyroid carcinoma and / or papillary thyroid carcinoma); HTIF1 (various); KIAA1217 (also known as SKT) (papillary thyroid carcinoma, lung adenocarcinoma, NSCLC); KIAA1468 (also known as PTC9 and RFG9) (papillary thyroid carcinoma, lung adenocarcinoma); KIF13A (NSCLC); KIF5B (NSCLC, ovarian cancer, spitzoid growths, lung adenocarcinoma, adenosquamous carcinoma); KTN1 (also known as PTC) 8)(Papillary thyroid carcinoma); MBD1 (also known as PCM1) (Papillary thyroid carcinoma); MPRIP (NSCLC); MYH10 (Infantile myofibromatosis); MYH13 (Medullary thyroid carcinoma); NCOA4 (also known as PTC3, ELE1, and RFG) (Papillary thyroid carcinoma, NSCLC, colon cancer, salivary gland cancer, metastatic colorectal cancer, lung adenocarcinoma, adenosquamous carcinoma of papillary thyroid carcinoma, diffuse sclerosing variant, breast cancer, acinar cell carcinoma, mammary gland-like secretory carcinoma); OLFM4 (Small bowel cancer); PARD3 (NSCLC); PCM1 (Papillary thyroid carcinoma);PIBF1 (Bronchioloalveolar Cell Carcinoma); PICALM (NSCLC); PPFIBP2 (Papillary Thyroid Carcinoma); PRKAR1A (also known as PTC2) (Papillary Thyroid Carcinoma); PTC1ex9 (a novel CCDC6 rearrangement) (Metastatic Papillary Thyroid Carcinoma); PTC4 (a novel NCO4 / ELE1 rearrangement) (Papillary Thyroid Carcinoma); RAB6 1P2 (Papillary Thyroid Carcinoma); RASAL2 (Sarcoma); RASGEF1A (Breast Cancer); RBPMS (NSCLC); RFG8 (Papillary Thyroid Carcinoma); RRBP1 (Colorectal Cancer); RUFY1 (Colorectal Cancer); RUFY2 (NSCLC; Papillary Thyroid Carcinoma); RUFY3 (Papillary Thyroid Carcinoma); SLC12A2 (NSCLC); SORBS2 (Papillary Thyroid Carcinoma); SPECC1L (Papillary Thyroid Carcinoma; Thyroid Cancer); SQS TM1 (papillary thyroid carcinoma); TAF3 (pancreatic cancer); TBL1XR1 (papillary thyroid carcinoma, thyroid cancer); TFG (pancreatic cancer); TIF1G (various types); TRIM24 (also known as PTC6) (papillary thyroid carcinoma); TRIM27 (also known as RFP) (papillary thyroid carcinoma); AKAP13 (papillary thyroid carcinoma); TRIM33 (also known as PTC7 and RFG7) (NSCLC, papillary thyroid carcinoma); and UEVLD (papillary thyroid carcinoma). Fusion proteins can be, for example, KIF5B-RET. Fusions identified in single tumors include CCDC186-RET, ERC1-RET, TN1-RET, and RUFY3-RET. Other RET fusion proteins may not be included in this list or are not yet known; however, compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, and their methods of use, are expected to be effective inhibitors.

[0147] Abnormal regulation of the expression, activity, or level of the RET gene, RET kinase, or any of them can be caused by one or more point mutations, insertions, or deletions in the RET gene (compared to wild-type RET). For reference, the sequence of the mature human RET protein (SEQ ID NO: 1) is provided herein:

[0148]

[0149] Non-restrictive examples of point mutations, insertions, or deletions in potentially activating RET kinase proteins compared to wild-type RET kinases can occur at the following amino acid positions: 2, 3, 4, 5, 6, 7, 8, 11, 12, 13, 20, 32 (e.g., S32L), 34 (e.g., D34S), 40 (e.g., L40P), 56 (e.g., L56M), 64 (e.g., P64L), 67 (e.g., R67H), 114 (e.g., R114H), 136 (e.g., glutamate to stop codon), 145 (e.g., V145G), 180 (e.g., arginine to stop codon), 200, 292 (e.g., V292M), 294, 321 (e.g., glutamate to stop codon), 200, 292 (e.g., V292M), 294, 321 (e.g., glutamate to stop codon), 200, 292 (e.g., glutamate to stop codon ... For example, G321R), 330 (e.g., R330Q), 338 (e.g., T338I), 360 (e.g., R360W), 373 (e.g., alanine to valine (p.A373V) frameshift), D378-G385delinsE, 393 (e.g., F393L), 423 (e.g., G423R), 432, 446 (e.g., G446R), 505-506 (6-base pair germline deletion within the framework in exon 7), 510 (e.g., A510V), 511 (e.g., E511K), 513 (e.g., G513D), 515 (e.g., C515R, C515S, C515W), 52 5 (e.g., R525W), 531 (e.g., C531R or 9-base pair replication), 532 (e.g., replication), 533 (e.g., G533C, G533S), 550 (e.g., G550E), 591 (e.g., V591I), 593 (e.g., G593E), 595 (e.g., E595D and E595A), 600 (e.g., R600Q), 602 (e.g., I602V), 603 (e.g., K603Q, K603E), 606 (e.g., Y606C), 609 (e.g., C609Y, C609S, C609G, C609R, C609F, C609W, C609C), 61 1 (e.g., C611R, C611S, C611G, C611Y, C611F, C611W), 616 (e.g., E616Q), 618 (e.g., C618S, C618Y, C618R, C618T, C618G, C618F, C618W), 620 (e.g., C620S, C620W, C620R, C620G, C620L, C620Y, C620F), 623 (e.g., E623K), 624 (e.g., D624N), 630 (e.g., C630A, C630R, C630S, C630Y, C630F, C630W, C630G), 631 (e.g.,D631N, D631Y, D631A, D631G, D631V, D631E), 632 (e.g., E632K, E632G), 632-633 (6-base pair deletion within the framework of exon 11), 633 (e.g., 9-base pair replication), 634 (e.g., C634W, C634Y, C634S, C634R, C634F, C634G, C634L, C634A, or C634T, or insertion of ELCR, or 12-base pair replication, or combination with A640G, A641A, or A641T) (e.g., causing MTC), 634 / 852 (e.g., C634R / I852M), 635 (e.g., R63... 5G), 636 (e.g., T636P, T636M), 648 (e.g., V648I), 649 (e.g., S649L), 664 (e.g., A664D), 665 (e.g., H665Q), 666 (e.g., K666E, K666M, K666N, K666R), 675 (T675T, silent nucleotide change), 686 (e.g., S686N), 689 (e.g., S689T), 691 (e.g., G691S), 694 (e.g., R694Q), 700 (e.g., M700L), 706 (e.g., V706M, V706A), 713 splice variant (e.g., E713K), 732 (e.g., E... 732K), 736 (e.g., G736R), 748 (e.g., G748C), 765 (e.g., S765P), 766 (e.g., P766S, P766M6), 768 (e.g., E768Q, E768D), 769 (e.g., L769L), 770 (e.g., R770Q), 771 (e.g., D771N), 777 (e.g., N777S), 778 (e.g., V778I), 781 (e.g., Q781R), 788 (e.g., I788I), 790 (e.g., L790F, L790T), 791 (e.g., Y791F, Y791N), 791 / 852 (e.g., Y791F / I852) M), 802, 804 (e.g., V804L, V804M, V804E, V804G, V804S) (e.g., causing MTC), 804 / 918 (e.g., V804M / M918T, V804L / M918T), 805 (e.g., E805K), 804 / 805 (e.g., V804M / E805K), 806 (e.g., Y806F, Y806C, Y806H, Y806Y), 810 (e.g., G810R, G810S, G810A, G810C, G810V), 818 (e.g., E818K), 819 (e.g., S819I), 823 (e.g., G823E), 826 (e.g.,Y826M, Y826S), 833 (e.g., R833C), 841 (e.g., P841L, P841P), 843 (e.g., E843D), 844 (e.g., R844W, R844Q, R844L), 848 (e.g., M848T), 852 (e.g., I852M), 865 (e.g., L865V), 870 (e.g., L870F), 873 (e.g., R873W), 876 (e.g., A876V), 881 (e.g., L881) V), 882, 883 (e.g., A883F, A883P, A883S, A883T, A883Y), 884 (e.g., E884K), 886 (e.g., R886W), 891 (e.g., S891A), 897 (e.g., R897Q), 898 (e.g., D898V), 900 (e.g., Y900F), 901 (e.g., E901K), 904 (e.g., S904F, S904C), 905 (e.g., Y905F), 907 (e.g., ... K907E, K907M), 908 (e.g., R908K), 911 (e.g., G911D), 912 (e.g., R912P, R912Q), 918 (e.g., M918T, M918V, M918L, M918R) (e.g., causing MTC), 919 (e.g., A919V), 921 (e.g., E921K), 922 (e.g., S922P, S922Y), 930 (e.g., T930M), 961 (e.g., F961L), 972 (e.g., R972G), 981 (e.g., Y981F), 982 (e.g., R982C), 1009 (e.g., M1009V), 1015 (e.g., Y1015F), 1017 (e.g., D1017N), 1041 (e.g., V1041G), 1064 (e.g., M1064T), 1096 (e.g., Y1096F), in-frame deletions in exons 6 and 11, a 3bp in-frame deletion in exon 15, nucleotide position 2136+2 (e.g.,2136+2T>G), del632-636ins6, and RET-extracellular cysteine ​​mutations (defined as mutations including at least one of the following cysteine ​​residues: 609, 611, 618, 620, 630, or 634). Other mutations include D631-liter633delinsE, E632-liter633del, A883F, D631-liter633delinsV, L790F, D898-E901del, D898_E901del+D903_S904delinsEP, K666 N, T636-V637insCRT, and D378-G385delinsE. Other mutations include D631-liter633delinsE, E632-liter633del, A883F, D631-liter633delinsV, L790F, D898-E901del, D898_E901del+D903S904delinsEP, K666 N, T636-V637insCRT, and D378-G385delinsE. The RET kinase protein point mutations / insertions / deletions can be, for example, M918T, M918V, C634W, V804L, or V804M. Other RET kinase protein point mutations / insertions / deletions may not be included in this list or are not yet known; however, compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, and their methods of use, are expected to be effective inhibitors.

[0150] Abnormal regulation of the expression, activity, or level of RET genes, RET kinases, or any of them may also include splicing variations in RET mRNA that result in the expression of an alternating splice variant of RET with at least one residue deletion (compared to wild-type RET kinase), said deletion resulting in the compositional activity of the RET kinase domain.

[0151] As defined herein, “RET kinase inhibitor” refers to a compound that inhibits RET activity using biological assays such as those described in the examples below.

[0152] In some cases, RET kinases containing mutations, insertions, or deletions are more resistant to inhibition of their phosphotransferase activity by one or more first RET kinase inhibitors compared to wild-type RET kinases or RET kinases not containing the same mutation. Such mutations may not reduce the sensitivity of cancer cells or tumors containing RET kinases to treatment with compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein (e.g., compared to cancer cells or tumors not containing a specific RET inhibitor resistance mutation). In these cases, when a first RET kinase inhibitor is present, RET inhibitor resistance mutations can result in increased V... max , decrease K m (Regarding ATP) and increased K D (For first RET kinase inhibitors) one or more RET kinases.

[0153] In other cases, RET kinases containing mutations, insertions, or deletions exhibit increased resistance to compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, compared to wild-type RET kinases or RET kinases without the same mutations. In such cases, in the presence of compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, RET inhibitor resistance mutations can result in increased resistance to V compared to wild-type RET kinases or RET kinases without the same mutations in the presence of the same compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein. max , decrease K m and the decrease of K D One or more RET kinases.

[0154] Examples of RET inhibitor resistance mutations can include point mutations, insertions, or deletions in or near the ATP-binding site in the tertiary structure of RET kinases, including but not limited to gatekeeper residues, P-loop residues, residues in or near the DFG motif, and amino acid residues at the solvent front of the ATP crack. Further examples of these types of mutations include changes in residues that may affect enzyme activity and / or drug binding, including but not limited to residues in the activation loop, residues near or interacting with the activation loop, residues that contribute to active or inactive enzyme conformations, and changes including mutations, deletions, and insertions in the loop preceding the C-helix and in the C-helix. Mutations at this location are known to produce specific residues or residue regions that generate RET inhibitor resistance, including but not limited to the following amino acids (based on the human wild-type RET protein sequence (SEQ ID NO: 1)): 732 (e.g., E732K); 788 (e.g., I788N); 804 (e.g., V804M, V804L, V804E); 804 / 805 (e.g., V804M / E805K); 806 (e.g., Y806C, Y806E, Y806S, Y806H, Y806N); 810 (e.g., G810A, G810C, G810R, G810S, G810V); and 865 (e.g., L865V). Other examples of RET inhibitor resistance mutation sites include, but are not limited to, the following amino acids (based on the human wild-type RET protein sequence (SEQ ID NO: 1)): 732 (e.g., E732K); 788 (e.g., I788N); 804 (e.g., V804M, V804L, V804E); 804 / 805 (e.g., V804M / E805K); 806 (e.g., Y806C, Y806E, Y806S, Y806H, Y806N); 810 (e.g., G810A, G810C, G810R, G810S, G810V); and 865 (e.g., L865V). NO: 1)): L730P, G731V, E732K, G733V, E734K, L760M, K761E, E762K, N763D, A76 4V, S765N, P766A, S767C, E768K, L779M, I788M, M868R, K869E, L870Q, V871M, H8 72R, R873P, D874Y, L881R, L895M, S896N, R897C, D898Y, V899G, Y900D, E901K, E902K, D903Y, S904C, Y905D, V906M, K907E, R908P, S909C, Q910R, G911C, and R912P. These mutations (which may also include single or multiple amino acid changes, insertions within or flanking the sequence, and deletions within or flanking the sequence) are thought to induce steric hindrance and / or active conformational effects that alter inhibitor binding characteristics.

[0155] Compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein can be used to treat patients with cancers that have certain RET inhibitor resistance mutations. For example, resistance mutations leading to increased resistance to RET inhibitors, such as substitutions at amino acid position 804 (e.g., V804M, V804L, or V804E), and / or one or more other RET inhibitor resistance mutations, as discussed above, can be used in combination therapy or as a follow-up to existing drug therapy. For example, if a patient is treated with a first RET kinase inhibitor and that patient has a RET inhibitor resistance mutation, the patient can subsequently be treated with compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, or pharmaceutically acceptable salts thereof (assuming that compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein are suitable inhibitors of the specific RET kinase inhibitor mutation present). As another example, if a patient is known to have a specific RET kinase inhibitor mutation (or multiple mutations), the patient can be treated simultaneously with multiple RET kinase inhibitors, including compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, or pharmaceutically acceptable salts thereof, which are effective against one or more present RET kinase inhibitor mutations. Examples of currently known RET kinase inhibitors include pazopanib, sitravatinib, regorafenib, motixabin, RXDX-105, alectinib, BLU6864, cabozantinib, dovitinib, fretinib, lenvatinib, ponatinib, praltinib, serpatinib, sorafenib, sunitinib, and vantanib.

[0156] The types of cancers that can be treated using compounds of formulas I, Ia, Ib, Ic, and Id, and their pharmaceutically acceptable salts, include hematologic cancers and solid tumor cancers. Examples of cancer types that can be treated using compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein include lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), thyroid cancer (e.g., anaplastic thyroid cancer, Hurthle cell thyroid cancer, follicular thyroid cancer, poorly differentiated thyroid cancer, papillary thyroid cancer, medullary thyroid cancer, differentiated thyroid cancer, recurrent thyroid cancer, or refractory differentiated thyroid cancer), thyroid adenoma, endocrine gland vegetations, lung adenocarcinoma, bronchioloalveolar cell carcinoma, type 2A or 2B multiple endocrine neoplasia (MEN2A or MEN2B, respectively), pheochromocytoma, parathyroid hyperplasia, breast cancer, mammary cancer, and breast cancer.Carcinoma, mammary neoplasm, colorectal cancer (e.g., metastatic colorectal cancer), papillary renal cell carcinoma, ganglioneuroma of the gastrointestinal mucosa, inflammatory myofibroblastoma, and cervical cancer. Specifically, the type of cancer can be lung cancer or thyroid cancer. More specifically, the cancer can be non-small cell lung cancer or medullary thyroid carcinoma. Other examples of cancer types that can be treated using compounds and methods of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), cancer in adolescents, adrenocortical carcinoma, anal cancer, appendiceal cancer, astrocytoma, atypical teratoid / rhabdoid tumor, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brainstem glioma, brain tumor, breast cancer, bronchial tumor, Burkitt lymphoma, carcinoid tumor, cancer of unknown primary origin, cardiac tumor, cervical cancer, childhood cancer, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative vegetation, vegetation classified by location, vegetation, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, cholangiocarcinoma, ductal carcinoma in situ, embryonal tumor, endometrial cancer. Ependymoma, esophageal cancer, olfactory neuroblastoma, Ewing sarcoma, extracranial extradermal cell tumor, gonadal extradermal cell tumor, extrahepatic bile duct carcinoma, ocular cancer, fallopian tube cancer, fibrous histiocytoma of bone, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, glioma, pilocytic tumor, pilocytic leukemia, head and neck cancer, thoracic tumors, head and neck growths, CNS tumors, primary CNS tumors. Heart cancer, hepatocellular carcinoma, histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors, pancreatic neuroendocrine tumors, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, lip and oral cancer, liver cancer, lung cancer, lymphoma, macroglobulinemia, malignant fibrous histiocytoma of bone, bone cancer, melanoma, Merkel cell carcinoma, mesothelioma, metastatic squamous neck cancer, midline tract carcinoma.Carcinoma, oral cancer, multiple endocrine neoplasia syndrome, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, myelodysplastic / myeloproliferative vegetations, vegetations classified by location, vegetations, myeloid leukemia, myeloid leukemia, multiple myeloma, myeloproliferative vegetations, nasal and paranasal sinus carcinoma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, lung vegetations, lung cancer, lung vegetations, lung cancer sarcoma, respiratory vegetations, bronchial protocarcinoma, bronchial vegetations, oral cancer, oral cavity cancer, lip cancer, oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, papilloma, paraganglioma, paranasal sinus and nasal cavity carcinoma, Parathyroid carcinoma, penile cancer, pharyngeal cancer, pheochromocytoma, pituitary cancer, plasma cell vegetations, pleural pulmonary germ cell tumors, pregnancy and breast cancer, primary central nervous system lymphoma, primary peritoneal cancer, prostate cancer, rectal cancer, colon cancer, colonic vegetations, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Cezari syndrome, skin cancer, small cell lung cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck cancer, gastric cancer, T-cell lymphoma, testicular cancer, laryngeal cancer, thymoma and thymic carcinoma, thyroid cancer, transitional cell carcinoma of the renal pelvis and ureter, cancer of unknown primary origin, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and Wilms' tumor.

[0157] Examples of types of hematologic cancers that can be treated using compounds and methods of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein include leukemia, lymphoma (non-Hodgkin lymphoma), Hodgkin's disease (also known as Hodgkin's lymphoma), and myeloma, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and chronic myelomonocytic leukemia. Cellular leukemia (CMML), chronic neutrophilic leukemia (CNL), acute undifferentiated leukemia (AUL), anaplastic large cell lymphoma (ALCL), prolymphocytic leukemia (PML), juvenile myelomonocytic leukemia (JMML), adult T-cell ALL, AML with trilineage dysplasia (AML / TMDS), mixed spectrum leukemia (MLL), myelodysplastic syndromes (MDSs), myelodysplastic disorders (MPD), and multiple myeloma (MM). Other examples of hematologic malignancies include myelodysplastic disorders (MPDs) such as polycythemia vera (PV), idiopathic thrombocytopenic purpura (ET), and idiopathic primary myelofibrosis (IMF / IPF / PMF). In one embodiment, the hematologic malignancy (e.g., a hematologic malignancy as a RET-related cancer) is AML or CMML.

[0158] Examples of solid tumor cancer types that can be treated using compounds and methods of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein include thyroid cancer (e.g., papillary thyroid carcinoma, medullary thyroid carcinoma), lung cancer (e.g., lung adenocarcinoma, small cell lung cancer), pancreatic cancer, pancreatic ductal carcinoma, xanthogranulomatous tumor, breast cancer, colon cancer, colorectal cancer, rectal neuroendocrine carcinoma, prostate cancer, renal cell carcinoma, head and neck tumors, neuroblastoma, and melanoma.

