Substituted pyrazolopyrimidines and substituted purines and their use as ubiquitin specific processing protease 1 (USP1) inhibitors

Abstract

The present disclosure provides compounds of Formula I, and pharmaceutically acceptable salts and solvates thereof, wherein X 1 , X 2 , X 11 , X 12 , R 1 , R 3 , R 5 , R 5 , R 6 , and R 7 are as defined in the specification. The present disclosure also relates to the use of compounds of Formula I to inhibit USP1 protein and / or to treat a disorder that is responsive to inhibition of USP1 protein and USP1 activity. The compounds of the present disclosure are particularly useful for treating cancer.

Description

[0001] Background of this disclosure Technical Field

[0002] This disclosure provides substituted pyrazolopyrimidines and substituted purines as ubiquitin-specific processing protein 1 (USP1) inhibitors, and treatment methods for diseases and ailments in which inhibition of USP1 provides benefit. Specifically, this disclosure provides a method for treating cancer by administering a USP1 inhibitor. Background Technology

[0003] Ubiquitin is a small (76 amino acid) protein that attaches to a target protein post-transcriptionally. The outcome of ubiquitination is determined by the number of ubiquitin molecules bound to the target protein and the bonding topology. For example, proteins exhibiting polyubiquitin chains linked by 48 lysine residues are generally targets for proteasome degradation, while monoubiquitination or polyubiquitin chains linked by other lysine residues regulate non-proteolytic functions such as cell cycle regulation, DNA damage repair, transcription, and endocytosis. Ubiquitination is a reversible process, and enzymes called deubiquitinating enzymes remove ubiquitin from the target protein.

[0004] USP1 is a deubiquitinating enzyme that plays a role in DNA damage repair. USP1 interacts with UAF1 (USP1-associated factor 1) to form a complex required for the deubiquitinating enzyme's activity. The USP1 / UAF1 complex deubiquitinates monoubiquitinated proliferating cell nuclear antigen (PCNA) and monoubiquitinated Fanconi anemia group complementation group D2 (FANCD2), proteins that play important roles in the translesion synthesis (TLS) and Fanconi anemia (FA) pathways, respectively. The USP1 / UAF1 complex also deubiquitinates Fanconi anemia complementation group I (FANCI). These two pathways are essential for repairing DNA damage induced by DNA cross-linking agents such as cisplatin and mitomycin C (MMC).

[0005] Safe and effective treatments targeting deubiquitinating enzymes are unclear, and they are not commercially available or have not yet been developed clinically. Summary of the Invention

[0006] In one respect, this disclosure relates to compounds having Formula I or pharmaceutically acceptable salts or solvates thereof (also referred to herein as compounds of this disclosure):

[0007]

[0008] in:

[0009] X 1 and X 2 Each is independently selected from N and CR 2 ;

[0010] R 1 and R 2 Each is independently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, and optionally substituted alkynyl group;

[0011] R 3 It is optionally substituted phenyl, optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyrazinyl, optionally substituted pyridazinyl, or optionally substituted pyrazolyl.

[0012] X 11 and X 12 Each is independently selected from N and CH;

[0013] R 5' Independently selected from hydrogen, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)alkynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)hydroxyalkyl, cyano, halogroup, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 -NR 31a SO2R 27 Optional replacement of (C6-C) 14 )Aryl, optionally substituted (C6-C) 14 ) aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl, optionally substituted -O-(C6-C14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic and optionally substituted -O-heterocyclic-(C1-C2)alkyl;

[0014] R 5 Independently selected from optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)alkynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)hydroxyalkyl, cyano, halogroup, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 -NR 31a SO2R 27 Optional replacement of (C6-C) 14 )Aryl, optionally substituted (C6-C) 14 ) aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl, optionally substituted -O-(C6-C 14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic and optionally substituted -O-heterocyclic-(C1-C2)alkyl; or

[0015] An R on an adjacent atom 5And an R 5' Together with the atoms it is attached to, it forms an optional substituted (C6-C) 14 ) aryl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optional substituted heteroaryl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optionally substituted (C3-C8) cycloalkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optionally substituted heterocyclic alkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the same atom it is attached to, it forms an optionally substituted spirocyclic alkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the same atom it is attached to, it forms an optional substituted spiroheterocyclic alkyl ring;

[0016] R 6 and R 7 Each is independently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, and optionally substituted alkynyl group;

[0017] R 23 Choose from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0018] R 31a and R 31b Each is independently selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, alkoxyalkyl, cycloalkyl, (cycloalkyl)alkyl, (heterocyclic)alkyl, aralkyl, and (heteroaryl)alkyl; and

[0019] R 24 R 25 R 27 R 32a and R 32bEach is independently selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, alkoxyalkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0020] In some embodiments, the compounds disclosed herein exhibit improved solubility, as measured, for example, by the ADME solubility assay disclosed herein.

[0021] In some embodiments, the compounds disclosed herein exhibit improved metabolic stability, as measured, for example, by the liver microsomal and hepatocyte metabolic stability assays disclosed herein.

[0022] In other embodiments, the compounds disclosed herein exhibit improved duration of action in vivo and oral exposure.

[0023] In some embodiments, the disclosed compound is a compound having formula I, wherein R 5' It is selected from hydrogen, halogenated and optionally substituted (C1-C6) alkyl groups.

[0024] In some embodiments, the disclosed compound is a compound having formula I, wherein X 11 and X 12 At least one of them is N. In some embodiments, X 11 It is N. In some embodiments, X 12 It is N.

[0025] In another embodiment, the disclosed compound is a compound having formula I, wherein X 11 and X 12 At least one of them is CH. In some embodiments, X 11 It is CH. In some embodiments, X 12 It is CH.

[0026] In one embodiment, the compound disclosed herein is a compound having formula I, wherein X 11 and X 12 Each is N.

[0027] In one embodiment, the compound disclosed herein is a compound having formula I, wherein X 11 and X 12 Each is CH.

[0028] In some embodiments, X11 and X 12 One of them is N and X 11 and X 12 The other one is CH. In one embodiment, X 11 It is N and X 12 It is CH. In another embodiment, X 11 It is CH and X 12 It is N.

[0029] In some embodiments, yes

[0030] In some embodiments, Selected from:

[0031]

[0032] In some embodiments, R 3 The optional substituents present on the group are independently selected from hydrogen, halogen, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, arylalkylarylalkoxy, alkylthio, carboxamide, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamide)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (cycloalkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl and (heteroaryl)alkyl; or

[0033] R 3 The two optional substituents together with the carbon or nitrogen atom to which they are attached form an optional substituted cycloalkyl, an optional substituted heterocyclic, an optional substituted aryl, or an optional substituted heteroaryl.

[0034] In some embodiments, R 3 It is a phenyl group that is optionally substituted, wherein the phenyl group is optionally substituted at the 2 position, optionally substituted at the 6 position, optionally double-substituted at the 2 and 6 positions, or optionally double-substituted at the 2 and 3 positions.

[0035] In some embodiments, R 3It is optionally substituted pyridin-3-yl or optionally substituted pyridin-4-yl, wherein the pyridin-3-yl is optionally substituted at the 2 position, optionally substituted at the 4 position, or optionally disubstituted at both the 2 and 4 positions; and wherein the pyridin-4-yl is optionally substituted at the 3 position, optionally substituted at the 5 position, or optionally disubstituted at both the 3 and 5 positions.

[0036] In some embodiments, R 3 It is a optionally substituted pyrimidin-5-yl group, wherein the pyrimidin-5-yl group is optionally substituted at the 4 position, optionally substituted at the 6 position, optionally disubstituted at the 4 and 6 positions, or optionally trisubstituted at the 2, 4 and 6 positions.

[0037] In some embodiments, R 3 It is a optionally substituted pyrazole-5-yl group, wherein the pyrazole-5-yl group is optionally substituted at the 1 position, optionally substituted at the 4 position, or optionally double-substituted at both the 1 and 4 positions.

[0038] In some embodiments, R 3 The substituted group is independently selected from methoxy, deuteroxy, ethoxy, isopropoxy, tert-butoxy, difluoromethoxy, 2-fluoroethoxy, 2-methoxyethoxy, cyclopropoxy, cyclobutoxy, (tetrahydrofuran-3-yl)oxy, benzoxy, methyl, ethyl, isopropyl, 2-fluoroisopropyl, tert-butyl, cyclopropyl, cyclobutyl, methylcyclopropyl, pyrrolidine-1-yl, azircyclobutane-1-yl, methylamino, dimethylamino, cyano, haloyl, methylthio, methylsulfonyl, and ethylsulfonyl.

[0039] In some embodiments, R 3 Choose from the following groups:

[0040]

[0041]

[0042] In some embodiments, R 5The optional substituents present on the [substituent name] are independently selected from hydrogen, halogen, nitro, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, alkoxy, hydroxy, carboxyl, carboxylalkyl, amino, alkylamino, dialkylamino, cycloalkylamino, heterocycloalkylamino, aralkylamino, heteroaralkylalkylamino, alkylthio, haloalkyl, haloalkoxy, hydroxyalkyl, hydroxyalkylamino, alkoxyalkyl, (alkoxyalkyl)amino, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (carboxamido)alkyl, mercaptoalkyl, (cyano)alkyl, (cycloalkyl)alkyl, aralkyl, aralkyl, aralkoxy, alkylcarbonyl, arylcarbonyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, (C 1-4 (haloalkoxy)alkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted cycloalkyl, carboxamide, sulfonyl, sulfonamide, sulfonyldiamine, alkylsulfonyl, alkylsulfonamide, alkylsulfonamide, arylsulfonyl, aryloxy, heteroaryloxy, -C(=O)R 23 -C(=O)OR 24 -C(=O)NR 31a R 31b -NR 31a C(=O)R 25 -NR 31a C(=O)OR 26 -NR 31a C(=O)NR 31a R 31b -NR 31a SO2R 27 -OC(=O)R 28 -OC(=O)OR 29 -OC(=O)NR 31a R 31b -OSO2R 30 and –NR 32a R 32b ;or

[0043] R 5 The two optional substituents, together with the carbon or nitrogen atom to which they are attached, form a cycloalkyl, heterocyclic, aryl, or heteroaryl group; and

[0044] R 24 R 25 R 26 R 27 R 28 R 29 R 30 R 32a and R32b Each is selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, alkoxyalkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0045] In some embodiments, R 5 Selected from optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)alkynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)hydroxyalkyl, cyano, halogroup, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 and -NR 31a SO2R 27 Optional replacement of —O—(C6-C) 14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic, optionally substituted -O-heterocyclic-(C1-C2)alkyl.

[0046] In some embodiments, R 5 Selected from the arbitrarily replaced (C6-C) 14 )Aryl, optionally substituted (C6-C) 14) aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl.

[0047] In some embodiments, R 5 It can be a substituted pyrrole group, a substituted imidazolyl group, a substituted pyrazolyl group, a substituted triazolyl group, or a substituted tetrazolyl group.

[0048] In some embodiments, R 5 It is the imidazole group that can be substituted.

[0049] In some embodiments, R 5 The pyrazol group can be substituted.

[0050] In some embodiments, R 5 It is the triazole group that can be substituted.

[0051] In some embodiments, R 5 It is an optionally substituted heteroaryl group, such as imidazolyl, pyrazolyl, or triazolyl, wherein the substituent is independently selected from halogen, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl, oxetane-3-yl, and methylazacyclobutane.

[0052] In some embodiments, R 5 Choose from the following groups:

[0053]

[0054]

[0055] In some embodiments, the compound has formula II:

[0056]

[0057] Where X 3 Selected from N and CR 10 ;X 4 Selected from N and CR 11 ;X 5 Selected from N and CR 12 ;and

[0058] R 8 R 9 R 10 R11 and R 12 Each is independently selected from the group consisting of: hydrogen, halogen, nitro, cyano, hydroxyl, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, arylalkylarylalkoxy, alkylthio, carboxamide, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamide)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (cycloalkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl or (heteroaryl)alkyl.

[0059] In some embodiments, the compounds disclosed herein have formula III, formula IV, formula V, formula VI, or formula VIa:

[0060]

[0061]

[0062] Where X 1 X 2 R 1 R 5 R 6 R 7 R 8 R 9 and R 11 Each is as defined above for Equation II.

[0063] In some embodiments, R 5 yes:

[0064]

[0065] Where X 6 Selected from NR 13 and CR 18 ;X 7 Selected from NR 14 and CR 19 ;X 8 Selected from NR 15 and CR 20 ;X 9 Selected from NR 16 and CR 21 ;X 10 Selected from NR 17 and CR 22 ;and

[0066] R 13 R 14 R 15 R 16 and R 17 Each of the following groups is absent or independently selected from hydrogen, halogenated, methyl, ethyl, isopropyl, cyclopropyl, methoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl and methylazacyclobutane.

[0067] R 18 R 19 R 20 R 21 and R 22 Each is independently selected from hydrogen, halogenated, methyl, ethyl, isopropyl, cyclopropyl, methoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl, and methylazacyclobutane.

[0068] In some embodiments, the compounds disclosed herein have formula VII, VIII, IX, X, XI, or XII:

[0069]

[0070]

[0071] Where X 1 X 2 R 1 R 3 R 6 R 7 R 15 R 16 R 17 R 20 R 21 and R 22 Each is as defined above for Equation II.

[0072] On the other hand, this disclosure relates to compounds of formula XIII, or pharmaceutically acceptable salts or solvates thereof:

[0073]

[0074] in:

[0075] X 1a and X 2a Each is independently selected from N and CR 2a ;

[0076] R 2aIndependently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, optionally substituted alkynyl group, alkoxy group, alkoxyalkyl group, alkylsulfonyl group, alkylthio group, aryl group, heteroaryl group, and heterocyclic group; and

[0077] The remaining substituents are defined as disclosed herein.

[0078] In some embodiments, the compound disclosed herein is one of the specific compounds listed in the embodiments, or a pharmaceutically acceptable salt or solvation thereof.

[0079] In some embodiments, the compounds disclosed herein inhibit the USP1 protein.

[0080] In some embodiments, the disclosed compounds showed an IC50 concentration of less than about 1 μM in a Ub-Rho deubiquitination assay. 50 It inhibits the USP1 protein.

[0081] In some embodiments, the Ub-Rho deubiquitination assay is the assay disclosed in Example 26.

[0082] In one aspect, this disclosure relates to a method of treating a patient with cancer, the method comprising administering to the patient a therapeutically effective amount of the disclosed compound, or a pharmaceutically acceptable salt or solvate thereof.

[0083] In one aspect, this disclosure relates to a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0084] In some embodiments, this disclosure relates to pharmaceutical compositions for treating cancer.

[0085] In some embodiments, this disclosure relates to compounds of this disclosure for the treatment of cancer.

[0086] In some embodiments, this disclosure relates to the use of the disclosed compounds in the manufacture of a medicament for treating cancer.

[0087] In one aspect, this disclosure relates to a kit containing a compound of the present disclosure, or a pharmaceutically acceptable salt or solvation thereof, or a pharmaceutical composition containing a compound of the present disclosure, and instructions for administering said compound, or a pharmaceutically acceptable salt or solvation thereof, or said pharmaceutical composition to a cancer patient.

[0088] In one aspect, this disclosure relates to a method of treating a patient with cancer, the method comprising administering to the patient a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a compound of the disclosure.

[0089] In some embodiments, cancer is selected from the group consisting of: blood cancers, lymphomas, and cancers with defective DNA damage repair pathways.

[0090] In some embodiments, the cancer comprises cancer cells with a mutation in the gene encoding p53. In some embodiments, the mutation in the gene encoding p53 is a germline mutation. In some embodiments, the mutation in the gene encoding p53 is a somatic mutation. In some embodiments, the cancer comprises cancer cells with a loss-of-function mutation in the gene encoding p53.

[0091] In some embodiments, the cancer is selected from the group consisting of: lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, and breast cancer.

[0092] In some embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0093] In some embodiments, the cancer is colon cancer.

[0094] In some embodiments, the cancer is bladder cancer.

[0095] In some embodiments, the cancer is ovarian cancer or breast cancer.

[0096] In some embodiments, the cancer is ovarian cancer.

[0097] In some embodiments, the cancer is breast cancer.

[0098] In some embodiments, the cancer is triple-negative breast cancer.

[0099] In some embodiments, the cancer comprises cancer cells with elevated RAD18 levels.

[0100] In some embodiments, elevated RAD18 levels refer to elevated RAD18 protein levels.

[0101] In some embodiments, elevated RAD18 levels refer to elevated RAD18 mRNA levels.

[0102] In some embodiments, elevated RAD18 levels were detected prior to administration.

[0103] On the other hand, this disclosure relates to a method that further includes detecting RAD18 levels in cancer samples obtained from a subject.

[0104] In some embodiments, the cancer is selected from the group consisting of: bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma, and meningioma; soft tissue cancer, including rhabdomyosarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer; uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.

[0105] In some embodiments, the cancer is a defective DNA damage repair pathway cancer.

[0106] In some embodiments, the cancer comprises cancer cells with a mutation in the gene encoding p53. In some embodiments, the mutation in the gene encoding p53 is a germline mutation. In some embodiments, the mutation in the gene encoding p53 is a somatic mutation. In some embodiments, the cancer comprises cancer cells with a loss-of-function mutation in the gene encoding p53.

[0107] In some embodiments, the cancer is a BRCA1-mutated cancer. In some embodiments, the BRCA1 mutation is a germline mutation. In some embodiments, the BRCA1 mutation is a somatic mutation. In some embodiments, the BRCA1 mutation causes a BRCA1 deficiency.

[0108] In some embodiments, the cancer is a BRCA2-mutant cancer. In some embodiments, the BRCA2 mutation is a germline mutation. In some embodiments, the BRCA2 mutation is a somatic mutation. In some embodiments, the BRCA2 mutation causes a BRCA2 deficiency.

[0109] In some embodiments, the cancer is a BRCA1-mutant cancer and a BRCA2-mutant cancer.

[0110] In some embodiments, the cancer is a BRCA1-deficient cancer.

[0111] In some embodiments, the cancer is a BRCA2-deficient cancer.

[0112] In some embodiments, the cancer is a BRCA1-deficient cancer and a BRCA2-deficient cancer.

[0113] In some embodiments, the cancer is a poly(ADP-ribose) polymerase (“PARP”) inhibitor-resistant or refractory cancer. In some embodiments, the cancer is a PARP inhibitor-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 mutant cancer. In some embodiments, the cancer is a PARP inhibitor-resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 deficient cancer.

[0114] In some embodiments, the cancer has a mutation in the gene encoding the ataxiate langiectasia mutated (ATM) protein kinase. In some embodiments, the ATM mutation is a germline mutation. In some embodiments, the ATM mutation is a somatic mutation. In some embodiments, the cancer is an ATM-deficient cancer.

[0115] In some embodiments, the cancer has mutations in at least two of the genes encoding p53, BRCA1, BRCA2, and ATM.

[0116] On the other hand, this disclosure relates to a method of treating a USP1 protein-mediated condition, comprising administering to a patient in need an amount of the disclosed compound, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising the disclosed compound.

[0117] In some embodiments, the USP1 protein comprises the amino acid sequence of SEQ ID NO:1.

[0118] On the other hand, this disclosure relates to a method for inhibiting the USP1 protein, the method comprising contacting the USP1 protein with a compound of the present disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a compound of the present disclosure.

[0119] In some embodiments, the contact occurs outside the body.

[0120] In some embodiments, the contact occurs within the body.

[0121] On the other hand, this disclosure relates to a method of treating a patient’s cancer, the method comprising administering a USP1 inhibitor to the subject, wherein the cancer comprises cancer cells with mutations in a gene encoding p53.

[0122] In some embodiments, mutations in the gene encoding p53 have been detected prior to administration.

[0123] In some embodiments, the method further includes detecting mutations in the gene encoding p53 in a cancer sample obtained from the patient.

[0124] On the other hand, this disclosure relates to a method for selecting cancer patients to be treated with a USP1 inhibitor, the method comprising detecting whether the cancer contains cancer cells with a mutation in a gene encoding p53, wherein if the cancer contains cancer cells with a mutation in a gene encoding p53, the patient is selected to be treated with a USP1 inhibitor.

[0125] On the other hand, this disclosure relates to a USP1 inhibitor for treating a patient’s cancer, wherein the cancer comprises cancer cells with mutations in the gene encoding p53.

[0126] In some embodiments, a patient is identified as responding to USP1 inhibitor treatment because cancer cells in a cancer sample obtained from the patient are detected to have a mutation in the gene encoding p53, wherein cancer cells in the cancer sample with a mutation in the gene encoding p53 identify the patient as responding to USP1 inhibitor treatment.

[0127] On the other hand, this disclosure relates to a USP1 inhibitor for treating cancer in a subject who is identified as responding to USP1 inhibitor treatment by the presence of cancer cells with a mutation in the gene encoding p53 in a cancer sample obtained from the subject, wherein the cancer cells with the mutation in the gene encoding p53 in the cancer sample identify the patient as responding to USP1 inhibitor treatment.

[0128] In some embodiments, if no mutation in the gene encoding p53 is detected in a cancer sample obtained from the subject, the USP1 inhibitor is not administered to the subject.

[0129] On the other hand, this disclosure relates to an in vitro method for identifying whether a cancer subject has responded to treatment with a USP1 inhibitor, the method comprising detecting mutations in a gene encoding p53 in a cancer sample obtained from the subject, wherein the mutation in the gene encoding p53 in the cancer sample indicates that the patient has responded to treatment with a USP1 inhibitor.

[0130] On the other hand, this disclosure relates to the in vitro use of at least one agent capable of specifically detecting mutations in genes encoding p53, the at least one agent being used to identify whether a cancer subject has responded to treatment with a USP1 inhibitor.

[0131] In some embodiments, mutations in the gene encoding p53 are loss-of-function mutations.

[0132] On the other hand, this disclosure relates to a method of treating a patient’s cancer, the method comprising administering a USP1 inhibitor to the subject, wherein the cancer comprises cancer cells having a mutation in a gene encoding BRCA1.

[0133] In some embodiments, mutations in the gene encoding BRCA1 have been detected prior to administration.

[0134] In some embodiments, the method further includes detecting mutations in the gene encoding BRCA1 in a sample obtained from the patient (e.g., a cancer sample or a blood sample).

[0135] On the other hand, this disclosure relates to a method for selecting cancer patients to be treated with a USP1 inhibitor, the method comprising detecting whether the cancer contains cancer cells with a mutation in a gene encoding BRCA1, wherein if the cancer contains cancer cells with a mutation in a gene encoding BRCA1, the patient is selected to be treated with a USP1 inhibitor.

[0136] On the other hand, this disclosure relates to a USP1 inhibitor for treating a patient’s cancer, wherein the cancer comprises cancer cells with mutations in the gene encoding BRCA1.

[0137] In some embodiments, a patient is identified as responding to USP1 inhibitor treatment by detecting a mutation in the gene encoding BRCA1 in a sample obtained from the patient (e.g., a cancer sample or a blood sample), wherein the mutation in the gene encoding BRCA1 in the sample identifies the patient as responding to USP1 inhibitor treatment.

[0138] On the other hand, this disclosure relates to a USP1 inhibitor for treating cancer in a subject who is identified as responding to USP1 inhibitor treatment by the presence of cancer cells with mutations in the gene encoding BRCA1 in a sample (e.g., a cancer sample or a blood sample) obtained from the subject, wherein the cancer cells with mutations in the gene encoding BRCA1 in the sample identify the patient as responding to USP1 inhibitor treatment.

[0139] In some embodiments, if a mutation in the gene encoding BRCA1 is not detected in a sample obtained from the subject (e.g., a cancer sample or a blood sample), the subject is not given a USP1 inhibitor.

[0140] On the other hand, this disclosure relates to an in vitro method for identifying whether a cancer subject has responded to treatment with a USP1 inhibitor, the method comprising detecting mutations in a gene encoding BRCA1 in a sample (e.g., a cancer sample or a blood sample) obtained from the subject, wherein a mutation in the gene encoding BRCA1 in the sample indicates that the patient has responded to treatment with a USP1 inhibitor.

[0141] On the other hand, this disclosure relates to the in vitro use of at least one agent capable of specifically detecting mutations in genes encoding BRCA1, the at least one agent being used to identify whether a cancer subject has responded to treatment with a USP1 inhibitor.

[0142] In some embodiments, the mutation in the gene encoding BRCA1 is a loss-of-function mutation.

[0143] On the other hand, this disclosure relates to a method of treating a patient’s cancer, the method comprising administering a USP1 inhibitor to the subject, wherein the cancer comprises cancer cells with mutations in a gene encoding BRCA2.

[0144] In some embodiments, mutations in the gene encoding BRCA2 are detected prior to administration.

[0145] In some embodiments, the method further includes detecting mutations in the gene encoding BRCA2 in a sample obtained from the patient (e.g., a cancer sample or a blood sample).

[0146] On the other hand, this disclosure relates to a method for selecting cancer patients to be treated with a USP1 inhibitor, the method comprising detecting whether the cancer contains cancer cells with a mutation in a gene encoding BRCA2, wherein if the cancer contains cancer cells with a mutation in a gene encoding BRCA2, the patient is selected to be treated with a USP1 inhibitor.

[0147] On the other hand, this disclosure relates to a USP1 inhibitor for treating a patient’s cancer, wherein the cancer comprises cancer cells with mutations in the gene encoding BRCA2.

[0148] In some embodiments, a patient is identified as responding to USP1 inhibitor treatment by detecting a mutation in the gene encoding BRCA2 in a sample obtained from the patient (e.g., a cancer sample or a blood sample), wherein the mutation in the gene encoding BRCA2 in the sample identifies the patient as responding to USP1 inhibitor treatment.

[0149] On the other hand, this disclosure relates to a USP1 inhibitor for treating cancer in a subject who is identified as responding to USP1 inhibitor treatment by the presence of cancer cells with mutations in a gene encoding BRCA2 in a sample (e.g., a cancer sample or a blood sample) obtained from the subject, wherein the cancer cells with mutations in a gene encoding BRCA2 in the sample identify the patient as responding to USP1 inhibitor treatment.

[0150] In some embodiments, if a mutation in the gene encoding BRCA2 is not detected in a sample obtained from the subject (e.g., a cancer sample or a blood sample), the USP1 inhibitor is not administered to the subject.