[0159] This document also provides compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx, or pharmaceutically acceptable salts thereof, as described herein for the treatment of RET-related diseases or disorders such as IBS or cancer. The foregoing describes cancers that can be treated using compounds of formulas I, Ia, Ib, Ic, and / or Id, or pharmaceutically acceptable salts thereof. Treatment of RET-related diseases or disorders may further include the following steps: performing an in vitro assay using a biological sample from a patient to determine the presence of an abnormality in the expression, activity, or level of a RET gene, RET kinase, or any of them; and, if an abnormality in the expression, activity, or level of a RET gene, RET kinase, or any of them is present, administering to the patient a therapeutically effective amount of a compound of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx, as described herein. In these uses, the biological sample may be a tumor sample, and the tumor sample may be analyzed using methods known to those skilled in the art, such as genome / DNA sequencing. Additionally, in these uses, the sample may be obtained from the patient prior to the first administration of a compound of formula I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein. These therapeutic uses of compounds of formula I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein may be patient-based, selected for treatment by at least one regulatory abnormality in the expression or activity or level of a RET gene, RET kinase, or any of these. Furthermore, in these therapeutic uses, a compound of formula I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein, or a pharmaceutically acceptable salt thereof, may be administered to the patient at a dose of about 1 mg / kg to 200 mg / kg (the effective dose range is as described above).

[0160] For clarity, certain stereochemical centers are left unspecified and certain substituents are eliminated in the following schemes, but this is not intended to limit the teachings of the schemes in any way. Furthermore, those skilled in the art can separate or resolve individual isomers, enantiomers, and diastereomers at any convenient point in the synthesis of the compounds of the present invention using methods such as selective crystallization or chiral chromatography (see, for example, J. Jacques et al., “Enantiomers, Racemates, and Resolutions,” John Wiley and Sons, Inc., 1981, and E.L. Leel and S.H. Wileh, “Stereochemistry of Organic Compounds,” Wiley-Interscience, 1994).

[0161] The designations “Isomer 1” and “Isomer 2” respectively denote the compounds eluted first and second from chiral chromatography under the conditions described herein, and the same designations apply to subsequent intermediates and examples if chiral chromatography is initiated first in the synthesis. Additionally, the intermediates described in the following schemes may contain a number of nitrogen or oxygen protecting groups. These variable protecting groups may be the same or different each time they appear, depending on the specific reaction conditions and the specific transformation to be performed. Protecting and deprotecting conditions are well known to those skilled in the art and are described in the literature (see, for example, “Greene’s Protective Groups in Organic Synthesis,” 4th edition, Peter GMWuts and Theodora W. Greene, John Wiley and Sons, Inc. 2007).

[0162] Some abbreviations are defined as follows: "ACN" represents acetonitrile; "ATP" represents adenosine triphosphate; "bis(pentanoyl)boron" represents 4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborhexacyclopentane; "Boc" represents tert-butoxycarbonyl; "Boc2O" represents ditert-butyl dicarbonate; "BSA" represents bovine serum albumin; "BTFFH" represents fluoro-N,N,N′,N′-bis(tetramethylene)formamidinium hexafluorophosphate; "Bu" represents butyl. "DCM" represents (1-cyano-2-ethoxy-2-oxoethyleneaminooxy)dimethylamino-morpholino-carbomony hexafluorophosphate; "DIPEA" represents N,N-diisopropylethylamine or N-ethyl-N-isopropyl-propane-2-amine; "DMAP" represents 4-dimethylaminopyridine; "DMF" represents N,N-dimethylformamide; "DMSO" represents dimethyl sulfoxide; "DTT" represents dithiothreitol; "EtOAc" represents ethyl acetate; "ee" indicates enantiomer excess; "Et2O" represents diethyl ether; "EtO" represents diethyl ether. "H" indicates ethanol; "FA" indicates formic acid; "GST" indicates glutathione S-transferase; "HATU" indicates 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; "HEK" indicates human embryonic kidney; "hr" or "hrs" indicates hours; "HOAc" indicates acetic acid; "HTRF" indicates homogeneous time-resolved fluorescence; "IgG" indicates immunoglobulin G; "IPA" indicates isopropanol; "IPAm" indicates isopropylamine; "iPr2O" indicates isopropyl ether; "[I "r(OMe)(1,5-cod)]2" represents di-μ-methoxy-1,5-diene or (1,5-cyclooctadiene)(methoxy)iridium(I) dimer or bis(1,5-cyclooctadiene)di-μ-methoxydiiridium(I); "KOAc" represents potassium acetate; "LDA" represents lithium diisopropylaminodimethyl; "MeI" represents iodomethane or iodomethane; "MeOH" represents methanol; "MeTHF" represents 2-methyltetrahydrofuran; "min" represents minutes; "Mn(dpm)3" represents manganese(3+)tri[(3Z)- 2,2,6,6-Tetramethyl-5-oxo-3-hepten-3-olide], Mn(TMHD)3, Shenvi hydrogenation catalyst, or tris(dipentanoylmethane)manganese; "NaOAc" indicates sodium acetate; "NaOMe" indicates sodium methoxide; "NMI" indicates 1-methylimidazole or N-methylimidazole; "Parkins catalyst" indicates hydrogenation of (dimethyltrivalent phosphonate-kP)[hydrobis(dimethyltrivalent phosphonate-kP)]platinum(II) and is CAS#173416-05-2; "PBS-T" indicates phosphate buffered saline + 20; “Pd(dppf)Cl2)” represents [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride; “Pd(dppf)Cl2)CH2Cl2” represents [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride coupled with DCM; “PE” represents petroleum ether; “PhSiH3” represents phenylsilane; “RT” represents room temperature; “SFC” represents supercritical fluid chromatography; “tert- "BuOH" represents tert-butanol; "TBTU" represents O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethylureonium tetrafluoroborate or N,N,N′,N′-tetramethyl-O-(benzotriazol-1-yl)ureonium tetrafluoroborate; "TCFH" represents N,N,N′,N′-tetramethylchloroformamidine hexafluorophosphate; "TEA" represents triethylamine; "TFA" represents trifluoroacetic acid; "THF" represents tetrahydrofuran; This indicates propanephosphonic anhydride, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphacyclohexane-2,4,6-trioxide, or PPACA; "t (R) "" indicates the retention period, and "WT" indicates wild type.

[0163] The compounds of the present invention or pharmaceutically acceptable salts thereof can be prepared according to the following preparations and examples using methods well known and understood in the art. Suitable reaction conditions for the steps of these preparations and examples are well known in the art, and appropriate substitutions of solvents and auxiliary reagents are within the scope of the art. Likewise, those skilled in the art will understand that synthetic intermediates can be isolated and / or purified as needed or desired by various well-known techniques, and often various intermediates may be used directly in subsequent synthetic steps without purification or with minimal purification. As an example, the compounds of the preparations and examples can be isolated, for example, by silica gel purification, or directly by filtration or crystallization. Furthermore, those skilled in the art will appreciate that, in some cases, the order of the introduced moieties is not important. The specific order of steps required to produce the compounds of the present invention depends on the specific compound being synthesized, the relative susceptibility of the starting compound and the substituted moieties, as is well known to skilled chemists. Unless otherwise indicated, all substituents are as defined above, and all reagents are well known and understood in the art.

[0164] In the following schemes, unless otherwise stated, all substituents are as defined above. Reagents and starting materials are generally readily available to those skilled in the art. Others can be prepared using standard techniques of organic and heterocyclic chemistry, similar to the synthesis of known structurally similar compounds and the procedures described in the subsequent preparations and examples, including any new procedures. It is intended that this specification include intermediates and methods that can be used to synthesize compounds described by formula (I).

[0165] Option 1

[0166]

[0167] Scheme 1 describes the preparation of compound (12). Those skilled in the art will recognize that the boronization of aryl bromide (1) under typical Miyaura reaction conditions can provide a borate ester (2). Treatment of the borate ester (2) with a suitable bromopyrazole (3) and a metal catalyst under Suzuki coupling conditions can provide an aminopyrazole (9).

[0168] Alternatively, palladium-catalyzed carbonylation of aryl bromide (1) with carbon monoxide and benzyl alcohol can provide benzyl carboxylate (4). Palladium-catalyzed debenzylation of benzyl carboxylate (4) with hydrogen can provide carboxylic acid (5). Preparation of the acyl chloride of acid (5) and subsequent reaction with malononitrile under basic conditions can provide malononitrile (6). Methylation of vinyl alcohol (6) can be achieved with dimethyl sulfate to provide methoxy group (7). Reaction of appropriately substituted hydrazine (8) with dionitrile Michael acceptor (7) can also provide aminopyrazole (9).

[0169] Under Suzuki coupling conditions, treatment of the borate ester (2) with a suitable bromide pyrazole (10) and a metal catalyst can provide an aminopyrazole (11). The carboxylic acid (12) can be obtained by saponification of the ester (11) with a suitable nucleophilic base.

[0170] Alternatively, carboxylic acid (12) can also be obtained from ester (9) in a two-step one-pot reaction of oxidation and saponification using peroxide and base.

[0171] Option 2

[0172]

[0173] Scheme 2 describes the preparation of compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein. Saponification of the ester (1) can provide a carboxylic acid (14) in substantially the same manner as described in Scheme 1 with respect to the carboxylic acid (12). Those skilled in the art will recognize that the carboxylic acid (14) and the primary amine (13) can be linked under suitable conditions for amide bond formation using a suitable amide coupling agent to provide an amide (15). Borylation of the aryl bromide (15) under typical Miyaura reaction conditions, followed by in-situ treatment of the borate ester with bromopyrazole (10) and a metal catalyst under Suzuki coupling conditions, can provide compounds of formulas I, Ia to IX, IIa to IIx and / or IIIa to IIIx.

[0174] Alternatively, amide coupling of carboxylic acids (12) and primary amines (13) can provide compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx in substantially the same manner as described above with respect to amides (15).

[0175] Option 3

[0176]

[0177] Scheme 3 describes an additional preparation of compounds of formulas I, Ia to I, IIa to IIx and / or IIIa to IIIx as described herein. Protecting a primary amine (16) with a suitable protecting group can provide a mono- or di-protected aminopyrazole (17). Iridium-catalyzed CH borylation of the pyrazole (17), followed by in-situ treatment of the borate ester with an aryl bromide (1) and a metal catalyst under Suzuki coupling conditions, can provide a biaryrheic ester (18). Saponification of the ester (18) can provide a carboxylic acid (19) in substantially the same manner as described in Scheme 1 regarding the carboxylic acid (12). Amide coupling of the carboxylic acid (19) and the primary amine (13) can provide an amide (20) in substantially the same manner as described in Scheme 2 regarding the amide (12). The nitrile moiety of the aminopyrazole (20) can be converted to an amide (21) under various conditions such as metal-catalyzed hydration, acidic hydrolysis, and oxidation. Removing the protecting group from the protected aminopyrazole (21) can provide additional compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx as described herein.

[0178] Option 4

[0179]

[0180] Scheme 4 describes an alternative preparation of compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein. Reaction of a suitably substituted hydrazine (8) with a dinitrile Michael acceptor (22) can provide aminopyrazole (23) in substantially the same manner as described in Scheme 1 with respect to aminopyrazole (9). Treatment of aryl bromide (23) with a suitable alkyl zinc halide and a metal catalyst under Negishi coupling conditions can provide tert-butyl ester (24). Acid deprotection of tert-butyl ester (24) can provide carboxylic acid (25).

[0181] Alternatively, the saponification of the ester (9) can provide the carboxylic acid (25) in substantially the same manner as described in Scheme 1 regarding the carboxylic acid (12). The amide coupling of the carboxylic acid (25) and the primary amine (13) can provide the amide (31) in substantially the same manner as described in Scheme 2 regarding the amide (12).

[0182] Alternatively, amide coupling of carboxylic acid (26) and primary amine (13) can provide amide (27) in substantially the same manner as described in Scheme 2 regarding amide (12). Saponification of ester (27) can provide carboxylic acid (28) in substantially the same manner as described in Scheme 1 regarding carboxylic acid (12). Preparation of acyl chloride of acid (28) and subsequent reaction with malononitrile under alkaline conditions can provide malononitrile (29). Methylation of vinyl alcohol (29) can be achieved with dimethyl sulfate to provide methoxy group (30). Reaction of appropriately substituted hydrazine (8) with dionitrile Michael acceptor (30) can provide aminopyrazole (31) in substantially the same manner as described in Scheme 1 regarding aminopyrazole (9).

[0183] Under various conditions such as metal-catalyzed hydration, acidic hydrolysis, and oxidation, the nitrile moiety of aminopyrazole (31) can be converted into other compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx as described herein.

[0184] In an optional step, pharmaceutically acceptable salts of compounds of formulas I, Ia to Ix, IIa to IIx and / or IIIa to IIIx can be formed by reacting a suitable free base of formula I, Ia to Ix, IIa to IIx and / or IIIa to IIIx with a suitable pharmaceutically acceptable acid in a suitable solvent under standard conditions. Furthermore, the formation of such salts can occur simultaneously with the deprotection of the nitrogen protecting group. The formation of such salts is well known and understood in the art. See, for example, Gould, PL, "Salt selection for basic drugs," International Journal of Pharmaceutics, 33:201-217 (1986); Bastin, RJ, et al., "Salt Selection and Optimization Procedures for Pharmaceutical New Chemical Entities," Organic Process Research and Development, 4:427-435 (2000); and Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Sciences, 66: 1-19, (1977). "Salt selection for basic drugs," International Journal of Pharmaceutics, 33201-217 (1986). Those skilled in the art will understand that compounds of formula I, Ia to Ix, IIa to IIx and / or IIIa to IIIx can be readily converted into pharmaceutically acceptable salts and can be isolated as pharmaceutically acceptable salts.

[0185] Preparation and Examples

[0186] Preparation 1

[0187] N-(tert-butoxycarbonyl)-N-(5-iodo-4-methyl-1,3-thiazo-2-yl)tert-butyl carbamate

[0188]

[0189] A mixture of 5-iodo-4-methyl-1,3-thiazolyl-2-amine (16.0 g, 66.7 mmol), Boc₂O (32.0 g, 146 mmol), and DMAP (0.81 g, 6.63 mmol) in THF (200 mL) was stirred at room temperature under N₂ for 2 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 12:1 to 5:1 PE:EtOAc to yield the title compound (20.0 g, 68%) as a yellow solid. ES / MS (m / z) 441.0 (M+H).

[0190] Preparation 2

[0191] 5-(2,2-dimethylpropyl)-4-methyl-1,3-thiazolyl-2-amine

[0192]

[0193] Zinc chloride (5.84 mL, 4.09 mmol, 0.7 M in THF) was added to a solution of magnesium bromine (2,2-dimethylpropyl)magnesium (6.50 mL, 3.25 mmol, 0.5 M in THF) at 0 °C, and the mixture was stirred at 0 °C under N2 for 30 min. N-(tert-butoxycarbonyl)-N-(5-iodo-4-methyl-1,3-thiazolyl-2-yl)carbamate tert-butyl ester (1.20 g, 2.73 mmol) and Pd(t-Bu3P)2 (0.28 g, 0.55 mmol) were added fractionally to the mixture over 1 min at room temperature. The mixture was stirred at 100 °C under N2 for an additional 2 h. The reaction was quenched by adding 20 mL of saturated aqueous NH4Cl solution at room temperature. The mixture was extracted with EtOAc (2 x 50 mL). The combined organic extracts were washed with brine (2 x 30 mL), dried over anhydrous Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 45% to 52% ACN in H₂O (0.1% NH₄HCO₃) to yield the title compound (80 mg, 16%) as a yellow solid. ES / MS (m / z) 185.1 (M+H).

[0194] Preparation 3

[0195] 2-[4-bromo-3-(bromomethyl)phenyl]acetic acid methyl ester

[0196]

[0197] Under N2 and at room temperature, 1-bromopyrrolidine-2,5-dione (4.23 g, 23.8 mmol) and 2-[(E)-(1-cyano-1-methyl-ethyl)azo]-2-methylpropionitrile (1.30 g, 7.92 mmol) were successively added to a stirred solution of methyl 2-(4-bromo-3-methyl-phenyl)acetate (5.50 g, 22.6 mmol) in CCl4 (200 mL). The reaction mixture was stirred at 80 °C for 5 hours. The mixture was poured into a solution of NaHCO3 and the aqueous layer was extracted with DCM (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The substance was purified by silica gel chromatography, eluting with a gradient of 2% to 10% EtOAc in cyclohexane to yield the title compound (1.69 g, 23%) as a colorless oil. ES / MS (m / z) 342 (M+Na).

[0198] Preparation 4

[0199] Methyl 2-[4-bromo-3-(methoxymethyl)phenyl]acetate

[0200]

[0201] NaOMe (0.41 mL, 2.19 mmol, 30 wt% in MeOH) was added to a stirred solution of methyl 2-[4-bromo-3-(bromomethyl)phenyl]acetate (0.60 g, 1.86 mmol) in MeOH (10 mL) at 5 °C under N2. The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated, H2O was added to the residue, and the mixture was extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over NaSO4, filtered, and concentrated under reduced pressure to yield the title compound (0.42 g, 83%) as a pale yellow oil. ES / MS (m / z) 274 (M+H).

[0202] Preparation 5

[0203] methyl 2-(4-bromo-3-chloro-2-fluoro-phenyl)acetate

[0204]

[0205] Thionyl chloride (0.39 mL, 5.33 mmol) was added dropwise to 2-(4-bromo-3-chloro-2-fluorophenyl)acetic acid (0.50 g, 1.78 mmol) in MeOH (15 mL) under N2 at 0 °C. The mixture was warmed to room temperature and stirred for 16 hours. The mixture was concentrated under reduced pressure and dried under vacuum. The substance was purified by silica gel chromatography, eluting with a gradient of 25% to 100% DCM in heptane to yield the title compound (0.50 g, 100%) as a colorless oil.

[0206] The following compounds in Table 1 were prepared using a suitable acid, essentially as described with respect to methyl 2-(4-bromo-3-chloro-2-fluoro-phenyl)acetate, with the reaction time adjusted to determine the end of the reaction, and purified by chromatography where appropriate, and, if necessary, with EtOH replaced with MeOH to prepare the ethyl ester.

[0207] Table 1

[0208]

[0209] Preparation 7

[0210] 2-[4-bromo-3-(tert-butoxymethyl)phenyl]methyl acetate

[0211]

[0212] A solution of methyl 2-[4-bromo-3-(bromomethyl)phenyl]acetate (0.50 g, 1.55 mmol) in dry DCM (5 mL) was added to a suspension of silver trifluoromethanesulfonate (0.44 g, 1.71 mmol) in dry tert-BuOH (10 mL) and dry DCM (20 mL) at room temperature under N2, avoiding visible light. The reaction mixture was stirred at room temperature for 16 hours and then poured into a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with DCM (2x). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure to yield the title compound (0.51 g, 95%) as a colorless oil. 1 H NMR (d6-DMSO, 400MHz) δ (ppm) 7.53 (d, J = 8.1Hz, 1H), 7.39 (d, J = 2.2Hz, 1H), 7. 12 (dd, J=8.1, 2.3Hz, 1H), 4.41 (s, 2H), 3.70 (s, 2H), 3.62 (s, 3H), 1.25 (s, 9H).

[0213] Preparation of 8

[0214] 2-[(tert-butoxycarbonyl)[(tert-butoxycarbonyl)amino]amino]-2-methylpropane-1-ol

[0215]

[0216] To a stirred solution of Mn(dpm)3 (0.42 g, 0.69 mmol) in IPA (150 mL), methyl allyl alcohol (5.00 g, 69.3 mmol), di-tert-butyl azodicarbonate (24.0 g, 103 mmol) and PhSiH3 (7.50 g, 69.3 mmol) were added dropwise at 0 °C under N2. The mixture was stirred at 0 °C under N2 for 1 h and then at room temperature under N2 for 12 h. The solvent was removed under reduced pressure, and the residue was purified by silica gel chromatography, eluting with a gradient of 10:1 to 4:1 PE:EtOAc to yield the title compound (15.0 g, 71%) as a white solid. 1 H NMR (CDCl3, 300MHz) δ4.30-4.02 (m, 2H), 3.42-3.19 (m, 1H), 1.59-1.42 (m, 24H).

[0217] Preparation 9

[0218] 2-Hydroxy-2-methylpropane-1-ol 2HCl

[0219]

[0220] To a solution of 2-[(tert-butoxycarbonyl)[(tert-butoxycarbonyl)amino]amino]-2-methylpropane-1-ol (15.0 g, 49.3 mmol) in 1,4-dioxane (100 mL), HCl (50 mL, 4 N in 1,4-dioxane) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The crude product was ground together with Et₂O (30 mL) to yield the title compound (4.4 g, 63%) as a white solid. 1 H NMR (d6-DMSO, 300MHz) δ6.41 (s, 5H), 3.40 (s, 2H), 1.14 (s, 6H).