[0151] On the other hand, this disclosure relates to an in vitro method for identifying whether a cancer subject has responded to treatment with a USP1 inhibitor, the method comprising detecting mutations in a gene encoding BRCA2 in a sample (e.g., a cancer sample or a blood sample) obtained from the subject, wherein a mutation in the gene encoding BRCA2 in the sample indicates that the patient has responded to treatment with a USP1 inhibitor.

[0152] On the other hand, this disclosure relates to the in vitro use of at least one agent capable of specifically detecting mutations in genes encoding BRCA2, the at least one agent being used to identify whether a cancer subject has responded to treatment with a USP1 inhibitor.

[0153] In some embodiments, the mutation in the gene encoding BRCA2 is a loss-of-function mutation.

[0154] On the other hand, this disclosure relates to a method of treating a patient’s cancer, the method comprising administering a USP1 inhibitor to the subject, wherein the cancer comprises cancer cells with mutations in a gene encoding ATM.

[0155] In some embodiments, mutations in the gene encoding ATM are detected prior to administration.

[0156] In some embodiments, the method further includes detecting mutations in the gene encoding ATM in a sample obtained from the patient.

[0157] On the other hand, this disclosure relates to a method for selecting cancer patients to be treated with a USP1 inhibitor, the method comprising detecting whether the cancer contains cancer cells with a mutation in a gene encoding ATM, wherein if the cancer contains cancer cells with a mutation in a gene encoding ATM, the patient is selected to be treated with a USP1 inhibitor.

[0158] On the other hand, this disclosure relates to a USP1 inhibitor for treating a patient’s cancer, wherein the cancer comprises cancer cells with mutations in the gene encoding ATM.

[0159] In some embodiments, a patient is identified as responding to treatment with a USP1 inhibitor because cancer cells in a cancer sample obtained from the patient are detected to have a mutation in the gene encoding ATM, wherein cancer cells in the cancer sample with a mutation in the gene encoding ATM identify the patient as responding to USP1 inhibitor treatment.

[0160] On the other hand, this disclosure relates to a USP1 inhibitor for treating cancer in a subject who is identified as responding to USP1 inhibitor treatment by the presence of cancer cells with mutations in a gene encoding ATM in a cancer sample obtained from the subject, wherein the cancer cells with mutations in the gene encoding ATM in the cancer sample identify the patient as responding to USP1 inhibitor treatment.

[0161] In some embodiments, if a mutation in the gene encoding ATM is not detected in a cancer sample obtained from the subject, the USP1 inhibitor is not administered to the subject.

[0162] On the other hand, this disclosure relates to an in vitro method for identifying whether a cancer subject is responding to USP1 inhibitor treatment, the method comprising detecting mutations in a gene encoding ATM in a cancer sample obtained from the subject, wherein the mutation in the gene encoding ATM in the cancer sample indicates that the patient is responding to USP1 inhibitor treatment.

[0163] On the other hand, this disclosure relates to the in vitro use of at least one agent capable of specifically detecting mutations in genes encoding ATM, the at least one agent being used to identify whether a cancer subject has responded to treatment with a USP1 inhibitor.

[0164] In some embodiments, the mutation in the gene encoding ATM is a loss-of-function mutation.

[0165] In some embodiments, the USP1 inhibitor is a compound disclosed herein.

[0166] In one aspect (A1), this disclosure provides a method of treating a subject's cancer, the method comprising administering a USP1 inhibitor to the subject, wherein the cancer comprises cancer cells having elevated RAD18 levels.

[0167] In one aspect (A2), this disclosure provides a method for treating a subject with triple-negative breast cancer, the method comprising administering a USP1 inhibitor to the subject. In another aspect (A3) of A1, the cancer comprises cancer cells having elevated RAD18 levels.

[0168] In one facet of A1 or A3 (A4), elevated RAD18 levels have been detected prior to application. In one facet of A4 (A5), the method further includes detecting RAD18 levels in a cancer sample obtained from the subject.

[0169] In one aspect (A5), this disclosure provides a method for selecting a subject with cancer to be treated with a USP1 inhibitor, the method comprising detecting whether the cancer contains cells with elevated RAD18 levels, wherein if the cancer contains cells with elevated RAD18 levels, the subject is selected to be treated with a USP1 inhibitor.

[0170] In one aspect (A6), this disclosure provides a USP1 inhibitor for treating a subject’s cancer, wherein the cancer comprises cancer cells having elevated RAD18 levels.

[0171] In one aspect (A7), this disclosure provides a USP1 inhibitor for treating a subject with triple-negative breast cancer.

[0172] In one aspect of A7 or A8 (A9), the subject is identified as responding to USP1 inhibitor treatment by detecting a level of RAD18 in a cancer sample obtained from the subject, wherein an elevated level of RAD18 in the cancer sample identifies the patient as responding to USP1 inhibitor treatment.

[0173] In one aspect (A10), this disclosure provides a USP1 inhibitor for treating cancer in a subject who is identified as responding to USP1 inhibitor treatment due to elevated RAD18 levels in a cancer sample obtained from the subject, wherein the elevated RAD18 levels in the cancer sample identify the patient as responding to USP1 inhibitor treatment.

[0174] In one aspect (A11), this disclosure provides a USP1 inhibitor for any one of A7-A10, wherein the USP1 inhibitor is not administered to the subject if no increase in RAD18 levels is detected in a cancer sample obtained from the subject.

[0175] In one aspect (A12), this disclosure relates to an in vitro method for identifying whether a cancer subject has responded to treatment with a USP1 inhibitor, the method comprising detecting RAD18 levels in a cancer sample obtained from the subject, wherein elevated RAD18 levels in the cancer sample indicate that the patient has responded to treatment with a USP1 inhibitor.

[0176] In one aspect (A13), this disclosure provides the in vitro use of at least one agent capable of specifically detecting RAD18, the at least one agent being used to identify whether a cancer subject has responded to treatment with a USP1 inhibitor.

[0177] In one aspect (A14), this disclosure provides a kit or component kit comprising: (a) a pharmaceutical composition comprising a USP1 inhibitor and one or more pharmaceutically acceptable excipients; and (b) a diagnostic kit comprising at least one agent capable of specifically detecting RAD18.

[0178] In one aspect of A13 or A14 (A15), the at least one agent capable of specifically detecting RAD18 is capable of specifically hybridizing with RAD18 mRNA. In one aspect of A13 or A14 (A16), the at least one agent capable of specifically detecting RAD18 is capable of specifically binding to the RAD18 protein.

[0179] In one aspect of A10, A12, or A13 (A17), the cancer is triple-negative breast cancer.

[0180] In one aspect of A10, A12, A13 or A17 (A18), the cancer comprises cancer cells having elevated RAD18 levels.

[0181] In one aspect (A19), this disclosure provides a method for classifying cancer in a subject, the method comprising detecting elevated RAD18 levels in a cancer sample obtained from the subject.

[0182] In any one aspect of A1-A19 (A20), the cancer is not a BRCA1-mutant cancer. In any one aspect of A1-A19 (A21), the cancer is not a BRCA2-mutant cancer. In any one aspect of A1-A19 (A22), the cancer is neither a BRCA1-mutant cancer nor a BRCA2-mutant cancer.

[0183] In any one aspect (A23) of A1-A19, the cancer is not a homologous recombination-deficient cancer. In any one aspect (A24) of A1-A19, the cancer is not a homologous recombination-deficient cancer.

[0184] In one aspect (A25) of any of A1-A23, the cancer comprises cancer cells having a mutation in a gene encoding p53, optionally wherein the mutation in the gene encoding p53 is a loss-of-function mutation.

[0185] In any one aspect of A1, A3-A7, A9-A12, and A18-A25 (A26), the elevated RAD18 level refers to an elevated RAD18 protein level. In one aspect of A26 (A27), the elevated RAD18 protein level is detected by Western blot. In one aspect of A26 (A28), the elevated RAD18 protein level is detected by fluorescence activated cell sorting (FACS). In one aspect of A26 (A29), the elevated RAD18 protein level is detected by immunohistochemistry.

[0186] In any one aspect (A30) of A1, A3-A7, A9-A12, and A18-A25, the elevated RAD18 level is an elevated RAD18 mRNA level. In one aspect (A31) of A30, the detection of the elevated RAD18 mRNA level is performed by quantitative reverse transcriptase (RT)-polymerase chain reaction (PCR), RNA-Seq, or microarray method.

[0187] In any one aspect of A1, A3-A7, A9-A12, and A18-A31 (A32), the elevated RAD18 level is at least as high as the RAD18 level in ES2 cells. In any one aspect of A1, A3-A7, A9-A12, and A18-A31 (A33), the elevated RAD18 level is higher than the RAD18 level in HEP3B217 cells.

[0188] In any one aspect of A1, A4-A7, A9-A16, and A18-A33 (A34), the cancer is ovarian cancer. In any one aspect of A1, A4-A7, A9-A16, and A18-A33 (A35), the cancer is breast cancer. In one aspect of A35 (A36), the breast cancer is triple-negative breast cancer.

[0189] In any one aspect of A1-A19 and A24-A36 (A37), the cancer is a BRCA1-mutant cancer. In any one aspect of A1-A19 and A24-A36 (A38), the cancer is a BRCA2-mutant cancer. In any one aspect of A1-A19 and A24-A36 (A39), the cancer is either a BRCA1-mutant cancer or a BRCA2-mutant cancer.

[0190] In any one aspect (A40) of A1-A18 and A20-A39, the USP1 inhibitor is selected from small molecules, siRNA, antisense oligonucleotides, peptides, or aptamers. In any one aspect (A41) of A1-A18 and A20-A40, the USP1 inhibitor specifically binds to the USP1 protein. In any one aspect (A42) of A41, the USP1 protein comprises an amino acid sequence (SEQ ID NO:1).

[0191] In one aspect (A43) of A1-A18 and A20-A42, the USP1 inhibitor inhibits the formation and / or activity of the USP1 / UAF1 complex.

[0192] In one aspect (A44) of any of A1-A18 and A20-A40, the USP1 inhibitor specifically binds to the USP1 / UAF1 complex.

[0193] In one aspect (A45) of any of A1-A18 and A20-A40, the USP1 inhibitor specifically binds to USP1 mRNA.

[0194] In one aspect (A46) of any of A1-A18 and A20-A45, the USP1 inhibitor increases monoubiquitination of PCNA.

[0195] In one aspect (A47) of any of A1-A18 and A20-A46, the USP1 inhibitor increases monoubiquitination of FANCD2.

[0196] In any one aspect of A1-A18 and A20-A39 (A48), the USP1 inhibitor is a purine. In any one aspect of A1-A18 and A20-A39 (A49), the USP1 inhibitor is GW7647. In any one aspect of A1-A18 and A20-A39 (A50), the USP1 inhibitor is pimozide. In any one aspect of A1-A18 and A20-A39 (A51), the USP1 inhibitor is ML323.

[0197] In any one aspect (A52) of A1-A18 and A20-A51, the subject is a human being.

[0198] Other embodiments and advantages of this disclosure will be set forth in part in the following embodiments and will be learned from the implementation, or may be learned by practicing this disclosure. The embodiments and advantages of this disclosure will be recognized and attained by means of the elements and combinations particularly pointed out in the appended claims.

[0199] It should be understood that the foregoing overview and the following embodiments are merely illustrative and explanatory, and not intended to limit the claimed invention. Attached Figure Description

[0200] Figure 1Provides a USP1 loss spectrum for approximately 500 cancer cell lines. Lower loss scores indicate higher sensitivity to USP1 loss. Breast cancer cell lines are shown in white, and ovarian cancer cell lines are shown in black. Symbols indicate BRCA1 / 2 mutation or triple-negative breast cancer (TNBC) status.

[0201] Figure 2 A schematic diagram of PCNA ubiquitination is provided from Jacquemont C. and Taniguchi T., BMC Biochemistry 8 (Suppl 1): S10 (2007). The E3 ligase Rad18 monoubiquitinates PCNA. USP1 deubiquitinates monoubiquitinated PCNA.

[0202] Figure 3 This plot shows the distribution of Rad18 mRNA expression in approximately 500 cancer cell lines. The x-axis displays normalized Rad18 mRNA expression values, and the y-axis shows the number of cell lines with the expression values ​​indicated on the x-axis. The 20 cell lines with the lowest USP1 loss fractions are shown as dark bars extending to the top of the plot.

[0203] Figure 4 The provided figures show the USP1 loss fraction of Rad 18 mRNA expression levels in breast cancer cell lines (top left), ovarian cancer cell lines (top right), and triple-negative breast cancer cell lines (bottom).

[0204] Figure 5 This shows the Rad18 mRNA levels in USP1 sensitive cell lines (59M and ES2) and insensitive cell lines (OVISE, JHH7, and HEP3B217) normalized for GAPDH expression and for OVISE expression, as measured by qRT-PCT.

[0205] Figure 6 This figure shows the RAD18 protein levels in USP1-sensitive cell lines (59M and ES2) and insensitive cell lines (OVISE, JHH7, and HEP3B217). The top plot shows the protein levels as determined by Western blot, and the bottom plot provides quantification of the protein levels as determined by Western blot. The cell line “HCC1395” (top plot) in the confirmed Western blot assay was contaminated by another cell line and therefore not used for quantification (bottom plot).

[0206] Figure 7The provided figure illustrates the rescue of USP1 deletion by Rad18 deletion. The figure shows relative cell viability in OR1A1 (negative control) knockout cells (left panel) or Rad18 knockout cells (right panel), with wizards added for OR1A1 negative control (“negative control”), EEF2 pan-lethal positive control (“lethal control”), or four different USP1 wizards (“USP1_1”, “USP1_2”, “USP1_3”, and “USP1_4”). Detailed Implementation

[0207] One aspect of this disclosure is based on the use of the disclosed compounds as inhibitors of ubiquitin-specific processing protein 1 (USP1). In view of this property, the disclosed compounds can be used to inhibit USP1 protein and can be used to treat diseases, conditions, or disorders that respond to inhibition of USP1 protein, such as cancer.

[0208] In some embodiments, the compounds disclosed herein exhibit improved solubility, as measured, for example, by the ADME solubility assay disclosed herein.

[0209] In some embodiments, the compounds disclosed herein exhibit improved metabolic stability, as measured, for example, by the liver microsomal metabolic stability assay disclosed herein.

[0210] In other embodiments, the compounds disclosed herein exhibit improved duration of action in vivo and oral exposure.

[0211] In one embodiment, the compound disclosed herein is a compound having Formula I:

[0212]

[0213] and its pharmaceutically acceptable salts or solvates, such as hydrates, wherein:

[0214] X 1 and X 2 Each is independently selected from N and CR 2 ;

[0215] R 1 and R 2 Each is independently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, and optionally substituted alkynyl group;

[0216] R 3 It is optionally substituted phenyl, optionally substituted pyridyl, optionally substituted pyrimidinyl, optionally substituted pyrazinyl, optionally substituted pyridazinyl, or optionally substituted pyrazolyl.

[0217] R 5'Selected from hydrogen, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)alkynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)hydroxyalkyl, cyano, halogroup, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 -NR 31a SO2R 27 Optional replacement of (C6-C) 14 )Aryl, optionally substituted (C6-C) 14 ) aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl, optionally substituted -O-(C6-C 14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic, optionally substituted -O-heterocyclic-(C1-C2)alkyl;

[0218] R 5 Selected from optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)alkynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy, (C1-C6)hydroxyalkyl, cyano, halogroup, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25-NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 -NR 31a SO2R 27 Optional replacement of (C6-C) 14 )Aryl, optionally substituted (C6-C) 14 ) aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl, optionally substituted -O-(C6-C 14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic, optionally substituted -O-heterocyclic-(C1-C2)alkyl; or

[0219] An R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optional substituted (C6-C) 14 ) aryl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optional substituted heteroaryl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optionally substituted (C3-C8) cycloalkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optionally substituted heterocyclic alkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the same atom it is attached to, it forms an optionally substituted spirocyclic alkyl ring; or an R on an adjacent atom 5 And an R 5' Together with the same atom it is attached to, it forms an optional substituted spiroheterocyclic alkyl ring;

[0220] R 6 and R7 Each is independently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, and optionally substituted alkynyl group;

[0221] X 11 and X 12 Each is independently selected from N and CH;

[0222] R 23 Choose from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0223] R 31a and R 31b Each is independently selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, alkoxyalkyl, cycloalkyl, (cycloalkyl)alkyl, (heterocyclic)alkyl, aralkyl, and (heteroaryl)alkyl; and

[0224] R 24 R 25 R 27 R 32a and R 32b Each is independently selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, alkoxyalkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0225] In one embodiment, the compound disclosed herein is a compound having formula I, wherein X 1 and X 2 Each is CR 2 .

[0226] In another embodiment, the disclosed compound is a compound having formula I, wherein X 1 and X 2At least one of them is N. In some embodiments, X 1 It is N. In some embodiments, X 2 It is N.

[0227] In another embodiment, the disclosed compound is a compound having formula I, wherein X 1 and X 2 At least one of them is CR 2 In some embodiments, X 1 It is CR 2 In some embodiments, X 2 It is CR 2 .

[0228] In another embodiment, the disclosed compound is a compound having formula I, wherein X 1 and X 2 Each is N.

[0229] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5' It is selected from hydrogen, halogenated and optionally substituted (C1-C6) alkyl groups.

[0230] In one embodiment, the compound disclosed herein is a compound having formula I, wherein Selected independently from:

[0231]

[0232] In another embodiment, yes

[0233] In another embodiment, yes

[0234] In another embodiment, yes

[0235] In another embodiment, yes

[0236] In another embodiment, Selected independently from:

[0237]

[0238] In another embodiment, yes

[0239] In another embodiment, yes

[0240] In another embodiment, yes

[0241] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 1 and R 2 Each group is independently selected from hydrogen, halogen, and cyano groups.

[0242] In another embodiment, the disclosed compound is a compound having formula I, wherein R 1 and R 2 Each is independently selected from the arbitrarily replaced (C) 1-4 )alkyl, optionally substituted (C 2-4 )alkenyl and optionally substituted (C 2-4 ) alkynyl group. In some embodiments, R 1 and R 2 Each is independently selected from the arbitrarily replaced (C) 1-4 alkyl. In some embodiments, R 1 and R 2 Each is independently selected from the arbitrarily replaced (C) 2-4 )alkenyl. In some embodiments, R 1 and R 2 Each is independently selected from the arbitrarily replaced (C) 2-4 ) yyn group.

[0243] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 It is an optional substituted phenyl group.

[0244] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 It is an optional substituted pyridinyl group.

[0245] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 It is an optional substituted pyrimidin group.

[0246] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 It is the pyrazin group that is optionally substituted.

[0247] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 It is an optional substituted pyridazinyl group.

[0248] In another embodiment, the disclosed compound is a compound having formula I, wherein R 3 The pyrazol group can be substituted.

[0249] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 The optional substituents present on the group are independently selected from hydrogen, halogen, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, arylalkylarylalkoxy, alkylthio, carboxamide, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamide)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (cycloalkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl and (heteroaryl)alkyl.

[0250] In another embodiment, R 3 The optional substituents present on the [top] are independently selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, halo-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, (C1-C4)alkoxy, halo-(C1-C4)alkoxy, (C6-C] 10 ) aryloxy, (C3-C6) heteroaryloxy, aryl-(C1-C4)alkyl, aryl-(C1-C4)alkoxy, (C1-C4)alkylthio, carboxamide, sulfonamide, (C1-C4)alkylcarbonyl, (C6-C 10 ) aryl carbonyl, (C1-C4) alkyl sulfonyl, (C6-C 10 ) arylsulfonyl, carboxyl, carboxyl-(C1-C4)alkyl, (C1-C4)alkyl, (C2-C4)alkenyl, (C2-C4)alkynyl, alkoxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, hydroxy-(C1-C4)alkylamino, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cyano)-(C1-C4)alkyl, (carboxamido)-(C1-C4)alkyl, mercapto-(C1-C4)alkyl, (heterocyclic)-(C1-C4)alkyl, (cycloalkylamino)-(C1-C4)alkyl, (C 1-4 (Haloalkoxy)-(C1-C4)alkyl and (heteroaryl)-(C1-C4)alkyl.

[0251] In another embodiment, R 3 The optional substituents present on the surface are independently selected from (C3-C8) cycloalkyl, (C6 ... and cycloalkyl groups.10 (C3-C6) heteroaryl and (C3-C8) heterocyclic.

[0252] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 The two optional substituents together with the carbon or nitrogen atom to which they are attached form an optional substituted cycloalkyl, an optional substituted heterocyclic, an optional substituted aryl, or an optional substituted heteroaryl.

[0253] In another embodiment, R 3 The two optional substituents, together with the carbon or nitrogen atom to which they are attached, form an optional substituted (C3-C8) cycloalkyl group, an optional substituted (C3-C8) heterocyclic group, or an optional substituted (C6-C8) cycloalkyl group. 10 ) aryl or optionally substituted (C3-C6) heteroaryl.

[0254] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 The phenyl group is optionally substituted, wherein the phenyl group is optionally substituted at the 2-position. In another embodiment, the phenyl group is optionally substituted at the 6-position. In another embodiment, the phenyl group is optionally disubstituted at both the 2- and 6-positions. In yet another embodiment, the phenyl group is optionally disubstituted at both the 2- and 3-positions.

[0255] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 It is optionally substituted pyridin-3-yl or optionally substituted pyridin-4-yl.

[0256] In another embodiment, R 3 The pyridin-3-yl group is optionally substituted. In another embodiment, the pyridin-3-yl group is optionally substituted at the 2-position. In another embodiment, the pyridin-3-yl group is optionally substituted at the 4-position. In yet another embodiment, the pyridin-3-yl group is optionally double-substituted at both the 2- and 4-positions.

[0257] In another embodiment, R 3 The pyridin-4-yl group is optionally substituted. In another embodiment, the pyridin-4-yl group is optionally substituted at the 3-position. In another embodiment, the pyridin-4-yl group is optionally substituted at the 5-position. In yet another embodiment, the pyridin-4-yl group is optionally double-substituted at both the 3- and 5-positions.

[0258] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3The pyrimidin-5-yl group is optionally substituted. In another embodiment, the pyrimidin-5-yl group is optionally substituted at position 2. In another embodiment, the pyrimidin-5-yl group is optionally substituted at position 4. In another embodiment, the pyrimidin-5-yl group is optionally substituted at position 6. In another embodiment, the pyrimidin-5-yl group is optionally disubstituted at positions 4 and 6. In another embodiment, the pyrimidin-5-yl group is optionally disubstituted at positions 2 and 6. In another embodiment, the pyrimidin-5-yl group is optionally disubstituted at positions 2 and 4. In another embodiment, the pyrimidin-5-yl group is optionally trisubstituted at positions 2, 4, and 6.

[0259] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 It is optionally substituted with pyrazole-3- or pyrazole-5-.

[0260] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 The pyrazole-5-yl group is optionally substituted. In another embodiment, the pyrazole-5-yl group is optionally substituted at position 1. In another embodiment, the pyrazole-5-yl group is optionally substituted at position 3. In another embodiment, the pyrazole-5-yl group is optionally substituted at position 4. In another embodiment, the pyrazole-5-yl group is optionally double-substituted at positions 1 and 4. In another embodiment, the pyrazole-5-yl group is optionally double-substituted at positions 1 and 3. In another embodiment, the pyrazole-5-yl group is optionally double-substituted at positions 3 and 4.

[0261] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 3 The substituted group is independently selected from methoxy, deuteroxy, ethoxy, isopropoxy, tert-butoxy, difluoromethoxy, 2-fluoroethoxy, 2-methoxyethoxy, cyclopropoxy, cyclobutoxy, (tetrahydrofuran-3-yl)oxy, benzoxy, methyl, ethyl, isopropyl, 2-fluoroisopropyl, tert-butyl, cyclopropyl, cyclobutyl, methylcyclopropyl, pyrrolidine-1-yl, azircyclobutane-1-yl, methylamino, dimethylamino, cyano, haloyl, methylthio, methylsulfonyl, and ethylsulfonyl.

[0262] In one embodiment, R 3 Choose from the following groups:

[0263]

[0264]

[0265] In another embodiment, the disclosed compound is a compound having formula I, wherein X 11 and X12 At least one of them is N. In some embodiments, X 11 It is N. In some embodiments, X 12 It is N.

[0266] In another embodiment, the disclosed compound is a compound having formula I, wherein X 11 and X 12 At least one of them is CH. In some embodiments, X 11 It is CH. In some embodiments, X 12 It is CH.

[0267] In one embodiment, the compound disclosed herein is a compound having formula I, wherein X 11 and X 12 Each is N.

[0268] In one embodiment, the compound disclosed herein is a compound having formula I, wherein X 11 and X 12 Each is CH.

[0269] In some embodiments, X 11 and X 12 One of them is N and X 11 and X 12 The other one is CH. In one embodiment, X 11 It is N and X 12 It is CH. In another embodiment, X 11 It is CH and X 12 It is N.

[0270] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 It is selected from optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, optionally substituted (C1-C4)alkoxy, (C1-C4)haloalkyl, (C1-C4)haloalkoxy, and (C1-C4)hydroxyalkyl.

[0271] In another embodiment, the disclosed compound is a compound having formula I, wherein R 5 Selected from cyano, halogenated, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR31a R 31b -S(O)2R 27 -NR 31a SO2R 27 .

[0272] In another embodiment, the disclosed compound is a compound having formula I, wherein R 5 Selected from the arbitrarily replaced (C6-C) 10 )Aryl, optionally substituted (C6-C) 10 ) aryl-(C1-C2)alkyl, optionally substituted (C3-C6)heteroaryl, optionally substituted (C3-C6)heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted (C3-C8)heterocyclic, optionally substituted (C3-C8)heterocyclic-(C1-C2)alkyl, optionally substituted —O—(C6-C 10 )Aryl, optionally substituted —O—(C6-C 10 ) aryl-(C1-C2)alkyl, optionally substituted -O-(C3-C6)heteroaryl, optionally substituted -O-(C3-C6)heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-(C3-C8)heterocyclic, optionally substituted -O-(C3-C8)heterocyclic-(C1-C2)alkyl.

[0273] In another embodiment, the disclosed compound is a compound having formula I, wherein R 5 It is an optional substituted heteroaryl group. In another embodiment, R 5 It is an optionally substituted 5- or 6-membered heteroaryl group containing one or more nitrogen atoms. In another embodiment, R 5 It is a 5- or 6-membered heteroaryl group containing only a nitrogen atom as one or more heteroatoms, which is optionally substituted.

[0274] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5' Together with the atoms it is attached to, it forms an optional substituted (C6-C) 10 )Aromatic ring.

[0275] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5'Together with the atoms to which it is attached, it forms an optional (C3-C6) heteroaryl ring.

[0276] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5' Together with the atoms to which it is attached, it forms an optional (C3-C8) cycloalkyl ring.

[0277] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5' Together with the atoms to which it is attached, it forms an optional (C3-C8) heterocyclic alkyl ring.

[0278] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5' Together with the atoms to which it is attached, it forms an optional spirocyclic alkyl ring.

[0279] In another embodiment, the compound disclosed herein is a compound having Formula I, wherein an R on an adjacent atom 5 And an R 5' Together with the atoms to which it is attached, it forms an optional spiroheterocyclic alkyl ring.

[0280] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 The optional substituents present on the [substituent name] are independently selected from hydrogen, halogen, nitro, cyano, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, alkoxy, hydroxy, carboxyl, carboxylalkyl, amino, alkylamino, dialkylamino, cycloalkylamino, heterocycloalkylamino, aralkylamino, heteroaralkylalkylamino, alkylthio, haloalkyl, haloalkoxy, hydroxyalkyl, hydroxyalkylamino, alkoxyalkyl, (alkoxyalkyl)amino, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (carboxamido)alkyl, mercaptoalkyl, (cyano)alkyl, (cycloalkyl)alkyl, aralkyl, aralkyl, aralkoxy, alkylcarbonyl, arylcarbonyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, optionally substituted cycloalkyl, carboxamide, sulfonyl, sulfonamide, sulfonyldiamine, alkylsulfonyl, alkylsulfonamide, alkylsulfonamide, arylsulfonyl, aryloxy, heteroaryloxy, -C(=O)R 23 -C(=O)OR24 -C(=O)NR 31a R 31b -NR 31a C(=O)R 25 -NR 31a C(=O)OR 26 -NR 31a C(=O)NR 31a R 31b -NR 31a SO2R 27 -OC(=O)R 28 -OC(=O)OR 29 -OC(=O)NR 31a R 31b -OSO2R 30 and –NR 32a R 32b .

[0281] In another embodiment, R 5 The optional substituents present on the [topic name] are independently selected from hydrogen, halogroup, nitro, cyano, optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, (C1-C4)alkoxy, hydroxyl, carboxyl, carboxyl-(C1-C4)alkyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, (C3-C8)cycloalkylamino, heterocyclic-(C1-C4)alkylamino, aryl-(C1-C4)alkylamino, heteroaryl-(C1-C4)alkylamino, (C1-C4)alkylthio, halo-(C1-C4)alkyl, halo-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkylamino, alkoxy -(C1-C4)alkyl, (alkoxyalkyl)amino, (amino)-(C1-C4)alkyl, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cycloalkylamino)-(C1-C4)alkyl, (carboxamido)-(C1-C4)alkyl, mercapto-(C1-C4)alkyl, (cyano)-(C1-C4)alkyl, (cycloalkyl)-(C1-C4)alkyl, aryl-(C1-C4)alkyl, aryl-(C1-C4)alkyl, aryl-(C1-C4)alkoxy, (C1-C4)alkylcarbonyl, arylcarbonyl, (heterocyclic)-(C1-C4)alkyl, (heteroaryl)-(C1-C4)alkyl, (amino)(hydroxy)-(C1-C4)alkyl, (arylalkylamino)-(C1-C4)alkyl and (C 1-4 (Haloalkoxy)-(C1-C4)alkyl.

[0282] In another embodiment, R 5The optional substituents present on the surface are independently selected from optional substituted (C3-C8) heterocyclic groups, optional substituted (C3-C6) heteroaryl groups, optional substituted (C6-C8) heterocyclic groups, and optional substituted (C3-C6) heteroaryl groups. 10 )Aryl, optionally substituted (C3-C8)cycloalkyl.

[0283] In another embodiment, R 5 The optional substituents present on the surface are independently selected from carboxamide, sulfonyl, sulfonamide, sulfonyldiamine, (C1-C4)alkylsulfonyl, (C1-C4)alkylsulfonamide, (C1-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, (C6-C4)alkylsulfonamide, ... 10 )arylsulfonyl, (C6-C 10 ) aryloxy group, (C3-C6) heteroaryloxy group, -C(=O)R 23 -C(=O)OR 24 -C(=O)NR 31a R 31b -NR 31a C(=O)R 25 -NR 31a C(=O)OR 26 -NR 31a C(=O)NR 31a R 31b -NR 31a S O2R 27 -OC(=O)R 28 -OC(=O)OR 29 -OC(=O)NR 31a R 31b -OSO2R 30 and –NR 32a R 32b .

[0284] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 The two optional substituents together with the carbon or nitrogen atom to which they are attached form an optional substituted cycloalkyl, an optional substituted heterocyclic, an optional substituted aryl, or an optional substituted heteroaryl.

[0285] In another embodiment, R 5 The two optional substituents, together with the carbon atoms they are attached to, form an optional substituted (C3-C8) cycloalkyl group, an optional substituted (C3-C8) heterocyclic group, or an optional substituted (C6-C8) cycloalkyl group. 10 ) aryl or optionally substituted (C3-C6) heteroaryl.

[0286] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5It is selected from optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)alkoxy, (C1-C6)haloalkyl, (C1-C6)haloalkoxy and (C1-C6)hydroxyalkyl.

[0287] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 Selected from cyano, halogenated, sulfonamide, -C(=O)R 23 -C(=O)OR 24 –NR 32a R 32b -NR 31a C(=O)R 25 -NR 31a C(=O)NR 31a R 31b -C(=O)NR 31a R 31b -S(O)2R 27 and -NR 31a SO2R 27 .

[0288] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 Selected from the arbitrarily replaced —O—(C6-C) 14 )Aryl, optionally substituted —O—(C6-C 14 ) aryl-(C1-C2)alkyl, optionally substituted -O-heteroaryl, optionally substituted -O-heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-heterocyclic, optionally substituted -O-heterocyclic-(C1-C2)alkyl. In another embodiment, R 5 Selected from the arbitrarily replaced —O—(C6-C) 10 )Aryl, optionally substituted —O—(C6-C 10 ) aryl-(C1-C2)alkyl, optionally substituted -O-(C3-C6)heteroaryl, optionally substituted -O-(C3-C6)heteroaryl-(C1-C2)alkyl, optionally substituted -O-(C3-C8)cycloalkyl, optionally substituted -O-((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted -O-(C3-C8)heterocyclic, optionally substituted -O-(C3-C8)heterocyclic-(C1-C2)alkyl.

[0289] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R5 Selected from the arbitrarily replaced (C6-C) 14 )Aryl, optionally substituted (C6-C) 14 Aryl-(C1-C2)alkyl, optionally substituted heteroaryl, optionally substituted heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted heterocyclic, optionally substituted heterocyclic-(C1-C2)alkyl. In another embodiment, R 5 Selected from the arbitrarily replaced (C6-C) 10 )Aryl, optionally substituted (C6-C) 10 ) aryl-(C1-C2)alkyl, optionally substituted (C3-C6)heteroaryl, optionally substituted (C3-C6)heteroaryl-(C1-C2)alkyl, optionally substituted (C3-C8)cycloalkyl, optionally substituted ((C3-C8)cycloalkyl)-(C1-C2)alkyl, optionally substituted (C3-C8)heterocyclic, optionally substituted (C3-C8)heterocyclic-(C1-C2)alkyl.

[0290] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 It can be an optionally substituted pyrrole group, an optionally substituted imidazolyl group, an optionally substituted pyrazolyl group, an optionally substituted triazolyl group, or an optionally substituted tetrazolyl group. In another embodiment, R 5 The pyrrole group is optionally substituted. In another embodiment, R 5 The substituted imidazole group is optional. In another embodiment, R 5 The pyrazol group is optionally substituted. In another embodiment, R 5 The triazole group is optionally substituted. In another embodiment, R 5 It is an optional substituted tetrazolium group.

[0291] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 The substituted group is independently selected from halogenated, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl, oxetane-3-yl, and methylazacyclobutane.

[0292] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 5 Choose from the following groups:

[0293]

[0294]

[0295] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 6 and R 7 Each group is independently selected from hydrogen, halogen, and cyano groups.

[0296] In another embodiment, the disclosed compound is a compound having formula I, wherein R 6 and R 7 Each is independently selected from optionally substituted (C1-C4) alkyl, optionally substituted (C2-C4) alkenyl, and optionally substituted (C2-C4) ynyl.

[0297] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 23 Choose from the group consisting of: hydrogen, optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, cyclo-(C1-C4)alkylamino, hydroxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cycloalkylamino)-(C1-C4)alkyl, (cycloalkyl)-(C1-C4)alkyl, aryl-(C6-C4)alkyl, etc. 10 Alkyl, (heterocyclic)-(C1-C4)alkyl, (heteroaryl)-(C1-C4)alkyl, (amino)(hydroxy)-(C1-C4)alkyl, (arylamino)-(C1-C4)alkyl, optionally substituted (C3-C8)heterocyclic, optionally substituted (C3-C6)heteroaryl, optionally substituted (C6-C8)heterocyclic 10 )Aryl and optionally substituted (C3-C8)cycloalkyl.

[0298] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 31a and R 31b Each is independently selected from the group consisting of: hydrogen, optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, hydroxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, alkoxy-(C1-C4)alkyl, (C3-C8)cycloalkyl, (cycloalkyl)-(C1-C4)alkyl, (heterocyclic)-(C1-C4)alkyl, aryl-(C1-C4)alkyl, and (heteroaryl)-(C1-C4)alkyl.

[0299] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 24 R 25 R 27 R 32a and R 32b Each is independently selected from the group consisting of: hydrogen, optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, (C3-C8)cycloalkylamino, hydroxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cyclo ... Alkylamino)-(C1-C4)alkyl, (cycloalkyl)-(C1-C4)alkyl, aryl-(C1-C4)alkyl, (heterocyclic)-(C1-C4)alkyl, (heteroaryl)-(C1-C4)alkyl, (amino)(hydroxy)-(C1-C4)alkyl, (arylalkylamino)-(C1-C4)alkyl, alkoxy-(C1-C4)alkyl, optionally substituted (C3-C8)heterocyclic, optionally substituted (C3-C6)heteroaryl, optionally substituted (C6-C4)heterocyclic 10 )Aryl and optionally substituted (C3-C8)cycloalkyl.

[0300] In one embodiment, the compound disclosed herein is a compound having formula I, wherein R 24 R 25 R 26 R 27 R 28 R 29 R 30 R 32a and R 32b Each is selected from the group consisting of: hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, amino, alkylamino, dialkylamino, cycloalkylamino, hydroxyalkyl, (amino)alkyl, (alkylamino)alkyl, (dialkylamino)alkyl, (cycloalkylamino)alkyl, (cycloalkyl)alkyl, aralkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, (amino)(hydroxy)alkyl, (aralkylamino)alkyl, alkoxyalkyl, optionally substituted heterocyclic, optionally substituted heteroaryl, optionally substituted aryl, and optionally substituted cycloalkyl.

[0301] In another embodiment, R 24 R 25 R 26 R 27 R 28 R 29 R30 R 32a and R 32b Each is selected from the group consisting of: hydrogen, optionally substituted (C1-C4)alkyl, optionally substituted (C2-C4)alkenyl, optionally substituted (C2-C4)alkynyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, (C3-C8)cycloalkylamino, hydroxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cycloalkyl) (C1-C4)alkyl, (cycloalkyl)-(C1-C4)alkyl, aryl-(C1-C4)alkyl, (heterocyclic)-(C1-C4)alkyl, (heteroaryl)-(C1-C4)alkyl, (amino)(hydroxy)-(C1-C4)alkyl, (arylalkylamino)-(C1-C4)alkyl, alkoxy-(C1-C4)alkyl, optionally substituted (C3-C8)heterocyclic, optionally substituted (C3-C6)heteroaryl, optionally substituted (C6-C4)heterocyclic 10 )Aryl and optionally substituted (C3-C8)cycloalkyl.

[0302] In another embodiment, the disclosed compound is a compound having Formula II:

[0303]

[0304] and its pharmaceutically acceptable salts or solvates, such as hydrates, wherein:

[0305] X 1 X 2 R 1 R 5 R 6 and R 7 Each is as defined above for Equation I;

[0306] X 3 Selected from N and CR 10 ;X 4 Selected from N and CR 11 And X 5 Selected from N and CR 12 ;and

[0307] R 8 R 9 R 10 R 11 and R 12Each is independently selected from the group consisting of: hydrogen, halogen, nitro, cyano, hydroxyl, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, arylalkylarylalkoxy, alkylthio, carboxamide, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamide)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (cycloalkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl or (heteroaryl)alkyl.

[0308] In another embodiment, R 8 R 9 R 10 R 11 and R 12 Each group is independently selected from hydrogen, halogen, nitro, cyano, hydroxyl, amino, (C1-C4)alkylamino, di-(C1-C4)alkylamino, halo-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, (C1-C4)alkoxy, halo-(C1-C4)alkoxy, (C6-C4)alkyl, (C6-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, (C6-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl ...6-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C6-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkoxy, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy-(C1-C4)alkyl, hydroxy 10 ) aryloxy, (C3-C6) heteroaryloxy, aryl-(C1-C4)alkylaryl-(C1-C4)alkoxy, (C1-C4)alkylthio, carboxamide, sulfonamide, (C1-C4)alkylcarbonyl, (C6-C 10 ) aryl carbonyl, (C1-C4) alkyl sulfonyl, (C6-C 10 ) arylsulfonyl, carboxyl, carboxyl-(C1-C4)alkyl, (C1-C4)alkyl, (C2-C4)alkenyl, (C2-C4)alkynyl, alkoxy-(C1-C4)alkyl, (amino)-(C1-C4)alkyl, hydroxy-(C1-C4)alkylamino, (alkylamino)-(C1-C4)alkyl, (dialkylamino)-(C1-C4)alkyl, (cyano)-(C1-C4)alkyl, (carboxamido)-(C1-C4)alkyl, mercapto-(C1-C4)alkyl, (heterocyclic)-(C1-C4)alkyl, (cycloalkylamino)-(C1-C4)alkyl, (C 1-4 (Haloalkoxy)-(C1-C4)alkyl or (heteroaryl)-(C1-C4)alkyl.

[0309] In another embodiment, R 8 R 9 R 10 R 11 and R12 Each is independently selected from optionally substituted (C3-C8) cycloalkyl groups, optionally substituted (C6-C8) cycloalkyl groups, and optionally substituted (C6-C8) cycloalkyl groups. 10 ) aryl, optionally substituted (C3-C6) heteroaryl and optionally substituted (C3-C8) heterocyclic group.

[0310] In one embodiment, the disclosed compound is a compound having formula III, IV, V, VI, or VIa:

[0311]

[0312]

[0313] Where X 1 X 2 R 1 R 5 R 6 R 7 R 8 R 9 and R 11 Each is as defined above for Equation II.

[0314] In another embodiment, R 5 yes:

[0315]

[0316] in:

[0317] X 6 Selected from NR 13 and CR 18 ;X 7 Selected from NR 14 and CR 19 ;X 8 Selected from NR 15 and CR 20 ;X 9 Selected from NR 16 and CR 21 ;X 10 Selected from NR 17 and CR 22 ;and

[0318] R 13 R 14 R 15 R 16 and R 17Each of the following is absent or independently selected from hydrogen, halogenated, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, methoxy, ethoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl, oxetane-3-yl and methylazacyclobutane.

[0319] R 18 R 19 R 20 R 21 and R 22 Each is independently selected from hydrogen, halogenated, methyl, ethyl, isopropyl, cyclopropyl, methoxy, triazolyl, cyano, optionally substituted alkyl, amino, alkylamino, dialkylamino, difluoromethyl, trifluoromethyl, methylsulfonyl, and methylazacyclobutane.

[0320] In one embodiment, the disclosed compound is a compound having formula VII, VIII, IX, X, XI, or XII:

[0321]

[0322]

[0323] Where X 1 X 2 R 1 R 3 R 6 R 7 R 15 R 16 R 17 R 20 R 21 and R 22 Each is as defined above for Equation II.

[0324] In one embodiment, the compound disclosed herein is a compound having formula XIII:

[0325]

[0326] in:

[0327] X 1a and X 2a Each is independently selected from N and CR 2a ;

[0328] R 2a Independently selected from hydrogen, halogroup, cyanogroup, optionally substituted alkyl group, optionally substituted alkenyl group, optionally substituted alkynyl group, alkoxy group, alkoxyalkyl group, alkylsulfonyl group, alkylthio group, aryl group, heteroaryl group, and heterocyclic group; and

[0329] The remaining substituents are defined as disclosed herein.

[0330] In one embodiment, the disclosed compound is a compound selected from the group consisting of:

[0331] 6-(3-methoxypyridin-4-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 8);

[0332] 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-(methylsulfonyl)-phenyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 3);

[0333] 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-methyl-6-(methylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 23);

[0334] 6-(2-(ethylsulfonyl)phenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 7);

[0335] 1-(4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)benzyl)-6-(2-isopropylpyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine (Example 20);

[0336] 6-(2-methoxy-4-methylpyridin-3-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 6);

[0337] 6-(4-methoxy-6-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 21);

[0338] 6-(2-isopropylpyridin-3-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 18);

[0339] 6-(2,6-dimethoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 4);

[0340] 6-(2-methoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 2);

[0341] 6-(2-isopropylphenyl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 15);

[0342] 6-(2-methoxy-6-methylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 5);

[0343] 2-Methoxy-3-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)isonicotinonitrile (Example 22);

[0344] 2-(2-Isopropylphenyl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine (Example 24);

[0345] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 12);

[0346] 6-(2-isopropylpyridin-3-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 9);

[0347] 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-methyl-6-(methylthio)phenyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 13);

[0348] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 19);

[0349] 6-(2-Cyclopropyl-6-methoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 11);

[0350] 6-(2-isopropylphenyl)-1-(4-(1-(1-methylazacyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 25);

[0351] 6-(2-isopropylphenyl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 16);

[0352] 6-(2-isopropylphenyl)-4-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 14);

[0353] 6-(3-fluoro-2-isopropylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 10);

[0354] 6-(2-isopropylphenyl)-3-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 17);

[0355] 6-(2-Isopropylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (Example 1);

[0356] 6-(4-(tert-butyl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0357] 6-(4-(2-fluoropropane-2-yl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0358] 6-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0359] 6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0360] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0361] (R)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0362] 6-(4-cyclopropyl-6-ethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0363] 6-(4-cyclopropyl-6-isopropoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0364] 6-Cyclopropyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-carboxynitrile;

[0365] 6-Cyclopropyl-N,N-Dimethyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine;

[0366] 6-(4,6-dimethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0367] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0368] 6-(4-isopropyl-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0369] 1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methyl-6-(methylthio)pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin;

[0370] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-methyl-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0371] 1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methyl-6-(methylsulfonyl)pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin;

[0372] 6-(4-(2-methoxyethoxy)-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0373] 6-(1-Isopropyl-4-methyl-1H-pyrazol-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0374] 6-(1-Isopropyl-4-methoxy-1H-pyrazol-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0375] 6-(4-cyclopropyl-1-isopropyl-1H-pyrazol-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0376] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7H-pyrrolo[2,3-d]pyrimidin; and

[0377] 5-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidinyl,

[0378] Or a pharmaceutically acceptable salt or solvate of any of the above, such as a hydrate.

[0379] In another embodiment, the disclosed compound is a compound selected from the group consisting of:

[0380] 6-(4,6-diethoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0381] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(trifluoromethyl)-5,6,7,8-tetrahydroimidazo[1,5-a]pyrazin-3-yl)benzyl)-1H-pyrazo[3,4-d]pyrimidinyl;

[0382] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-5-(trifluoromethyl)-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0383] 6-(1-Cyclopropyl-4-methoxy-1H-pyrazol-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0384] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-((6-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0385] (R)-6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0386] (S)-6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0387] (R)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0388] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0389] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0390] 6-(4-(2-fluoropropane-2-yl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0391] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0392] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-((6-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-3-yl)methyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0393] 1-(4-((6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-5-methyl-1H-pyrazol-3-carboxylonitrile;

[0394] 6-(4-cyclopropyl-1-ethyl-1H-pyrazol-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0395] 6-(4-cyclopropyl-6-(methoxy-d3)pyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0396] 6-Cyclopropyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-carboxynitrile;

[0397] 6-(4-cyclopropoxy-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0398] 6-(4-cyclobutoxy-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0399] 6-(4-cyclopropyl-6-(difluoromethoxy)pyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0400] 6-(4-cyclopropyl-6-methoxy-2-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0401] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(oxecyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0402] 6-(4-(tert-butyl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0403] 6-(4-(tert-butoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0404] 6-(4-cyclopropyl-6-(2-fluoroethoxy)pyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0405] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0406] 1-(4-(1-cyclobutyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidine;

[0407] 6-(4,6-dimethoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0408] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0409] 1-(4-(1-cyclopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0410] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0411] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0412] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0413] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(2-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0414] 6-(2,4-Dimethoxy-6-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0415] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0416] 6-(4-cyclopropyl-6-(2-methoxyethoxy)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0417] 6-(4-cyclopropyl-6-(2-methoxyethoxy)pyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0418] 6-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0419] 6-(1-Isopropyl-4-methyl-1H-pyrazol-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0420] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin;

[0421] (R)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0422] 6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0423] 6-(4-cyclopropyl-6-isopropoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0424] 6-(4-isopropyl-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0425] 6-(4-cyclopropyl-6-ethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0426] N,N,6-Trimethyl-5-(1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine;

[0427] 1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methyl-6-(methylthio)pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin;

[0428] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0429] 6-(4,6-dimethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0430] 6-(4-(2-methoxyethoxy)-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0431] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(1-methylazacyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0432] 6-(4-methoxy-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0433] 3-(1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyridinecarboxylonitrile;

[0434] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0435] 6-Cyclopropyl-N-methyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine;

[0436] 1-(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methoxy-6-methylpyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidine;

[0437] 6-(4-cyclopropyl-6-((tetrahydrofuran-3-yl)oxy)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0438] 6-(4-(benzoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0439] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0440] 6-(4-(benzoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0441] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methoxy-1-methyl-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0442] 2-(2-(4-((6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)-N,N-dimethylacetamide;

[0443] 6-Cyclopropyl-N,N-Dimethyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine;

[0444] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0445] 6-(4-cyclopropyl-6-(pyrrolidone-1-yl)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0446] 6-(4-(azacyclobutan-1-yl)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; and

[0447] 5-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidinyl,

[0448] Or a pharmaceutically acceptable salt or solvate of any of the above, such as a hydrate.

[0449] In another embodiment, the disclosed compound is a compound selected from the group consisting of:

[0450] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3-(methylsulfonyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0451] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3-(methylthio)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0452] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3-((4-methoxybenzyl)thio)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0453] (S)-1-(1-(4-(5-bromo-1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidin;

[0454] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7H-pyrrolo[2,3-d]pyrimidinyl;

[0455] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-8-(methoxymethyl)-9H-purine;

[0456] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-isopropyl-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine;

[0457] 3-Cyclobutoxy-6-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0458] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidinyl;

[0459] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-ethyl-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine;

[0460] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidin;

[0461] 3-(azacyclobutane-1-yl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine;

[0462] 1-(4-(5-bromo-1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidin;

[0463] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidinyl;

[0464] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidin;

[0465] (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidin;

[0466] 6-(4-cyclopropyl-6-ethoxypyrimidin-5-yl)-3-ethoxy-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0467] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-ethoxy-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl;

[0468] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidinyl;

[0469] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine;

[0470] 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3-methoxy-1H-pyrazolo[3,4-d]pyrimidinyl;

[0471] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-methyl-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine;

[0472] 8-Cyclopropyl-2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0473] 8-Cyclobutyl-2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0474] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-isopropyl-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0475] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-ethyl-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0476] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7H-pyrrolo[2,3-d]pyrimidinyl;

[0477] 2-(4-Cyclopropyl-6-methoxypyrimidin-5-yl)-8-methyl-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0478] 5-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidinyl;

[0479] 2-(4-methoxy-6-methylpyrimidin-5-yl)-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0480] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-purine;

[0481] 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine; and

[0482] 2-(2-isopropylphenyl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine.

[0483] definition

[0484] For the purposes of this disclosure, the term "alkyl" as used alone or as part of another group refers to a group containing one to twelve carbon atoms (i.e., C16, C26, C36, C46, ​​C56, C6 ... 1-12 Alkyl groups are straight-chain or branched aliphatic hydrocarbons with a specified number of carbon atoms (i.e., C1 alkyl such as methyl, C2 alkyl such as ethyl, C3 alkyl such as propyl or isopropyl, etc.). Alkyl groups may suitably be selected from straight-chain C1 alkyl groups. 1-10 Alkyl, branched C 3-10 Alkyl, straight-chain C 1-6 Alkyl, branched C 3-6 Alkyl, straight-chain C1-4 Alkyl, branched C 3-4 Alkyl, straight-chain or branched C 3-4 Alkyl group. Alkyl groups can be partially or fully deuterated, i.e., one or more hydrogen atoms of the alkyl group are replaced by deuterium atoms. Non-limiting illustrative C 1-10 Alkyl groups include methyl (including -CD3), ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, isobutyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Non-limiting illustrative C 1-4 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, and isobutyl.