[0221] Preparation 10

[0222] N-(1-Methylcyclopropyl)-N-nitroso-tert-butyl carbamate

[0223]

[0224] Tert-butyl nitrite (2.40 g, 23.3 mmol) was added fractionally to a stirred solution of N-(1-methylcyclopropyl)carbamate (2.00 g, 11.7 mmol) in DCM (20 mL) at room temperature under N2, and the mixture was stirred at room temperature under N2 for 2 hours. The mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography, eluting with PE:EtOAc at a ratio of 25:1 to 20:1 to yield the title compound (1.30 g, 55%) as a brown liquid. 1 H NMR (d6-DMSO) δ1.59 (s, 9H), 1.10 (s, 3H), 0.86-0.84 (m, 2H), 0.70-0.65 (m, 2H).

[0225] Preparation 11

[0226] (1-Methylcyclopropyl)hydrazine hydrochloride

[0227]

[0228] Zn (0.78 g, 11.9 mmol) was added fractionally to a stirred solution of N-(1-methylcyclopropyl)-N-nitroso-carbamate (1.20 g, 5.99 mmol) in HCl (8 mL, 4 N in 1,4-dioxane) at 0 °C under N2, and the mixture was stirred at room temperature under N2 for 12 hours. The mixture was filtered, the filter cake was washed with H2O (3 x 10 mL), and the filtrate was concentrated under reduced pressure to yield the title compound (2.0 g, crude product) as a grayish-white solid, which was used directly without further purification. 1H NMR (d6-DMSO) δ1.30 (s, 3H), 0.77-0.73 (m, 2H), 0.54-0.50 (m, 2H).

[0229] Preparation 12

[0230] methyl spiro[2.2]pentane-2-carboxylate

[0231]

[0232] 2M diazonylmethyl (trimethyl)silane (6.9 mL, 13.8 mmol) in hexane was added dropwise to a stirred solution of spiro[2.2]pentane-1-carboxylic acid (95%, 1.25 g, 10.6 mmol) in Et₂O (37.5 mL) and MeOH (7.5 mL). The reaction mixture was stirred overnight at room temperature. Et₂O was removed under reduced pressure and the reaction mixture was diluted with pentane (125 mL). The solution was washed with saturated aqueous NaHCO₃ solution (2 x), H₂O (3 x), and brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to produce the title compound (1.53 g, crude) as a yellow oil, which was used directly without further purification.

[0233] Preparation 13

[0234] 2-(trifluoromethyl)spiro[3.3]heptane-2-carbonyl chloride

[0235]

[0236] 2-(trifluoromethyl)spiro[3.3]heptane-2-carboxylic acid (1.02 g, 4.90 mmol) was suspended in DCM (40 mL) and DMF (0.10 mL). The slurry was cooled to 0 °C and oxalyl chloride (3.0 mL, 6.00 mmol, 2 M in DCM) was added. The reaction mixture was stirred at 0 °C for 5 min, then warmed to room temperature and stirred for 1 h. The solvent was removed under reduced pressure to produce a yellow residue. The residue was used without further purification.

[0237] The compounds listed in Table 2 were prepared essentially as described with respect to 2-(trifluoromethyl)spiro[3.3]heptane-2-carbonyl chloride using a suitable acid, oxalyl chloride (1-1.5 equivalents), and solvents (DCM, DMF) in the range of 0-50 mL, and the reaction time was adjusted to determine the end of the reaction. The reaction temperature could be in the range of 0 °C-25 °C.

[0238] Table 2

[0239]

[0240] Preparation of 17

[0241] Benzyl 2,2-dimethylcyclobutanecarboxylate

[0242]

[0243] Oxaloyl chloride (1.0 mL, 11.7 mmol) was added dropwise to a stirred solution of 2,2-dimethylcyclobutane-1-carboxylic acid (1.50 g, 11.7 mmol) in DCM (20 mL) at 0 °C under N2, followed by the addition of DMF (30 μL). The mixture was slowly warmed to room temperature and stirred for 5 hours. Benzyl alcohol (1.4 mL, 13.5 mmol) was added dropwise at 0 °C, and the reaction mixture was slowly warmed to room temperature and stirred for 20 hours. The mixture was poured into a saturated aqueous solution of NaHCO3. The aqueous layer was extracted with pentane (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, eluting with a gradient of 5% to 50% DCM in pentane to yield the title compound (2.6 g, 100%) as a colorless oil. ES / MS (m / z) 219 (M+H).

[0244] Preparation of 18

[0245] 2-[2-(3,3-dimethylcyclobutyl)-1,3-dioxacyclopentan-2-yl]acetonitrile

[0246]

[0247] Ethane-1,2-diol (0.67 mL, 12.0 mmol) and chloro(trimethyl)silane (1.5 mL, 12.0 mmol) were added to a stirred solution of 3-(3,3-dimethylcyclobutyl)-3-oxopropionitrile (0.60 g, 4.00 mmol) in DCM (8 mL) at 25 °C under N2. The reaction mixture was refluxed and stirred for 12 hours and cooled to room temperature. NaHCO3 (aqueous solution) was added. The aqueous layer was extracted with DCM. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and dried under vacuum. The residue was purified by silica gel chromatography, eluting with a gradient of 10% to 30% EtOAc in cyclohexane to yield the title compound (0.56 g, 72%) as a colorless oil. 1 H NMR (d6-DMSO, 400MHz) δ (ppm) 4.09-3.98 (m, 2H), 4.01-3.90 (m, 2H), 2.79 (s, 2H), 2.61(s, 2H), 1.79-1.57(m, 6H), 1.41(s, 2H), 1.12(s, 3H), 1.01(s, 3H).

[0248] Preparation of 19

[0249] 4-(1-Methylcyclopropyl)-3-oxo-butyronitrile

[0250]

[0251] ACN (0.36 mL, 6.89 mmol) and 2-(1-methylcyclopropyl)benzyl acetate (1.27 g, 6.22 mmol) were combined in THF (15 mL) under N2 and cooled to -78 °C. LDA (11.0 mL, 22.0 mmol, 2 M in THF) was added dropwise and the reaction mixture was stirred at -78 °C for 10 min. The reaction mixture was then warmed to 21 °C over 1 hour. The reaction mixture was acidified to pH 1 with concentrated HCl and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated to yield the title compound as an orange oil (1.4 g, crude). The substance was used without further purification.

[0252] Preparation 20

[0253] 3-Oxo-3-[2-(trifluoromethyl)spiro[3.3]heptane-2-yl]propionitrile

[0254]

[0255] ACN (0.28 mL, 5.36 mmol) and benzyl 2-(trifluoromethyl)spiro[3.3]heptane-2-carboxylate (1.40 g, 4.69 mmol) were combined in THF (15 mL) under N2 and cooled to -78 °C. LDA (2.82 mL, 5.64 mmol, 2 M in THF) was added dropwise and the reaction mixture was stirred at -78 °C for 10 min. The reaction mixture was then warmed to 21 °C over 1 hour. The reaction mixture was acidified to pH 1 with concentrated HCl and the mixture was extracted with EtOAc (3 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated to yield the title compound (1.05 g, 97%) as an orange oil. This substance was used directly without further purification.

[0256] The compounds listed in Table 3 were prepared essentially as described with respect to 3-oxo-3-[2-(trifluoromethyl)spiro[3.3]heptane-2-yl]propionitrile using 1.2–4.2 equivalents of a suitable base (LDA or nBuLi in hexane or THF) and the reaction time was adjusted to determine the end of the reaction.

[0257] Table 3

[0258]

[0259] 1 nBuLi is used as a replacement for LDA.

[0260] Preparation 25

[0261] 3-[3-methylbicyclo[1.1.1]pentan-1-yl]-3-oxopropionitrile

[0262]

[0263] n-BuLi (1.71 mL, 4.28 mmol, 2.5 M in THF) was added to THF (5 mL) at 0 °C under N2. ACN (0.24 mL, 4.59 mmol) in THF (5 mL) was added dropwise to the mixture at -78 °C under N2 for 5 min. The mixture was stirred at -78 °C for 1 h. Methyl 3-methylbicyclo[1.1.1]pentane-1-carboxylate (0.40 g, 2.85 mmol) in THF (5 mL) was added dropwise to the mixture at -78 °C under N2. The mixture was stirred at room temperature for 1 h. The reaction was quenched by adding 20 mL of saturated NH4Cl aqueous solution at 0 °C. The mixture was acidified to pH 4 with 1 N HCl aqueous solution and extracted with DCM (3 x 50 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to produce the title compound (0.36 g, 85%) as a brown liquid. The crude product was used without further purification.

[0264] Preparation 26

[0265] 3-(3-fluorobicyclo[1.1.1]pentan-1-yl)-3-oxopropionitrile

[0266]

[0267] A solution of ACN (0.28 mL, 5.36 mmol) in THF (4 mL) was added dropwise to a stirred solution of n-BuLi (2.0 mL, 2.5 M in n-hexane, 5.00 mmol) in THF (20 mL) at -78 °C under N2. The mixture was stirred at -78 °C for 30 min. A solution of methyl 3-fluorobicyclo[1.1.1]pentane-1-carboxylate (0.48 g, 3.33 mmol) in THF (4 mL) was added dropwise to the mixture at -78 °C for 10 min. The mixture was warmed to room temperature and stirred for another 3 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution, acidified to pH 4 with 2N HCl aqueous solution, and extracted with DCM (3 x 50 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to produce the title compound (0.50 g, crude product) as a brown oil. The crude product was used without further purification. 1 H NMR (CDCl3, 300MHz) δ3.57 (s, 2H), 2.50 (d, 6H).

[0268] Preparation 27

[0269] 3-Oxo-3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]propionitrile

[0270]

[0271] n-BuLi (0.83 mL, 2.08 mmol, 2.5 M solution in hexane) was added to THF (10 mL) at 0 °C under N2. ACN (0.12 mL, 2.30 mmol) in THF (2 mL) was added dropwise over 2 min at -78 °C under N2, and the reaction mixture was stirred at -78 °C for 1 h. Methyl 3-(trifluoromethyl)bicyclo[1.1.1]pentane-1-carboxylate (0.27 g, 1.39 mmol) in THF (2 mL) was added dropwise to the reaction mixture at -78 °C, and the reaction mixture was stirred at room temperature for 12 h. The reaction was quenched by adding saturated NH4Cl aqueous solution (20 mL), acidified to pH 4 with concentrated HCl, and extracted with DCM (3 x 50 mL). The combined organic extracts were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to yield the title compound (0.29 g, crude). The crude product is used without further purification. 1 H NMR (CDCl3, 300MHz) δ3.54 (s, 2H), 2.39 (s, 6H).

[0272] The following compounds in Table 4 were prepared essentially as described with respect to 3-oxo-3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]propionitrile, with the reaction time adjusted to determine the end of the reaction and the purification conditions adjusted as appropriate. After the addition of the carboxylic ester, the reactants can be warmed to room temperature. n-BuLi can be in hexane or THF.

[0273] Table 4

[0274]

[0275] Preparation 39

[0276] 5-(2,5-dimethyl-1H-pyrrolo-1-yl)-3-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole

[0277]

[0278] A solution of 3-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole-5-amine (0.50 g, 2.59 mmol), hexane-2,5-dione (0.91 mL, 7.76 mmol), and HOAc (0.015 mL, 0.26 mmol) in toluene (10 mL) was stirred at 100 °C under N2 for 2 hours. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 gel column) with 30% to 50% H2. ２ Gradient elution of ACN in O (0.1% FA) yielded the title compound (0.61 g, 87%) as a yellow solid. ES / MS (m / z) 272.1 (M+H).

[0279] Preparation of 40

[0280] 3-(2,5-dimethyl-1H-pyrrolo-1-yl)-1-methyl-5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole

[0281]

[0282] A solution of 5-(2,5-dimethyl-1H-pyrrolo-1-yl)-3-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole (0.50 g, 1.84 mmol), MeI (0.46 mL, 7.39 mmol), and K₂CO₃ (0.51 g, 3.69 mmol) in ACN (10 mL) was stirred at 80 °C under N₂ for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative TLC, eluting with 5:1 PE:EtOAc to yield the title compound (95 mg, 18%) as a white solid. ES / MS (m / z) 286.2 (M+H).

[0283] Preparation 41

[0284] 1-Methyl-5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole-3-amine

[0285]

[0286] A solution of KOH (53 mg, 0.94 mmol) in EtOH (1 mL) and H₂O (1 mL) was added to a solution of hydroxylamine hydrochloride (0.13 g, 1.87 mmol) in EtOH (2 mL). 3-(2,5-dimethyl-1H-pyrrolo-1-yl)-1-methyl-5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole (90 mg, 0.32 mmol) was added, and the mixture was stirred at 100 °C under N₂ for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 gel column) using a gradient elution of 10% to 30% ACN in H₂O (0.1% FA) to yield the title compound (45 mg, 68%) as a white solid. ES / MS (m / z) 208.1 (M+H).

[0287] Preparation 42

[0288] 3-[(1-methylcyclopropyl)methyl]isoxazole-5-amine

[0289]

[0290] 4-(1-methylcyclopropyl)-3-oxo-butyronitrile (1.40 g, 10.2 mmol) was dissolved in H₂O (15 mL). NaOH (0.45 g, 11.3 mmol) was added, followed by hydroxylamine sulfate (1.84 g, 11.2 mmol). The pH was adjusted to ~10 with 2M NaOH aqueous solution (5.0 mL, 10.0 mmol). The mixture was heated to 100 °C and held for 1.5 h. Concentrated HCl (1 mL) was added and the mixture was stirred for 15 min. The mixture was diluted with H₂O (30 mL) and extracted with EtOAc (2 x 30 mL). The combined organic extracts were dried over anhydrous NH₂SO₄ and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 10% to 70% EtOAc in hexanes to yield the title compound (0.54 g, 35%). ES / MS (m / z) 153.0 (M+H).

[0291] Preparation 43

[0292] 3-(3-Fluorobico[1.1.1]pentan-1-yl)isoxazol-5-amine

[0293]

[0294] Hydroxylamine hydrochloride (0.68 g, 9.79 mmol) and KOAc (0.96 g, 9.78 mmol) were added to a stirred solution of 3-(3-fluorobicyclo[1.1.1]pentan-1-yl)-3-oxopropionitrile (0.50 g, 3.26 mmol) in MeOH (5 mL) at 0 °C under N2. The mixture was stirred at room temperature under N2 for 5 hours. The mixture was poured into H2O (50 mL) and extracted with DCM (3 x 50 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 25% to 45% ACN in H2O (0.1% NH4HCO3) to yield the title compound (0.22 g, 40%) as a pale yellow solid. ES / MS (m / z) 169.1 (M+H).

[0295] Preparation of 44

[0296] 3-(bicyclo[1.1.1]pentan-1-yl)isoxazol-5-amine

[0297]

[0298] Hydroxylamine hydrochloride (1.34 g, 19.3 mmol) and KOAc (1.89 g, 19.3 mmol) were added to a stirred solution of 3-(bicyclo[1.1.1]pentan-1-yl)-3-oxopropionitrile (0.87 g, 6.44 mmol) in MeOH (10 mL) at 0 °C under N2. The mixture was stirred at room temperature under N2 for 5 hours. The mixture was poured into H2O (50 mL) and extracted with DCM (3 x 50 mL). The combined organic extracts were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was washed with hexane (2 x 5 mL) to yield the title compound (0.73 g, 75%) as a pale yellow solid. ES / MS (m / z) 151.3 (M+H).

[0299] Preparation of 45

[0300] 3-(3-methyl-1-bicyclo[1.1.1]pentyl)isoxazole-5-amine

[0301]

[0302] NaOH (0.19 g, 4.75 mmol) was added fractionally to a stirred mixture of 3-[3-methylbicyclo[1.1.1]pentan-1-yl]-3-oxopropionitrile (0.35 g, 2.35 mmol) and hydroxylamine hydrochloride (0.18 g, 2.59 mmol) in H2O (8 mL) at room temperature under N2. The mixture was stirred at 100 °C under N2 for 1 hour. The mixture was then cooled to room temperature. The precipitated solid was collected by filtration and washed with H2O (3 x 20 mL) to yield the title compound (0.30 g, 78%) as a pale yellow solid. ES / MS (m / z) 165.1 (M+H).

[0303] Preparation of 46

[0304] 3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)isoxazol-5-amine

[0305]

[0306] NaOH (0.11 g, 2.75 mmol) was added fractionally to a mixture of 3-oxo-3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]propionitrile (0.28 g, 1.38 mmol), hydroxylamine hydrochloride (0.11 g, 1.58 mmol), and H₂O (10 mL) at room temperature under N₂. The mixture was heated to 100 °C and held for 1 hour. The mixture was cooled to room temperature and extracted with DCM (3 x 50 mL). The combined organic extracts were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The resulting solid was ground with Et₂O (10 mL). The solid was collected by filtration and washed with Et₂O to yield the title compound (0.22 g, 73%). ES / MS (m / z) 219.2 (M+H).

[0307] Using approximately 1-6 equivalents of a suitable reagent (hydroxylamine sulfate or hydroxylamine hydrochloride) and approximately 1-6 equivalents of a base (NaOH, NaOAc, or KOAc), in a suitable solvent (EtOH, MeOH, H2O, or 1,4-dioxane), prepare the following compounds in Table 5 essentially as described with respect to 3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)isoxazol-5-amine, adjusting the reaction time to determine the end of the reaction and purifying using appropriate chromatographic conditions or precipitation. The reaction temperature can be in the range of 0-100°C, and the reaction mixture can be acidified in post-treatment if appropriate. The reagents can be added part by part or all at once at 0°C.

[0308] Table 5

[0309]

[0310]

[0311]

[0312] 1 No alkali is used, and the product is used without further purification after post-treatment.

[0313] 2 After heating at 70-100℃ for 2 hours, add concentrated HCl (c HCl) (0.9 equivalents) and heat the mixture at 60-100℃ for 30 min-24 hours. Cool the mixture to 0℃ and adjust the pH to 9-11 with 30% NaOH or NaHCO3 aqueous solution, and then perform the post-treatment as described above.

[0314] Preparation of 74

[0315] 3-(3,3-dimethylcyclobutyl)-4-fluoro-isoxazo-5-amine

[0316]

[0317] In a sealed tubular flask, 12 mL of ACN was added to 1-(chloromethyl)-4-fluoro-1,4-diazabicyclo[2.2.2]octane; di(tetrafluoroborate) (0.46 g, 1.30 mmol) and 3-(3,3-dimethylcyclobutyl)isoxazole-5-amine (0.20 g, 1.20 mmol). The mixture was irradiated at 100 °C under microwave conditions for 20 min. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude material was purified by reversed-phase chromatography (C18Aq column) using a gradient elution of 0% to 100% ACN in H2O (0.1% HOAc). The desired fractions were combined and concentrated to remove ACN. Saturated NaHCO3 aqueous solution was added to the remaining aqueous solution until a pH of 9 was reached. The mixture was extracted with DCM (2 x 10 mL). The organic extracts were combined, dried over anhydrous Na2SO4, diatomaceous earth was added, and the mixture was concentrated under vacuum. The substance was further purified by silica gel chromatography, eluting with a gradient of 5% to 40% EtOAc in cyclohexane to yield the title compound (48 mg, 22%). ES / MS (m / z) 185.3 (M+H).

[0318] The following compounds in Table 6 were prepared using appropriate reagents, essentially as described with respect to 3-(3,3-dimethylcyclobutyl)-4-fluoro-isoxazole-5-amine, with the reaction time adjusted to determine the end of the reaction and the purification conditions adjusted as appropriate.

[0319] Table 6

[0320]

[0321] Preparation of 77

[0322] 5-(2-methylbutan-2-yl)-1,2-oxazol-3-amine

[0323]

[0324] NaHCO3 (4.53 g, 53.9 mmol) was added fractionally to a stirred mixture of 4,4-dimethyl-3-oxohexanonitrile (3.00 g, 21.6 mmol) and hydroxylamine sulfate (3.89 g, 23.7 mmol) in MeOH (5 mL) and H2O (45 mL) at room temperature under N2. The mixture was stirred at 65 °C under N2 for 5 hours. The mixture was then cooled to room temperature. The mixture was acidified to pH 1 with concentrated HCl and refluxed under N2 with stirring for 20 min. The mixture was then cooled to room temperature and the pH was adjusted to 8 with NaOH. The mixture was extracted with DCM (3 x 200 mL). The combined organic extracts were washed with brine (2 x 200 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 30% to 40% of ACN in H2O (0.1% NH4HCO3) to produce the title compound (1.20 g, 36%) as a pale yellow solid. 1 H NMR (CDCl3) δ5.53 (s, 1H), 1.66 (q, 2H), 1.27 (s, 6H), 0.82 (t, 3H).

[0325] Preparation of 78

[0326] 5-(3,3-Dimethylcyclobutyl)isoxazole-3-amine

[0327]

[0328] Add hydroxylamine hydrochloride (0.79 g, 11.4 mmol) in MeOH (0.5 mL) and NH3 (2.0 mL, 14.0 mmol) in MeOH (7 M) to a sealed tubular flask. Stir the suspension at 25 °C for 30 min. Add a solution of 2-[2-(3,3-dimethylcyclobutyl)-1,3-dioxacyclopentan-2-yl]acetonitrile (0.56 g, 2.87 mmol) in MeOH (0.5 mL) and quinoline-8-ol (42 mg, 0.29 mmol), and stir the mixture at 65 °C for 12 h and cool to room temperature. Filter the mixture, concentrate the filtrate under reduced pressure, and reconcentrate it from toluene (3x) to produce the intermediate 2-[2-(3,3-dimethylcyclobutyl)-1,3-dioxacyclopentan-2-yl]-N′-hydroxyacetamidine as a yellow solid, which is used without further purification. In a sealed tubular flask, 2-[2-(3,3-dimethylcyclobutyl)-1,3-dioxacyclopentan-2-yl]-N′-hydroxy-acetamidinium was dissolved in EtOH (2 mL) and acidified to pH 1 with concentrated HCl. The reaction mixture was stirred at 80 °C for 12 hours. The mixture was concentrated under reduced pressure and the residue was diluted with DCM. Saturated NaHCO3 aqueous solution was added until the solution was alkaline (pH 11). The organic layer was separated and the aqueous phase was extracted with DCM. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude substance was purified by silica gel chromatography, eluting with a gradient of 0% to 5% MeOH in DCM to yield the title compound (30 mg, 6.3%) as a yellow solid. ES / MS (m / z) 167 (M+H).

[0329] Preparation 79

[0330] 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl]methyl acetate

[0331]

[0332] Methyl 2-(4-bromo-2-methoxy-phenyl)acetate (0.15 g, 0.579 mmol), dipentanoyl diborone (0.18 g, 0.709 mmol), KOAc (95%, 0.18 g, 1.74 mmol), Pd(dppf)Cl2·CH2Cl2 (95%, 22 mg, 0.029 mmol), and 1,4-dioxane (2.7 mL) were added together. The reaction mixture was stirred overnight at 80 °C. The reaction was quenched with H2O and DCM was added. The two layers were separated and the aqueous phase was extracted with DCM (3 x 10 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under vacuum to produce the title compound as a brown oil, which was used as a crude product without further purification. ES / MS (m / z) 307.2 (M+H).

[0333] The following compounds in Table 7 were prepared essentially as described with respect to methyl 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl]acetate using appropriate reagents. The reaction time was adjusted to determine the end of the reaction, and the mixture was purified by chromatography as appropriate. The reaction temperature can be in the range of about 80-100 °C. The mixture was filtered through talc and purified as appropriate.

[0334] Table 7

[0335]

[0336] Preparation of 81

[0337] 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)-3-chlorophenyl]methyl acetate

[0338]

[0339] A solution of methyl 2-(3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl)acetate (1.00 g, 3.22 mmol), 5-amino-3-bromo-1-isopropylpyrazole-4-carboxynitrile (0.73 g, 3.22 mmol), Pd(dppf)Cl2 (0.47 g, 0.64 mmol), and K3PO4 (2.05 g, 9.66 mmol) in 1,4-dioxane:H2O (12 mL, 5:1) was stirred at 120 °C under N2 for 2 hours and then cooled to room temperature. The solution was filtered through diatomaceous earth, washed with EtOAc (200 mL), filtered again, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 10% to 50% ACN in H2O to yield the title compound (0.30 g, 28%) as a white solid. ES / MS (m / z) 333.1 (M+H).

[0340] The compounds listed in Table 8 were prepared essentially as described with respect to methyl 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)-3-chlorophenyl]acetate, with the reaction time adjusted to determine the end of the reaction and purification by titration or chromatography as appropriate. The reaction temperature can range from room temperature to 120°C.

[0341] Table 8

[0342]

[0343] Preparation of 83

[0344] 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)-2,3-difluorophenyl]methyl acetate

[0345]

[0346] KOAc (3.70 g, 37.7 mmol) and Pd(dppf)Cl2·CH2Cl2 (1.54 g, 1.88 mmol) were added to a stirred mixture of methyl 2-(4-bromo-2,3-difluorophenyl)acetate (5.00 g, 18.9 mmol) and dipentanoyl diborone (5.75 g, 22.6 mmol) in 1,4-dioxane (30 mL) at room temperature. The mixture was stirred at 80 °C under N2 for 1 hour, filtered, and the filter cake was washed with 1,4-dioxane (5 mL). 5-Amino-3-bromo-1-isopropylpyrazole-4-carboxynitrile (4.75 g, 20.7 mmol), Pd(dppf)Cl2·CH2Cl2 (1.54 g, 1.88 mmol), K2CO3 (5.21 g, 37.7 mmol), and H2O (7 mL) were added to the filtrate at room temperature. The mixture was stirred at 80°C for 1 hour. The mixture was cooled to room temperature and the residue was purified by silica gel chromatography, eluting with PE:EtOAc at a ratio of 3:1 to 2:1 to yield the title compound as a yellow solid (4.90 g, 78%). ES / MS (m / z) 335.1 (M+H).

[0347] Preparation of 84

[0348] 2-Fluoro-4-(2-methoxy-2-oxoethyl)benzyl benzoate

[0349]

[0350] In a pressure vessel, Pd(dppf)Cl2 (0.32 g, 0.44 mmol) was added to a solution of methyl 2-(4-bromo-3-fluorophenyl)acetate (5.45 g, 22.1 mmol), TEA (9.2 mL, 66.0 mmol), and benzyl alcohol (2.5 mL, 24.0 mmol) in toluene (150 mL). The mixture was purified with N2 for 1 min and then pressurized to 30 atm with CO. The mixture was stirred at 115 °C under a CO atmosphere for 24 h. The mixture was cooled to room temperature, filtered, and the filter cake was washed with EtOAc (2 x 50 mL). The filtrate was concentrated under reduced pressure, and the residue was purified by silica gel chromatography with a gradient elution of 10:1 to 3:1 PE:EtOAc to yield the title compound (2.63 g, 39%) as a colorless oil. ES / MS (m / z) 320.05 (M+NH3+H).

[0351] Preparation of 85

[0352] 2-Fluoro-4-(2-methoxy-2-oxoethyl)benzoic acid

[0353]

[0354] Pd / C (10%, 0.90 g, 0.85 mmol) was added to a solution of 2-fluoro-4-(2-methoxy-2-oxoethyl)benzoate (2.63 g, 8.70 mmol) in MeOH (25 mL) at room temperature under N2. The mixture was stirred at room temperature under H2 for 2 hours. The mixture was filtered, the filter cake was washed with MeOH (2 x 10 mL), and the filtrate was concentrated under reduced pressure to yield the title compound (1.51 g, 82%) as a white solid, which was used without further purification.

[0355] Preparation of 86

[0356] 2-[4-(2,2-dicyano-1-methoxyethyl-1-en-1-yl)phenyl]methyl acetate

[0357]

[0358] Oxaloyl chloride (21.2 mL, 247 mmol) was added dropwise to a stirred solution of 4-(2-methoxy-2-oxoethyl)benzoic acid (40.0 g, 206 mmol) and a few drops of DMF in DCM (300 mL) at 0 °C. The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to produce the intermediate methyl 2-(4-(chlorocarbonyl)phenyl)acetate. In another container, a solution of malononitrile (13.6 g, 206 mmol) in THF (100 mL) was added dropwise to a stirred suspension of NaH (16.5 g, 412 mmol, 60% in mineral oil) in THF (100 mL) at 0–10 °C under N2. The mixture was stirred at room temperature for 20 min. methyl 2-(4-(chlorocarbonyl)phenyl)acetate in THF (200 mL) was added dropwise to this mixture at 0–10 °C. The mixture was stirred at room temperature for 1 hour. Dimethyl sulfate (23.4 mL, 247 mmol) was added, and the mixture was refluxed overnight at 80 °C under N2. H2O (300 mL) was added, and the mixture was extracted with EtOAc (3 x 200 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 4:1 to 1:1 PE:EtOAc to yield the title compound (42.0 g, 80%) as a yellow solid. 1 H NMR (CDCl3, 400MHz) δ7.51-7.40 (m, 4H), 3.96 (s, 3H), 3.75 (s, 3H), 3.74 (s, 2H).

[0359] Preparation of 87

[0360] 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)phenyl]methyl acetate

[0361]

[0362] A solution of methyl 2-[4-(2,2-dicyano-1-methoxyeth-1-en-1-yl)phenyl]acetate (2.50 g, 9.76 mmol), isopropylhydrazine HCl (1.29 g, 11.7 mmol), and TEA (4.1 mL, 29.4 mmol) in EtOH (40 mL) was stirred at room temperature under N2 for 1 h. The mixture was concentrated under reduced pressure and extracted with EtOAc (3 x 100 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by grinding with Et2O (50 mL) to yield the title compound (2.00 g, 69%) as a yellow solid. ES / MS (m / z) 299.0 (M+H).

[0363] Preparation of 88

[0364] 2-[4-(5-amino-4-carbamoyl-1-isopropyl-pyrazol-3-yl)-2-methoxy-phenyl]methyl acetate

[0365]

[0366] Methyl 2-[2-methoxy-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl]acetate (67%, 0.21 g, 0.460 mmol), 5-amino-3-bromo-1-isopropyl-pyrazole-4-carboxamide (0.14 g, 0.567 mmol), K₂CO₃ (0.13 g, 0.941 mmol), and Pd(dppf)Cl₂·CH₂Cl₂ (19 mg, 0.0231 mmol) were added to a mixture of 1,4-dioxabane (3.7 mL) and H₂O (0.73 mL). The reaction mixture was heated at 140 °C for 1 hour under microwave irradiation. The reaction mixture was filtered through diatomaceous earth, washed with EtOAc, and evaporated to dryness. The crude material was purified by silica gel chromatography, eluting with a gradient of 0% to 8% MeOH in DCM to yield the title compound (0.17 g, quantitative yield). ES / MS (m / z) 347.2 (M+H).

[0367] The following compounds in Table 9 were prepared essentially as described with respect to methyl 2-[4-(5-amino-4-carbamoyl-1-isopropyl-pyrazol-3-yl)-2-methoxy-phenyl]acetate, with the reaction time adjusted to determine the end of the reaction and purification performed where appropriate.

[0368] Table 9

[0369]

[0370] Preparation of 90

[0371] N-tert-butoxycarbonyl-N-(4-cyano-2-isopropyl-pyrazole-3-yl)carbamate tert-butyl ester

[0372]

[0373] Preparation of 91

[0374] N-(4-cyano-2-isopropyl-pyrazole-3-yl)tert-butyl carbamate

[0375]

[0376] To a stirred solution of 5-amino-1-isopropylpyrazole-4-carboxynitrile (4.80 g, 32.0 mmol) in THF (205 mL), N,N-dimethylpyridin-4-amine (0.38 g, 3.11 mmol), TEA (13 mL, 93.3 mmol), and tert-butoxycarbonyl tert-butyl carbonate (14.7 g, 67.4 mmol) were successively added. The reaction mixture was stirred overnight at room temperature. The reaction mixture was quenched with saturated aqueous NH4Cl solution (15 mL) and extracted with EtOAc (3 x 20 mL). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 2% to 30% EtOAc in cyclohexane to produce N-tert-butoxycarbonyl-N-(4-cyano-2-isopropyl-pyrazole-3-yl)carbamate tert-butyl ester (7.67 g, 68%) as an off-white solid, ES / MS (m / z) 373 (M+Na) and N-(4-cyano-2-isopropyl-pyrazole-3-yl)carbamate tert-butyl ester (1.12 g, 14%) as an off-white solid, ES / MS (m / z) 251 (M+H).

[0377] Preparation of 92

[0378] 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazole-3-yl]-2,3-difluoro-phenyl]methyl acetate

[0379]

[0380] THF (4.8 mL) was added under N2 atmosphere to N-tert-butoxycarbonyl-N-(4-cyano-2-isopropyl-pyrazole-3-yl)carbamate tert-butyl ester (0.79 g, 2.25 mmol), dipentanoyl diboron (0.46 g, 1.81 mmol), [Ir(OMe)(1,5-cod)]2 (31 mg, 0.047 mmol), and 4-tert-butyl-2-(4-tert-butyl-2-pyridinyl)pyridine (26 mg, 0.093 mmol). The mixture was refluxed and stirred for 4 hours. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in a 4:1 DCM:EtOAc mixture (20 mL), and the solution was passed through a silica gel pad (2 g). The pad was washed with a 3:1 DCM:EtOAc mixture (2 x 10 mL). The filtrates were combined, concentrated under reduced pressure, and dried under vacuum to produce a crude residue. Under N2, Cs2CO3 (2.14 g, 6.57 mmol) and methyl 2-(4-bromo-2,3-difluoro-phenyl)acetate (0.50 g, 1.89 mmol) in 1,4-dioxane (10 mL) were added to the residue. Then, under Ar, Pd(dppf)Cl2·CH2Cl2 (0.10 g, 0.123 mmol) and... Molecular sieves. The mixture was heated to 90°C overnight. The mixture was cooled to room temperature, diluted with EtOAc, and filtered through diatomaceous earth. The filtrate was diluted with H2O and the aqueous layer was extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, eluting with a gradient of 2% to 80% DCM in EtOAc to yield the title compound as a beige solid (0.64 g, 94% purity, 73%). ES / MS (m / z) 435 (M+H).

[0381] Preparation of 93

[0382] 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazole-3-yl]phenyl]methyl acetate

[0383]

[0384] THF (25 mL) was added to N-tert-butoxycarbonyl-N-(4-cyano-2-isopropyl-pyrazole-3-yl)carbamate (4.50 g, 12.8 mmol), dipentanoyl diboron (2.61 g, 10.3 mmol), [Ir(Ome)(1,5-cod)]2 (0.18 g, 0.267 mmol), and 4-tert-butyl-2-(4-tert-butyl-2-pyridyl)pyridine (0.15 g, 0.535 mmol) under N2. The mixture was refluxed and stirred for 4 hours. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in a 4:1 DCM:EtOAc mixture (20 mL), and the solution was passed through a silica gel pad and washed with 3:1 DCM:EtOAc (2 x 10 mL). The filtrates were combined, concentrated under reduced pressure, and dried under vacuum to produce a crude residue. Under N2, Cs2CO3 (10.5 g, 32.2 mmol) and methyl (4-bromophenyl)acetate (2.50 g, 10.9 mmol) in 1,4-dioxane (50 mL) were added to the residue. Then, under Ar, Pd(dppf)Cl2·CH2Cl2 (0.53 g, 0.642 mmol) and Molecular sieves. The mixture was heated to 90°C and held for 12 hours, cooled to room temperature, diluted with EtOAc, and filtered through diatomaceous earth. H2O was added to the filtrate, and the aqueous layer was extracted with EtOAc, washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude brown oil was purified by silica gel chromatography, eluting with a gradient of 2% to 80% DCM in EtOAc to yield the title compound (3.80 g, 87%) as a white solid. ES / MS (m / z) 399 (M+H).

[0385] The compounds listed in Table 10 were prepared essentially as described with respect to methyl 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]phenyl]acetate using appropriate reagents. The reaction time was adjusted to determine the end of the reaction, and the compounds were purified by chromatography as appropriate. K₂CO₃ can be substituted for Cs₂CO₃. For the second coupling reaction, the temperature can be in the range of about 50-90 °C. The mono-tert-butoxycarbonylamino and di-tert-butoxycarbonylamino compounds can be separated and used to obtain the common product in subsequent steps.

[0386] Table 10

[0387]

[0388]

[0389]

[0390] Preparation of 103

[0391] 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazole-3-yl]phenyl]acetic acid

[0392]

[0393] To a stirred solution of methyl 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]phenyl]acetate (3.80 g, 9.54 mmol) in MeOH (40 mL), 2 M NaOH (14 mL, 28 mmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The organic layer was concentrated. H₂O was added, followed by dropwise addition of KHSO₄ (3.92 g, 28.8 mmol) in H₂O. The precipitate formed was filtered, washed with H₂O, and dried under vacuum to yield the title compound (2.88 g, 79%), which was used without further purification. ES / MS (m / z) 385 (M+H).

[0394] The compounds listed in Table 11 were prepared essentially as described with respect to 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]phenyl]acetic acid, with the reaction time adjusted to determine the end of the reaction. 1,4-Dioxane could be substituted for MeOH, NH4Cl for KHSO4, and the product was extracted from H2O if appropriate. The mono-tert-butoxycarbonylamino and di-tert-butoxycarbonylamino compounds could be separated and used to obtain a common product in subsequent steps.

[0395] Table 11

[0396]

[0397]

[0398] Preparation 112

[0399] 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]-2,3-difluoro-phenyl]acetic acid

[0400]

[0401] 1 M LiOH hydrate (4.4 mL, 4.40 mmol) was added to methyl 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]-2,3-difluoro-phenyl]acetate (0.64 g, 1.47 mmol) in 1,4-dioxane (6.5 mL) and H₂O (2 mL). The mixture was stirred overnight at room temperature. The reaction mixture was concentrated and poured into H₂O and washed with EtOAc. The aqueous layer was acidified to pH 2–3 with 1 M KHSO₄ aqueous solution and then extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to yield the title compound as a beige solid (0.54 g, 94% purity, 82%). ES / MS (m / z) 421 (M+1).

[0402] Preparation of 113

[0403] 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)methyl benzoate

[0404]

[0405] Added partically to a stirred solution of [4-(methoxycarbonyl)phenyl]acetic acid (20.0 g, 103 mmol), 3-(2,2-dimethylpropyl)-1,2-oxazol-5-amine (17.5 g, 113 mmol), and DIPEA (89.5 mL, 514 mmol) in DMF (200 mL) at 0 °C. (328 g, 515 mmol, 50% in DMF). The mixture was stirred at 50 °C for 2 hours. EtOAc (1 L) was added to the mixture. The mixture was washed with H2O (3 x 300 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 8:1 to 4:1 PE:EtOAc to yield the title compound (31.0 g, 91%) as a pale yellow solid. ES / MS (m / z) 331.2 (M+H).

[0406] The compounds listed in Table 12 are prepared essentially as described with respect to methyl 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoate, with appropriate reagents. The reaction time is adjusted to determine the end of the reaction, and the mixture is purified under appropriate chromatographic conditions or used without further purification. The reaction temperature can be in the range of room temperature to 100°C. Add it to DCM instead of DMF.

[0407] Table 12

[0408]

[0409] Preparation of 117

[0410] 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoic acid

[0411]

[0412] LiOH (11.2 g, 468 mmol) was added fractionally to a stirred solution of methyl 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoate (31.0 g, 93.8 mmol) in THF (240 mL) and H₂O (80 mL). The mixture was stirred overnight. The mixture was acidified to pH 5 with 2N HCl and extracted with EtOAc (3 x 200 mL). The combined organic extracts were washed with brine (3 x 50 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to yield the title compound (28.5 g, 96%) as a white solid. ES / MS (m / z) 317.2 (M+1).

[0413] Prepare the following compounds in Table 13 using appropriate esters, essentially as described with respect to 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoic acid. Adjust the reaction time to determine the end of the reaction, and after completion, adjust the pH to 1-5. Collect the solution as appropriate by extraction or precipitation. If solubility is required, a solvent mixture of 1,4-dioxane / H2O or MeOH / H2O can be used, and the temperature can be in the range of about room temperature to 50°C. 1M NaOH can replace LiOH, and citric acid or KHSO4 can replace HCl.

[0414] Table 13

[0415]

[0416] The compounds listed in Table 14 are prepared essentially as described with respect to methyl 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoate, with appropriate reagents. The reaction time is adjusted to determine the end of the reaction, and the mixture is purified under appropriate chromatographic conditions or used without further purification. The reaction temperature can range from room temperature to 100°C. Add it to DCM or EtOAc instead of DMF.

[0417] Table 14

[0418]

[0419] Preparation 129

[0420] 2-[4-bromo-2-(hydroxymethyl)phenyl]-N-[5-(1-methylcyclopentyl)isoxazo-3-yl]acetamide

[0421]

[0422] Under N2 conditions, 2-[4-bromo-2-(tert-butoxymethyl)phenyl]-N-[5-(1-methylcyclopentyl)isoxazol-3-yl]acetamide (0.29 g, 0.645 mmol) in DCM (8.29 mL) was added to TFA (16.6 mL), 2,2,2-trifluoroacetic acid (2,2,2-trifluoroacetyl) ester (5.42 g, 25.8 mmol), and triethylsilane (1.0 mL, 6.26 mmol). The reaction mixture was stirred for 4 hours. The reaction mixture was concentrated under reduced pressure and dried under vacuum. The substance was dissolved in THF (9 mL) and H2O (4.5 mL). A saturated aqueous solution of NaHCO3 (4 mL) was added, and the mixture was stirred at room temperature for 1 hour. THF was removed under vacuum. The mixture was diluted with DCM (10 mL), the organic layer was separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 1% to 10% MeOH in DCM to yield the title compound (0.20 g, 79%). ES / MS (m / z) 394 (M+H).