[0485] For the purposes of this disclosure, the term "optionally substituted alkyl" used alone or as part of another group means that the alkyl group as defined above is unsubstituted or substituted by one, two, or three substituents independently selected from: halogroup, nitrogroup, cyanogroup, hydroxyl group, amino group, alkylamino group, dialkylamino group, haloalkyl group, hydroxyalkyl group, alkoxy group, haloalkoxy group, aryloxy group, aralkyl group, arylalkoxy group, alkylthio group, carboxamido group, sulfonamide group, alkylcarbonyl group, arylcarbonyl group, alkylsulfonyl group, arylsulfonyl group, carboxyl group, carboxylalkyl group, alkoxyalkyl group, (amino)alkyl group, hydroxyalkylamino group, (alkylamino)alkyl group, (dialkylamino)alkyl group, (cyano)alkyl group, (carboxamido)alkyl group, mercaptoalkyl group, (heterocyclic)alkyl group, or (heteroaryl)alkyl group. The alkyl group may be an optionally substituted C14 group. 1-4 Alkyl group. Optionally substituted alkyl groups may be replaced by two substituents or one substituent. Non-limiting illustrative examples of optionally substituted alkyl groups include -CH2CH2NO2, -CH2CH2CO2H, -CH2CH2SO2CH3, -CH2CH2COPh, and -CH2C6H. 11 .

[0486] For the purposes of this disclosure, the term "alkylene" refers to a divalent alkyl group. Any of the monovalent alkyl groups mentioned above can be an alkylene group obtained by removing a second hydrogen atom from an alkyl group. Alkylene groups can also be C1-C6 or C1-C4 alkylene groups. Non-limiting illustrative alkylene groups include -CH2-, -CH(CH3)-, -C(CH3)2-, -CH2CH2-, -CH2CH(CH3)-, -CH2C(CH3)2-, -CH2CH2CH2-, and -CH2CH2CH2CH2-.

[0487] For the purposes of this disclosure, the term "cycloalkyl" as used alone or as part of another group refers to a group containing one to three rings and having three to twelve carbon atoms (i.e., C16, C26, C36, C46, ​​C56, C6 ... 3-12A cycloalkyl group is a saturated or partially unsaturated (containing one or two double bonds) cyclic aliphatic hydrocarbon with a specified number of carbon atoms. A cycloalkyl group may have two rings or one ring. The cycloalkyl group may be selected from C10. 3-8 cycloalkyl and C 3-6 Cycloalkyl. Cycloalkyl may contain one or more carbon-carbon double bonds or one carbon-carbon double bond. Non-limiting illustrative cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decahydronaphthalene, adamantyl, cyclohexenyl and spiro[3.3]heptane.

[0488] For the purposes of this disclosure, the term “optionally substituted cycloalkyl” used alone or as part of another group means that the cycloalkyl group as defined above is not substituted or is substituted by one, two or three substituents independently selected from the following: halogroup, nitrogroup, cyanogroup, hydroxyl group, amino group, alkylamino group, dialkylamino group, haloalkyl group, hydroxyalkyl group, alkoxy group, haloalkoxy group, aryloxy group, aralkyl group, arylalkoxy group, alkylthio group, carboxamide group, sulfonamide group, alkyl carbonyl group, aryl carbonyl group, alkyl sulfonyl group, aryl sulfonyl group, carboxyl group, carboxylalkyl group, alkyl group, optionally substituted cycloalkyl group, alkenyl group, alkynyl group, optionally substituted aryl group, optionally substituted heteroaryl group, optionally substituted heterocyclic group, alkoxyalkyl group, (amino)alkyl group, hydroxyalkylamino group, (alkylamino)alkyl group, (dialkylamino)alkyl group, (cyano)alkyl group, (carboxamide)alkyl group, mercaptoalkyl group, (heterocyclic)alkyl group or (heteroaryl)alkyl group. The substituted cycloalkyl group may be replaced by two substituents or one substituent.

[0489] For the purposes of this disclosure, the term "alkenyl" used alone or as part of another group refers to an alkyl group as defined above, containing one, two, or three carbon-carbon double bonds. The alkenyl group may be selected from C... 2-6 alkenyl and C 2-4 Alkenyl. Non-limiting illustrative alkenyl groups include vinyl, propenyl, isopropenyl, butenyl, sec-butenyl, pentenyl, and hexenyl.

[0490] For the purposes of this disclosure, the term “optionally substituted alkenyl” used alone or as part of another group means that the alkenyl group as defined above is not substituted or is substituted by one, two or three substituents independently selected from: halogroup, nitrogroup, cyanogroup, hydroxyl group, amino group, alkylamino group, dialkylamino group, haloalkyl group, hydroxyalkyl group, alkoxy group, haloalkoxy group, aryloxy group, arylalkoxy group, alkylthio group, carboxamide group, sulfonamide group, alkyl carbonyl group, aryl carbonyl group, alkyl sulfonyl group, aryl sulfonyl group, carboxyl group, carboxylalkyl group, optionally substituted cycloalkyl group, optionally substituted aryl group, optionally substituted heteroaryl group or optionally substituted heterocyclic group.

[0491] For the purposes of this disclosure, the term "alkynyl" used alone or as part of another group refers to an alkyl group as defined above containing one to three carbon-carbon triple bonds. An alkynyl group may have one carbon-carbon triple bond. The alkynyl group may be selected from C... 2-6 alkynyl group and C 2-4 Alkynyl. Non-limiting illustrative alkynyl groups include ethynyl, propynyl, butynyl, 2-butynyl, pentynyl, and hexynyl.

[0492] For the purposes of this disclosure, the term “optionally substituted alkynyl” used alone or as part of another group means that the alkynyl group as defined above is not substituted or is substituted by one, two or three substituents independently selected from the following: halogroup, nitrogroup, cyanogroup, hydroxyl group, amino group, alkylamino group, dialkylamino group, haloalkyl group, hydroxyalkyl group, alkoxy group, haloalkoxy group, aryloxy group, arylalkoxy group, alkylthio group, carboxamide group, sulfonamide group, alkyl carbonyl group, aryl carbonyl group, alkyl sulfonyl group, aryl sulfonyl group, carboxyl group, carboxylalkyl group, cycloalkyl group, aryl group, heteroaryl group or heterocyclic group.

[0493] For the purposes of this disclosure, the term "haloalkyl" used alone or as part of another group refers to an alkyl group substituted with one or more fluorine, chlorine, bromine, and / or iodine atoms. The alkyl group may be substituted with one, two, or three fluorine and / or chlorine atoms. The haloalkyl group may be selected from C10. 1-4 Halogenated alkyl groups. Non-limiting illustrative halogenated alkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, pentafluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl, and trichloromethyl.

[0494] For the purposes of this disclosure, the term "hydroxyalkyl" as used alone or as part of another group refers to an alkyl group substituted with one or more, such as one, two, or three hydroxyl groups. Hydroxyalkyl may be selected from monohydroxyalkyl, i.e., substituted with one hydroxyl group; dihydroxyalkyl, i.e., substituted with two hydroxyl groups; and C 1-4 Hydroxyalkyl. Non-limiting illustrative hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl, and hydroxybutyl, such as 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 3-hydroxybutyl, 4-hydroxybutyl, 2-hydroxy-1-methylpropyl, and 1,3-dihydroxypropyl-2-yl.

[0495] For the purposes of this disclosure, the term "alkoxy" as used alone or as part of another group refers to an optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclic, optionally substituted alkenyl, or optionally substituted alkynyl group attached to a terminal oxygen atom. The alkoxy group may be selected from C... 1-4alkoxy groups and C atoms attached to the terminal oxygen atom 1-4 Alkyl groups, such as methoxy, ethoxy, and tert-butoxy.

[0496] For the purposes of this disclosure, the term "alkylthio" used alone or as part of another group refers to a sulfur atom that has been optionally substituted with an alkyl group. The alkylthio group may be selected from C... 1-4 Alkyl thio group. Non-limiting illustrative alkyl thio groups include -SCH3 (i.e., methyl thio) and -SCH2CH3.

[0497] For the purposes of this disclosure, the term "alkoxyalkyl" as used alone or as part of another group refers to an alkyl group substituted with an alkoxy group. Non-limiting illustrative alkoxyalkyl groups include methoxymethyl, methoxyethyl, methoxypropyl, methoxybutyl, ethoxymethyl, ethoxyethyl, ethoxypropyl, ethoxybutyl, propoxymethyl, isopropoxymethyl, propoxyethyl, propoxypropyl, butoxymethyl, tert-butoxymethyl, isobutoxymethyl, sec-butoxymethyl, and pentoxymethyl.

[0498] For the purposes of this disclosure, the term "halogenated" as used alone or as part of another group refers to a halogen atom. Non-limiting illustrative halogenations include fluorine, chloro, bromine, and iodine groups.

[0499] For the purposes of this disclosure, the term "haloalkoxy" used alone or as part of another group refers to a haloalkyl group attached to a terminal oxygen atom. Non-limiting illustrative haloalkoxy groups include fluoromethoxy, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy.

[0500] For the purposes of this disclosure, the term "heteroalkyl" as used alone or as part of another group refers to a stable straight-chain or branched hydrocarbon group containing 1 to 10 carbon atoms and at least two heteroatoms, which may be the same or different and selected from O, N, and S, wherein: 1) the nitrogen atom and sulfur atom may optionally be oxidized; and / or 2) the nitrogen atom may optionally be quaternized. Heteroatoms may be located at any internal position of the heteroalkyl group or at a position where the heteroalkyl group is attached to the rest of the molecule. Heteroalkyl groups may contain two oxygen atoms, one oxygen atom and one nitrogen atom, or two nitrogen atoms. Non-limiting illustrative heteroalkyl groups include -CH2OCH2CH2OCH3, -OCH2CH2OCH2CH2OCH3, -CH2NHCH2CH2OCH2, -OCH2CH2NH2, -NHCH2CH2N(H)CH3, -NHCH2CH2OCH3, and -OCH2CH2OCH3.

[0501] For the purposes of this disclosure, the term "aryl" used alone or as part of another group refers to an aryl group having six to fourteen carbon atoms (i.e., C64 ...6-14 A monocyclic or bicyclic aromatic ring system of aryl groups. The aryl group can be selected from C... 6-14 Aryl and C 6-10 Aryl. Non-limiting illustrative aryl groups include phenyl (abbreviated "Ph"), naphthyl, phenanthryl, anthraceneyl, indene, chamomilecycloyl, biphenyl, biphenylene, and fluorene. The aryl group may be selected from phenyl or naphthyl. The aryl group may be phenyl.

[0502] For the purposes of this disclosure, the term "optionally substituted aryl" used alone or as part of another group means, as defined above, that the aryl group is unsubstituted or substituted by one to five substituents independently selected from the group consisting of: halogen, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, heteroaryloxy, aralkylarylalkoxy, alkylthio, carboxamide, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamide)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (cycloalkylamino)alkyl, (C 1-4 (Haloalkoxy)alkyl, (heteroaryl)alkyl. The optionally substituted aryl group can be an optionally substituted phenyl group. The optionally substituted phenyl group can have four, three, two, or one substituents. The optionally substituted phenyl group can have one amino, alkylamino, dialkylamino, (amino)alkyl, (alkylamino)alkyl, or (dialkylamino)alkyl substituent. Non-limiting illustrative substituted aryl groups include 2-methylphenyl, 2-methoxyphenyl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-methylphenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 4-methylphenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 2,6-di-fluorophenyl, 2,6-di-chlorophenyl, 2-methyl, 3-methoxyphenyl, 2-ethyl, 3-methoxyphenyl, 3,4-di-methoxyphenyl, 3,5-di-fluorophenyl, 3,5-di-methylphenyl, 3,5-dimethoxy, 4-methylphenyl, 2-fluoro-3-chlorophenyl, 3-chloro-4-fluorophenyl, and 2-phenylpropane-2-amine. The term "optionally substituted aryl" is intended to include groups having a fused optional substituted cycloalkyl ring and a fused optional substituted heterocyclic ring. Examples include:

[0503]

[0504] For the purposes of this disclosure, the term "aryloxy group," used alone or as part of another group, refers to an optionally substituted aryl group attached to a terminal oxygen atom. A non-limiting illustrative aryloxy group is PhO-.

[0505] For the purposes of this disclosure, the term "heteroaryloxy" used alone or as part of another group refers to an optionally substituted heteroaryl group attached to a terminal oxygen atom.

[0506] For the purposes of this disclosure, the term "arylalkoxy" or "arylalkoxy" used alone or as part of another group refers to an aryl alkyl group attached to a terminal oxygen atom. A non-limiting illustrative arylalkoxy group is PhCH₂O-.

[0507] For the purposes of this disclosure, the term "heteroaryl" or "heteroary group" refers to a group having 5 to 14 ring atoms (i.e., C14, C24, C34, C44, C54, C6 ... 5-14 A heteroaryl group is a monocyclic or bicyclic aromatic ring system consisting of one, two, three, or four heteroatoms independently selected from oxygen, nitrogen, or sulfur. The heteroaryl group may be selected from C... 5-14 heteroaryl and C 3-6Heteroaryl groups. Heteroaryl groups can have three, two, or one heteroatom. Heteroaryl groups can be C5 or C6 heteroaryl groups. Non-limiting illustrative heteroaryl groups include thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthyl, furanyl, benzofuranyl, pyranyl, isobenzofuranyl, benzoxazolone, chromenyl, oxanthyl, 2H-pyrroleyl, pyrroleyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, isoindolyl, 3H-indolyl, indolyl, inzolyl, purinel, isoquinolinyl, quinolinyl, phthalazinyl, naphridinyl, cinolinyl, quinazolinyl, pteridinyl, 4aH-carbazolyl, carbazolyl, β-carbazolyl, phenadiazinyl, acridineyl, phenolinyl, phenazinyl, thiazolyl, isothiazolyl, phenathiazolyl, isoxazolyl, furazinyl, triazolyl, tetrazolyl, and phenoxazinyl. The heteroaryl group can be selected from thienyl (e.g., thien-2-yl and thien-3-yl), furanyl (e.g., 2-furanyl and 3-furanyl), pyrroleyl (e.g., 1H-pyrrole-2-yl and 1H-pyrrole-3-yl), imidazolyl (e.g., 2H-imidazolyl-2-yl and 2H-imidazolyl-4-yl), pyrazolyl (e.g., 1H-pyrazol-3-yl, 1H-pyrazol-4-yl and 1H-pyrazol-5-yl), pyridyl (e.g., pyridinyl-2-yl, pyridinyl-3-yl and pyridinyl-4-yl), pyrimidinyl (e.g., ... Examples of N-aryl groups include pyrimidin-2-yl, pyrimidin-4-yl, and pyrimidin-5-yl; thiazolyl (e.g., thiazolyl-2-yl, thiazolyl-4-yl, and thiazolyl-5-yl); isothiazolyl (e.g., isothiazol-3-yl, isothiazol-4-yl, and isothiazol-5-yl); oxazolyl (e.g., oxazol-2-yl, oxazol-4-yl, and oxazol-5-yl); isoxazolyl (e.g., isoxazol-3-yl, isoxazol-4-yl, and isoxazol-5-yl); and triazolyl (e.g., 1,2,4-triazolyl and 1,2,3-triazolyl). The term "heteroaryl" is also intended to include possible N-oxides. Exemplary N-oxides include pyridinyl N-oxides.

[0508] For the purposes of this disclosure, the term "optionally substituted heteroaryl" used alone or as part of another group means that the heteroaryl group as defined above is unsubstituted or substituted by one to four substituents, for example, one or two substituents, independently selected from halogen, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aralkylaryloxy, aralkylalkoxy, alkylthio, carboxamido, sulfonamide, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxyl, carboxylalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclic, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamido)alkyl, mercaptoalkyl, (heterocyclic)alkyl, (heteroaryl)alkyl, -N(R) 33 (R) 34 ) or -N(H)C(=O)-R 35 , where R 33 Is it hydrogen or C? 1-4 Alkyl; R 34 It is an alkoxyalkyl, (heterocyclic)alkyl, (amino)alkyl, (alkylamino)alkyl, or (dialkylamino)alkyl; and R 35 It is an alkyl group, an optionally substituted aryl group, or an optionally substituted heteroaryl group. An optionally substituted heteroaryl group may have one substituent. The substituent may be amino, alkylamino, dialkylamino, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (heterocyclic)alkyl, -N(R) 33 (R) 34 ) or -N(H)C(=O)-R 35 The optionally substituted heteroaryl group can be an optionally substituted pyridyl group, namely 2-pyridyl, 3-pyridyl, or 4-pyridyl. Any available carbon or nitrogen atom can be substituted.

[0509] For the purposes of this disclosure, the term "heterocycle" or "heterocyclic group," used alone or as part of another group, refers to a saturated or partially unsaturated (e.g., containing one or two double bonds) cyclic group containing one, two, or three rings and having three to fourteen ring members (i.e., 3- to 14-membered heterocyclic groups) and at least one heteroatom. The heterocyclic group may be selected from C... 3-14 Heterocyclic groups and C 3-8Heterocyclic group. Each heteroatom is independently selected from the group consisting of: an oxygen atom; a sulfur atom, including sulfoxides and sulfones; and / or a nitrogen atom, which may be quaternized. The term "heterocyclic group" is intended to include cyclic urea groups, such as imidazoalkyl-2-ones; cyclic amide groups, such as β-lactam, γ-lactam, δ-lactam, and ε-lactam groups; and cyclic carbamate groups, such as oxazolidinyl-2-ones. The term "heterocyclic group" is also intended to include groups having fused optional substituted aryl groups, such as indololinyl, indololinyl-2-one, and benzo[d]oxazolyl-2(3H)-one. The term "heterocyclic group" is also intended to include groups having fused optional substituted heteroaryl groups, such as 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine. The heterocyclic group may be selected from 4-, 5-, 6-, 7-, or 8-membered cyclic groups containing one ring and one or more oxygen and / or nitrogen atoms; 5- or 6-membered cyclic groups containing one ring and one or two nitrogen atoms; or 8-, 9-, 10-, 11-, or 12-membered cyclic groups containing two rings and one or two nitrogen atoms. The heterocyclic group may optionally be connected to the remainder of the molecule via a carbon or nitrogen atom. Non-limiting illustrative heterocyclic groups include 2-oxopyrrolidone-3-yl, 2-imidazolidineone, piperidinyl, morpholinyl, piperazine, pyrrolylyl, azirrobutyl, 8-azibicyclo[3.2.1]octane (norhysterosine), 6-azispiro[2.5]octane, 6-azispiro[3.4]octane, indololinyl, indololin-2-one, and 1,3-dihydro-2H-benzo[d]imidazol-2-one.

[0510] For the purposes of this disclosure, the term "optionally substituted heterocyclic group" as used herein, either alone or as part of another group, means that the heterocyclic group as defined above is either unsubstituted or substituted by one to four independently selected substituents from the following: halogroup, nitrogroup, cyanogroup, hydroxyl group, amino group, alkylamino group, dialkylamino group, haloalkyl group, hydroxyalkyl group, alkoxy group, haloalkoxy group, aryloxy group, aralkylarylalkoxy group, alkylthio group, carboxamido group, sulfonamide group, alkylcarbonyl group, arylcarbonyl group, alkylsulfonyl group, arylsulfonyl group, carboxyl group, carboxylalkyl group, alkyl group, cycloalkyl group, alkenyl group, alkynyl group, aryl group, heteroaryl group, heterocyclic group, alkoxyalkyl group, (amino)alkyl group, hydroxyalkylamino group, (alkylamino)alkyl group, (dialkylamino)alkyl group, (cyano)alkyl group, (carboxamido)alkyl group, mercaptoalkyl group, (heterocyclic)alkyl group, and (heteroaryl)alkyl group. Substitution may occur on any available carbon or nitrogen atom and may form a spirocyclic ring.

[0511] For the purposes of this disclosure, the term "amino" as used alone or as part of another group refers to -NH2.

[0512] For the purposes of this disclosure, the term "alkylamino" used alone or as part of another group refers to –NHR 36, where R 36 It is C 1-6 Alkyl group. R 36 It can be C 1-4 Alkyl groups. Non-limiting illustrative alkylamino groups include -N(H)CH3 and -N(H)CH2CH3.

[0513] For the purposes of this disclosure, the term "dialkylamino" used alone or as part of another group refers to –NR 37a R 37b , where R 37a and R 37b Each is C independently 1-6 Alkyl group. R 37a and R 37b Each can be C independently 1-4 Alkyl groups. Non-limiting illustrative dialkylamino groups include -N(CH3)2 and -N(CH3)CH2CH(CH3)2.

[0514] For the purposes of this disclosure, the term "hydroxyalkylamino" used alone or as part of another group refers to –NHR 38 , where R 38 It is a hydroxyalkyl group.

[0515] For the purposes of this disclosure, the term "cycloalkylamino" used alone or as part of another group refers to –NR 39a R 39b , where R 39a It is an optional substituted cycloalkyl group and R 39b Is it hydrogen or C? 1-4 alkyl.

[0516] For the purposes of this disclosure, the term "arylamino" used alone or as part of another group refers to –NR 40a R 40b , where R 40a It is an aryl group and R 40b Is it hydrogen or C? 1-4 Alkyl groups. Non-limiting illustrative arylalkylamino groups include -N(H)CH2Ph ​​and -N(CH3)CH2Ph.

[0517] For the purposes of this disclosure, the term "(amino)alkyl" used alone or as part of another group refers to an alkyl group substituted with an amino group. The alkyl group can be C10, C20, C30, C40, C50, C6 ... 1-4Alkyl groups. Non-limiting illustrative (amino)alkyl groups include -CH2NH2, -C(NH2)(H)CH3, -CH2CH2NH2, -CH2C(NH2)(H)CH3, -CH2CH2CH2NH2, -CH2CH2CH2CH2NH2 and -CH2C(CH3)2CH2NH2.

[0518] For the purposes of this disclosure, the term "(alkylamino)alkyl" used alone or as part of another group refers to an alkyl group substituted with an alkylamino group. The alkyl group can be C10, C20, C30, C40, C50, C6 ... 1-4 Alkyl. Non-limiting illustrative (alkylamino) alkyl is -CH2CH2N(H)CH3.

[0519] For the purposes of this disclosure, the term "(dialkylamino)alkyl" used alone or as part of another group refers to an alkyl group substituted with a dialkylamino group. The alkyl group can be C10 ... 1-4 Alkyl group. Non-limiting illustrative (dialkylamino) alkyl group is -CH2CH2N(CH3)2.

[0520] For the purposes of this disclosure, the term "(cycloalkylamino)alkyl" used alone or as part of another group refers to an alkyl group substituted with a cycloalkylamino group. The alkyl group can be C10, C20, C30, C40, C50, C6 ... 1-4 Alkyl. Non-limiting illustrative (cycloalkylamino) alkyl groups include -CH2N(H)cyclopropyl, -CH2N(H)cyclobutyl and -CH2N(H)cyclohexyl.

[0521] For the purposes of this disclosure, the term "(aralkylamino)alkyl" used alone or as part of another group refers to an alkyl group substituted with an aralkylamino group. The alkyl group can be C10. 1-4 Alkyl. Non-limiting illustrative (arylamino) alkyl is -CH2CH2CH2N(H)CH2Ph.

[0522] For the purposes of this disclosure, the term "(cyano)alkyl" used alone or as part of another group refers to an alkyl group substituted with one or more cyano groups, such as -CN groups. The alkyl group can be C... 1-4 Alkyl. Non-limiting illustrative (cyano) alkyl groups include -CH2CH2CN, -CH2CH2CH2CN and -CH2CH2CH2CH2CN.

[0523] For the purposes of this disclosure, the term "(amino)(hydroxy)alkyl" used alone or as part of another group refers to an alkyl group substituted with one amino, alkylamino, or dialkylamino group and one hydroxyl group. An alkyl group is a C14 group. 1-6 Alkyl or C 1-4 alkyl.

[0524] For the purposes of this disclosure, the term "(amino)(aryl)alkyl" used alone or as part of another group refers to an alkyl group substituted with an amino, alkylamino, or dialkylamino group and an optionally substituted aryl group. The alkyl group can be C10, C20, C30, C40, C50, C60, C60, C70, C6 ... 1-6 Alkyl group. The optionally substituted aryl group may be an optionally substituted phenyl group.

[0525] For the purposes of this disclosure, the term "(cycloalkyl)alkyl" used alone or as part of another group refers to an alkyl group substituted with one optionally substituted cycloalkyl group. The alkyl group can be C10, C20, C30, C40, C50, C6 ... 1-4 Alkyl or C 3-6 Cycloalkyl. Optionally, the substituted cycloalkyl group may be replaced by an amino or (amino)alkyl group.

[0526] For the purposes of this disclosure, the term "(hydroxy)(aryl)alkyl" used alone or as part of another group refers to an alkyl group substituted with a hydroxyl group and an optionally substituted aryl group. The alkyl group can be C10. 1-6 Alkyl group. The optionally substituted aryl group may be an optionally substituted phenyl group. Non-limiting illustrative (hydroxy)(aryl)alkyl groups include:

[0527]

[0528] For the purposes of this disclosure, the term "carboxamide group" used alone or as part of another group refers to the formula -C(=O)NR 41a R 41b The group, wherein R 41a and R 41b Each is independently hydrogen, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl, or R 41a and R 41b Together with the nitrogen atom it is attached to, it forms 3- to 8-membered heterocyclic groups. R 41a and R 41b Each group may be hydrogen or optionally substituted alkyl group. Non-limiting illustrative carboxamide groups include -CONH2, -CON(H)CH3, -CON(CH3)2 and -CON(H)Ph.

[0529] For the purposes of this disclosure, the term "(carboxamido)alkyl" used alone or as part of another group refers to an alkyl group substituted with a carboxamido group. Non-limiting illustrative (carboxamido)alkyl groups include -CH2CONH2, -C(H)CH3-CONH2, and -CH2CON(H)CH3.

[0530] For the purposes of this disclosure, the term "sulfonamide group" used alone or as part of another group refers to the formula -SO2NR. 42a R42b The group, wherein R 42a and R 42b Each is independently hydrogen, optionally substituted alkyl or optionally substituted aryl, or R 42a and R 42b Together with the nitrogen to which it is attached, it forms a 3- to 8-membered heterocyclic group. Non-limiting illustrative sulfonamide groups include -SO2NH2, -SO2N(H)CH3 and -SO2N(H)Ph.

[0531] For the purposes of this disclosure, the term "alkyl carbonyl" used alone or as part of another group refers to a carbonyl group substituted with an alkyl group, i.e., -C(=O)-. A non-limiting illustrative alkyl carbonyl group is -COCH3.

[0532] For the purposes of this disclosure, the term "aryl carbonyl" used alone or as part of another group refers to a carbonyl group that is optionally substituted with an aryl group, i.e., -C(=O)-. A non-limiting illustrative aryl carbonyl group is -COPh.

[0533] For the purposes of this disclosure, the term "alkylsulfonyl" used alone or as part of another group refers to a sulfonyl group substituted with any of the optional substituted alkyl groups described above, i.e., -SO2-. Non-limiting illustrative alkylsulfonyl groups are -SO2CH3 (i.e., methylsulfonyl) and -SO2CH2CH3 (i.e., ethylsulfonyl).

[0534] For the purposes of this disclosure, the term "arylsulfonyl" used alone or as part of another group refers to a sulfonyl group substituted with any of the optional aryl groups described above, i.e., -SO2-. A non-limiting illustrative arylsulfonyl group is -SO2Ph.

[0535] For the purposes of this disclosure, the term “mercaptoalkyl” used alone or as part of another group refers to any of the above-mentioned alkyl groups substituted with a –SH group.

[0536] For the purposes of this disclosure, the term "carboxyl" used alone or as part of another group refers to a group of the formula -COOH.

[0537] For the purposes of this disclosure, the term "carboxyalkyl" as used alone or as part of another group refers to any of the above-mentioned alkyl groups substituted with -COOH. A non-limiting illustrative carboxyalkyl group is -CH2CO2H.

[0538] For the purposes of this disclosure, the term "alkoxycarbonyl" used alone or as part of another group refers to a carbonyl group substituted with an alkoxy group, i.e., -C(=O)-. Non-limiting illustrative alkoxycarbonyl groups are -CO2Me and -CO2Et.

[0539] For the purposes of this disclosure, the term "aralkyl" or "arylalkyl" used alone or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted aryl groups. An aralkyl group may be a C-aryl group substituted with one optionally substituted aryl group. 1-4 Alkyl groups. Non-limiting illustrative aralkyl groups include benzyl, phenethyl, -CHPh2, -CH2(4-OH-Ph) and -CH(4-F-Ph)2.

[0540] For the purposes of this disclosure, the term "(heterocyclic)alkyl" used alone or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted heterocyclic groups. The (heterocyclic)alkyl group may be a C-aryl group substituted with one optionally substituted heterocyclic group. 1-4 Alkyl groups. Heterocyclic groups can be attached to alkyl groups via carbon or nitrogen atoms. Non-limiting illustrative (heterocyclic) alkyl groups include:

[0541]

[0542] For the purposes of this disclosure, the term "heteroaryl" or "(heteroaryl)alkyl" used alone or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted heteroaryl groups. (Heteroaryl)alkyl can be a C-aryl group substituted with one optionally substituted heteroaryl group. 1-4 Alkyl groups. Non-limiting illustrative (heteroaryl) alkyl groups include:

[0543]

[0544] For the purposes of this disclosure, the term "alkylcarbonylamino" used alone or as part of another group refers to an alkyl carbonyl group attached to an amino group. A non-limiting illustrative alkylcarbonylamino is -NHCOCH3.

[0545] This disclosure covers any of the compounds of this disclosure that are isotopically labeled (i.e., radioactively labeled) by substituting one or more atoms with atoms of different atomic masses or mass numbers. Examples of isotopes that may be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, respectively, as follows: 2 H (or deuterium (D)), 3 H, 11 C 13 C 14 C 15 N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl, for example3 H, 11 C and 14 C. This disclosure also provides a composition in which substantially all atoms at a certain position within the compound of this disclosure are replaced by atoms with different atomic masses or mass numbers. This disclosure also provides a composition in which a subset of atoms at a certain position within the compound of this disclosure are replaced by atoms with different atomic masses or mass numbers, i.e., the compound of this disclosure is enriched at a certain position with atoms of different atomic masses or mass numbers. In one embodiment, this disclosure provides a composition in which 1 to 8 hydrogen atoms of the compound of this disclosure are replaced by deuterium. Isotopically labeled compounds of this disclosure can be prepared by methods known in this art.

[0546] The compounds disclosed herein may contain one or more asymmetric centers and may produce enantiomers, diastereomers, and other stereoisomers. This disclosure is intended to cover the use of all such possible forms, as well as their racemic and resolved forms and mixtures thereof. In view of this disclosure, individual enantiomers can be isolated according to methods known in this art. When the compounds described herein contain alkene double bonds or other geometrically asymmetric centers, and unless otherwise stated, it is contemplated that the compounds include E and Z geometric isomers. All tautomers are also contemplated to be covered in this disclosure.

[0547] As used herein, the term "stereoisomer" is a general term for all isomers of an individual molecule that differ only in the spatial orientation of their atoms. The term includes enantiomers and isomers (diasteromers) of compounds having more than one chiral center that are not mirror images of each other.

[0548] The term "chiral center" or "asymmetric carbon atom" refers to a carbon atom that is connected to four different groups.

[0549] The terms “enantiomer” and “enantiomer” refer to molecules that cannot be superimposed on their mirror image and are therefore optically active, wherein the enantiomer is rotated in one direction along a polarizing plane and its mirror compound is rotated in the opposite direction along a polarizing plane.

[0550] The term "racemic" refers to a mixture of equal parts enantiomers that is not optically active.

[0551] The term "absolute configuration" refers to the spatial arrangement of atoms in a chiral molecular entity (or group) and its stereochemical description, such as R or S.

[0552] Unless otherwise indicated, the stereochemical terminology and conventions used in this specification are intended to be consistent with those described in Pure & Appl. Chem 68:2193 (1996).

[0553] The term "enantiomer excess" or "ee" refers to a measure of the amount of one enantiomer present relative to another. For a mixture of R and S enantiomers, the enantiomer excess percentage is defined as |RS|*100, where R and S are the corresponding molar or weight fractions of the enantiomers in the mixture, thus R+S=1. Based on the knowledge of the optical rotation of chiral substances, the enantiomer excess percentage is defined as ([α]... obs / [α] max )*100, where [α] obs It is the optical rotation of the enantiomer mixture and [α] max This is the optical rotation of the pure enantiomer. Enantiomer excess can be determined using various analytical techniques, including NMR spectroscopy, chiral column chromatography, or optical rotation determination.

[0554] The term "enantiomerically pure" or "enantiomerically pure" means that all molecules in a chiral substance sample (within the detection limit) have the same chirality.

[0555] The term "enantiomer enrichment" or "enantiomer concentration" refers to a chiral sample where the enantiomer ratio is greater than 50:50. The enantiomerically enriched compounds can be enantiomerically pure.

[0556] It should be understood that the embodiments of the invention described herein include "consisting of the embodiments" and / or "substantially consisting of the embodiments". Unless otherwise indicated, as used herein, the singular forms "a / an" and "described" include the plural forms. The term "or" as used herein is not intended to imply that alternatives are mutually exclusive.

[0557] In this application, unless expressly stated or understood by one of those skilled in the art, the word "or" means "and / or". In cases of multiple appendices, "or" refers to one or more of the aforementioned independent or appendiceal items.

[0558] As used herein, the term “about” includes the numerical value ±10%. Therefore, “about 10” means 9 to 11. Those skilled in the art will understand that references to “about” a value or parameter herein include (and describe) instances of the value or parameter itself. For example, a description of “about X” includes a description of “X”.

[0559] This disclosure covers the preparation and use of salts of the compounds disclosed herein, including non-toxic and pharmaceutically acceptable salts. Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts and basic salts. Pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium, potassium, cesium, and similar salts; alkaline earth metal salts such as calcium, magnesium, and similar salts; organic amine salts such as triethylamine, pyridine, methylpyridine, ethanolamine, triethanolamine, dicyclohexylamine, N,N'-diphenylmethylethylenediamine, and similar salts; inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulfate, and similar salts; organic acid salts such as citrate, lactate, tartrate, maleate, fumarate, mandelate, acetate, dichloroacetate, trifluoroacetate, oxalate, formate, and similar salts; sulfonates such as methanesulfonate, benzenesulfonate, p-toluenesulfonate, and similar salts; and amino acid salts such as arginine, aspartate, glutamate, and similar salts. As used herein, the term "pharmaceutically acceptable salt" means, for example, any salt that is physiologically tolerable to a target patient (e.g., a mammal, such as a human) by reacting the compounds of this disclosure with an acid or base.

[0560] Acid addition salts can be formed by mixing a solution of a specific compound of the present disclosure with a solution of a pharmaceutically acceptable, non-toxic acid (such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, dichloroacetic acid, or similar acids). Basic salts can be formed by mixing a solution of a compound of the present disclosure with a solution of a pharmaceutically acceptable, non-toxic alkali (such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, and similar alkalis).

[0561] This disclosure covers the preparation and use of solvates of the compounds disclosed herein. Solvates typically do not significantly alter the physiological activity or toxicity of the compound and therefore may function as a pharmacological equivalent. As used herein, the term "solvate" is a combination, physical association, and / or solvation of the compound of this disclosure with solvent molecules, such as a disolvate, monosolvate, or hemisolvate, wherein the ratio of solvent molecules to the compound of this disclosure is about 2:1, about 1:1, or about 1:2, respectively. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the lattice of a crystalline solid, the solvate can be separated. Therefore, "solvate" encompasses the solution phase and the separable solvate. The compounds of this disclosure may be in a form solvated with pharmaceutically acceptable solvents such as water, methanol, ethanol, and the like, and this disclosure is intended to include both solvated and unsolvated forms of the compounds of this disclosure. One type of solvate is a hydrate. "Hydrate" refers to a specific solvate in which the solvent molecules are a subgroup of water. Solvates typically function as pharmacological equivalents. Preparation of solvates is known in this art. See, for example, M. Caira et al., J. Pharmaceut. Sci., 93(3):601-611 (2004), which describes the preparation of solvates of fluconazole with ethyl acetate and with water. Similar preparations of solvates, semisolvents, hydrates, and analogs are described in EC van Tonder et al., AAPS Pharm. Sci. Tech., 5(1): Article 12 (2004); and ALBingham et al., Chem. Commun. 603-604 (2001). Typical, non-limiting methods for preparing solvates involve dissolving the disclosed compound in a desired solvent (organic solvent, water, or a mixture thereof) at a temperature above 20°C to about 25°C, followed by cooling the solution at a rate sufficient to form crystals, and separating the crystals by known methods, such as by filtration. Analytical techniques, such as infrared spectroscopy, can be used to determine the presence of solvent in solvate crystals.

[0562] Since the compounds disclosed herein are inhibitors of the USP1 protein, this disclosure provides a method for inhibiting the USP1 protein, the method comprising contacting the USP1 protein or a composition containing the USP1 protein with one or more of the compounds disclosed herein.

[0563] Because the compounds disclosed herein are inhibitors of the USP1 protein, they can be used to treat a variety of diseases, disorders, or conditions mediated by the USP1 protein. Therefore, this disclosure generally relates to a method for treating animals suffering from or at risk of developing a disease, disorder, or condition that responds to inhibition of the USP1 protein, the method comprising administering an effective amount of one or more of the compounds disclosed herein to the animal.

[0564] This disclosure further relates to a method for inhibiting the USP1 protein in a desired animal, the method comprising administering to the animal a therapeutically effective amount of at least one compound of this disclosure.

[0565] As used herein, “treatment” is a method for achieving a beneficial or desired clinical outcome. As used herein, “treatment” encompasses any administration or application of a therapeutic agent for a disease in mammals, including humans. For the purposes of this disclosure, a beneficial or desired clinical outcome includes, but is not limited to, any one or more of the following: relief of one or more symptoms; reduction of the severity of the disease; prevention or delay of disease spread (e.g., metastasis); prevention or delay of disease recurrence; delay or slowing of disease progression; improvement of the disease state; inhibition of the disease or disease progression; inhibition or slowing of the disease or its progression; cessation of its development; and remission (whether partial or complete remission). “Treatment” also encompasses the reduction of the pathological consequences of proliferative diseases. The methods provided herein cover any one or more of the aforementioned aspects of treatment. Consistent with the foregoing, the term treatment does not require the complete removal of all aspects of the condition.

[0566] In the context of cancer, the term "treatment" includes, but is not limited to, inhibiting cancer cell growth; inhibiting cancer cell replication; reducing the overall tumor burden; and delaying, stopping, or slowing tumor growth, progression, or metastasis.

[0567] As used herein, “delay” means to postpone, halt, slow, delay, stabilize, suppress, and / or postpone the development or progression of a disease (such as cancer). Depending on the patient’s medical history and / or the individual being treated, the delay may have different durations.

[0568] The "therapeutic effective dose" of a substance can vary depending on a variety of factors, such as disease state; individual age, sex, and weight; and the substance's ability to elicit the desired response in the individual. A therapeutic effective dose is also the amount by which the beneficial therapeutic effect outweighs any toxic or harmful effects of the substance. A therapeutic effective dose can be delivered in one or more administrations. A therapeutic effective dose refers to the amount that effectively achieves the desired therapeutic outcome at the necessary dosage and within the necessary time period.

[0569] The terms “administer” and similar forms refer to methods that can be used to deliver a therapeutic agent to a desired biological site of action. Administration techniques that can be used with the agents and methods described herein can be found, for example, in Goodman and Gilman, *The Pharmacological Basis of Therapeutics*, current edition, Pergamon; and Remington's, *Pharmaceutical Sciences* (current edition), Mack Publishing Co., Easton, Pa.

[0570] The terms "pharmaceutical formulation" and "pharmaceutical composition" refer to preparations that are in a form that enables the bioactivity of the active ingredient and do not contain other components that would cause unacceptable toxicity to a subject to whom the formulation will be administered. The formulation may be sterile.

[0571] "Pharmaceutically acceptable carrier" refers to a non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid, or carrier commonly used in this technology with therapeutic agents, which together constitute a "pharmaceutical composition" for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to the recipient at the dose and concentration used and is compatible with other components of the formulation. The pharmaceutically acceptable carrier is suitable for the formulation used.

[0572] "Sterilized" formulations are sterile or substantially free of live microorganisms and their spores.

[0573] The term "container" means any container and its cap suitable for storing, transporting, distributing and / or disposing of pharmaceutical products.

[0574] The term "insert" or "packaging insert" refers to information accompanying a pharmaceutical product that provides a description of how the product should be used, as well as safety and efficacy data necessary for physicians, pharmacists, and patients to make informed decisions regarding the use of the product. Packaging inserts are generally considered the "label" of pharmaceutical products.

[0575] As used herein, the terms "disease" or "disorder" or "symptom" refer to a condition that requires and / or desires treatment and indicate a disorder and / or abnormality of function that is generally considered pathological and may manifest as specific signs, symptoms, and / or functional impairments. As shown below, the compounds disclosed herein inhibit the USP1 protein and can be used to treat diseases and disorders such as proliferative disorders, wherein inhibition of the USP1 protein will provide benefits.

[0576] The term "peptide" and "protein" are used interchangeably to refer to a polymer of amino acid residues and are not limited to the shortest length. The polymer of amino acid residues may contain native or non-native amino acid residues and includes, but is not limited to, peptides, oligopeptides, dimers, trimers, and polymers of amino acid residues. The definition covers full-length proteins and fragments thereof. The term also includes post-expression modifications of peptides, such as glycosylation, sialylation, acetylation, phosphorylation, and similar modifications. Additionally, for the purposes of this disclosure, "peptide" refers to a protein that includes modifications of its native sequence (such as deletions, additions, and substitutions (which are generally conserved)) as long as the protein maintains the desired activity. The modifications may be intentional, such as modifications performed by site-directed mutagenesis; or they may be incidental, such as modifications caused by mutations in the host producing the protein, or errors caused by PCR amplification.

[0577] As used herein, “USP1” and “ubiquitin-specific processing protease 1” refer to any natural polypeptide or polynucleotide encoding USP1. The term “USP1” covers the “full-length” unprocessed USP1 polypeptide as well as any form of USP1 obtained by intracellular processing (e.g., removal of the signal peptide). The term also covers naturally occurring variants of USP1, such as those encoded by splice variants and allelic variants. The USP1 polypeptide described herein can be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. Human USP1 sequences are known and include sequences (including isotypes) disclosed, for example, in UniProt No. 094782. As used herein, the term “human USP1 protein” refers to a USP1 protein comprising the amino acid sequence shown in SEQ ID NO:1 of U.S. Provisional Patent Application No. 62 / 857,986, filed June 6, 2019.

[0578] USP1 is a deubiquitinating enzyme that acts as part of a complex with UAF1. The “deubiquitinating enzyme activity” of USP1 includes its ability to deubiquitinate as part of the USP1-UAF1 complex.

[0579] The term "specific binding" to a protein or a protein domain is well-known in the art, as are the methods for determining such specific binding. A molecule is considered to exhibit "specific binding" or "preferential binding" if its reaction or association with a particular protein or protein domain is more frequent, faster, longer-lasting, and / or has a higher affinity than its reaction or association with alternative proteins or domains. It should be understood that a molecule that specifically or preferentially binds to a first protein or domain may or may not specifically or preferentially bind to a second protein or domain. Therefore, "specific binding" or "preferential binding" does not necessarily require (but may include) exclusive binding. Generally, but not necessarily, the mention of binding implies preferential binding. For example, USP1 inhibitors that specifically bind to USP1, UAF1, and / or the USP1-UAF1 complex may not bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (USP46-UAF1), or may bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with a lower affinity than those binding to USP1.

[0580] The terms “reduction” or “inhibition” refer to a decrease or cessation of any phenotypic trait, or a decrease or cessation of the occurrence, extent, or likelihood of said trait. “Reduction” or “inhibition” means to reduce, decrease, or stop the activity, function, and / or amount of a substance relative to a reference. In some embodiments, “reduction” or “inhibition” means the ability to cause an overall reduction of 20% or higher percentage. In some embodiments, “reduction” or “inhibition” means the ability to cause an overall reduction of 50% or higher percentage. In some embodiments, “reduction” or “inhibition” means the ability to cause an overall reduction of 75%, 85%, 90%, 95%, or higher percentage. In some embodiments, the aforementioned amounts are inhibited or reduced over the same time period relative to a control over the same time period.

[0581] In some embodiments, inhibition of the USP1 protein means inhibiting one or more activities or functions of the USP1 protein. It should be understood that the activity or function of said one or more USP1 proteins can be inhibited in vitro or in vivo. Non-limiting examples of USP1 activity and function include deubiquitinating enzyme activity and complex formation with UAF1, which are described herein. Exemplary levels of inhibition of the activity of one or more USP1 proteins include at least 10% inhibition, at least 20% inhibition, at least 30% inhibition, at least 40% inhibition, at least 50% inhibition, at least 60% inhibition, at least 70% inhibition, at least 80% inhibition, at least 90% inhibition, and up to 100% inhibition.

[0582] The terms “individual” or “subject” are used interchangeably herein and refer to an animal, such as a mammal, like a human. In some cases, methods of treating mammals are provided, including but not limited to humans, rodents, apes, cats, dogs, horses, cattle, pigs, sheep, goats, laboratory mammals, farm mammals, sporting mammals, and pet mammals. In some instances, “individual” or “subject” refers to an individual or subject who requires treatment for a disease or condition. In some cases, a subject receiving treatment may be a patient, indicating that the subject has been identified as having a treatment-related condition or is at particular risk of contracting an infectious disease.

[0583] As used herein, the terms “cancer” and “tumor” refer to or describe a physiological disorder in which a population of cells in a mammal is characterized by uncontrolled cell growth. The terms encompass solid tumors and hematologic / lymphomas. Examples of cancer include, but are not limited to, cancers with defective DNA damage repair pathways. Other examples of cancer include, but are not limited to, ovarian cancer, breast cancer (including triple-negative breast cancer), non-small cell lung cancer (NSCLC), and osteosarcoma. Cancer can be BRCA1 or BRCA2 wild-type. Cancer can also be BRCA1 or BRCA2 mutant. Cancer can also be PARP inhibitor-resistant or refractory cancer, or PARP inhibitor-resistant or refractory BRCA1 or BRCA2 mutant cancer.

[0584] As used herein, the term "loss-of-function" mutation refers to a mutation that results in the absence of a gene, reduced gene expression, or the production of a less active or inactive gene product (e.g., a protein). Loss-of-function mutations include, for example, missense mutations, nucleotide insertions, nucleotide deletions, and gene deletions. Loss-of-function mutations also include dominant-negative mutations. Therefore, cancer cells with loss-of-function mutations in the gene encoding p53 include cancer cells containing missense mutations in the gene encoding p53 as well as cancer cells lacking the gene encoding p53.

[0585] USP1 inhibitors

[0586] In various embodiments, the compounds disclosed herein are USP1 inhibitors that reduce the level of the USP1 protein and / or inhibit or reduce at least one biological activity of the USP1 protein.

[0587] In some embodiments, the disclosed compounds specifically bind to the USP1 protein. In some embodiments, the disclosed compounds specifically bind to the USP1 protein in the USP1-UAF1 complex. In some embodiments, the disclosed compounds specifically bind to USP1 mRNA. In some embodiments, the disclosed compounds specifically bind to the USP1 protein (USP1 protein alone or USP1 protein in the USP1-UAF1 complex) or USP1 mRNA. In some embodiments, the disclosed compounds specifically bind to UAF1 (UAF1 alone or UAF1 in the USP1-UAF1 complex) and inhibit or reduce the formation or activity of the USP1-UAF1 complex.

[0588] In some embodiments, the compounds disclosed herein reduce the formation of the USP1-UAF1 complex. In some embodiments, the compounds disclosed herein reduce the activity of the USP1-UAF1 complex. In some embodiments, the compounds disclosed herein reduce the deubiquitinating enzyme activity of USP1. In some embodiments, the compounds disclosed herein increase monoubiquitinated PCNA. In some embodiments, the compounds disclosed herein increase monoubiquitinated FANCD2. In some embodiments, the compounds disclosed herein increase monoubiquitinated FANCI.

[0589] In some embodiments, the disclosed compounds do not bind to other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) or bind to deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with an affinity at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold lower than their affinity for USP1 (i.e., the USP1 inhibitor has a lower Kelvin-weighted affinity for other deubiquitinating enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1)). D K is for USP1 D (At least 5 times, at least 10 times, at least 20 times, or at least 100 times).

[0590] In some embodiments, the disclosed compounds inhibit USP1 deubiquitinase activity, for example, with an IC50 less than about 50 nM, between about 50 nM and about 200 nM, between about 200 nM and about 2 pM, or greater than 2 pM, as measured using the assay disclosed in U.S. Patent Application Publication No. 2017 / 0145012, or for example, with an IC50 of 50 nM to 1000 nM, as measured using the assay disclosed in Liang et al., Nat Chem Biol 10:289-304 (2014). In some embodiments, the disclosed compounds inhibit USP1 deubiquitinase activity with an IC50 as measured using the assay disclosed in Chen et al., Chem Biol., 18(11):1390-1400 (2011). In some embodiments, the disclosed compounds do not inhibit the activity of other deubiquitinizing enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1), or inhibit the activity of other deubiquitinizing enzymes, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with an IC50 of at least 5, at least 10, at least 20, or at least 100 times the IC50 of the inhibition of USP1 deubiquitinizing enzyme activity.

[0591] In some embodiments, the affinity of the disclosed compounds for the USP1 protein is in the range of 1 pM to 100 μM, or 1 pM to 1 μM, or 1 pM to 500 nM, or 1 pM to 100 nM. In some embodiments, the affinity of the disclosed compounds for the USP1 protein is about 1 pM to about 100 μM, about 1 nM to about 100 μM, about 1 μM to about 100 μM, about 1 μM to about 50 μM, about 1 μM to about 40 μM, about 1 μM to about 30 μM, about 1 μM to about 20 μM, or about 1 μM to about 10 μM, about 1 μM, about 5 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, about 35 μM, about 40 μM, about 45 μM, about 50 μM, about 60 μM, about 70 μM, about 80 μM, about 90 μM, or about 100 μM. In some embodiments, the affinity of the disclosed compounds for the USP1 protein is about 100 nM to about 1 μM, about 100 nM to about 900 nM, about 100 nM to about 800 nM, about 100 nM to about 700 nM, about 100 nM to about 600 nM, about 100 nM to about 500 nM, about 100 nM to about 400 nM, about 100 nM to about 300 nM, about 100 nM to about 200 nM, about 200 nM, etc. 0 nM to about 1 μM, about 300 nM to about 1 μM, about 400 nM to about 1 μM, about 500 nM to about 1 μM, about 600 nM to about 1 μM, about 700 nM to about 1 μM, about 800 nM to about 1 μM, about 900 nM to about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM. In some embodiments, the affinity of the disclosed compounds for the USP1 protein is about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100nM, about 50nM to about 100nM, about 60nM to about 100nM, about 70nM to about 100nM, about 80nM to about 100nM, about 90nM to about 100nM, about 1nM, about 2nM, about 3nM, about 4nM, about 5nM, about 6nM, about 7nM, about 8nM, about 9nM, about 10nM, about 20nM, about 30nM, about 40nM, about 50nM, about 60nM, about 70nM, about 80nM, about 90nM, or about 100nM.In some embodiments, the compounds of this disclosure have an affinity for the USP1 protein of less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the compounds of this disclosure have an affinity for the USP1 protein of less than 1 nM.