[0423] Preparation of 130

[0424] 5-Amino-3-[2-(tert-butoxymethyl)-4-[2-[[5-(1-methylcyclopentyl)isoxazol-3-yl]amino]-2-oxo-ethyl]phenyl]-1-isopropyl-pyrazole-4-carboxamide

[0425]

[0426] Dry THF (4 mL) was added to dipentanoyl diboron (93 mg, 0.366 mmol), 2-[4-bromo-3-(tert-butoxymethyl)phenyl]-N-[5-(1-methylcyclopentyl)isoxazol-3-yl]acetamide (0.15 g, 0.334 mmol), KOAc (0.11 g, 1.12 mmol), and Pd(dppf)Cl2·CH2Cl2 (28 mg, 0.033 mmol) under N2. The reaction mixture was degassed with Ar for 15 min and then heated to 50 °C. The reaction mixture was stirred overnight at 50 °C, then stirred a second night at 65 °C, and then stirred at 80 °C for 24 h. The reaction mixture was cooled to room temperature. 5-Amino-3-bromo-1-isopropyl-pyrazole-4-carboxamide (0.11 g, 0.445 mmol) was added to THF (1.3 mL) and K3PO4 (0.22 g, 1.04 mmol) to H2O (1.3 mL). The reaction mixture was degassed with Ar and XantPhos Pd G3 (17 mg, 0.0167 mmol) was added. The mixture was heated to 50 °C overnight. The mixture was filtered through a talc pad; H2O was added to the filtrate, and the mixture was extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 1% to 10% MeOH in DCM to yield the title compound (67 mg, 37%) as a brown powder. ES / MS (m / z) 537 (M+H).

[0427] Preparation of 131

[0428] 2-[4-(2,2-dicyano-1-hydroxyethyl-1-en-1-yl)phenyl]-N-[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]acetamide

[0429]

[0430] Oxaloyl chloride (10.6 mL, 124 mmol) was added dropwise to a stirred solution of 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoic acid (32.0 g, 101 mmol) and DMF (0.16 mL, 2.14 mmol) in DCM (350 mL). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure to produce crude 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoyl chloride. NaH (7.65 g, 191 mmol, 60% in mineral oil) was added to a solution of malononitrile (6.31 g, 95.5 mmol) in THF (100 mL) at 0 °C. The mixture was stirred for 15 min. 4-([[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)benzoyl chloride (32.0 g, 95.6 mmol) was added dropwise to a solution at 0 °C in THF (150 mL), and the mixture was stirred for 2 hours and warmed to room temperature. The mixture was diluted with H₂O (300 mL), acidified to pH 5 with 2N HCl, and extracted with EtOAc (3 x 200 mL). The combined organic extracts were washed with brine (3 x 100 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to yield the title compound (30.0 g, 82%). The crude product was used directly without further purification. ES / MS (m / z) 365.1 (M+H).

[0431] The compounds listed in Table 15 were prepared essentially as described with respect to 2-[4-(2,2-dicyano-1-hydroxyethyl-1-en-1-yl)phenyl]-N-[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]acetamide, with the reaction time adjusted under N2 if necessary to determine the end of the reaction. A solution of malononitrile in THF and NaH can be stirred at 0°C to room temperature for 15-30 min. The reactants can be acidified or concentrated to dryness and purified by chromatography as appropriate.

[0432] Table 15

[0433]

[0434]

[0435] Preparation of 136

[0436] 2-[4-(2,2-dicyano-1-methoxyethyl-1-en-1-yl)phenyl]-N-[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]acetamide

[0437]

[0438] Dimethyl sulfate (39.0 mL, 411 mmol) was added dropwise to a stirred solution of 2-[4-(2,2-dicyano-1-hydroxyethyl-1-en-1-yl)phenyl]-N-[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]acetamide (25.0 g, 68.6 mmol) and TEA (28.7 mL, 206 mmol) in ACN (240 mL). The mixture was stirred at 50 °C under N2 for 14 hours. The mixture was diluted with EtOAc (500 mL), washed with brine (3 x 50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 8:1 to 2:1 PE:EtOAc to yield the title compound (9.0 g, 35%) as a pale yellow solid. ES / MS(m / z) 379.1(M+H).

[0439] The compounds listed in Table 16 were prepared essentially as described with respect to 2-[4-(2,2-dicyano-1-methoxyethyl-1-en-1-yl)phenyl]-N-[3-(2,2-dimethylpropyl)-1,2-oxazol-5-yl]acetamide, with the reaction time adjusted to determine the end of the reaction and purification performed where appropriate. The temperature can be in the range of about 50°C to 80°C.

[0440] Table 16

[0441]

[0442]

[0443] Preparation of 141

[0444] 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazole-3-yl)phenyl)methyl acetate

[0445]

[0446] Methyl 2-[4-(2,2-dicyano-1-methoxyeth-1-en-1-yl)phenyl]acetate (0.48 g, 1.87 mmol) was added fractionally to a stirred mixture of (1-methylcyclopropyl)hydrazine HCl (crude, 2.00 g, 16.3 mmol) and TEA (1.6 mL, 11.5 mmol) in EtOH (20 mL) at room temperature under N2. The mixture was stirred at 50 °C under N2 for 2 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 30% to 50% ACN in H2O (0.1% NH4HCO3) to yield the title compound (0.50 g, 86%) as a white solid. ES / MS (m / z) 311.3 (M+H).

[0447] The compounds listed in Table 17 were prepared essentially as described with respect to methyl 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetate using appropriate reagents. The reaction time was adjusted to determine the end of the reaction, and the compounds were purified by chromatography as appropriate. The reaction temperature can be in the range of room temperature to 80°C, and the solvent can be THF if suitable, and the base can be DIPEA or TEA.

[0448] Table 17

[0449]

[0450]

[0451]

[0452] Preparation of 155

[0453] 2-(4-(5-amino-4-cyano-1-(1-hydroxy-2-methylpropane-2-yl)-1H-pyrazol-3-yl)phenyl)-N-(3-neopentylisoxazol-5-yl)acetamide

[0454]

[0455] 2-Hydroxy-2-methylpropane-1-ol HCl (43 mg, 0.306 mmol), EtOH (1.5 mL), TEA (0.12 mL, 0.861 mmol), and 2-(4-(2,2-dicyano-1-methoxyvinyl)phenyl)-N-(3-neopentylisoxazol-5-yl)acetamide (0.11 g, 0.291 mmol) were added together and stirred at 45 °C for 18 hours. The mixture was diluted with H2O (1 mL) and extracted with EtOAc (3 × 3 mL). The organic extract was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 0% to 100% EtOAc in hexanes to yield the title compound (39 mg, 30%) as a white solid. ES / MS (m / z) 451.3 (M+H).

[0456] The following compounds in Table 18 were prepared essentially as described with respect to 2-(4-(5-amino-4-cyano-1-(1-hydroxy-2-methylpropane-2-yl)-1H-pyrazol-3-yl)phenyl)-N-(3-neopentylisoxazol-5-yl)acetamide, using 1-3 equivalents of appropriate hydrazine, adjusting the reaction time to determine the end of the reaction, and adjusting the purification conditions as appropriate. The reaction temperature can be in the range of room temperature to 45°C, and THF can be replaced with THF as the solvent based on solubility.

[0457] Table 18

[0458]

[0459]

[0460]

[0461] Preparation of 173

[0462] 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazole-3-yl)phenyl)acetic acid

[0463]

[0464] H2O2 (38.0 mL, 372 mmol, 30% in H2O) was added to a mixture of methyl 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)phenyl]acetate (7.40 g, 24.8 mmol), NaOH (5.96 g, 149 mmol), H2O (24 mL), DMSO (12 mL), and EtOH (60 mL) at room temperature. The mixture was heated to 40 °C and stirred for 6 hours. The mixture was quenched with a saturated aqueous solution of Na2S2O3 and adjusted to pH 4 with a dilute aqueous solution of HCl. The mixture was extracted with EtOAc (3 x 200 mL). The combined organic extracts were concentrated under reduced pressure and the resulting solid was ground together with EtOAc (20 mL). The precipitated solid was collected by filtration, washed with water (3 x 60 mL), and dried under reduced pressure to yield the title compound (4.30 g, 57%) as a yellow solid. ES / MS(m / z) 303.1(M+H).

[0465] Preparation of 174

[0466] 2-(4-(5-amino-4-carbamoyl-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetic acid

[0467]

[0468] Methyl 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetate (0.50 g, 1.61 mmol) and NaOH (0.32 g, 8.00 mmol) in a stirred mixture of EtOH / H2O / DMSO (5 mL / 2 mL / 1 mL) was added dropwise to H2O2 (2.43 mL, 23.8 mmol, 30% in H2O). The mixture was stirred overnight at 50 °C. The reaction was quenched at room temperature with saturated aqueous Na2SO3 solution (10 mL). The residue was adjusted to pH 4 with 1N aqueous HCl solution and extracted with EtOAc (5 x 50 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The obtained solid was ground together with EtOAc (10 mL), and the precipitated solid was collected by filtration and washed with EtOAc (3 x 5 mL) to yield the title compound (0.20 g, 40%) as a white solid. ES / MS (m / z) 315.2 (M+H).

[0469] The compounds listed in Table 19 are prepared essentially as described with respect to 2-(4-(5-amino-4-carbamoyl-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetic acid. The reaction time is adjusted to determine the end of the reaction, and purification is performed by titration or chromatography as appropriate. The reaction temperature can be in the range of about room temperature to 50°C, and about 15-30 equivalents of H2O2 can be used. Reagents can also be added together at about 0°C.

[0470] Table 19

[0471]

[0472] Preparation of 180

[0473] 2-(4-(5-amino-4-carbamoyl-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)phenyl)acetic acid, isomer 1

[0474]

[0475] Preparation of 181

[0476] 2-(4-(5-amino-4-carbamoyl-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)phenyl)acetic acid, isomer 2

[0477] 2-(4-(5-amino-4-carbamoyl-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)phenyl)acetic acid (0.500 g, 1.40 mmol) was separated by Chiral-HPLC under the following conditions: column: Chiralpak IG, 2*25 cm, 5 μm; mobile phase A: hexane (0.1% TFA), mobile phase B: IPA, flow rate 20 mL / min, eluted with 25% B over 12.5 min; to produce the title compound as isomer 1, a pale yellow solid. (R) 6.7min, (0.240g, 48%), ES / MS (m / z) 357.1 (M+H), [α] D 25 = -15.789 (C = 0.304, MeOH), and the title compound as isomer 2, a pale yellow solid, t (R) 9.5min (0.210g, 42%), ES / MS (m / z) 357.1 (M+H), [α] D 25 =17.197 (C=0.314, MeOH).

[0478] Preparation of 182

[0479] 2-[4-(1-isopropyl-6,6-dimethyl-4-oxo-5,7-dihydropyrazolo[3,4-d]pyrimidin-3-yl)phenyl]acetic acid

[0480]

[0481] TFA (0.005 mL) was added to 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (0.100 g, 0.331 mmol) in acetone (5 mL). The mixture was stirred at 50 °C for 2 hours, concentrated under reduced pressure, and dried under vacuum to yield the title compound (0.113 g, 100%) as a yellow solid, which was used without further purification. ES / MS (m / z) 343.2 (M+H).

[0482] The compounds listed in Table 20 were prepared essentially as described with respect to 2-[4-(1-isopropyl-6,6-dimethyl-4-oxo-5,7-dihydropyrazolo[3,4-d]pyrimidin-3-yl)phenyl]acetic acid, with the reaction time adjusted to determine the end of the reaction.

[0483] Table 20

[0484]

[0485] 1 Replace TFA with ethoxyethane HCl and stir the mixture at room temperature for 30 min.

[0486] Preparation of 185

[0487] 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazole-3-yl)phenyl)acetic acid

[0488]

[0489] A solution of methyl 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetate (4.30 g, 13.9 mmol) and LiOH (1.66 g, 69.3 mmol) in THF (40 mL) and H₂O (10 mL) was stirred at room temperature under N₂ for 3 h. The mixture was concentrated under reduced pressure, diluted with H₂O (10 mL), and acidified to pH 3 with 1N HCl aqueous solution. The precipitated solid was collected by filtration and washed with H₂O (3 x 30 mL) to yield the title compound (4.00 g, 97%) as a white solid. ES / MS (m / z) 297.3 (M+H).

[0490] Prepare the following compounds in Table 21 using appropriate esters, essentially as described with respect to 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetic acid. Adjust the reaction time to determine the end of the reaction, and after completion, adjust the pH to 1-5. Collect the solution by extraction or precipitation as appropriate. If solubility is required, a solvent mixture of 1,4-dioxane / H₂O or MeOH / H₂O can be used, and the temperature can be in the range of about room temperature to 50°C. 1M NaOH can replace LiOH, and citric acid or KHSO₄ can replace HCl.

[0491] Table 21

[0492]

[0493]

[0494] Preparation of 200

[0495] 2-[(4-bromo-2,3-difluorophenyl)(methoxy)methylene]malononitrile

[0496]

[0497] NaH (1.67 g, 41.8 mmol, 60% in mineral oil) and malononitrile (1.37 g, 20.7 mmol) were added to THF (50 mL) at 0 °C. The mixture was stirred at 0 °C under N2 for 30 min. 4-Bromo-2,3-difluorobenzoyl chloride (5.30 g, 20.7 mmol) in THF (10 mL) was added dropwise to the mixture at 0 °C. The mixture was stirred at room temperature for 1 h. The intermediate mixture of 2-[(4-bromo-2,3-difluoro-phenyl)-hydroxy-methylene]malononitrile was used directly without further purification. ES / MS (m / z) ( 79 Br / 81Br) 282.9 / 284.9 (MH). Dimethyl sulfate (2.4 mL, 25.3 mmol) was added to a solution of 2-[(4-bromo-2,3-difluoro-phenyl)-hydroxy-methylene]malonium (theoretical 5.90 g, 20.7 mmol) in THF (60 mL). The mixture was refluxed under N2 with stirring overnight. The solution was cooled to room temperature and concentrated under reduced pressure. The crude product was dissolved in EtOAc (200 mL) and diluted with H2O (200 mL), and extracted with EtOAc (3 x 300 mL). The organic extract was washed with brine (2 x 100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography, eluting with a gradient of 3:1 to 2:1 PE:EtOAc to yield the title compound (3.00 g, 48%) as a yellow solid.

[0498] Preparation of 201

[0499] 5-Amino-3-(4-bromo-2,3-difluorophenyl)-1-(1-methylcyclopropyl)pyrazole-4-carboxynitrile

[0500]

[0501] A solution of 2-[(4-bromo-2,3-difluorophenyl)(methoxy)methylene]malononitrile (2.50 g, 8.36 mmol), (1-methylcyclopropyl)hydrazine HCl (3.07 g, 25.0 mmol), and TEA (5.8 mL, 41.6 mmol) in THF (30 mL) was stirred at room temperature under N2 for 1 hour. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel chromatography using a gradient elution of 3:1 to 2:1 PE:EtOAc to yield the title compound (2.50 g, 85%) as a pale yellow solid. ES / MS (m / z) ( 79 Br / 81 Br)353.1 / 355.1(M+H).

[0502] Preparation of 202

[0503] 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazole-3-yl]-2,3-difluorophenyl]tert-butyl acetate

[0504]

[0505] A solution of 5-amino-3-(4-bromo-2,3-difluorophenyl)-1-(1-methylcyclopropyl)pyrazole-4-carboxynitrile (1.80 g, 5.10 mmol), tert-butyl 2-(zinc bromide)acetate (3.98 g, 15.3 mmol), and Pd(t-Bu3P)2 (0.26 g, 0.509 mmol) in THF (20 mL) was stirred at 80 °C under N2 for 1 hour and then cooled to room temperature. The solution was concentrated under reduced pressure, and the residue was purified by silica gel chromatography using a gradient elution of 2:1 to 1:1 PE:EtOAc to yield the title compound (0.50 g, 25%) as a yellow solid. ES / MS (m / z) 389.2 (M+H).

[0506] Preparation of 203

[0507] 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluoro-phenyl]acetic acid

[0508]

[0509] A solution of tert-butyl 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluorophenyl]acetate (0.49 g, 1.26 mmol) and TFA (5 mL) in DCM (10 mL) was stirred at room temperature under N2 for 2 hours and concentrated under reduced pressure. The mixture was diluted with Et2O (20 mL) and concentrated under reduced pressure to yield the title compound (0.40 g, 96%) as a yellow solid. ES / MS (m / z) 333.1 (M+H).

[0510] Preparation of 204

[0511] 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluorophenyl]-N-(3-[3-fluorobicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)acetamide

[0512]

[0513] The following were added: 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluoro-phenyl]acetic acid (80 mg, 0.241 mmol), 3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazole-5-amine (49 mg, 0.291 mmol), DIPEA (0.25 mL, 1.44 mmol), and A solution of (0.46 g, 0.723 mmol, 50% in EtOAc) in DCM (3.00 mL) was stirred at 50 °C under N2 for 1 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 40% to 60% ACN in H2O (0.1% FA) to yield the title compound (60 mg, 52%) as a yellow solid. ES / MS (m / z) 483.1 (M+H).

[0514] Preparation of 205

[0515] 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluorophenyl]-N-(3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)acetamide

[0516]

[0517] Add to a solution of 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluoro-phenyl]acetic acid (80 mg, 0.241 mmol), 3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-amine (47 mg, 0.286 mmol) and DIPEA (0.13 mL, 0.746 mmol) in DCM (3 mL) (0.46 g, 0.723 mmol, 50% in EtOAc). The reactants were stirred in a sealed tube at 50 °C for 2 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 40% to 60% ACN in H2O (0.1% NH4HCO3) to yield the title compound (70 mg, 61%) as a white solid. ES / MS (m / z) 479.1 (M+H).

[0518] Preparation of 206

[0519] 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)-N-(3-(2-chloro-4-fluorophenyl)isoxazo-5-yl)acetamide

[0520]

[0521] 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetic acid (60 mg, 0.202 mmol) was dissolved in DCM (0.41 mL) at 0 °C and then mixed with 3-(2-chloro-4-fluoro-phenyl)isoxazole-5-amine (47 mg, 0.221 mmol). Treatment with (0.77 g, 1.21 mmol, 50% in EtOAc) and DIPEA (0.21 mL, 1.18 mmol). The mixture was stirred for about 20 min, evaporated to dryness, and used without further purification. ES / MS (m / z) 491.1 (M+H).

[0522] Preparation of 207

[0523] 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)phenyl]-N-[3-(pyridin-2-yl)-1,2-oxazol-5-yl]acetamide

[0524]

[0525] Add [4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)phenyl]acetic acid (0.100 g, 0.352 mmol) and 3-(pyridin-2-yl)-1,2-oxazol-5-amine (62 mg, 0.385 mmol) to a stirred mixture in DCM (3 mL) (0.337 g, 0.530 mmol, 50% in EtOAc) and DIPEA (0.31 mL, 1.78 mmol). The mixture was stirred at room temperature for 2 hours and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 10% to 50% ACN in H2O (0.1% FA) to yield the title compound (70 mg, 47%) as a pale yellow solid. ES / MS (m / z) 428.2 (M+H).

[0526] Using appropriate amines and carboxylic acids, with about 1-6 equivalents of coupling agent, about 1-4 equivalents of base, and about 0.3-6 mL of solvent, prepare the following compounds in Table 22 essentially as described with respect to 2-[4-(5-amino-4-cyano-1-isopropylpyrazol-3-yl)phenyl]-N-[3-(pyridin-2-yl)-1,2-oxazol-5-yl]acetamide, adjusting the reaction time to determine the end of the reaction, and adjusting the purification conditions as appropriate. The reaction temperature can be in the range of -20°C to 50°C. Add to 50% EtOAc, 50% DMF, or 50% MeTHF. TCFH can be added in portions, and the reaction can be completed under N2 and in a sealed test tube.

[0527] *Coupling agent: 1 2TCFH, 3BTFFH, 4 5TBTU, 6TFFH, 7HATU 8EDCI / HOBT

[0528] Bases: a. NMI, b. pyridine, c. DIPEA

[0529] Solvents: I EtOAc, II ACN, III DMF, IV DCM, V MeTHF

[0530] Table 22

[0531]

[0532]

[0533]

[0534]

[0535]

[0536]

[0537]

[0538]

[0539]

[0540]

[0541]

[0542] Preparation 278

[0543] N-[4-cyano-5-[4-[2-[[3-(2,2-dimethylpropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]-2,3-difluoro-phenyl]-2-isopropyl-pyrazole-3-yl]tert-butyl carbamate

[0544]

[0545] Under nitrogen (N2), 3-(2,2-dimethylpropyl)-1,2-oxazol-5-amine (58 mg, 0.376 mmol), 2-[4-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]-2,3-difluoro-phenyl]acetic acid (0.100 g, 0.238 mmol), and DIPEA (98 μL, 0.563 mmol) were added together to DMF (0.69 mL). (0.454 g, 0.713 mmol, 50% in MeTHF) and the reaction mixture was stirred overnight at room temperature. The mixture was quenched with a saturated aqueous NH4Cl solution and the aqueous layer was extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 10% to 50% EtOAc in cyclohexane to yield the title compound as a white solid (45 mg, 95% purity, 32%). ES / MS (m / z) 557 (M+H).

[0546] Preparation 279

[0547] N-[4-carbamoyl-5-[4-[2-[[3-(2,2-dimethylpropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]-2,3-difluoro-phenyl]-2-isopropyl-pyrazole-3-yl]tert-butyl carbamate

[0548]

[0549] A mixture of tert-BuOH (1.8 mL) and H₂O (0.9 mL) was added to N₂ with N₂ containing N-[4-cyano-5-[4-[2-[[3-(2,2-dimethylpropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]-2,3-difluoro-phenyl]-2-isopropyl-pyrazole-3-yl]carbamate (75 mg, 0.135 mmol) and Parkin' catalyst (5.8 mg, 0.0135 mmol). The mixture was heated to 65 °C and held for 16 hours. The reaction mixture was cooled to room temperature and diluted with EtOAc. The organic layer was washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, eluting with a gradient of 1% to 10% DCM in MeOH to yield the title compound (48 mg, 62%) as a white solid. ES / MS (m / z) 597 (M+Na).

[0550] Preparation 280

[0551] N-[4-carbamoyl-5-[4-[2-[[3-(2,2-dimethylcyclopropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]phenyl]-2-isopropyl-pyrazole-3-yl]tert-butyl carbamate

[0552]

[0553] Parkin's catalyst (15 mg, 0.0352 mmol) was added to a stirred solution of N-[4-cyano-5-[4-[2-[[3-(2,2-dimethylcyclopropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]phenyl]-2-isopropyl-pyrazole-3-yl]carbamate (0.365 g, 0.704 mmol) in tert-BuOH (10 mL) and H2O (5 mL). The reaction mixture was stirred at 60 °C for 4 h. The reaction mixture was quenched with a saturated aqueous solution of NH4Cl. The aqueous layer was extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The substance was purified by silica gel chromatography, eluting with a gradient of 30% to 80% EtOAc in cyclohexane to yield the title compound (0.363 g, 96%) as a white solid. ES / MS (m / z) 559.4 (M+Na).

[0554] The following compounds in Table 23 were prepared essentially as described with respect to N-[4-carbamoyl-5-[4-[2-[[3-(2,2-dimethylcyclopropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]phenyl]-2-isopropyl-pyrazole-3-yl]tert-butyl carbamate. The reaction time was adjusted to determine the end of the reaction. t-BuOH or 4:1 EtOH:H2O was used as the solvent, and purification was performed using appropriate chromatographic conditions. The temperature could be in the range of approximately 60-70°C.

[0555] Table 23

[0556]

[0557]

[0558]

[0559]

[0560] Preparation of 314

[0561] 3-(2,4-Dichloro-3-fluoro-phenyl)-3-oxo-propionitrile

[0562]

[0563] ACN (0.13 mL, 2.47 mmol) was added dropwise to a stirred solution of LDA (1.35 mL, 2.69 mmol) in THF (4 mL) at -78 °C under N2. The reaction mixture was stirred at -78 °C for 1 hour, followed by the dropwise addition of a solution of methyl 2,4-dichloro-3-fluorobenzoate in THF. Once the addition was complete, the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with a saturated aqueous NH4Cl solution. The pH was adjusted to 5 with HCl (1N), followed by extraction with EtOAc. The organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated to provide the title compound (0.83 g). This crude material was used without further purification to prepare 3-(2,4-dichloro-3-fluoro-phenyl)isoxazole-5-amine.

[0564] Preparation of 315

[0565] 3-(2,4-Dichloro-3-fluoro-phenyl)isoxazole-5-amine

[0566]

[0567] A solution of 3-(2,4-dichloro-3-fluoro-phenyl)-3-oxo-propionitrile (0.27 g, 1.15 mmol) in EtOH (3.7 mL) and H₂O (3.7 mL) was treated with hydroxylamine hydrochloride (0.40 g, 5.74 mmol) and KOAc (0.68 g, 6.89 mmol). The mixture was stirred at 60 °C for 1 h. The reactants were extracted with EtOAc and the organic layer was concentrated under vacuum. The residue was purified by normal-phase silica gel chromatography, eluting with 0% to 100% EtOAc in hexanes, followed by 0% to 20% MeOH in EtOAc, to provide the title compound (0.18 g, 65%). ES / MS (m / z) 247.0 (M+H).

[0568] Preparation of 316

[0569] 2-[5-(5-amino-4-cyano-1-ethyl-pyrazol-3-yl)-2-pyridyl]acetate

[0570]

[0571] A mixture of 5-amino-3-bromo-1-ethyl-pyrazole-4-carboxynitrile (233.58 mg, 1.09 mmol), K₂CO₃ (299.78 mg, 2.17 mmol), Pd(dppf)Cl₂ (79.48 mg, 0.11 mmol), and methyl 2-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-2-pyridyl]acetate (301 mg, 1.09 mmol) in a reaction flask containing 1,4-dioxane (9.05 mL) and H₂O (1.81 mL) was bubbled with nitrogen and heated to 80 °C for 90 min. The reactants were partitioned between DCM and H₂O and extracted with DCM (3x) through a phase separator. The organic phases were combined and concentrated under vacuum. The residue was purified by silica gel chromatography, eluting with 0% to 100% EtOAc in hexanes to yield the title compound (172 mg, 55.5%). ES / MS (m / z) 286.1 (M+H).

[0572] Preparation of 317

[0573] 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]methyl acetate

[0574]

[0575] 2-(6-chloropyridin-3-yl)methyl acetate (2.00 g, 10.78 mmol), hexa-n-butylditin (9.38 g, 16.16 mmol) and Pd(PPh3)4 (7.47 g, 6.46 mmol) were combined in 1,4-dioxane (40 mL) under N2 and the reaction mixture was stirred at 65 °C for three days.

[0576] 5-Amino-3-bromo-1-isopropylpyrazole-4-carboxynitrile (0.75 g, 3.30 mmol) was added to a portion (25%) of the above reaction mixture under N2, and the reaction mixture was stirred at 75 °C for 12 h. The solvent was removed under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 2:1 to 1:1 PE:EtOAc to provide the crude product. The product was further purified by reversed-phase chromatography (C18 gel column), eluting with a gradient of 20% to 50% ACN in H2O (0.1% FA) to give the title compound (0.12 g, 15%) as a yellow solid. ES / MS (m / z) 300.2 (M+H).

[0577] Preparation of 318

[0578] 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-2-pyridyl]methyl acetate

[0579]

[0580] Methyl 2-(5-bromopyridin-2-yl)acetate (1.00 g, 4.35 mmol), dipentanoyl diboron (1.33 g, 5.22 mmol), KOAc (0.85 g, 8.70 mmol), Pd(dppf)Cl2 (0.32 g, 0.43 mmol), and 1,4-dioxane (30 mL) were added together. The reaction mixture was stirred at 100 °C for 2 hours under N2. The reaction mixture was filtered, and the filtrate was concentrated under vacuum to produce an intermediate residue.

[0581] The residue, 5-amino-3-bromo-1-isopropylpyrazole-4-carboxynitrile (0.99 g, 4.33 mmol), potassium carbonate (1.19 g, 8.60 mmol), Pd(dppf)Cl2 (0.32 g, 0.43 mmol), 1,4-dioxane (30 mL), and H2O (7 mL) were added together. The reaction mixture was stirred at 100 °C for 2 hours under N2. The reaction mixture was diluted with H2O (50 mL) and extracted with EtOAc (3 x 100 mL). The combined organic extracts were washed with brine (2 x 100 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography, eluting with a gradient of 6:1 to 2:1 PE:EtOAc to yield the title compound (0.65 g, 50%) as a yellow solid. ES / MS (m / z) 300.1 (M+H).

[0582] The compounds listed in Table 24 were prepared essentially as described with respect to methyl 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-2-pyridyl]acetate, with the reaction time adjusted to determine the end of the reaction.

[0583] Table 24

[0584]

[0585] Preparation of 321

[0586] 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]acetic acid

[0587]

[0588] LiOH (0.024 g, 1.00 mmol) was added to methyl 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]acetate (0.10 g, 0.33 mmol) in methanol (2 mL) and H₂O (2 mL). The mixture was stirred at 60 °C for 2 h. The reaction mixture was acidified to pH 5 with 1 M HCl aqueous solution and then extracted with EtOAc (2 x 30 mL). The combined organic extracts were washed with brine (2 x 20 mL), dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 gel column) by elution with a gradient of 5% to 60% ACN in H₂O (0.1% FA) to yield the title compound (0.080 g, 80%) as a yellow solid. ES / MS (m / z) 286.4 (M+H).

[0589] The compounds listed in Table 25 were prepared essentially as described with respect to 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]acetic acid, with the reaction time adjusted to determine the end of the reaction.

[0590] Table 25

[0591]

[0592] Preparation of 324

[0593] 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-2-pyridyl]lithium acetate

[0594]

[0595] LiOH (0.17 g, 7.20 mmol) was added to methyl 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-2-pyridyl]acetate (2.40 g, 8.00 mmol) in THF (20 mL) and H₂O (4 mL). The mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with H₂O (20 mL) and then washed with EtOAc (3 x 50 mL). The aqueous layer was concentrated under reduced pressure to yield the title compound (1.50 g, 65%) as a yellow solid. ES / MS (m / z) 286.1 (M+H).

[0596] The compounds listed in Table 26 were prepared essentially as described with respect to lithium 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-2-pyridyl]acetate, with the reaction time adjusted to determine the end of the reaction.

[0597] Table 26

[0598]

[0599] Preparation of 327

[0600] 5-Amino-3-(5-chloropyrazin-2-yl)-1-isopropyl-pyrazole-4-carboxynitrile

[0601]

[0602] DIPEA (0.30 g, 2.32 mmol) was added fractionally to a stirred solution of 5-chloropyrazine-2-carboxaldehyde (0.30 g, 2.11 mmol) and isopropylhydrazine hydrochloride (0.26 g, 2.32 mmol) in DMF (4 mL) at room temperature under N2. The mixture was stirred at 80 °C for 1 hour and then cooled to room temperature. A solution of N-bromosuccinimide (0.41 g, 2.32 mmol) in DMF (4 mL) was added fractionally over 5 min. The mixture was stirred at room temperature for 2 hours.

[0603] Meanwhile, sodium ethoxide (0.39 g, 5.80 mmol) was added to a stirred solution of malononitrile (0.15 g, 2.32 mmol) in EtOH (2 mL) at 0 °C under N2. The mixture was stirred at room temperature for 30 min.

[0604] The first mixture was added dropwise to the second mixture at 0°C under N2 for 30 min. The reactants were stirred at 80°C for 2 h. The mixture was diluted with EtOAc and washed with H2O. The organic layer was concentrated under vacuum. The residue was purified by silica gel chromatography, eluting with a gradient of 3:1 to 2:1 PE:EtOAc to yield the title compound (0.26 g, 47%) as a white solid. 1 H NMR (d6-DMSO, 300MHz) δ (ppm) 8.88 (d, 1H), 8.82 (d, 1H), 6.80 (s, 2H), 4.56 (m, 1H), 1.37 (d, 6H).

[0605] The compounds listed in Table 27 were prepared essentially as described with respect to 5-amino-3-(5-chloropyrazin-2-yl)-1-isopropyl-pyrazole-4-carboxynitrile, with the reaction time adjusted to determine the end of the reaction.

[0606] Table 27

[0607]

[0608] Preparation 329

[0609] N-[5-(5-bromopyrimidin-2-yl)-4-cyano-2-isopropylpyrazol-3-yl]-N-tert-butoxycarbonyl-tert-butyl carbamate

[0610]

[0611] A mixture of 5-amino-3-(5-bromopyrimidin-2-yl)-1-isopropyl-pyrazole-4-carboxynitrile (0.35 g, 1.14 mmol), di-tert-butyl dicarbonate (0.50 g, 2.28 mmol), and DMAP (0.029 g, 0.23 mmol) in THF (10 mL) was stirred at room temperature for 4 hours. The mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 10:1 to 1:1 PE:EtOAc to yield the title compound (0.40 g, 69%) as a yellow solid. ES / MS (m / z) 507.0, 509.1 (M+H).

[0612] Preparation of 330

[0613] 2-[2-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]pyrimidin-5-yl]tert-butyl acetate

[0614]

[0615] A solution of N-[5-(5-bromopyrimidin-2-yl)-4-cyano-2-isopropyl-pyrazol-3-yl]-N-tert-butoxycarbonyl-tert-butyl carbamate (0.39 g, 0.77 mmol), tert-butyl 2-(zinc bromide)acetate (0.40 g, 1.54 mmol), and Pd(t-Bu3P)2 (0.079 g, 0.15 mmol) in THF (20 mL) was stirred at 80 °C under N2 for 4 h and then cooled to room temperature. The solution was concentrated under reduced pressure and the residue was purified by silica gel chromatography, eluting with EtOAc to provide the crude product. The product was further purified by reversed-phase chromatography (C18 column), eluting with a gradient of 30% to 50% ACN in H2O (0.1% FA) to yield the title compound (0.24 g, 71%) as a yellow solid. ES / MS(m / z) 443.2(M+H).

[0616] The compounds listed in Table 28 were prepared essentially as described with respect to 2-[2-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]pyrimidin-5-yl]tert-butyl acetate, with the reaction time adjusted to determine the end of the reaction.

[0617] Table 28

[0618]

[0619] Preparation of 332

[0620] 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)pyrazin-2-yl]acetic acid

[0621]

[0622] TFA (10 mL) was added to 2-[5-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)pyrazin-2-yl]tert-butyl acetate (0.26 g, 0.76 mmol). The mixture was stirred at room temperature for 30 min and concentrated under reduced pressure to produce the title compound (0.20 g, 91%) as a yellow solid, which was used without further purification.

[0623] Preparation of 333

[0624] 2-[2-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)pyrimidin-5-yl]acetic acid

[0625]

[0626] To tert-butyl 2-[2-[5-(tert-butoxycarbonylamino)-4-cyano-1-isopropyl-pyrazol-3-yl]pyrimidin-5-yl]acetate (0.22 g, 0.50 mmol), 4 M HCl (10 mL) in 1,4-dioxane was added. The mixture was stirred at room temperature for 1 h and concentrated under reduced pressure. The reaction mixture was adjusted to pH 6 with saturated aqueous NaHCO3 solution and then extracted with EtOAc (3 x 50 mL). The combined organic extracts were concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 30% to 40% ACN in H2O (0.1% FA) to yield the title compound (0.075 g, 53%) as a yellow solid. ES / MS (m / z) 287.1 (M+H).

[0627] Example 1

[0628] 5-Amino-1-isopropyl-3-(4-(2-((3-(((1-methylcyclopropyl)methyl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1H-pyrazole-4-carboxamide

[0629]

[0630] 3-[(1-methylcyclopropyl)methyl]isoxazole-5-amine (62 mg, 0.407 mmol) and 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (0.10 g, 0.331 mmol) were combined in EtOAc (0.38 mL), pyridine (0.64 mL), and ACN (0.64 mL). The mixture was stirred for 5 min and a turbid suspension was observed. The mixture was cooled to -20 °C and added dropwise. (0.58 mL, 0.99 mmol, 50% in DMF). The solution was stirred at -20 °C for 45 min. The mixture was diluted with H₂O (50 mL) and extracted with EtOAc (2 x 50 mL). The combined organic extracts were dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 10% to 100% EtOAc in hexanes to yield the title compound (28 mg, 19%). ES / MS (m / z) 437.2 (M+H).

[0631] Example 2

[0632] 5-Amino-1-isopropyl-3-(4-(2-oxo-2-((5-(1,1,1-trifluoro-2-methylpropane-2-yl)isoxazol-3-yl)amino)ethyl)phenyl)-1H-pyrazole-4-carboxamide

[0633]

[0634] 5-(2,2,2-trifluoro-1,1-dimethyl-ethyl)isoxazol-3-amine (0.394 g, 2.03 mmol) and 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (0.500 g, 1.65 mmol) were combined in ACN (20 mL), EtOAc (16 mL), and pyridine (2 mL). The mixture was stirred for 2 min until a clear and colorless solution was produced. The solution was cooled to -20 °C and added dropwise. (1.89 mL, 3.23 mmol, 50% in DMF). The solution was stirred at -20 °C for 30 min. The reactants were diluted with H₂O (8 mL) and extracted with EtOAc (2 x 10 mL). The combined organic extracts were dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 0% to 100% EtOAc in hexanes, followed by a gradient of 0% to 10% MeOH in EtOAc (containing 0.1% NH₄OH) to yield the title compound (0.363 g, 46%). ES / MS (m / z) 478.8 (M+H).

[0635] Example 3

[0636] 5-Amino-1-isopropyl-3-(4-(2-((3-(1-methylcyclopentyl)isoxazo-5-yl)amino)-2-oxoethyl)phenyl)-1H-pyrazole-4-carboxamide

[0637]

[0638] DIPEA (0.034 mL, 0.195 mmol) was added to a mixture of 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (20 mg, 0.066 mmol), 3-(1-methylcyclopentyl)isoxazole-5-amine (11 mg, 0.066 mmol), and [[(E)-(1-cyano-2-ethoxy-2-oxo-ethylidene)amino]oxy-morpholino-methylene]-dimethyl-ammonium hexafluorophosphate (34 mg, 0.079 mmol) in DMF (3 mL), and the mixture was stirred at room temperature for 6 hours. The mixture was diluted with H2O (20 mL) and extracted with EtOAc (2 x 20 mL). The combined organic extracts were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 0% to 100% EtOAc in hexanes to yield the title compound (3.4 mg, 11%). ES / MS (m / z) 451.2 (M+H).

[0639] Example 4

[0640] 5-Amino-3-(4-(2-((3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-isopropyl-1H-pyrazole-4-carboxamide

[0641]

[0642] In a sealed test tube, 0.100 g (0.331 mmol) of 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid and 61 mg (0.363 mmol) of 3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazole-5-amine were added. (0.28 mL, 0.470 mmol, 50% in EtOAc) and DIPEA (0.17 mL, 0.976 mmol) were added together to DCM (5 mL) and stirred at room temperature under N2 for 10 hours. The mixture was concentrated under reduced pressure and the residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 20% to 50% ACN in H2O (0.1% NH4HCO3) to yield the title compound (67 mg, 45%) as a white solid. ES / MS (m / z) 453.1 (M+H).

[0643] Example 5

[0644] 5-Amino-3-(4-[[(3-[bicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)carbamoyl]methyl]phenyl)-1-isopropylpyrazole-4-carboxamide

[0645]

[0646] 2-(4-(5-amino-4-carbamoyl-1-isopropyl1H-pyrazol-3-yl)phenyl)acetic acid (0.100 g, 0.331 mmol), 3-[bicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-amine (60 mg, 0.400 mmol), and DIPEA (0.17 mL, 0.976 mmol) were added together to DCM (10 mL). (0.30 mL, 0.504 mmol, 50% in EtOAc) and the mixture was stirred at room temperature under N2 for 2 hours. The solution was concentrated under reduced pressure and redissolved in ACN (5 mL). The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 20% to 60% ACN in H2O (0.1% NH4HCO3) to yield the title compound (24 mg, 17%) as a white solid. ES / MS (m / z) 435.3 (M+H).

[0647] Example 6

[0648] 5-Amino-1-(1-methylcyclopropyl)-3-[4-([[5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1,2-oxazol-3-yl]carbamoyl]methyl)phenyl]pyrazole-4-carboxamide

[0649]

[0650] 5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1,2-oxazol-3-yl)phenyl)acetic acid (70 mg, 0.223 mmol) and NMI (55 mg, 0.670 mmol) were added fractionally to a stirred mixture of 2-(4-(5-amino-4-carbamoyl-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)acetic acid (70 mg, 0.223 mmol) and NMI (55 mg, 0.670 mmol) in ACN (5 mL) at room temperature under N2. The mixture was stirred at 50 °C for 2 hours under N2. The mixture was cooled to room temperature and concentrated under reduced pressure to a residue. The residue was purified by preparative TLC, eluted with 100% EtOAc, and further purified by reversed-phase chromatography (C18 column) with a gradient elution of 40% to 50% ACN in H2O to yield the title compound (20 mg, 18%) as a white solid. ES / MS (m / z) 491.2 (M+H).