[0592] In some embodiments, the disclosed compounds inhibit the activity of USP1 by IC 50 The concentrations are 1 pM to 100 μM, or 1 pM to 1 μM, or 1 pM to 500 nM, or 1 pM to 100 nM. In some embodiments, the disclosed compounds inhibit USP1 activity at IC50. 50 The concentrations are approximately 1 pM to approximately 100 μM, approximately 1 nM to approximately 100 μM, approximately 1 μM to approximately 100 μM, approximately 1 μM to approximately 50 μM, approximately 1 μM to approximately 40 μM, approximately 1 μM to approximately 30 μM, approximately 1 μM to approximately 20 μM, or approximately 1 μM to approximately 10 μM, approximately 1 μM, approximately 5 μM, approximately 10 μM, approximately 15 μM, approximately 20 μM, approximately 25 μM, approximately 30 μM, approximately 35 μM, approximately 40 μM, approximately 45 μM, approximately 50 μM, approximately 60 μM, approximately 70 μM, approximately 80 μM, approximately 90 μM, or approximately 100 μM. In some embodiments, the compounds disclosed herein inhibit the activity of USP1 by an IC50. 50 From about 100 nM to about 1 μM, from about 100 nM to about 900 nM, from about 100 nM to about 800 nM, from about 100 nM to about 700 nM, from about 100 nM to about 600 nM, from about 100 nM to about 500 nM, from about 100 nM to about 400 nM, from about 100 nM to about 300 nM, from about 100 nM to about 200 nM, from about 200 nM to about 1 μM, from about 300 nM M to about 1 μM, about 400 nM to about 1 μM, about 500 nM to about 1 μM, about 600 nM to about 1 μM, about 700 nM to about 1 μM, about 800 nM to about 1 μM, about 900 nM to about 1 μM, about 100 nM, about 200 nM, about 300 nM, about 400 nM, about 500 nM, about 600 nM, about 700 nM, about 800 nM, or about 900 nM. In some embodiments, the compounds disclosed herein inhibit the activity of USP1 by an IC 50For about 1 nM to about 100 nM, 1 nM to about 90 nM, 1 nM to about 80 nM, 1 nM to about 70 nM, 1 nM to about 60 nM, 1 nM to about 50 nM, 1 nM to about 40 nM, 1 nM to about 30 nM, 1 nM to about 20 nM, 1 nM to about 10 nM, about 10 nM to about 100 nM, about 20 nM to about 100 nM, about 30 nM to about 100 nM, about 40 nM to about 100 nM, about 50 nM to about 1 00 nM, about 60 nM to about 100 nM, about 70 nM to about 100 nM, about 80 nM to about 100 nM, about 90 nM to about 100 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 20 nM, about 30 nM, about 40 nM, about 50 nM, about 60 nM, about 70 nM, about 80 nM, about 90 nM, or about 100 nM. In some embodiments, the compounds disclosed herein inhibit the activity of USP1 by an IC 50 Less than 1 μM, less than 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM. In some embodiments, the compounds disclosed herein inhibit USP1 activity at IC50 concentrations. 50 Less than 1 nM.

[0593] Example assay

[0594] Any suitable assay in this technique can be used to determine activity, detect results or effects, determine efficacy, etc. Some non-limiting illustrative assays that can be used in the methods provided herein have been described.

[0595] In some cases, methods for determining whether the disclosed compounds inhibit USP1 deubiquitinase activity measure the change in mass following the binding of the deubiquitinase to cleave diubiquitin. For example, ubiquitin aldehydes and ubiquitin vinyl sulfones form irreversible covalent bonds with the deubiquitinase, thereby causing an observable change in the mass of the deubiquitinase. Similarly, diubiquitin cleavage causes an observable change in mass.

[0596] In some cases, methods for determining whether the disclosed compounds inhibit the activity of USP1 deubiquitinating enzymes involve luminescence or increased fluorescence after cleavage, which can be detected, for example, on a plate reader. Such assays can use ubiquitin linked to a fluorophore via a linker bond, such as ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) or ubiquitin-rosin 110. Such assays can also use diubiquitins containing isopeptide bonds. An exemplary diubiquitin may contain a fluorophore on one ubiquitin and a quencher on the other, thereby increasing fluorescence upon diubiquitin cleavage. Such assays can also use enzyme-coupled systems in which ubiquitin is coupled to an enzyme that is active only when a luciferase product is produced and released from the ubiquitin.

[0597] An exemplary deubiquitination assay for USP1 / UAF1 activity and inhibitor testing Deubiquitinating enzyme activity can be measured using ubiquitin-rose 110 as a substrate. The cleavage of the amide bond between rose 110 and the C-terminal glycine of ubiquitin increases the fluorescence signal. The assay can be performed in 20 μL of assay buffer (50 mM Tris-HCl, pH 7.8, 0.5 mM EDTA, 0.01% bovine serum albumin, 1 mM DTT, 0.01% Tween-20) and 0.05 nM USP1 / UAF1 enzyme. The reaction can be initiated by adding 150 nM ubiquitin-rose 110 (Boston Biochem) substrate.

[0598] The disclosed compound can be dissolved in DMSO and tested in a dose-response manner, starting at 10 μM.

[0599] The disclosed compound can be added to an enzyme / assay buffer mixture and incubated for 10 minutes. A substrate mixture can be added, and the reaction mixture can be read in kinetic mode at Ex480 / Em540 for 30 minutes, and an IC50 reaction curve can be plotted. See, for example, Chen et al., Chem Biol., 18(11):1390-1400 (2011).

[0600] How to use

[0601] In some embodiments, the compounds disclosed herein can be used to inhibit the activity of the USP1 protein. For example, in some embodiments, the method of inhibiting the USP1 protein includes contacting the USP1 protein with the compounds disclosed herein. This contact may occur in vitro or in vivo.

[0602] In some embodiments, the disclosed compounds can be used to treat “USP1 protein-mediated conditions”. USP1 protein-mediated conditions are any pathological disorders in which the USP1 protein is known to play a role. In some embodiments, USP1 protein-mediated conditions are proliferative diseases, such as cancer.

[0603] This document provides various methods for treating diseases and conditions using the compounds disclosed herein. Exemplary diseases and conditions that can be treated with the compounds disclosed herein include, but are not limited to, cancer.

[0604] In some embodiments, a method of treating cancer with the compounds disclosed herein is provided. The method includes administering a therapeutically effective amount of the compounds disclosed herein to a subject suffering from cancer.

[0605] In some embodiments, the cancer to be treated with the compounds disclosed herein is selected from hematologic malignancies, lymphomas, and cancers with DNA damage repair pathway defects. In some embodiments, the cancer to be treated with the compounds disclosed herein is a cancer containing cancer cells with mutations in the gene encoding p53. In some embodiments, the cancer to be treated with the compounds disclosed herein is a cancer containing cancer cells with loss-of-function mutations in the gene encoding p53. In some embodiments, the cancer to be treated with the compounds disclosed herein is a cancer containing cancer cells with mutations in the gene encoding BRCA1. In some embodiments, the cancer to be treated with the compounds disclosed herein is a cancer containing cancer cells with mutations in the gene encoding BRCA2. In some embodiments, the cancer to be treated with the compounds disclosed herein is a cancer containing cancer cells with loss-of-function mutations in the gene encoding ATM.

[0606] In some embodiments, the cancer intended to be treated with the compounds disclosed herein is selected from non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer. In some embodiments, the cancer is ovarian cancer or breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is triple-negative breast cancer.

[0607] In some embodiments, the cancers intended to be treated with the compounds disclosed herein are selected from the group consisting of: bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma, and meningioma; soft tissue cancer, including rhabdomyosarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer; uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.

[0608] This document provides various methods of treating cancer with the compounds disclosed herein. In some embodiments, a therapeutically effective amount of the compounds disclosed herein is administered to a subject suffering from cancer comprising cancer cells having elevated RAD18 levels. In some embodiments, the elevated RAD18 level is an elevated RAD18 protein level. In some embodiments, the elevated RAD18 level is an elevated RAD18 mRNA level. In some embodiments, the elevated RAD18 (e.g., RAD18 protein and / or RAD18 mRNA) level has been detected prior to administration (e.g., in a cancer sample obtained from the subject). That is, in some embodiments, the RAD18 protein or mRNA in the subject's cancer has been tested before initiating treatment with a USP1 inhibitor.

[0609] In some embodiments, the method includes (a) identifying the subject's cancer as a USP1 inhibitor-sensitive cancer, and then (b) administering a therapeutically effective amount of the disclosed compound to the subject.

[0610] In some embodiments, the method includes (a) detecting the level of RAD18 (e.g., RAD18 protein and / or RAD18 mRNA) in cancer cells (e.g., in a cancer sample obtained from a subject), followed by (b) administering a therapeutically effective amount of the disclosed compound to a subject with cancer comprising cells having elevated RAD18 levels.

[0611] In some embodiments, the method includes administering a therapeutically effective amount of the disclosed compound to a subject suffering from triple-negative breast cancer.

[0612] In some embodiments, the compounds of this disclosure are used to treat cancer, wherein the cancer is a homologous recombination-deficient cancer. In some embodiments, the compounds of this disclosure are used to treat cancer, wherein the cancer comprises cancer cells with mutations in a gene encoding p53. In some embodiments, the compounds of this disclosure are used to treat cancer, wherein the cancer comprises cancer cells with loss-of-function mutations in a gene encoding p53. In some embodiments, the compounds of this disclosure are used to treat cancer, wherein the cancer is not defective in the homologous recombination pathway.

[0613] In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is a BRCA1-mutant cancer. In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is a BRCA2-mutant cancer. In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is both a BRCA1-mutant cancer and a BRCA2-mutant cancer. In some embodiments, the cancer is neither a BRCA1-mutant cancer nor a BRCA2-mutant cancer. In some embodiments, the cancer is a BRCA1-deficient cancer. In some embodiments, the cancer is a BRCA2-deficient cancer. In some embodiments, the cancer is both a BRCA1-deficient cancer and a BRCA2-mutant cancer.

[0614] In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is an ATM-mutant cancer. In some embodiments, the cancer is not an ATM-mutant cancer. In some embodiments, the cancer is an ATM-deficient cancer.

[0615] In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is PARP inhibitor-resistant or refractory cancer. In some embodiments, the compounds disclosed herein are used to treat cancer, wherein the cancer is PARP inhibitor-resistant or refractory BRCA1-deficient cancer.

[0616] In some embodiments, the cancer is a BRCA1 and / or BRCA2 mutant cancer, wherein the cancer comprises cells with elevated RAD18 levels, for example, wherein the elevated RAD18 levels are at least as high as the RAD18 protein and / or mRNA levels in ES2 cells, or wherein the elevated RAD18 levels are higher than the RAD18 protein and / or mRNA levels in HEP3B217 cells. In some embodiments, triple-negative breast cancer is a BRCA1 and / or BRCA2 mutant cancer.

[0617] In some cases, the cancer is a solid tumor. In some cases, the cancer is a blood cancer / lymphoma. In some cases, the cancer is a cancer with a defective DNA damage repair pathway. In some cases, the cancer is a cancer with a defective homologous recombination pathway. In some cases, the cancer comprises cancer cells with mutations in the gene encoding p53. In some cases, the cancer comprises cancer cells with loss-of-function mutations in the gene encoding p53. In some cases, the cancer is selected from the group consisting of: non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer (including triple-negative breast cancer). In some cases, the cancer is ovarian cancer or breast cancer (including triple-negative breast cancer). In some cases, the cancer is ovarian cancer. In some cases, the cancer is breast cancer (including triple-negative breast cancer).

[0618] In some embodiments, the disclosed compounds are used in combination with one or more additional therapeutic agents for the treatment of cancer. It has been reported that p53 status determines PARP inhibitor sensitization (Sa et al., Genome Biology, (2019) 20:253), and BRCA1 / 2 status predicts the clinical efficacy of PARP inhibitors (Audeh et al., Lancet (2010) 376(9737), 245-51). As shown below, p53-mutant cancers and BRCA-mutant cancers exhibit increased sensitivity to USP1 inhibitors. Therefore, in some embodiments, the disclosed compounds are used in combination with PARP inhibitors for the treatment of cancer.

[0619] In some embodiments, the presently disclosed compounds are provided for use as pharmaceutical agents or for the preparation of pharmaceutical agents, for example, for treating cancer. In some embodiments, the presently disclosed compounds are provided for use in methods of treating cancer.

[0620] Pharmaceutical Composition

[0621] The disclosed compounds may be administered to mammals as a raw material chemical reagent free of any other components, or as part of a pharmaceutical composition containing the disclosed compounds in combination with a pharmaceutically acceptable carrier (see, for example, Gennaro, Remington: The Science and Practice of Pharmacy with Facts and Comparisons: Drugfacts Plus, 20th edition (2003); Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th edition, Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook of Pharmaceutical Excipients, 3rd edition, Pharmaceutical Press (2000)). Such carriers may be selected from pharmaceutically acceptable excipients and adjuvants. The terms “pharmaceutically acceptable carrier” or “pharmaceutically acceptable medium” cover any standard pharmaceutical carrier, solvent, surfactant, or medium. Standard drug carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 19th edition, 1995.

[0622] The pharmaceutical compositions disclosed herein can be prepared in the form of liquid suspensions or solutions using liquids such as oils, water, alcohols, and combinations thereof.

[0623] The pharmaceutical compositions disclosed herein can be prepared as sterile, injectable formulations, which can be aqueous or oily suspensions. The suspensions can be formulated according to techniques known in this art.

[0624] The pharmaceutical compositions disclosed herein can be administered orally in any orally acceptable dosage form, including capsules, tablets, aqueous suspensions, or solutions.

[0625] The pharmaceutical composition disclosed herein can be administered in the form of a suppository for rectal use.

[0626] The pharmaceutical compositions disclosed herein can also be applied topically, particularly when the therapeutic target includes areas or organs easily accessible by topical application, including diseases of the eye, skin, or lower intestine. Topical application to the lower intestine can be achieved with rectal suppository formulations (see above) or with suitable enema formulations. Transdermal patches may also be used. For topical application, the pharmaceutical compositions can be formulated as suitable ointments, lotions, or creams containing an active ingredient suspended or dissolved in one or more carriers.

[0627] The pharmaceutical compositions disclosed herein can also be applied ocularly and formulated as micronized suspensions in isotonic, pH-adjusted sterile saline, or preferably as solutions in isotonic, pH-adjusted sterile saline with or without preservatives (such as benzalkonium chloride). Alternatively, for ocular application, the pharmaceutical compositions can be formulated as ointments, such as petrolatum.

[0628] The pharmaceutical compositions disclosed herein can also be administered via nasal aerosol or inhalation. The compositions are prepared according to techniques well-known in the field of pharmaceutical formulation and can be prepared as a solution in saline using benzyl alcohol or other suitable preservatives, absorption enhancers to improve bioavailability, fluorocarbons, and / or other conventional solubilizers or dispersants.

[0629] Pharmaceutical compositions intended for internal administration can be sterile. This can be easily achieved by filtration, for example, through a sterile filter membrane.

[0630] Pharmaceutical compositions within the scope of this disclosure include all compositions combining the disclosed compounds with one or more pharmaceutically acceptable carriers. In one embodiment, the presence of the disclosed compound in the composition effectively achieves its intended therapeutic purpose.

[0631] The pharmaceutical compositions of this disclosure may be administered to any patient who may experience the beneficial effects of the compounds of this disclosure. Most importantly, the patient is a mammal, such as a human or a companion animal, though this disclosure is not intended to be limited thereto. In one embodiment, the patient is a human. In another embodiment, the pharmaceutical compositions of this disclosure may be administered to a patient with PARP inhibitor-resistant or refractory cancer. In another embodiment, the pharmaceutical compositions of this disclosure may be administered to a patient with PARP inhibitor-resistant or refractory BRCA1-deficient cancer. In yet another embodiment, the pharmaceutical compositions of this disclosure may be administered to a patient in combination with a PARP inhibitor.

[0632] In another embodiment, this disclosure provides a kit containing a compound of the present disclosure (or a composition containing a compound of the present disclosure) packaged in a manner conducive to practicing the methods of the present disclosure. In one embodiment, the kit includes a compound of the present disclosure (or a composition containing a compound of the present disclosure) packaged in a container, such as a resealable bottle or container, and a label affixed to or included in the kit describing a method of practicing the present disclosure using the compound or composition. In one embodiment, the compound or composition is packaged in a unit dosage form. The kit may additionally include means adapted to administer the composition according to the intended route of administration. In some embodiments, this disclosure provides a kit containing a compound of the present disclosure, or a pharmaceutically acceptable salt or solvation thereof, and instructions for administering the compound, or a pharmaceutically acceptable salt or solvation thereof, to a patient with cancer.

[0633] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0634] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having Formula I, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0635] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula II, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0636] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula III, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0637] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula IV, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0638] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula V, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0639] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula VI, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0640] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula VIa, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0641] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula VII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0642] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula VIII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0643] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula IX, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0644] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula X, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0645] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having the formula XI, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0646] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound having formula XII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0647] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the compound binds to a protein encoded by the USP1 gene.

[0648] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is used to treat cancer.

[0649] In some embodiments, this disclosure provides a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is used to manufacture an agent for treating cancer.

[0650] Example

[0651] General synthesis methods

[0652] The compounds disclosed herein are prepared according to this disclosure using methods known to those skilled in the art, or by the illustrative methods shown in the following general schemes. In any of the general schemes, a suitable protecting group may be employed in the synthesis. (See Wuts, PGM; Greene, TW, “Greene’s Protective Groups in Organic Synthesis”, 4th edition, J. Wiley & Sons, NY, 2007).

[0653] Unless otherwise instructed, all reagents are used without further purification. 1 ¹H NMR spectra were obtained at room temperature on a Bruker 300MHz instrument in DMSO-d₆ or CD₃OD. When more than one conformational isomer was detected, the chemical shift of the most abundant conformational isomer was reported. 1 Chemical shifts in ¹H NMR spectra are recorded in parts per million (ppm) relative to an internal standard in the residual solvent, based on a delta scale. Splitting patterns are designed as s (singleton), d (doublet), t (triplet), q (quartet), m (multiplex), and br (broad peak). LC-MS or preparative HPLC conditions are described below:

[0654] LCMS Method A

[0655] LCMS column: Agilent Zorbax XDB C18 4.6×50mm, 3.5μm

[0656] Mobile phase: Solvent A: Water (containing 0.1% formic acid)

[0657] Solvent B: MeOH

[0658] Flow rate: 1.0 mL / min

[0659] Runtime: 2 minutes of gradient (20%-90% B), followed by 3 minutes of 90% B.

[0660] Temperature: 30℃

[0661] LCMS Method B

[0662] LCMS column: X-Select CSH C18 3.0×50mm, 2.5μm

[0663] Mobile phase: Solvent A: Water (containing 0.05% formic acid) / MeCN (95:5)

[0664] Solvent B: 0.05% formic acid in MeCN

[0665] Flow rate: 1.2 mL / min

[0666] Runtime: 2-minute gradient (0%-98% B), hold for 1 minute, then 0% B, up to 4 minutes.

[0667] Temperature: 50℃

[0668] LCMS Method C

[0669] LCMS column: X-Select CSH C18 3.0×50mm, 2.5μm

[0670] Mobile phase: Solvent A: 5mM ammonium bicarbonate

[0671] Solvent B: MeCN

[0672] Flow rate: 1.2 mL / min

[0673] Runtime: 2-minute gradient (0%-98% B), hold for 1 minute, then 0% B, up to 4 minutes.

[0674] Temperature: 50℃

[0675] HPLC column A : Agilent SB-C18 4.6×150mm, 3.5μm

[0676] Mobile phase: Solvent A: Water (containing 0.02% TFA)

[0677] Solvent B: MeOH

[0678] Flow rate: 1.0 mL / min

[0679] Runtime: 0.5 minutes, 10% B; 9.5 minutes gradient (10%-90% B); then 10 minutes, 90% B.

[0680] Temperature: 30℃

[0681] Preparative column A : Phenomenex Luna 5u 100A, 21.2×250mm, 5μm

[0682] Mobile phase: Solvent A: Water

[0683] Solvent B: MeOH

[0684] Flow rate: 10 mL / min

[0685] Runtime: 1 minute, 20% B; 30-minute gradient (20%-80% B); then 10 minutes, 90% B.

[0686] Temperature: Ambient temperature

[0687] Preparative column B : X-Select C18, 30×250mm, 5μm

[0688] Mobile phase: Solvent A: 10 mm formic acid in water

[0689] Solvent B: MeCN

[0690] Flow rate: 30 mL / min

[0691] Running time: 50 min, at the same concentration of 5% A: 95% B

[0692] Temperature: Ambient temperature

[0693] Preparative column C : Nucleodur C18, 30×250mm, 5μm

[0694] Mobile phase: Solvent A: 10 mM ammonium bicarbonate in water

[0695] Solvent B: MeCN

[0696] Flow rate: 30 mL / min

[0697] Running time: 50 min, at the same concentration of 5% A: 95% B

[0698] Temperature: Ambient temperature

[0699] Preparative Column D :X-Select C18, 30×250mm, 5μm

[0700] Mobile phase: Solvent A: 10 mM ammonium bicarbonate in water

[0701] Solvent B: MeCN

[0702] Flow rate: 30 mL / min

[0703] Running time: 50 min, at the same concentration of 5% A: 95% B

[0704] Temperature: Ambient temperature

[0705] Preparative Column E : Nucleodur C18, 30×250mm, 5μm

[0706] Mobile phase: Solvent A: 10 mM NH3 in water

[0707] Solvent B: MeCN

[0708] Flow rate: 30 mL / min

[0709] Running time: 50 min, at the same concentration of 5% A: 95% B

[0710] Temperature: Ambient temperature

[0711] SFC method : Green Sep ES Diol, 250×4.6mm, 5μm

[0712] Mobile phase: Solvent A: CO2

[0713] Solvent B: MeCN

[0714] Flow rate: 80 mL / min; Diluent: MeCN

[0715] Running time: 10-20% B, 3 minutes; 20-25% B, 9 minutes; 25-30% B, 3 minutes; 30-50% B, 6 minutes Clock; 50%B 50 minutes

[0716] Chiral Method A : DIACEL CHIRAL PAK-IC (250×4.6mm, 5μm); Wavelength: 260nm

[0717] Mobile phase solvent A: CO2

[0718] Solvent B: MeOH + DCM (80:20)

[0719] Flow rate: 3 mL / min

[0720] Run time: 5 minutes, 25-50% B; 50% B, maintain for 4 minutes; 10 minutes, 50-25% B; 25% B, maintain 2 minutes

[0721] Chiral method B : DIACEL CHIRAL PAK-IG (250×4.6mm, 5μm); Wavelength: 280nm

[0722] Mobile phase solvent A: CO2

[0723] Solvent B: MeOH + 0.1% NH3

[0724] Flow rate: 3 mL / min

[0725] Run time: 5 minutes, 10-40% B; 40% B, maintain for 4 minutes; 10 minutes, 40-10% B; 10% B, maintain 2 minutes

[0726] Use the following abbreviations in the main text:

[0727] PE = Petroleum Ether

[0728] EA or EtOAc = Ethyl acetate

[0729] DMSO = dimethyl sulfoxide

[0730] DMF = N,N-dimethylformamide

[0731] DMA = N,N-dimethylacetamide

[0732] MeOH = methanol

[0733] MTBE = Methyl tert-butyl ether

[0734] DCM = dichloromethane

[0735] TEA = Triethylamine

[0736] DIPEA = diisopropylethylamine

[0737] TFA = Trifluoroacetic acid

[0738] MeCN or ACN = Acetonitrile

[0739] TLC = Thin-layer chromatography

[0740] (BPin)2 = Bis(pinacol)diboron

[0741] HFIP = 1,1,1,3,3,3-hexafluoropropane-2-ol

[0742] DIBAL-H = Diisobutylaluminum hydride

[0743] MeI = methyl iodide

[0744] hex or n-hex = n-hexane

[0745] DCE = 1,2-Dichloroethane

[0746] TBSCl = tert-butyldimethylchlorosilane

[0747] Tf₂O = Trifluoromethanesulfonic anhydride

[0748] n-BuLi = n-Butyllithium

[0749] DMAP = 4-Dimethylaminopyridine

[0750] KOAc = Potassium acetate

[0751] NaOAc = Sodium acetate

[0752] TFAA = Trifluoroacetic anhydride

[0753] m-CPBA = m-chloroperoxybenzoic acid

[0754] DME = 1,2-Dimethoxyethane

[0755] TPP = Triphenylphosphine

[0756] DTAD = di-tert-butyl azodicarbonate

[0757] DEAD = Diethyl azodicarbonate

[0758] DIAD = diisopropyl azodicarbonate

[0759] PS-TPP = Polystyrene-supported triphenylphosphine

[0760] NBS = N-bromosuccinimide

[0761] Preparation of general intermediate I-1:

[0762]

[0763] Step 1: Synthesis of methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate:

[0764]

[0765] To a mixture of 3,3-dibromo-1,1,1-trifluoropropane-2-one (3.1 g, 11.49 mmol) in water (6 mL), NaOAc (951.8 mg, 11.60 mmol) was added. The mixture was heated at 100 °C for 1 hour, then cooled to room temperature. Next, a solution of methyl 4-formylbenzoate (1.7 g, 10.34 mmol) in MeOH (47 mL) and NH4OH (11 mL) was added to the reaction mixture at room temperature. After 40 minutes, the reaction mixture was heated to 100 °C and stirred for 2 hours. The mixture was quenched with water (50 mL) and then extracted with EA (100 mL × 2). The organic layer was dried over Na2SO4 (30 g), filtered, and concentrated. The concentrated residue was purified by silica gel chromatography (PE:EA = 20:1 to 5:1) to give 1.9 g of the title compound. LC-MS (Method A) (ESI+): m / z 271 (M+H) + ; 1H-NMR (300MHz, CD3OD) δ8.12 (d, J = 8.7 Hz, 2H), 8.01 (d, J = 8.7 Hz, 2H), 7.72 (s, 1H), 3.93 (s, 3H).

[0766] Step 2: Synthesis of methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate:

[0767]

[0768] At 0 °C, NaH (60% dispersion) (338 mg, 8.44 mmol) was added fractionally over 5 minutes to a solution of methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (1.9 g, 7.03 mmol) in THF (50 mL). After stirring the mixture at 0 °C for 30 minutes, MeI (1.2 g, 8.44 mmol) was added dropwise to the reaction mixture over 2 minutes. The reaction mixture was allowed to warm to room temperature over 3 hours, followed by quenching with ice water (30 g). The resulting solution was extracted with EA (50 mL × 3), and the combined organic layers were dried over Na₂SO₄ (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 20:1 to 5:1) to give 1.4 g of the title compound. LC-MS (Method A) (ESI+): m / z 285 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ8.17(d,J=8.4Hz,2H),7.80(d,J=8.4Hz,2H),7.75(s,1H),3.94(s,3H),3.83(s,3H).