[0651] Example 7

[0652] 5-Amino-1-isopropyl-3-[4-([[3-(1-methylcyclopropyl)-1,2-oxazol-5-yl]carbamoyl]methyl)phenyl]pyrazole-4-carboxamide

[0653]

[0654] DIPEA (0.26 mL, 1.49 mmol) was added portionwise to a stirred mixture of 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (0.300 g, 0.992 mmol) and 3-(1-methylcyclopropyl)-1,2-oxazol-5-amine (0.151 g, 1.09 mmol) in DCM (10 mL) at room temperature under N2. (0.71 mL, 1.19 mmol, 50% in EtOAc). The mixture was stirred at room temperature under N2 for 3 hours and concentrated to dryness. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 30% to 40% ACN in H2O (0.1% NH4HCO3) to yield the title compound (69 mg, 16%) as a white solid. ES / MS (m / z) 423.3 (M+H).

[0655] Example 8

[0656] 5-Amino-3-[4-([[1-Methyl-5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazol-3-yl]carbamoyl]methyl)phenyl]-1-(propane-2-yl)-1H-pyrazol-4-carboxamide]

[0657]

[0658] A solution of 1-methyl-5-(1,1,1-trifluoro-2-methylpropane-2-yl)-1H-pyrazole-3-amine (35 mg, 0.169 mmol), 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazole-3-yl)phenyl)acetic acid (56 mg, 0.185 mmol), HATU (96 mg, 0.252 mmol), and DIPEA (0.088 mL, 0.505 mmol) in DMF (2 mL) was stirred at room temperature under N2 for 2 hours. The mixture was purified by reversed-phase chromatography (C18 gel column) with a gradient elution of 50% to 70% ACN in H2O (0.1% FA) to yield the title compound (28 mg, 34%) as a white solid. ES / MS (m / z) 492.3 (M+H).

[0659] Example 9

[0660] 5-Amino-1-isopropyl-3-[4-([[3-(1,1,1-trifluoro-2-methylpropane-2-yl)-1,2-oxazol-5-yl]carbamoyl]methyl)phenyl]pyrazole-4-carboxamide

[0661]

[0662] 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazole-3-yl)phenyl)acetic acid (0.150 g, 0.496 mmol), 3-(1,1,1-trifluoro-2-methylpropane-2-yl)-1,2-oxazol-5-amine (0.106 g, 0.546 mmol), DIPEA (0.35 mL, 2.01 mmol) and A solution of (0.59 mL, 0.991 mmol, 50% in EtOAc) in DCM (5 mL) was stirred at room temperature under N2 for 2 hours. The mixture was concentrated and the residue was purified by reversed-phase chromatography (C18 gel column) with a gradient elution of 30% to 50% ACN in H2O (0.1% FA). The product was further purified by reversed-phase preparative HPLC (Phenomenex Gemini C6-Phenyl column) with a gradient elution of 37% to 45% ACN in H2O (0.05% FA) to yield the title compound (81 mg, 34%) as a white solid. ES / MS (m / z) 479.3 (M+H).

[0663] Example 10

[0664] 5-Amino-3-(4-(2-((5-(tert-butyl)isothiazo-3-yl)amino)-2-oxoethyl)phenyl)-1-isopropyl-1H-pyrazole-4-carboxamide

[0665]

[0666] 5-tert-butylisothiazol-3-amine (47 mg, 0.301 mmol) and 2-[4-(5-amino-4-carbamoyl-1-isopropyl-pyrazol-3-yl)phenyl]acetic acid (60 mg, 0.198 mmol) were combined in ACN (1.05 mL), EtOAc (0.65 mL), and pyridine (0.31 mL). The mixture was stirred for 2 min until a clear and colorless solution was produced. The solution was cooled to -20 °C and added dropwise. (0.26 mL, 0.437 mmol, 50% in EtOAc). The solution was stirred at -20 °C for 30 min. The reactants were diluted with H2O (8 mL) and extracted with DCM (2 x 10 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 0% to 100% EtOAc in hexanes, followed by a gradient of 0% to 20% MeOH in EtOAc (containing 1% NH4OH), to yield the title compound (21 mg, 24%) as a pale yellow solid. ES / MS (m / z) 441.2 (M+H).

[0667] Example 11

[0668] 5-Amino-1-isopropyl-3-(4-[[(3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)carbamoyl]methyl]phenyl)pyrazole-4-carboxamide

[0669]

[0670] DIPEA (0.52 mL, 2.99 mmol) was added to a stirred mixture of 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (0.300 g, 0.992 mmol) and 3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-amine (0.163 g, 0.993 mmol) in DCM (20 mL) at room temperature. (0.89 mL, 1.50 mmol, 50% in EtOAc). The mixture was stirred in a sealed tube at 50 °C under N2 for 1 h. The mixture was concentrated under reduced pressure and purified by reversed-phase chromatography (C18 column) with a gradient elution of 30% to 40% ACN in H2O (0.1% NH4HCO3) to yield the title compound (90 mg, 20%) as a white solid. ES / MS (m / z) 449.2 (M+H).

[0671] Example 12

[0672] 5-Amino-1-isopropyl-3-(4-(2-oxo-2-((3-(2-(trifluoromethyl)phenyl)isoxazo-5-yl)amino)ethyl)phenyl)-1H-pyrazole-4-carboxamide

[0673]

[0674] The 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazole-3-yl)phenyl)acetic acid (75 mg, 0.248 mmol) and 3-[2-(trifluoromethyl)phenyl]isoxazole-5-amine (53 mg, 0.232 mmol) in EtOAc (0.14 mL) and ACN (0.14 mL) were mixed with pyridine (0.14 mL) and (0.28 mL, 0.470 mmol, 50% in EtOAc) was treated. The mixture was stirred at 0 °C under N2 for 20 min, and then partitioned between H2O and EtOAc. The organic layer was concentrated and purified by reversed-phase preparative HPLC, eluting with a gradient of 5% to 95% ACN in H2O (0.2% TFA) to yield the title compound (4.1 mg, 3.4%). ES / MS (m / z) 513.1 (M+H).

[0675] Using appropriate amines and carboxylic acids, with about 1-6 equivalents of coupling agent, about 1-4 equivalents of base, and about 0.3-20 mL of solvent, the following compounds in Table 29 were prepared essentially as described with respect to 5-amino-1-isopropyl-3-(4-(2-oxo-2-((3-(2-(trifluoromethyl)phenyl)isoxazol-5-yl)amino)ethyl)phenyl)-1H-pyrazole-4-carboxamide, adjusting the reaction time to determine the end of the reaction, and adjusting the purification conditions as appropriate. The reaction temperature can be in the range of -20°C to 50°C. Add 50% EtOAc and 50% DMF, and TCFH can be added in portions.

[0676] *Coupling agent: 1 2TCFH, 3BTFFH, 4 5TBTU, 6TFFH, 7HATU, 8EDCI / HOBT

[0677] Bases: a. NMI, b. pyridine, c. DIPEA

[0678] Solvents: I EtOAc, II ACN, III DMF, IV DCM, V THF

[0679] Table 29

[0680]

[0681]

[0682]

[0683]

[0684]

[0685]

[0686]

[0687]

[0688]

[0689]

[0690]

[0691]

[0692]

[0693]

[0694]

[0695]

[0696]

[0697]

[0698]

[0699]

[0700]

[0701] Example 153

[0702] 5-Amino-1-isopropyl-3-(4-(2-oxo-2-((3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)isoxazol-5-yl)amino)ethyl)phenyl)-1H-pyrazole-4-carboxamide

[0703]

[0704] 2-(4-(5-amino-4-carbamoyl-1-isopropyl-1H-pyrazol-3-yl)phenyl)acetic acid (196 mg, 0.65 mmol) and 3-(3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl)isoxazol-5-amine (140.00 mg, 0.65 mmol) were combined in DCM. DIPEA (251.36 mg, 1.95 mmol) was added dropwise at room temperature and under N2. (536.32 mg, 0.84 mmol, 50 wt% in EtOAc), and the reactants were stirred in a sealed tube at 50 °C for 1 hour. The reactants were concentrated under reduced pressure and the residue was purified by reversed-phase rapid chromatography (C18 column) with a gradient elution of 40% to 50% ACN in H2O (0.1% NH4HCO3) to yield the title compound (120 mg). The substance was ground together with hexane (3 mL). The precipitated solid was collected by filtration and washed with hexane (3 x 3 mL) to yield the title product (85.1 mg, 34%) as a white solid. ES / MS (m / z) 503.2 (M+H).

[0705] Example 154

[0706] 5-Amino-3-[4-[2-[[3-(2,2-dimethylpropyl)isoxazo-5-yl]amino]-2-oxo-ethyl]-2,3-difluoro-phenyl]-1-isopropyl-pyrazole-4-carboxamide

[0707]

[0708] TFA (0.23 mL, 3.12 mmol) was added to N-[4-carbamoyl-5-[4-[2-[[3-(2,2-dimethylpropyl)isoxazol-5-yl]amino]-2-oxo-ethyl]-2,3-difluoro-phenyl]-2-isopropyl-pyrazole-3-yl]carbamate (45 mg, 0.078 mmol) in DCM (0.55 mL). The mixture was stirred for 3 hours. The reaction mixture was concentrated under reduced pressure, the residue was pHed to alkaline with saturated NaHCO3 aqueous solution, and extracted with EtOAc. The organic extract was washed with brine and concentrated under reduced pressure. The residue was purified by silica gel chromatography, eluting with a gradient of 1% to 10% MeOH in DCM to produce a white solid. The solid was ground in iPr₂O at room temperature, filtered, washed with iPr₂O, and dried overnight under vacuum at 80 °C to produce the title compound (26 mg, 70%) as a white solid. ES / MS (m / z) 475 (M+H).

[0709] Example 155

[0710] 5-Amino-3-(2,3-difluoro-4-(2-((3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-isopropyl-1H-pyrazole-4-carboxamide

[0711]

[0712] TFA (0.39 mL, 5.10 mmol) was added to (30 mg, 0.051 mmol) tert-butyl (4-carbamoyl-3-(2,3-difluoro-4-(2-((3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-isopropyl-1H-pyrazole-5-yl)carbamate in DCM (2 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with cold, saturated aqueous NaHCO3 solution, and the aqueous layer was extracted with EtOAc. The organic extract was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, eluting with a gradient of 10% to 100% EtOAc in heptane to yield the title compound (10 mg, 40%). ES / MS (m / z) 489.4 (M+H).

[0713] The compounds listed in Table 30 are prepared essentially as described with respect to 5-amino-3-(2,3-difluoro-4-(2-((3-(3-fluorobicyclo[1.1.1]pentan-1-yl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-isopropyl-1H-pyrazole-4-carboxamide, with the reaction time adjusted to determine the end of the reaction, purification performed using appropriate chromatographic conditions, and / or grinding in a suitable solvent if necessary. TFA can also be added dropwise to the reactants. THF can be used instead of DCM if appropriate for solubility.

[0714] Table 30

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721]

[0722] Example 196

[0723] 5-Amino-1-isopropyl-3-[2-(methoxymethyl)-4-[2-[[5-(1-methylcyclopentyl)isoxazol-3-yl]amino]-2-oxo-ethyl]phenyl]pyrazole-4-carboxamide

[0724]

[0725] Dipentanoyl diboron (89 mg, 0.350 mmol) and KOAc (95 mg, 0.968 mmol) were added to 2-[4-bromo-3-(methoxymethyl)phenyl]-N-[5-(1-methylcyclopentyl)isoxazol-3-yl]acetamide (0.130 g, 0.319 mmol) in THF (3 mL). The reaction mixture was degassed with Ar for 10 min, and Pd(dpPf)Cl2·CH2Cl2 (27 mg, 0.032 mmol) was added. The mixture was stirred overnight at 50 °C and cooled to room temperature. 5-Amino-3-bromo-1-isopropyl-pyrazole-4-carboxamide (95 mg, 0.384 mmol) was added to THF (1.2 mL), followed by K3PO4 (0.206 g, 0.970 mmol) in H2O (1.2 mL). The mixture was degassed with Ar for 10 min and XantPhos Pd G3 (95%, 16 mg, 0.016 mmol) was added. The mixture was heated to 50 °C and held for 20 h, cooled to room temperature, and filtered through talc. The filtrate was diluted with EtOAc / H2O and the aqueous layer was extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by silica gel chromatography, eluting with a gradient of 1% to 10% MeOH in DCM, followed by a second chromatography eluting with a gradient of 90% to 100% EtOAc in DCM, and then reversed-phase chromatography (C18Aq column) eluting with a gradient of 0% to 100% ACN (0.1% HOAc) in H2O to produce the title compound. The residue was dissolved in DCM and added dropwise to pentane. The suspension was stirred at room temperature for 2 h and filtered. The solid was dried under vacuum at 38°C for 3 days to produce the title compound (35 mg, 22%). ES / MS (m / z) 495 (M+H).

[0726] The compounds listed in Table 31 were prepared essentially as described with respect to 5-amino-1-isopropyl-3-[2-(methoxymethyl)-4-[2-[[5-(1-methylcyclopentyl)isoxazol-3-yl]amino]-2-oxo-ethyl]phenyl]pyrazole-4-carboxamide, with the reaction time adjusted to determine the end of the reaction and purification performed under appropriate chromatographic conditions. The reaction temperature can be in the range of about 50-55°C.

[0727] Table 31

[0728]

[0729] Example 199

[0730] 5-Amino-3-(4-(2-((3-(2-chloro-4-fluorophenyl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-(1-methylcyclopropyl)-1H-pyrazole-4-carboxamide

[0731]

[0732] 2-(4-(5-amino-4-cyano-1-(1-methylcyclopropyl)-1H-pyrazol-3-yl)phenyl)-N-(3-(2-chloro-4-fluorophenyl)isoxazo-5-yl)acetamide (66 mg, 0.134 mmol) and Parkins catalyst (69.2 mg, 0.161 mmol) were stirred at 60 °C for 4 h in EtOH (1.0 mL) and H₂O (1.0 mL), followed by stirring at 50 °C for 4 h and then stirring overnight at room temperature. The reaction mixture was extracted with EtOAc and washed with H₂O (3x). The combined organic extracts were concentrated and purified by silica gel chromatography, eluting with a gradient of 0% to 100% EtOAc in hexanes, followed by a gradient of 0% to 20% MeOH (containing 1% NH₄OH) in EtOAc, to yield the title compound (35 mg, 51%). ES / MS (m / z) 509.1

[0733] The following compounds in Table 32 were prepared essentially as described with respect to 5-amino-3-(4-(2-((3-(2-chloro-4-fluorophenyl)isoxazol-5-yl)amino)-2-oxoethyl)phenyl)-1-(1-methylcyclopropyl)-1H-pyrazole-4-carboxamide, with the reaction time adjusted to determine the end of the reaction and purification performed under appropriate chromatographic conditions.

[0734] Table 32

[0735]

[0736]

[0737]

[0738]

[0739] Example 221

[0740] 5-Amino-3-[2,3-difluoro-4-[2-[[3-(3-fluoro-1-bicyclo[1.1.1]pentyl)isoxazol-5-yl]amino]-2-oxo-ethyl]phenyl]-1-(1-methylcyclopropyl)pyrazole-4-carboxamide

[0741]

[0742] 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluorophenyl]-N-(3-[3-fluorobicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)acetamide (60 mg, 0.124 mmol), NaOH (25 mg, 0.625 mmol), and H2O2 (0.38 mL, 3.72 mmol, 30% in H2O) were added together in an EtOH:DMSO mixture (6 mL, 5:1) and stirred at 50 °C for 1 hour. The solution was cooled to room temperature, quenched with a saturated aqueous solution of Na2SO3 (10 mL), and extracted with EtOAc (3 x 20 mL). The organic extract was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 40% to 60% ACN in H2O (0.1% FA) to yield the title compound (45 mg, 73%). ES / MS (m / z) 501.2 (M+H).

[0743] Example 222

[0744] 5-Amino-3-(2,3-difluoro-4-[[(3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)carbamoyl]methyl]phenyl)-1-(1-methylcyclopropyl)pyrazole-4-carboxamide

[0745]

[0746] H₂O₂ (0.22 mL, 2.15 mmol, 30% in H₂O) was added to a solution of 2-[4-[5-amino-4-cyano-1-(1-methylcyclopropyl)pyrazol-3-yl]-2,3-difluorophenyl]-N-(3-[3-methylbicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl)acetamide (70 mg, 0.146 mmol) and NaOH (29 mg, 0.725 mmol) in EtOH (3 mL) and DMSO (0.60 mL). The reaction mixture was stirred at 50 °C for 1 hour. The mixture was cooled to room temperature and quenched with a saturated aqueous solution of Na₂SO₃ (10 mL) at 0 °C. The aqueous layer was extracted with EtOAc (2 x 30 mL). The combined organic extracts were washed with brine (2 x 20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 column) with a gradient elution of 40% to 60% of ACN in H2O (0.1% NH4HCO3) to yield the title compound (36 mg, 50%) as a white solid. ES / MS (m / z) 497.3 (M+H).

[0747] Example 223

[0748] 5-Amino-1-isopropyl-3-[4-([[3-(pyridin-2-yl)-1,2-oxazol-5-yl]carbamoyl]methyl)phenyl]pyrazole-4-carboxamide

[0749]

[0750] H2O2 (0.22 mL, 2.15 mmol, 30% in H2O) was added to a stirred solution of 2-[4-(5-amino-4-cyano-1-isopropylpyrazole-3-yl)phenyl]-N-[3-(pyridin-2-yl)-1,2-oxazol-5-yl]acetamide (60 mg, 0.140 mmol) and NaOH (28 mg, 0.700 mmol) in EtOH (1.5 mL), DMSO (0.30 mL), and H2O (0.60 mL). The mixture was stirred at 40 °C for 3 hours. The reaction was quenched with saturated aqueous Na2SO3 solution (10 mL). The mixture was extracted with EtOAc (3 x 100 mL). The combined organic extracts were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase chromatography, eluting with a gradient of 10% to 30% ACN in H2O (0.1% NH3H2O) to yield the title compound (26 mg, 40%) as a white solid. ES / MS (m / z) 446.2 (M+H).

[0751] The compounds listed in Table 33 were prepared essentially as described with respect to 5-amino-1-isopropyl-3-[4-([[3-(pyridin-2-yl)-1,2-oxazol-5-yl]carbamoyl]methyl)phenyl]pyrazole-4-carboxamide, 0.3 formic acid. The reaction time was adjusted to determine the end of the reaction, and purification was performed using appropriate chromatographic conditions. Salt formation depends on the purification conditions. H2O2 (about 15-30 equivalents) can be added dropwise or fractionally, and H2O can be added if necessary, with the temperature ranging from about room temperature to 50°C.

[0752] Table 33

[0753]

[0754]

[0755]

[0756]

[0757]

[0758] Example 258

[0759] 5-Amino-3-[4-[([3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl]carbamoyl)methyl]phenyl]-1-[1,1,1-trifluoropropane-2-yl]pyrazole-4-carboxamide, isomer 1

[0760]

[0761] Example 259

[0762] 5-Amino-3-[4-[([3-[3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl]carbamoyl)methyl]phenyl]-1-[1,1,1-trifluoropropane-2-yl]pyrazol-4-carboxamide, isomer 2, was purified by preparative chiral HPLC (Prep-CHIRAL-HPLC) under the following conditions: column: Lux 5μCellulose-4, AXIA Packed, 2.12*25cm, 5μm, eluted with 80% hexane (0.1% TFA) and 20% IPA; flow rate 20mL / min, over 37min, 205 / 250nm, to produce the title compound t as a light brown solid, isomer 1. (R) 18.6 min (15.4 mg, 22.0%, ee = 100%), ES / MS (m / z) 557.2 (M+H), and the title compound t as a light brown solid isomer 2. (R) 29.8 min (14.9 mg, 21.2%, ee=99.5%), ES / MS (m / z) 557.2 (M+H).

[0763] The compounds listed in Table 34 were prepared essentially as described with respect to 5-amino-3-[4-[([3-[3-(trifluoromethyl)bicyclo[1.1.1]pentan-1-yl]-1,2-oxazol-5-yl]carbamoyl)methyl]phenyl]_1-[1,1,1-trifluoropropane-2-yl]pyrazole-4-carboxamide isomers 1 and 2, with the purification system adjusted as appropriate.

[0764] Table 34

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771] 1 CHIRAL ART Amylose-SA, 2*25cm, 5μm, 70% hexanes (10mM NH3 in MeOH) and 30% EtOH.

[0772] 2 CHIRALPAK IA 2*25cm, 5μm, 70% hexanes (10mM NH3 in MeOH) and 30% EtOH.

[0773] 3 CHIRALPAK AD 250mm x 4.6, 5μm, with 70% to 30% CO2 of EtOH (0.5% IPAm).

[0774] 4 Waters Prep SFC200 Chiralpak AD-H 5μm, 250x30mm 70 30CO2 / EtOH (0.5% IPAm).

[0775] 5 CHIRALPAK AD-H 5μm, 250x30mm, 78% CO2 / MeOH (0.5% IPAm).

[0776] 6 Chiralcel OD 500x76.5mm, 20μm, 100% ACN (0.05% IPA).

[0777] 7 Waters Prep SFC80 (Pirkle (R, R) Whelk-015μm, 250x21.1mm, 70% CO2 / MeOH (0.5% IPAm).

[0778] 8 ChiralPak IC, 4.6x150mm, 30% MeOH / CO2.

[0779] 9 ChiralPak AD-H, 4.6x150mm, 35% EtOH / CO2.

[0780] 10 ChiralCel OJ-H, 4.6x150mm, 15% MeOH / CO2.

[0781] 11 ChiralPak IA, 4.6x150mm, 30% EtOH / CO2.

[0782] Example 292

[0783] 5-Amino-1-isopropyl-3-[5-[2-oxo-2-[3-(trifluoromethyl)anilino]ethyl]-2-pyridyl]pyrazole-4-carboxamide

[0784]

[0785] Add to a stirred solution of 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]acetic acid (0.070 g, 0.24 mmol), 3-(trifluoromethyl)aniline (0.047 g, 0.29 mmol), and DIPEA (0.16 g, 1.22 mmol) in DMF (2 mL) (0.46 g, 0.73 mmol, 50% in DMF). The mixture was stirred at room temperature under N2 for 2 h, and then concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 gel column) with a gradient elution of 40% to 60% ACN in H2O (0.1% FA) to yield 2-[6-(5-amino-4-cyano-1-isopropylpyrazol-3-yl)-3-pyridyl]-N-[3-(trifluoromethyl)phenyl]acetamide (0.050 g, 45%) as a white solid. ES / MS (m / z) 429.1 (M+H).

[0786] H₂O₂ (0.13 g, 1.16 mmol, 30% in H₂O) was added to a stirred solution of 2-[6-(5-amino-4-cyano-1-isopropyl-pyrazol-3-yl)-3-pyridyl]-N-[3-(trifluoromethyl)phenyl]acetamide (0.050 g, 0.11 mmol) and NaOH (0.13 g, 0.58 mmol) in EtOH:DMSO (1.2 mL, 5:1). The mixture was stirred at 40 °C for 4 hours. The solution was cooled to 0 °C, quenched with a saturated aqueous solution of Na₂SO₃, and extracted with EtOAc (2 x 10 mL). The combined organic extracts were washed with brine (2 x 30 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by reversed-phase chromatography (C18 gel column) with a gradient elution of 35% to 45% ACN in H2O (0.1% NH3H2O) to yield the title compound (0.021 g, 40%) as a white solid. ES / MS (m / z) 447.1 (M+H).

[0787] The compounds in Table 35 were prepared essentially as described with respect to 5-amino-1-isopropyl-3-[5-[2-oxo-2-[3-(trifluoromethyl)anilino]ethyl]-2-pyridyl]pyrazole-4-carboxamide using appropriate amines and carboxylic acids (or lithium salts), with the reaction time adjusted as needed to complete the reaction.

[0788] Table 35

[0789]

[0790]

[0791]

[0792]

[0793]

[0794]

[0795]

[0796]

[0797] Biological assay

[0798] The following assays confirm that the compound described in this article is a RET kinase inhibitor.

[0799] Assay A: RET enzyme assay

[0800] Using CisBio KinEASE TM -TK assay technology screens compounds of formulas I, Ia to IX, IIa to IIx and / or IIIa to IIIx for their ability to inhibit wild-type, V804M and G810S mutant RET kinases. Recombinant human RET cytoplasmic domain (aa 658-terminus) (1.25 nM RET; catalog number 14-570M) N-terminal GST-labeled recombinant human V804M mutant RET cytoplasmic domain (amino acid 658-terminus) (1.25 nM enzyme; catalog number 14-760) or N-terminal GST-labeled recombinant human G810S (aa-658-terminus) (1.25 nM enzyme; produced in insect cells) were prepared with 62.5 nM TK-substrate biotin (CisBio, part of catalog number 62TK0PEC) and 1 mM ATP, along with the test compound (0.4% final DMSO in this assay) in 1X Cisbio enzyme buffer (containing 1 nM DTT, 5 mM ATP, and 62.5 nM TK-substrate biotin (part of CisBio; catalog number 62TK0PEC) and 1 mM ATP, and with the test compound (0.4% final DMSO in this assay). The reaction was incubated in 10 μL of MgCl2, 0.04% BSA, and 0.05% Tween 20. The compound was typically prepared by a three-fold serial dilution in DMSO and added to the assay to produce the appropriate final concentration. After incubation at 22°C for 40–60 minutes (40 minutes for V804M, 60 minutes for WT and G810S), the reaction was quenched by adding 10 μL of a quenching solution containing 7.8 nM streptavidin-XL665 and 0.5X TK-ab-Cryptate (both from CisBio, catalog number 62TK0PEC) to the HTRF detection buffer. After incubation at 22°C for 60–80 minutes (60 minutes for WT, 80 minutes for V804M and G810S), the extent of the reaction was determined using a Pherastar microplate reader by HTRF detection at excitation / emission at 337 / 665 nm. Calculate the percentage of inhibition, where 0% inhibition refers to the control condition without the compound (0.4% DMSO only), and 100% inhibition represents the condition without the enzyme. Fit the percentage inhibition values ​​to a 4-parameter logic curve, and determine the IC50. 50 The value was defined as the estimated concentration of the inhibitor at the inflection point of the fitted curve. Compounds from Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 153, 154, 199, 221, 222, 292, 293, 294, 299, 300, 301, 302, 303, 306, 308, 310, 311, 316, 317, 318, 321, 324, 329, 330, and 335 all showed IC50 values ​​less than 150 nM for at least one of the wild-type, V804M, and G810S mutant RET kinases in these assays.50 The data confirms that these compounds are RET inhibitors, as are the other compounds disclosed in this article.

[0801] Assay B: RET cell assay

[0802] The ability of compounds of formulas I, Ia to Ix, IIa to IIx, and / or IIIa to IIIx to inhibit intracellular autophosphorylation of RET at tyrosine 1062 was screened, as detected by In-Cell Western blotting. HEK293 cells containing doxycycline inducible plasmids (for inducible expression of KIF5B-RET WT and RET mutant forms V804M and G810S) were seeded at 25,000 cells / well in black poly-D-lysine-coated 384-well plates (Corning, catalog number 356697). KIF5B-RET expression was induced with 1 μg / mL doxycycline, and cells were incubated overnight at 37°C. Compounds were prepared by three-fold serial dilutions in DMSO, followed by a further 1:100 dilution in culture medium, and then added to cells (0.1% final DMSO concentration). The cells were treated with the compound at 37°C for 1 hour, followed by fixation with 4% formaldehyde at room temperature for 20 minutes, and then permeabilized with ice-cold MeOH at room temperature for 10 minutes. The cells were then... Incubate with blocking buffer (Li-COR, catalog number 927-70010) at room temperature for 1 hour. Incubate overnight at 4°C with primary antibody against human phosphorylated-RET (Y1062) (R&D, catalog number AF5009, 1 / 250 dilution) and human glyceraldehyde-3-phosphate dehydrogenase (clone 6C5, Merck, catalog number MAB374, 1 / 1000 dilution). Incubate with secondary antibody... 800CW goat anti-rabbit IgG (Li-COR, catalog number 926-32211, 1 / 1000 dilution) and 680CW goat anti-mouse IgG (Li-COR, catalog number 926-68070, 1 / 1000 dilution) was incubated together at room temperature for 1 hour. In a solution containing 0.05% Tween 20... All antibody dilutions were performed in blocking buffer. After washing cells with PBS-T (Cell Signaling technology, catalog number 9809), images were acquired at 700 and 800 nm on a Li-COROdyssey LCx. The percentage of DMSO control was calculated, where 100% signal indicates the control condition without the compound (0.4% DMSO only), and 0% signal indicates the condition containing the control compound. The percentage values ​​of DMSO control were fitted to a 4-parameter logistic curve, and the measured EC50 values ​​were analyzed. 50The value was defined as the estimated concentration of the inhibitor at the inflection point of the fitted curve. Compounds in Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 153, 154, 199, 221, 222, 292, 293, 294, 299, 300, 301, 302, 303, 306, 308, 310, 311, 316, 317, 318, 321, 324, 329, 330, and 335 all inhibited intracellular autophosphorylation of RET at tyrosine 1062 in at least one of the wild-type, V804M, and G810S RET mutant cell assays, exhibiting EC50 concentrations less than 150 nM. 50 The data confirms that these compounds are RET inhibitors, as are the other compounds disclosed in this article. sequence list <110> Eli Lilly and Company <120> RET kinase inhibitors <130> X22787 <150> 63 / 110,643 <151> 2020-11-06 <160> 1 <170> PatentIn version 3.5 <210> 1 <211> 1114 <212> PRT <213> Homo sapiens <400> 1 Met Ala Lys Ala Thr Ser Gly Ala Ala Gly Leu Arg Leu Leu Leu Leu 1 5 10 15 Leu Leu Leu Pro Leu Leu Gly Lys Val Ala Leu Gly Leu Tyr Phe Ser 20 25 30 Arg Asp Ala Tyr Trp Glu Lys Leu Tyr Val Asp Gln Ala Ala Gly Thr 35 40 45 Pro Leu Leu Tyr Val His Ala Leu Arg Asp Ala Pro Glu Glu Val Pro 50 55 60 Ser Phe Arg Leu Gly Gln His Leu Tyr Gly Thr Tyr Arg Thr Arg Leu 65 70 75 80 His Glu Asn Asn Trp Ile Cys Ile Gln Glu Asp Thr Gly Leu Leu Tyr 85 90 95 Leu Asn Arg Ser Leu Asp His Ser Ser Trp Glu Lys Leu Ser Val Arg 100 105 110 Asn Arg Gly Phe Pro Leu Leu Thr Val Tyr Leu Lys Val Phe Leu Ser 115 120 125 Pro Thr Ser Leu Arg Glu Gly Glu Cys Gln Trp Pro Gly Cys Ala Arg 130 135 140 Val Tyr Phe Ser Phe Phe Asn Thr Ser Phe Pro Ala Cys Ser Ser Leu 145 150 155 160 Lys Pro Arg Glu Leu Cys Phe Pro Glu Thr Arg Pro Ser Phe Arg Ile 165 170 175 Arg Glu Asn Arg Pro Pro Gly Thr Phe His Gln Phe Arg Leu Leu Pro 180 185 190 Val Gln Phe Leu Cys Pro Asn Ile Ser Val Ala Tyr Arg Leu Leu Glu 195 200 205 Gly Glu Gly Leu Pro Phe Arg Cys Ala Pro Asp Ser Leu Glu Val Ser 210 215 220 Thr Arg Trp Ala Leu Asp Arg Glu Gln Arg Glu Lys Tyr Glu Leu Val 225 230 235 240 Ala Val Cys Thr Val His Ala Gly Ala Arg Glu Glu Val Val Met Val 245 250 255 Pro Phe Pro Val Thr Val Tyr Asp Glu Asp Asp Ser Ala Pro Thr Phe 260 265 270 Pro Ala Gly Val Asp Thr Ala Ser Ala Val Val Glu Phe Lys Arg Lys 275 280 285 Glu Asp Thr Val Val Ala Thr Leu Arg Val Phe Asp Ala Asp Val Val 290 295 300 Pro Ala Ser Gly Glu Leu Val Arg Arg Tyr Thr Ser Thr Leu Leu Pro 305 310 315 320 Gly Asp Thr Trp Ala Gln Gln Thr Phe Arg Val Glu His Trp Pro Asn 325 330 335 Glu Thr Ser Val Gln Ala Asn Gly Ser Phe Val Arg Ala Thr Val His 340 345 350 Asp Tyr Arg Leu Val Leu Asn Arg Asn Leu Ser Ile Ser Glu Asn Arg 355 360 365 Thr Met Gln Leu Ala Val Leu Val Asn Asp Ser Asp Phe Gln Gly Pro 370 375 380 Gly Ala Gly Val Leu Leu Leu His Phe Asn Val Ser Val Leu Pro Val 385 390 395 400 Ser Leu His Leu Pro Ser Thr Tyr Ser Leu Ser Val Ser Arg Arg Ala 405 410 415 Arg Arg Phe Ala Gln Ile Gly Lys Val Cys Val Glu Asn Cys Gln Ala 420 425 430 Phe Ser Gly Ile Asn Val Gln Tyr Lys Leu His Ser Ser Gly Ala Asn 435 440 445 Cys Ser Thr Leu Gly Val Val Thr Ser Ala Glu Asp Thr Ser Gly Ile 450 455 460 Leu Phe Val Asn Asp Thr Lys Ala Leu Arg Arg Pro Lys Cys Ala Glu 465 470 475 480 Leu His Tyr Met Val Val Ala Thr Asp Gln Gln Thr Ser Arg Gln Ala 485 490 495 Gln Ala Gln Leu Leu Val Thr Val Glu Gly Ser Tyr Val Ala Glu Glu 500 505 510 Ala Gly Cys Pro Leu Ser Cys Ala Val Ser Lys Arg Arg Leu Glu Cys 515 520 525 Glu Glu Cys Gly Gly Leu Gly Ser Pro Thr Gly Arg Cys Glu Trp Arg 530 535 540 Gln Gly Asp Gly Lys Gly Ile Thr Arg Asn Phe Ser Thr Cys Ser Pro 545 550 555 560 Ser Thr Lys Thr Cys Pro Asp Gly His Cys Asp Val Val Glu Thr Gln 565 570 575 Asp Ile Asn Ile Cys Pro Gln Asp Cys Leu Arg Gly Ser Ile Val Gly 580 585 590 Gly His Glu Pro Gly Glu Pro Arg Gly Ile Lys Ala Gly Tyr Gly Thr 595 600 605 Cys Asn Cys Phe Pro Glu Glu Glu Lys Cys Phe Cys Glu Pro Glu Asp 610 615 620 Ile Gln Asp Pro Leu Cys Asp Glu Leu Cys Arg Thr Val Ile Ala Ala 625 630 635 640 Ala Val Leu Phe Ser Phe Ile Val Ser Val Leu Leu Ser Ala Phe Cys 645 650 655 Ile His Cys Tyr His Lys Phe Ala His Lys Pro Pro Ile Ser Ser Ala 660 665 670 Glu Met Thr Phe Arg Arg Pro Ala Gln Ala Phe Pro Val Ser Tyr Ser 675 680 685 Ser Ser Gly Ala Arg Arg Pro Ser Leu Asp Ser Met Glu Asn Gln Val 690 695 700 Ser Val Asp Ala Phe Lys Ile Leu Glu Asp Pro Lys Trp Glu Phe Pro 705 710 715 720 Arg Lys Asn Leu Val Leu Gly Lys Thr Leu Gly Glu Gly Glu Phe Gly 725 730 735 Lys Val Val Lys Ala Thr Ala Phe His Leu Lys Gly Arg Ala Gly Tyr 740 745 750 Thr Thr Val Ala Val Lys Met Leu Lys Glu Asn Ala Ser Pro Ser Glu 755 760 765 Leu Arg Asp Leu Leu Ser Glu Phe Asn Val Leu Lys Gln Val Asn His 770 775 780 Pro His Val Ile Lys Leu Tyr Gly Ala Cys Ser Gln Asp Gly Pro Leu 785 790 795 800 Leu Leu Ile Val Glu Tyr Ala Lys Tyr Gly Ser Leu Arg Gly Phe Leu 805 810 815 Arg Glu Ser Arg Lys Val Gly Pro Gly Tyr Leu Gly Ser Gly Gly Ser 820 825 830 Arg Asn Ser Ser Ser Leu Asp His Pro Asp Glu Arg Ala Leu Thr Met 835 840 845 Gly Asp Leu Ile Ser Phe Ala Trp Gln Ile Ser Gln Gly Met Gln Tyr 850 855 860 Leu Ala Glu Met Lys Leu Val His Arg Asp Leu Ala Ala Arg Asn Ile 865 870 875 880 Leu Val Ala Glu Gly Arg Lys Met Lys Ile Ser Asp Phe Gly Leu Ser 885 890 895 Arg Asp Val Tyr Glu Glu Asp Ser Tyr Val Lys Arg Ser Gln Gly Arg 900 905 910 Ile Pro Val Lys Trp Met Ala Ile Glu Ser Leu Phe Asp His Ile Tyr 915 920 925 Thr Thr Gln Ser Asp Val Trp Ser Phe Gly Val Leu Leu Trp Glu Ile 930 935 940 Val Thr Leu Gly Gly Asn Pro Tyr Pro Gly Ile Pro Pro Glu Arg Leu 945 950 955 960 Phe Asn Leu Leu Lys Thr Gly His Arg Met Glu Arg Pro Asp Asn Cys 965 970 975 Ser Glu Glu Met Tyr Arg Leu Met Leu Gln Cys Trp Lys Gln Glu Pro 980 985 990 Asp Lys Arg Pro Val Phe Ala Asp Ile Ser Lys Asp Leu Glu Lys Met 995 1000 1005 Met Val Lys Arg Arg Asp Tyr Leu Asp Leu Ala Ala Ser Thr Pro 1010 1015 1020 Ser Asp Ser Leu Ile Tyr Asp Asp Gly Leu Ser Glu Glu Glu Thr 1025 1030 1035 Pro Leu Val Asp Cys Asn Asn Ala Pro Leu Pro Arg Ala Leu Pro 1040 1045 1050 Ser Thr Trp Ile Glu Asn Lys Leu Tyr Gly Met Ser Asp Pro Asn 1055 1060 1065 Trp Pro Gly Glu Ser Pro Val Pro Leu Thr Arg Ala Asp Gly Thr 1070 1075 1080 Asn Thr Gly Phe Pro Arg Tyr Pro Asn Asp Ser Val Tyr Ala Asn 1085 1090 1095 Trp Met Leu Ser Pro Ser Ala Ala Lys Leu Met Asp Thr Phe Asp 1100 1105 1110 To be

Claims

1. A compound of the following formula or a pharmaceutically acceptable salt thereof: Where A-(R1) n yes in Indicates the connection to the main chain; Each R1 is independently a C1-C6 alkyl, -(C0-C4 alkyl)(C3-C7 cycloalkyl), -(C0-C4 alkyl)(C3-C7 heteroalkyl), or bridged C4-C 10 Bicycloalkyl or phenyl, wherein each R1 is unsubstituted or, when it is substituted, is substituted by one or more substituents, said substituents being independently selected from the group consisting of halogens, methyl, ethyl and mono-, di- or tri-halomethyl, and n is 1, 2 or 3; X1 and X3 are each independently CH or C-halogens; X2 and X4 are each independently an N, CH, or C halogen; and R2 is 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein at least one of X1, X2, X3 and X4 is CH.

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein X1, X2, X3 and X4 are each CH.

4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein at least one of X2 and X4 is N.

5. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein X2 is N and each of X1, X3 and X4 is CH.

6. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein A-(R1) n yes And n is 1 or 2.

7. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein A-(R1) n yes 8. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein A-(R1) n yes Where n is 1 or 2.

9. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

10. The compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, wherein n is 1 and R1 is 2,2-dimethylpropyl or 11. The compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, wherein R2 is 12. The compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, wherein R2 is 13. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is:

14. A compound or a pharmaceutically acceptable salt thereof, wherein said compound is 15. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-14 in the preparation of a medicament for treating cancer in a patient, wherein the cancer is RET-related cancer.

16. The use according to claim 15, wherein the RET-related cancer is selected from the group consisting of: lung cancer, thyroid cancer, type 2A or 2B multiple endocrine neoplasia, pheochromocytoma, parathyroid hyperplasia, breast cancer, colorectal cancer, papillary renal cell carcinoma, ganglioneuroma of the gastrointestinal mucosa, and cervical cancer.

17. The use according to claim 16, wherein the thyroid cancer is medullary thyroid carcinoma, differentiated thyroid carcinoma, or recurrent thyroid carcinoma.

18. The use according to claim 16, wherein the thyroid cancer is papillary thyroid carcinoma or refractory differentiated thyroid carcinoma.

19. The use according to claim 16, wherein the lung cancer is small cell lung cancer, non-small cell lung cancer, bronchioloalveolar cell carcinoma, or lung adenocarcinoma.

20. A pharmaceutical composition comprising a compound according to any one of claims 1-14 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier, diluent or excipient.

Images