[0769] Step 3: Synthesis of (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol:

[0770]

[0771] At 0 °C, DIBAL-H (24 mL, 24.6 mmol, 1 M toluene solution) was added via syringe to a solution of methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (1.4 g, 4.93 mmol) in THF (40 mL). After addition, the mixture was stirred at 0 °C for 20 min, followed by a 30 min warming to room temperature. TLC analysis indicated the reaction was complete. The mixture was quenched with ice water (10 mL) and the pH was adjusted to 5 with 1 N HCl solution. The mixture was extracted with EA (50 mL × 2), and the combined organic layers were washed with brine (30 mL), dried over Na₂SO₄ (30 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 20:1 to 5:1) to give 1.2 g of the title compound. LC-MS (Method A) (ESI+): m / z 257 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ7.69 (s, 1H), 7.63 (d, J = 8.1 Hz, 2H), 7.52 (d, J = 8.1 Hz, 2H), 4.69 (s, 2H), 3.78 (s, 3H).

[0772] Step 4: Synthesis of 2-(4-(chloromethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium:

[0773]

[0774] SOCl2 (1.7 g, 14.1 mmol) was added in a single batch to a mixture of (1.2 g, 4.68 mmol) of (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol in a DCE (40 mL). After addition, the mixture was stirred at 50 °C for 20 min, then concentrated to dryness to give 1.28 g of the crude title compound. LC-MS (Method A) (ESI+): m / z 275, 277 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ8.04 (s, 1H), 7.74 (d, J = 8.4Hz, 2H), 7.68 (d, J = 8.4Hz, 2H), 4.75 (s, 2H), 3.87 (s, 3H).

[0775] Step 5: Synthesis of 6-chloro-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-1):

[0776]

[0777] To a solution of 2-(4-(chloromethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium (1 g, 3.64 mmol) in DMF (25 mL), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (675 mg, 4.37 mmol) and K₂CO₃ (1.26 g, 9.10 mmol) were added. The resulting mixture was stirred at 35 °C for 20 h, quenched with water (50 mL), and extracted with EA (100 mL × 2). The combined organic phases were washed with water and brine, dried over Na₂SO₄ (50 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 20:1 to 10:1) to give 580 mg of the title compound containing a small amount of regioisomers. LC-MS (Method A) (ESI+): m / z 393,395 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.06 (s, 1H), 8.18 (s, 1H), 7.61 (d, J = 8.1Hz, 2H), 7.47 (d, J = 8.1Hz, 2H), 7.30 (s, 1H), 5.69 (s, 2H), 3.74 (s, 3H).

[0778] The following intermediates are prepared from appropriate heterocyclic compounds and alkylating agents according to the method for preparing I-1.

[0779]

[0780]

[0781] Preparation of general intermediate I-8:

[0782]

[0783] Step 1: Synthesis of 3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde:

[0784]

[0785] The solution of 2-chloro-1-methyl-4-(trifluoromethyl)-1H-imidazolium (1.00 g, 5.40 mmol), (2-fluoro-4-formylphenyl)boronic acid (1.00 g, 6.50 mmol), and K3PO4 (3.40 g, 16.30 mmol) in dioxane (20 mL) and H2O (5 mL) was degassed with argon for 5 min. Xphos-Pd-G2 (0.42 g, 0.54 mmol) and X-Phos (1.03 g, 2.10 mmol) were added sequentially to the resulting reaction mixture, and the reaction mixture was heated at 100 °C for 16 h in a sealed tube. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with water and extracted with EA. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound obtained was purified by silica gel chromatography using 0–30% EA in hexane to give 1.40 g of the title compound. LC-MS (Method B) (ESI+): m / z 272.90 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ10.09(s,1H),8.08(s,1H),7.83-7.94(m,3H),3.66(s,3H).

[0786] Step 2: Synthesis of (3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol:

[0787]

[0788] To an ice-cooled solution of 3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde (1.40 g, 5.14 mmol) in MeOH (30 mL), NaBH4 (0.293 g, 7.70 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude reaction mixture was dissolved in water and extracted with EA. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1.40 g of the title compound, which was used directly without further purification. LC-MS (Method B) (ESI+): m / z 274.85 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ7.99 (s, 1H), 7.54 (t, J = 7.8 Hz, 1H), 7.28-7.34 (m, 2H), 5.46 (t, J = 5.6 Hz, 1H), 4.60 (d, J = 5.4 Hz, 2H), 3.59 (s, 3H).

[0789] Step 3: Synthesis of 2-(4-(chloromethyl)-2-fluorophenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium:

[0790]

[0791] SOCl2 (1.80 g, 15.0 mmol) was added to an ice-cooled solution of (3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-methanol (1.40 g, 5.10 mmol) in DCM (15 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with saturated NaHCO3 solution and extracted with DCM. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1.42 g of the title compound, which was used directly without further purification. LC-MS (Method B) (ESI+): m / z 292.85 (M+H) + .

[0792] Step 4: Synthesis of 6-chloro-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-8):

[0793]

[0794] A solution of 2-(4-(chloromethyl)-2-fluorophenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium (1.41 g, 4.80 mmol), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine 5 (0.65 g, 4.20 mmol), and K₂CO₃ (1.45 g, 10.50 mmol) in DMF (12 mL) was stirred for 16 hours at 35 °C. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water and extracted with EA. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–15% EA in hexane to give 0.70 g of the title compound. LC-MS (Method B) (ESI+): m / z 410.85 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ9.33(s,1H),8.54(s,1H),7.99(s,1H),7.57(t,J=7.6 Hz, 1H), 7.31 (d, J = 10.8 Hz, 1H), 7.19 (d, J = 7.8 Hz, 1H), 5.76 (s, 2H), 3.58 (s, 3H).

[0795] The following intermediates are prepared from appropriate aldehydes and alkylating agents according to the procedure for intermediate I-8.

[0796]

[0797]

[0798] Preparation of general intermediate I-15:

[0799]

[0800] Step 1: Synthesis of 1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazolium:

[0801]

[0802] Sodium hydride (60% dispersion, 1.294 g, 32.35 mmol) was added to a stirred solution of 4-(trifluoromethyl)-1H-imidazole (4.0 g, 29.4 mmol) in anhydrous THF (100 mL) at 0 °C. The resulting mixture was stirred for 15 min, followed by the addition of iodomethane-d3 (4.26 g, 29.4 mmol) at 0 °C and stirring for 30 min. The reaction mixture was then heated to room temperature and stirred for 4 h. After the reaction was complete (monitored by TLC), the reaction mixture was quenched with saturated ammonium chloride solution (50 mL) and extracted with EA (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and dried under vacuum. The crude product was purified by silica gel chromatography (50–100% EA in n-hexane) to give 3.0 g of the title compound. LC-MS (Method B) (ESI+): m / z 154.45 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ7.48 (s, 1H), 7.27 (s, 1H).

[0803] Step 2: Synthesis of 2-chloro-1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazolium:

[0804]

[0805] At -78 °C, a solution of 2.5 M n-butyllithium in n-hexane (7 mL, 17.64 mmol) was added to a stirred solution of 1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazole (2.70 g, 17.6 mmol) in THF (50 mL). The resulting mixture was stirred at -78 °C for 30 min, followed by treatment with hexachloroethane (2.08 g, 8.82 mmol). After 1 hour, the reaction mixture was warmed to room temperature and stirred for another 3 hours. After the reaction was complete (monitored by TLC), the reaction mixture was quenched with saturated ammonium chloride solution (25 mL) and extracted with EA (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (30–100% EA in n-hexane) to give 1.30 g of the title compound. LC-MS (Method B) (ESI+): m / z 188.30 (M+H) + .

[0806] Step 3: Synthesis of 4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde:

[0807]

[0808] X-Phos-Pd-G2 catalyst (0.294 g, 0.374 mmol) was added to a solution of 2-chloro-1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazolium (1.40 g, 7.49 mmol) in 1,4-dioxane (60 mL). The resulting mixture was purged with argon for 10 min, followed by the addition of solid K3PO4 (4.76 g, 22.5 mmol) and (4-formylphenyl)boronic acid (2.24 g, 14.97 mmol), and then purged with argon for another 5 min. The reaction mixture was then heated at 90 °C for 16 h. After the reaction was complete (monitored by TLC), the mixture was diluted with water (60 mL) and EA (60 mL). The organic layer was separated, and the aqueous layer was extracted with EA (2 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography (0-30% EA in n-hexane) to give 1.62 g of the title compound. LC-MS (Method B) (ESI+): m / z 258.00 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ10.09 (s, 1H), 8.00 (d, J = 7.88Hz, 2H), 7.86 (d, J = 7.88Hz, 2H), 7.37 (s, 1H).

[0809] Step 4: Synthesis of (4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol:

[0810]

[0811] At 0 °C, NaBH4 (0.118 g, 3.11 mmol) was added to a stirred solution of 1.60 g (6.23 mmol) of 4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde in 50 mL of methanol. After the reaction was complete (monitored by TLC), the mixture was quenched with ice water (30 mL) and extracted with 10% methanol in DCM solution (4 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (0–60% EA / n-hexane) to give 1.30 g of the title compound. LC-MS (Method B) (ESI+): m / z 259.80 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ7.60 (d, J = 8.31 Hz, 2H), 7.45 (d, J = 7.83 Hz, 2H), 7.31 (s, 1H), 4.76 (s, 2H).

[0812] Step 5: Synthesis of 6-chloro-1-(4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-15):

[0813]

[0814] Triphenylphosphine (0.657 g, 2.509 mmol) was added to a mixture of (4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (0.500 g, 1.930 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidine 7 (0.335 g, 2.181 mmol) in anhydrous THF (5 mL) at 0 °C. The resulting mixture was stirred for 5 min, followed by dropwise treatment with diethyl azodicarbonate (0.434 g, 2.509 mmol). The mixture was then stirred at 0 °C for 2 h. After the reaction was complete (monitored by TLC), the mixture was diluted with water (30 mL) and extracted with EA (3 × 30 mL). The combined organic layers were washed with brine (3 × 30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude compound was purified by silica gel chromatography (0-3% methanol in DCM) to give 0.400 g of the title compound. LC-MS (Method B) (ESI+): m / z 396.00 (M+H)+ ; 1 H-NMR (400MHz, CDCl3) δ9.05 (s, 1H), 8.18 (s, 1H), 7.61 (d, J = 8.31Hz, 2H), 7.47 (d, J = 7.82Hz, 2H), 7.30 (s, 1H), 5.69 (s, 2H).

[0815] Preparation of general intermediate I-16:

[0816]

[0817] Step 1: Synthesis of methyl 4-(2-bromoacetyl)benzoate:

[0818]

[0819] Br2 (1.00 mL, 19.6 mmol) in 10 mL of AcOH was added dropwise to a stirred solution of methyl 4-acetylbenzoate (5.00 g, 28.0 mmol) in AcOH (40 mL), and the reaction mixture was stirred at room temperature for 6 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was poured into ice water (100 mL), filtered, and the resulting solid was dried. The crude compound was purified by silica gel chromatography using 5–7% EA in hexane to give 3.50 g of the title compound. LC-MS (Method C) (ESI+): m / z 278.95 (M+Na) + ; 1 H-NMR (400MHz, DMSO-d6) δ8.04-8.15(m,4H),5.00(s,2H),3.89(s,3H).

[0820] Step 2: Synthesis of methyl 4-(2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate:

[0821]

[0822] Methyl 4-(2-bromoacetyl)benzoate (2.00 g, 7.80 mmol) was added to a stirred solution of 2,2,2-trifluoroacetamidine (4.37 g, 39.0 mmol) in THF (30 mL), and the reaction mixture was stirred at room temperature for 5 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with EA (20 mL) and washed with brine (2 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 10–15% EA in hexane to give 0.81 g of the title compound. LC-MS (Method B) (ESI+): m / z 270.90 (M+H)+ ; 1 H-NMR (400MHz, DMSO-d6) δ13.89(s,1H),8.14(s,1H),7.98(s,4H),3.86(s,3H).

[0823] Step 3: Synthesis of methyl 4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate:

[0824]

[0825] At 0 °C, NaH (60% dispersion in mineral oil, 0.178 g, 4.40 mmol) was added fractionally to a stirred solution of methyl 4-(2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate (0.80 g, 2.9 mmol) in THF (15 mL), and the reaction mixture was stirred for 15 min at the same temperature. Iodomethane (0.624 g, 4.40 mmol) was added to the resulting reaction mixture, and the mixture was stirred for 16 h at room temperature. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with ice water (50 mL) and extracted with EA (2 × 30 mL). The combined organic layers were washed with brine (2 × 30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 0.80 g of the title compound. LC-MS (Method B) (ESI+): m / z 284.95 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ8.12-8.17(m,1H),7.96-8.02(m,2H),7.88-7.94(m,2H),3.86(s,6H).

[0826] Step 4: Synthesis of (4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)phenyl)methanol:

[0827]

[0828] At 0 °C, methyl 4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate (0.80 g, 2.8 mmol) was added dropwise to a stirred solution of methyl 4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate in 10 mL of THF, and the reaction mixture was stirred at room temperature for 1 hour. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with 2N HCl (20 mL) and extracted with EA (2 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 0.65 g of the title compound, which was used in subsequent reactions without further purification. LC-MS (Method B) (ESI+): m / z 256.85 (M+H) + .

[0829] Step 5: Synthesis of 4-(4-(chloromethyl)phenyl)-1-methyl-2-(trifluoromethyl)-1H-imidazolium:

[0830]

[0831] SOCl2 (0.27 mL, 3.80 mmol) was added to a stirred solution of (0.65 g, 2.5 mmol) of (4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)phenyl)methanol in 10 mL of DCM at 0 °C, and the reaction mixture was stirred at room temperature for 1 hour. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with 50 mL of DCM and washed with saturated NaHCO3 solution (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 0.65 g of the title compound, which was used in subsequent reactions without further purification. LC-MS (Method B) (ESI+): m / z 274.90 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ 8.00 (s, 1H), 7.77 (d, J = 7.8Hz, 2H), 7.46 (d, J = 7.8Hz, 2H), 4.78 (s, 2H), 3.84 (s, 3H).

[0832] Step 6: Synthesis of 6-chloro-1-(4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-16):

[0833]

[0834] K₂CO₃ (0.517 g, 3.70 mmol) was added in a single addition to a stirred solution of 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.40 g, 2.5 mmol) in DMF (10 mL). After stirring at room temperature for 10 min, 4-(4-(chloromethyl)phenyl)-1-methyl-2-(trifluoromethyl)-1H-imidazolium (0.638 g, 2.30 mmol) was added, and the reaction mixture was stirred at room temperature for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was poured onto ice water (50 mL) and extracted with EA (2 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–50% EA / hexane to give 0.15 g of the title compound. LC-MS (Method B) (ESI+): m / z 392.95 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ9.30(s,1H),8.49(s,1H),7.94(s,1H),7.72(d,J=8.3Hz,2H),7.29(d,J=8.3Hz,2H),5.63(s,2H),3.82(s,3H).

[0835] The following intermediates are prepared from appropriate reagents according to the procedure for intermediate I-16.

[0836]

[0837] Preparation of general intermediate I-18:

[0838]

[0839] Step 1: Synthesis of ethyl 4-acetyl-3-methoxybenzoate:

[0840]

[0841] Triethylamine (3.67 mL, 26.2 mmol) was added to a stirred solution of 1-(4-bromo-2-methoxyphenyl)ethyl-1-one (3.00 g, 13.1 mmol) in ethanol (60 mL), and the mixture was degassed with argon for 10 min. At room temperature, PdCl2 (dppf) (0.957 g, 0.131 mmol) was added to the reaction mixture, and the resulting mixture was heated at 80 °C for 3 h under carbon monoxide (20 psi pressure). The reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–5% EA in hexane as eluent to give the title compound (2.50 g). LC-MS (Method B) (ESI+): m / z 223.00 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ7.72-7.76(m,1H),7.64-7.70(m,2H),4.42(dq,J=3.99,7.15Hz,2H),4.00(br s,1H),3.99(s,2H),2.65(s,1H),2.64(s,2H),1.43(dt,J=3.99,6.98Hz,3H).

[0842] Step 2: Synthesis of ethyl 4-(2,2-dibromoacetyl)-3-methoxybenzoate:

[0843]

[0844] Bromine (0.16 mL, 3.153 mmol) was added to a stirred solution of ethyl 4-acetyl-3-methoxybenzoate 2 (1.00 g, 4.50 mmol) in acetic acid (10 mL) at room temperature. The resulting mixture was stirred for 3 hours, and the reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude residue was diluted with water (30 mL) and EA (50 mL), and then neutralized with saturated Na₂CO₃ solution. The organic layer was washed with (30 mL) brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the title compound (1.20 g). 1 H-NMR (400MHz, CDCl3) δ7.83(d,J=7.98Hz,1H),7.69(d,J=8.48Hz,1H),7.67(s ,1H),4.59(s,1H),4.41(q,J=7.15Hz,2H),4.01(s,3H),1.42(t,J=6.98Hz,3H).

[0845] Step 3: Synthesis of ethyl 4-(2-bromoacetyl)-3-methoxybenzoate:

[0846]

[0847] At room temperature, triethylamine (0.44 mL, 3.17 mmol) and diethyl phosphite (0.433 g, 3.166 mmol) were added to a stirred solution of ethyl 4-(2,2-dibromoacetyl)-3-methoxybenzoate 3 (1.20 g, 3.17 mmol) in THF (15 mL). The resulting mixture was stirred for 3 hours, and the reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with EA (30 mL) and washed with brine (3 × 10 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 10–15% EA in hexane as the eluent to give the title compound (0.900 g). LC-MS (Method B) (ESI+): m / z 300.80 (M) + ; 1 H-NMR (400MHz, CDCl3) δ7.84(d,J=7.98Hz,1H),7.69(d,J=7.98Hz,1H),7.67(s ,1H),4.60(s,2H),4.41(q,J=7.15Hz,2H),4.02(s,3H),1.42(t,J=6.98Hz,3H).

[0848] Step 4: Synthesis of ethyl 3-methoxy-4-(2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate:

[0849]

[0850] At room temperature, 2,2,2-trifluoroacetamidine (1.68 g, 15.0 mmol) was added to a stirred solution of ethyl 4-(2-bromoacetyl)-3-methoxybenzoate 4 (0.900 g, 3.00 mmol) in THF (20 mL). The reaction mixture was then heated at 60 °C for 16 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with EA (50 mL) and washed with saturated NaHCO3 solution (2 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography using 20–30% EA in hexane as eluent to give the title compound (0.300 g). LC-MS (Method B) (ESI+): m / z 315.10 (M+H) + ; 1H-NMR(400MHz,DMSO-d6)δ13.82(brs,1H),8.17(d,J=7.48Hz,1H),7.92(br s, 1H), 7.64 (d, J = 7.98Hz, 1H), 7.59 (s, 1H), 4.33 (q, J = 7.31Hz, 2H), 4.00 (s, 3H), 1.34 (t, J = 6.98Hz, 3H).

[0851] Step 5: Synthesis of ethyl 3-methoxy-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate:

[0852]

[0853] At 0 °C, a dispersion of 60% sodium hydride in oil (0.057 g, 1.43 mmol) and methyl iodide (0.089 mL, 1.43 mmol) were added to a stirred solution of ethyl 3-methoxy-4-(2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate 5 (0.300 g, 0.955 mmol) in DMF (10 mL). The reaction mixture was heated to room temperature and stirred for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was poured into ice-cold water (50 mL) and extracted with EA (2 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–15% EA in hexane as eluent to give the title compound (0.240 g). LC-MS (Method B) (ESI+); m / z 328.90 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ8.14(d,J=8.48Hz,1H),8.04(s,1H),7.64(dd,J=1.50,7.98Hz,1 H), 7.59 (s, 1H), 4.33 (q, J = 7.15Hz, 2H), 4.00 (s, 3H), 3.87 (s, 3H), 1.33 (t, J = 7.23Hz, 3H).

[0854] Step 6: Synthesis of (3-methoxy-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)phenyl)methanol:

[0855]

[0856] At 0 °C, DIBAL-H (1 M solution in toluene, 1.46 mL, 1.46 mmol) was added to a stirred solution of ethyl 3-methoxy-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate 6 (0.320 g, 0.975 mmol) in anhydrous THF (10 mL). The reaction mixture was heated to room temperature and then stirred for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with 1 N HCl (20 mL) and extracted with EA (3 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude title compound (0.260 g). LC-MS (Method B) (ESI+): m / z 287.30 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ7.94(d,J=7.83Hz,1H),7.84(s,1H),7.05(s,1H),6.95(d,J= 7.82Hz, 1H), 5.21 (t, J = 4.89Hz, 1H), 4.51 (d, J = 4.89Hz, 2H), 3.92 (s, 3H), 3.84 (s, 3H).

[0857] Step 7: Synthesis of 4-(4-(chloromethyl)-2-methoxyphenyl)-1-methyl-2-(trifluoromethyl)-1H-imidazolium:

[0858]

[0859] At 0 °C, thionyl chloride (0.065 mL, 0.91 mmol) was added to a stirred solution of (3-methoxy-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)phenyl)methanol 7 (0.260 g, 0.909 mmol) in DCM (10 mL). The mixture was stirred for 1 hour, and the reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with DCM (20 mL) and washed with saturated Na2CO3 solution (3 × 20 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude title compound (0.260 g). LC-MS (Method B) (ESI+): m / z 304.80 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ8.00(d,J=7.98Hz,1H),7.90(s,1H),7.18(s,1H),7.08(d,J=7.98Hz,1H),4.78(s,2H),3.94(s,3H),3.85(s,3H).

[0860] Step 8: Synthesis of 6-chloro-1-(3-methoxy-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-18):

[0861]

[0862] Potassium carbonate (0.170 g, 1.23 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.126 g, 0.822 mmol) were added to a stirred solution of 4-(4-(chloromethyl)-2-methoxyphenyl)-1-methyl-2-(trifluoromethyl)-1H-imidazolium 8 (0.250 g, 0.822 mmol) in DMF (5 mL) at room temperature. The resulting mixture was stirred for 16 hours, and the reaction progress was monitored by TLC. After completion, the reaction mixture was poured into ice-cold water (30 mL) and extracted with EA (2 × 30 mL). The combined organic layers were washed with (2 × 30 mL) brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 30–50% EA in hexane as the eluent to give the title compound (0.150 g). LC-MS (Method B) (ESI+): m / z 423.00 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ9.31 (s, 1H), 8.50 (s, 1H), 7.92 (d, J = 7.98Hz, 1H), 7.86 ( s,1H),7.11(s,1H),6.82(d,J=7.98Hz,1H),5.64(s,2H),3.89(s,3H),3.83(s,3H).

[0863] Preparation of universal intermediate I-19:

[0864]

[0865] Step 1: Synthesis of methyl 4-(5-methyl-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[0866]

[0867] TEA (994 mg, 9.84 mmol) was added dropwise to a solution of methyl 4-hydrazinobenzoate hydrochloride (1.0 g, 4.92 mmol) and 1,1,1-trifluoropentane-2,4-dione (758 mg, 4.92 mmol) in HFIP (5 mL) cooled to 0 °C over 5 minutes. After addition, the mixture was stirred at room temperature for 1 hour, quenched with water (20 mL), and extracted with DCM (100 mL × 3). The combined organic layers were dried over anhydrous Na₂SO₄ (30 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (EA:n-Hex = 1:20) to give 1.1 g of the title compound. LC-MS (Method A) (ESI+): m / z 285 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ8.18 (d, J = 8.7 Hz, 2H), 7.57 (d, J = 8.7 Hz, 2H), 6.50 (s, 1H), 3.96 (s, 3H), 2.41 (s, 3H).

[0868] Step 2: Synthesis of (4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol:

[0869]

[0870] At 0 °C, methyl 4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (1.0 g, 3.52 mmol) in anhydrous THF (20 mL) was added to DIBAL-H (10.5 mL, 10.56 mmol, 1 M toluene solution) over 10 minutes. After addition, the reaction mixture was allowed to warm to room temperature. TLC analysis indicated the reaction was complete. The reaction was quenched with saturated NH4Cl solution (20 mL) and extracted with EA (100 mL × 3). The combined organic layers were dried over anhydrous Na2SO4 (50 g), filtered, and concentrated. The crude product was purified by silica gel chromatography (PE:EA = 20:1 to 10:1) to give 1.1 g of the title compound. 1 H-NMR (300MHz, CDCl3) δ7.40-7.49(m,4H),6.46(s,1H),4.76(s,2H),2.34(s,3H),2.17(br s,1H).

[0871] Step 3: Synthesis of 1-(4-(chloromethyl)phenyl)-5-methyl-3-(trifluoromethyl)-1H-pyrazole:

[0872]

[0873] SOCl2 (1.14 g, 9.61 mmol) was added in a single addition to a solution of (0.82 g, 3.20 mmol) of (4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol in DCE (20 mL). After stirring at 50 °C for 30 min, the mixture was cooled to 0 °C and quenched with water (10 mL). The resulting solution was neutralized with saturated NaHCO3 (aqueous solution) and extracted with DCM (3 × 100 mL). The combined organic layers were dried over anhydrous Na2SO4 (50 g), filtered, and concentrated to dryness to give 480 mg of crude product, which was used directly in the next step. LC-MS (Method A) (ESI+): m / z 275 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.53 (d, J = 8.7Hz, 2H), 7.46 (d, J = 8.7Hz, 2H), 6.47 (s, 1H), 4.64 (s, 2H), 2.37 (s, 3H).

[0874] Step 4: Synthesis of 6-chloro-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-19):

[0875]

[0876] To a solution of 1-(4-(chloromethyl)phenyl)-5-methyl-3-(trifluoromethyl)-1H-pyrazole (0.48 g, 1.75 mmol) in DMF (8 mL), 6-chloro-1H-pyrazole[3,4-d]pyrimidine (0.27 g, 1.75 mmol) and K₂CO₃ (0.60 g, 4.38 mmol) were added. The reaction was stirred at room temperature for 2 h, followed by quenching with water (20 mL) and extraction with EA (50 mL × 3). The combined organic layers were dried over anhydrous Na₂SO₄ (30 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 20:1 to 10:1) to give 370 mg of the title product containing trace regioisomers. LC-MS (Method A) (ESI+): m / z 393, 395 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.06 (s, 1H), 8.19 (s, 1H), 7.51 (d, J = 8.7Hz, 2H), 7.42 (d, J = 8.7Hz, 2H), 6.44 (s, 1H), 5.70 (s, 2H), 2.33 (s, 3H).

[0877] The following general intermediates are prepared from appropriate reagents according to the procedure in I-19:

[0878]

[0879] Preparation of general intermediate I-22:

[0880]

[0881] Step 1: Synthesis of 6-chloro-4-methoxy-1H-pyrazolo[3,4-d]pyrimidine:

[0882]

[0883] Potassium carbonate (240 mg, 1.74 mmol) was added to a solution of 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine 1 (330 mg, 1.74 mmol) in methanol (10 mL). The mixture was stirred at room temperature for 24 hours, then quenched with saturated ammonium chloride (30 mL) and extracted with EA (20 mL × 3). The combined organic layers were dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: EA / n-Hex = 1 / 10) to give 120 mg of the title compound. LC-MS (Method A) (ESI+): m / z 185 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ10.69(br s,1H),8.09(s,1H),4.19(s,3H).

[0884] Step 2: Synthesis of 6-chloro-4-methoxy-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-22):

[0885]

[0886] The compound was synthesized according to the procedure for preparing the general intermediate I-19. LC-MS (Method A) (ESI+): m / z 423 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ8.05(s,1H),7.45(m,4H),6.44(s,1H),5.63(s,2H),4.15(s,3H),2.31(s,3H).

[0887] The following general intermediates are prepared from appropriate reagents according to the procedure in I-22:

[0888]

[0889] Preparation of general intermediate I-23:

[0890]

[0891] Step 1: Synthesis of methyl 4-hydrazino-3-methoxybenzoate:

[0892]

[0893] At -10 °C, an aqueous solution of NaNO2 (1.99 g, 29.0 mmol) in water (5 mL) was added dropwise to a stirred solution of methyl 4-amino-3-methoxybenzoate 1 (5.0 g, 27.6 mmol) in concentrated HCl (55 mL). A solution of SnCl2 (26.0 g, 138 mmol) in concentrated HCl (35 mL) was added dropwise to the resulting reaction mixture, and the mixture was stirred for 2 hours. The reaction progress was monitored by TLC. After completion, the resulting solid was filtered and dried under reduced pressure. The crude compound was washed with diethyl ether (50 mL), filtered, and dried under reduced pressure to give the crude title compound (8.0 g), which was used in the next step without purification. 1 H-NMR (400MHz, DMSO-d6) δ10.43(br s,2H),7.95-8.40(m,1H),7.57(d,J=8.31Hz,1H),7.44(s,1H),7.09(d,J=8.31Hz,1H),3.88(s,3H).

[0894] Step 2: Synthesis of methyl 3-methoxy-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[0895]

[0896] Methyl 4-hydrazino-3-methoxybenzoate 2 (2.0 g, 12.98 mmol) and 1,1,1-trifluoropentane-2,4-dione (6.09 g, 25.97 mmol) in ethanol (20 mL) were heated at 90 °C with stirring for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude compound was purified by size chromatography using 0-30% EA in hexane to give the title compound (2.5 g). LC-MS (Method B) (ESI+): m / z 315.05 (M+H) + .

[0897] Step 3: Synthesis of (3-methoxy-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol:

[0898]

[0899] At 0 °C, DIBAL-H (16 mL, 15.9 mmol, 1.0 M in THF) was added dropwise to a stirred solution of methyl 3-methoxy-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate 3 (2.5 g, 7.96 mmol) in THF (40 mL). The mixture was stirred at 0 °C for 2 h, and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with 1 N HCl (4 mL) at 0 °C and extracted with EA (100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude product 4 (1.5 g). The crude product was used as is without purification. LC-MS (Method B) (ESI+): m / z 287.45 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ7.31(d,J=7.98Hz,1H),7.21(s,1H),7.05(d,J=7.98Hz,1H),6.67(s,1H),5.40(br s,1H),4.59(br s,2H),3.79(s,3H),2.09(s,3H).

[0900] Step 4: Synthesis of 6-chloro-1-(3-methoxy-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-23):

[0901]

[0902] To an ice-cold solution of (3-methoxy-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol 4 (1.0 g, 3.494 mmol) in THF (10 mL), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.486 g, 3.144 mmol), DTAD (1.20 g, 5.240 mmol), and TPP (2.747 g, 10.48 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water (30 mL) and extracted with EA (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by size chromatography using 0–40% EA in hexane as eluent to give the title compound (1.0 g). LC-MS (Method B) (ESI+): m / z 423 (M+H) + .

[0903] Preparation of general intermediate I-24:

[0904]

[0905] Step 1: Synthesis of (4-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol:

[0906]

[0907] A solution of 3,3-dibromo-1,1,1-trifluoropropane-2-one (1.0 g, 7.3 mmol) and NaOAc (533.5 mg, 8.1 mmol) in H₂O (7.3 mL) was stirred for 1 hour at 100 °C. The reaction mixture was then cooled to 0 °C and a solution of 4-(hydroxymethyl)benzaldehyde (2.18 g, 8.08 mmol) and NH₄OH (9.3 mL) in MeOH (37 mL) was added over 10 minutes. After the addition, the reaction mixture was stirred overnight at ambient temperature. The resulting mixture was concentrated under reduced pressure to remove most of the solvent. Water (20 mL) was added to the concentrated residue, and the mixture was extracted with EA (100 mL × 3). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄ (50 g), filtered, and concentrated to dryness under vacuum. The resulting crude product formed a slurry in DCM (30 mL), which was collected by filtration and dried under vacuum to give 1.1 g of the title compound. 1 H-NMR (300MHz, CD3OD) δ7.88 (d, J = 8.4 Hz, 2H), 7.64 (s, 1H), 7.48 (d, J = 8.4 Hz, 2H), 4.66 (s, 2H).

[0908] Step 2: Synthesis of 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde:

[0909]

[0910] At 0 °C, Dess-Martin reagent (2.6 g, 6.2 mmol) was added sequentially over 5 minutes to a solution of (4-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (1 g, 4.1 mmol) in EA (80 mL). After addition, the reaction was stirred overnight at ambient temperature. The resulting suspension was filtered, and the filter cake was washed with EA (20 mL). The filtrate was washed with saturated NH4Cl aqueous solution (20 mL), dried over anhydrous Na2SO4 (20 g), filtered, and concentrated under vacuum. The concentrated residue was purified by silica gel chromatography (PE:EA = 20:1 to 5:1) to give 0.9 g of the title compound. 1H-NMR (300MHz, CD3OD) δ10.04 (s, 1H), 8.12 (d, J = 8.4Hz, 2H), 8.03 (d, J = 8.4Hz, 2H), 7.74 (s, 1H).

[0911] Step 3: Synthesis of 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde:

[0912]

[0913] At room temperature, Cs₂CO₃ (3.7 g, 11.2 mmol) and MeI (1.1 g, 7.5 mmol) were added in a single addition to a solution of 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde (0.9 g, 3.8 mmol) in DMF (40 mL). After stirring at ambient temperature for 1 hour, the reaction was quenched with water (100 mL) and extracted with EA (100 mL × 2). The combined organic layers were washed with water (50 mL × 2), dried over anhydrous Na₂SO₄ (50 g), and concentrated under vacuum. The concentrated residue was purified by silica gel chromatography (PE:EA = 20:1 to 5:1) to give 0.8 g of the title compound. 1 H-NMR (300MHz, CDCl3) δ10.09 (s, 1H), 8.00 (d, J = 8.4Hz, 2H), 7.86 (d, J = 8.4Hz, 2H), 7.38 (s, 1H), 3.85 (s, 3H).

[0914] Step 4: Synthesis of 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl-1-ol:

[0915]

[0916] At 0 °C, CH3MgBr (4.7 mL, 4.7 mmol, 1 M in THF) was added dropwise over 10 minutes to a solution of 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde (0.8 g, 3.2 mmol) in 30 mL of THF. After addition, the reaction was stirred at 0 °C for 1.5 h, followed by quenching with 20 mL of saturated NH4Cl aqueous solution. The resulting mixture was extracted with EA (100 mL × 2), and the combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4 (30 g), and concentrated under vacuum to give 0.9 g of the crude title compound. 1H-NMR (300MHz, CDCl3) δ7.60 (d, J = 8.1 Hz, 2H), 7.47 (d, J = 8.1 Hz, 2H), 7.31 (s, 1H), 4.97 (q, J = 6.3, 1H), 3.81 (s, 3H), 1.52 (d, J = 6.3 Hz, 3H).

[0917] Step 5: Synthesis of 2-(4-(1-chloroethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium:

[0918]

[0919] SOCl2 (753.8 mg, 6.3 mmol) was added in a single addition to a solution of 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl-1-ol (570 mg, 2.1 mmol) in DCE (20 mL). After addition, the mixture was stirred at 60 °C for 1 hour, followed by vacuum concentration to dryness. The residue was dissolved in EA (100 mL) and then treated with saturated NaHCO3 solution. After separation, the organic layer was dried over anhydrous Na2SO4 (30 g) and concentrated under vacuum. The concentrated crude product was purified by silica gel chromatography (PE:EA = 20:1) to give 330 mg of the title compound. 1 H-NMR (300MHz, CDCl3) δ7.64 (d, J = 8.4Hz, 2H), 7.53 (d, J = 8.4Hz, 2H), 7.32 (s, 1H), 5.13 (q, J = 6.6Hz, 1H), 3.79 (s, 3H), 1.87 (d, J = 6.6Hz, 3H).

[0920] Step 6: Synthesis of 6-chloro-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine (I-24):

[0921]

[0922] To a solution of 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (321.2 mg, 2.1 mmol) in DMF (14 mL), 2-(4-(1-chloroethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazolium (400 mg, 1.4 mmol) and K₂CO₃ (574.5 mg, 4.2 mmol) were added. After addition, the reaction mixture was stirred overnight at 65 °C, quenched with water (30 mL), and extracted with EA (50 mL × 2). The combined organic layers were washed with water (20 mL × 2), dried over anhydrous Na₂SO₄ (30 g), filtered, and concentrated to dryness under vacuum. The concentrated residue was purified by silica gel chromatography (PE:EA = 5:1 to 3:1) to give 40 mg of the title compound. The regiomeric isomers were then obtained as the major products after purification. 1 H-NMR (300MHz, CDCl3) δ9.03 (s, 1H), 8.19 (s, 1H), 7.50-7.64 (m, 4H), 7.29 (s, 1H), 6.26 (q, J = 7.2, 1H), 3.75 (s, 3H), 2.04 (d, J = 7.2Hz, 3H).

[0923] The following general intermediates are prepared from appropriate reagents according to the procedure in I-24:

[0924]

[0925] Preparation of general intermediate I-27:

[0926]

[0927] Step 1: Synthesis of 2-bromo-1-ethyl-4-(trifluoromethyl)-1H-imidazolium:

[0928]

[0929] At -78 °C, n-BuLi (1.6 M solution in hexane, 68.5 mL, 110 mmol) was added dropwise to a stirred solution of 1-ethyl-4-(trifluoromethyl)-1H-imidazolium 1 (12.0 g, 73.2 mmol) in THF (75 mL). The resulting mixture was stirred at -78 °C for 30 min, followed by treatment with a solution of CBr4 (36.3 g, 109.75 mmol) in anhydrous THF (75 mL). The reaction mixture was then heated to room temperature and stirred for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with saturated NH4Cl solution and extracted with EA (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel column chromatography with 0–10% EA elution in hexane to give the title compound (8.00 g). 1 H-NMR (400MHz, DMSO-d6) δ8.09 (s, 1H), 4.01 (q, J = 7.34Hz, 2H), 1.33 (t, J = 7.34Hz, 3H).

[0930] Step 2: Synthesis of 1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl-1-one:

[0931]

[0932] K3PO4 (3.94 g, 18.6 mmol) and (4-acetyl-2-fluorophenyl)boronic acid (1.48 g, 8.18 mmol) were added to a stirred solution of 2-bromo-1-ethyl-4-(trifluoromethyl)-1H-imidazolium 2 (1.80 g, 7.44 mmol) in dioxane:H2O (8:2 mL). The resulting mixture was degassed with argon for 10 min, and then treated in a sealed tube at room temperature with X-Phos (0.708 g, 1.49 mmol) and X-Phos-Pd-G2 (0.292 g, 0.372 mmol). The reaction mixture was then degassed with argon for another 10 min, followed by heating at 100 °C for 2 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography with 0–20% EA elution in hexane to give the title compound (1.85 g). LC-MS (Method B) (ESI+): m / z 301.15 (M+H) + ; 1H-NMR (400MHz, DMSO-d6) δ8.14-8.20 (m, 1H), 7.93 (d, J = 9.29Hz, 2H), 7.76 (t ,J=7.34Hz,1H),3.93(q,J=7.01Hz,2H),2.66(s,3H),1.29(t,J=7.34Hz,3H).

[0933] Step 3: Synthesis of (R)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl-1-ol:

[0934]

[0935] At room temperature, (S)-2-methyl-CBS-oxazolylborane (0.42 mL, 1.53 mmol) was added to a stirred solution of 1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl-1-one 4 (2.30 g, 7.67 mmol) in THF (18 mL). The resulting mixture was then heated at 45 °C for 1 hour. Borane-DMS (1.09 mL, 11.5 mmol) was added to the resulting reaction mixture, and the mixture was heated at 60 °C for 16 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, quenched with methanol (10 mL), and concentrated under reduced pressure. The crude compound was purified by silica gel column chromatography with 0–30% EA elution in hexane to give the title compound (2.10 g). LC-MS (Method B) (ESI+): m / z 303.00 (M+H) + ; 1 H-NMR(400MHz,DMSO-d6)δ8.10(s,1H),7.50(t,J=7.82Hz,1H),7.30-7.37(m,2H),5.43(d,J=4.40Hz ,1H),4.81(m,J=5.87Hz,1H),3.88(q,J=7.17Hz,2H),1.37(d,J=6.36Hz,3H),1.27(t,J=7.09Hz,3H).

[0936] Step 3: Synthesis of (S)-6-chloro-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine (I-27):

[0937]

[0938] At 0 °C, 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (1.07 g, 6.95 mmol), DEAD (2.41 g, 13.9 mmol), and TPP (2.25 g, 13.9 mmol) were added to a stirred solution of (R)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl-1-ol 5 (2.10 g, 6.95 mmol) in THF (12 mL). The reaction mixture was stirred at room temperature for 2 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water (10 mL) and extracted with EA (3 × 10 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography using 0–30% EA in hexane to give the title compound (2.20 g). LC-MS (Method B) (ESI+): m / z 439.22 (M+H) + .

[0939] The following general intermediates are prepared according to procedure I-27 from a suitable ketone and a CBS reducing agent:

[0940]

[0941] Preparation of general intermediate I-31:

[0942]

[0943] Step 1: Synthesis of 2-(4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)-4-(trifluoromethyl)-1H-imidazolium:

[0944]

[0945] At room temperature, TBSCl (1.42 g, 9.42 mmol) was added in a single addition to a solution of (4-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (1.90 g, 7.85 mmol), DMAP (95.8 mg, 0.79 mmol), and TEA (1.19 g, 11.78 mmol) in THF (40 mL). After addition, the reaction mixture was stirred at room temperature for 1.5 hours. The reaction was then quenched with water (20 mL) and extracted with EA (150 mL × 3). The combined organic layers were dried over anhydrous Na₂SO₄ (50 g), filtered, and concentrated under vacuum. The concentrated residue was purified by silica gel chromatography (EA:n-Hex = 1:10 to 1:1) to give 2.69 g of the title compound. LC-MS (Method A) (ESI+): m / z 357 (M+H) + ; 1H-NMR (300MHz, CDCl3) δ9.82 (br s, 1H), 7.68 (d, J = 8.7Hz, 2H), 7.26-7.31 (m, 3H), 4.66 (s, 2H), 0.84 (s, 9H), 0.00 (s, 6H).

[0946] Step 2: Synthesis of 3-(2-(4-(hydroxymethyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)azacyclobutane-1-carboxylate:

[0947]

[0948] To a solution of 2-(4-(((tert-butyldimethylsilyl)oxy)methyl)phenyl)-4-(trifluoromethyl)-1H-imidazolium (2.50 g, 7.02 mmol) in DMF (40 mL), tert-butyl 3-iodozacyclobutane-1-carboxylate (2.58 g, 9.13 mmol) and Cs₂CO₃ (6.87 g, 21.06 mmol) were added. The reaction mixture was stirred at 120 °C for 4 hours. The reaction was then quenched with water (80 mL) and extracted with EA (150 mL × 3). The combined organic layers were washed with water (50 mL × 2), dried over anhydrous Na₂SO₄ (100 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (DCM:MeOH = 100:1 to 20:1) to give 917 mg of the title compound. LC-MS (Method A) (ESI+): m / z 398 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.73(s,1H),7.37-7.44(m,4H),5.06(m,1H),4.74(s,2H),4.37-4.43(m,2H),4.07-4.16(m,2H),2.86(br s,1H),1.46(s,9H).

[0949] Step 3: Synthesis of tert-butyl 3-(2-(4-(chloromethyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)azacyclobutane-1-carboxylate:

[0950]

[0951] To a solution of tert-butyl 3-(2-(4-(hydroxymethyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)azacyclobutane-1-carboxylate (0.92 g, 2.32 mmol) in a DCE (22 mL), pyridine (1.10 g, 13.92 mmol) and SOCl2 (0.83 g, 6.95 mmol) were added. The mixture was then stirred overnight at room temperature, followed by concentration to dryness under vacuum. The resulting residue was purified by column chromatography (EA:n-Hex = 1:10 to 1:4) to give 480 mg of the title compound. LC-MS (Method A) (ESI+): m / z 416 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.75 (s, 1H), 7.52 (d, J = 8.1Hz, 2H), 7.46 (d, J = 8.1Hz, 2H), 5. 07(m,1H),4.64(s,2H),4.41-4.47(t,J=9.0Hz,2H),4.09-4.14(m,2H),4.47(s,9H).

[0952] Step 4: Synthesis of tert-butyl 3-(2-(4-((6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)azacyclobutane-1-carboxylate (I-31):

[0953]

[0954] A solution of tert-butyl 3-(2-(4-(chloromethyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)azacyclobutane-1-carboxylate (0.45 g, 1.07 mmol), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.18 g, 1.18 mmol), and K₂CO₃ (0.37 g, 2.68 mmol) in DMF (13 mL) was stirred for 3 hours at room temperature. The mixture was then quenched with water (30 mL) and extracted with EA (100 mL × 3). The combined organic layers were washed with water (20 mL), dried over anhydrous Na₂SO₄ (50 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (EA:Hex = 1:10 to 1:0) to give 280 mg of the title compound. LC-MS (Method A) (ESI+): m / z 534 (M+H) + ; 1H-NMR (300MHz, CDCl3) δ9.07(s,1H),8.19(s,1H),7.72(s,1H),7.41-7.49(m,4H),5. 70(s,2H),5.01(m,1H),4.36-4.42(t,J=8.7Hz,2H),4.06-4.13(m,2H),1.42(s,9H).

[0955] Preparation of general intermediate I-32:

[0956]

[0957] Step 1: Synthesis of ethyl 4-(3-(difluoromethyl)-5-methyl-1H-pyrazole-1-yl)benzoate:

[0958]

[0959] At 0 °C, a solution of TEA (299.6 mg, 3.0 mmol) in HFIP (1 mL) was added dropwise over 2 minutes to a solution of ethyl 4-hydrazinobenzoate hydrochloride (300 mg, 1.5 mmol) and 1,1-difluoropentane-2,4-dione (201.5 mg, 1.5 mmol) in HFIP (2 mL). After addition, the reaction was stirred at room temperature for 2.5 h. The reaction was then quenched with water (10 mL) and extracted with DCM (20 mL × 2). The combined organic layers were washed with water (10 mL), dried over anhydrous Na₂SO₄ (20 g), and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 15:1 to 3:1) to give 300 mg of the title compound. 1 H-NMR (300MHz, CDCl3) δ8.17 (dd, J = 6.9, 1.8 Hz, 2H), 7.56 (dd, J = 6.9, 1.8 Hz, 2H), 6.71 (t, J = 54.9 Hz, 1H), 6.48 (s, 1H), 3.96 (s, 3H), 2.42 (s, 3H).

[0960] Step 2: Synthesis of (4-(3-(difluoromethyl)-5-methyl-1H-pyrazole-1-yl)phenyl)methanol:

[0961]

[0962] The title compound was synthesized according to step 3 of the general intermediate I-1. 1H-NMR (300MHz, CDCl3) δ7.49 (dd, J=6.3, 2.1Hz, 2H), 7.42 (dd, J=6.3, 2.1Hz, 2H), 6.70 (t ,J=55.2Hz,1H),6.44(s,1H),4.78(d,J=5.7Hz,2H),2.35(s,3H),1.90(t,J=5.7Hz,1H).

[0963] Step 3: Synthesis of 1-(4-(chloromethyl)phenyl)-3-(difluoromethyl)-5-methyl-1H-pyrazole:

[0964]

[0965] SOCl2 (426.4 mg, 3.6 mmol) was added in a single batch to a solution of (290 mg, 1.2 mmol) of (4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)phenyl)methanol in 10 mL of DCE. After addition, the mixture was stirred at 60 °C for 1 hour, followed by vacuum concentration to give 300 mg of the crude title compound. 1 H-NMR (300MHz, CDCl3) δ7.52 (dd, J = 6.3, 2.1 Hz, 2H), 7.44 (dd, J = 6.3, 2.1 Hz, 2H), 6.70 (t, J = 54.9 Hz, 1H), 6.44 (s, 1H), 4.65 (s, 2H), 2.37 (s, 3H).

[0966] Step 4: Synthesis of 6-chloro-1-(4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-32):

[0967]

[0968] A solution of 1-(4-(chloromethyl)phenyl)-3-(difluoromethyl)-5-methyl-1H-pyrazole (300 mg, 1.2 mmol), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (270.9 mg, 1.8 mmol), and K₂CO₃ (484.6 mg, 351 mmol) in DMF (12 mL) was stirred for 1.5 h at 90 °C. The reaction was then quenched with water (25 mL) and extracted with EA (50 mL × 2). The combined organic layers were washed with water (20 mL × 2), dried over anhydrous Na₂SO₄ (30 g), and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 5:1 to 3:1) to give 150 mg of the title compound containing a small amount of regioisomers. 1H-NMR (300MHz, CDCl3) δ9.06 (s, 1H), 8.19 (s, 1H), 7.50 (dd, J = 6.6, 1.8Hz, 2H), 7.41 ( dd,J=6.6,1.8Hz,2H),6.67(t,J=54.9Hz,1H),6.42(s,1H),5.69(s,2H),2.33(s,3H).

[0969] Preparation of universal intermediates I-33 and I-34:

[0970]

[0971] Step 1: Synthesis of 4-bromophenyliminocyanide ethyl ester hydrochloride:

[0972]

[0973] At 0 °C, gaseous HCl was bubbled through a vigorously stirred solution of 4-bromobenzonitrile (4.2 g, 23.07 mmol) in anhydrous MeOH (40 mL) for 3 hours. The mixture was then stirred overnight at room temperature. The resulting reaction mixture was poured into Et₂O (40 mL), and the resulting solid precipitate was collected by filtration. The solid was suspended in cold DCM (80 mL) and neutralized with a saturated aqueous solution of NaHCO₃ (80 mL). After separation of the organic phase, the aqueous phase was extracted with DCM (40 mL × 2). The combined organic phases were dried over sodium sulfate (40 g), filtered, and concentrated to give 5.0 g of the title compound. LC-MS (Method A) (ESI+): m / z 214 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.81 (br s, 1H), 7.51-7.68 (m, 4H), 3.92 (s, 3H).

[0974] Step 2: Synthesis of 4-bromobenzoiminohydrazide:

[0975]

[0976] At 0 °C, hydrazine hydrate (1.6 g, 25.9 mmol) was added dropwise to a solution of 4-bromophenyliminocyanide (4.6 g, 21.6 mmol) in IPA (80 mL) over 30 minutes in 20 mL of IPA. The reaction mixture was stirred in an ice-water bath for 1 hour, then heated to room temperature overnight. TLC analysis indicated the reaction was complete, and the reaction mixture was concentrated to dryness. The residue was diluted with Et₂O (35 mL) to precipitate a large amount of solid. The suspension was stirred at room temperature for 1 hour, and the solid was collected by filtration to give 3.8 g of the crude title compound. LC-MS (Method A) (ESI+): m / z 214 (M+H) + ; 1 H-NMR (300MHz, DMSO) δ7.63 (d, J = 8.7 Hz, 2H), 7.52 (d, J = 8.7 Hz, 2H), 5.66 (br s, 2H), 5.08 (br s, 2H).

[0977] Step 3: Synthesis of 3-(4-bromophenyl)-5-(difluoromethyl)-1H-1,2,4-triazole:

[0978]

[0979] At 0 °C, a solution of 2,2-difluoroacetic anhydride (1.63 g, 9.38 mmol) in DCM (20 mL) was added dropwise over 5 minutes to a mixture of 4-bromobenzoyliminohydrazine (2.0 g, 9.38 mmol) and TEA (2.84 g, 28.14 mmol) in DCM (70 mL). After addition, the reaction mixture was heated to room temperature and stirred overnight. TLC analysis indicated the reaction was complete. The mixture was quenched with brine (80 mL) and extracted with DCM (80 mL × 2). The combined organic layers were dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 8:1 to 4:1) to give 1.2 g of the title compound. LC-MS (Method A) (ESI+): m / z 274 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.85 (d, J = 8.4Hz, 2H), 7.66 (d, J = 8.4Hz, 2H), 6.81 (t, J = 53.4Hz, 1H).

[0980] Step 4: Synthesis of 5-(4-bromophenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole and 3-(4-bromophenyl)-5-(difluoromethyl)-1-methyl-1H-1,2,4-triazole:

[0981]

[0982] Sodium hydride (200 mg, 5.0 mmol) was added fractionally over 5 minutes to a solution of 3-(4-bromophenyl)-5-(difluoromethyl)-1H-1,2,4-triazole (1.05 g, 3.85 mmol) in DMF (25 mL). The resulting mixture was stirred at 0 °C for 20 minutes, followed by dropwise addition of MeI (1.09 g, 7.69 mmol) over 1 minute. The reaction mixture was heated to room temperature and stirred for 1 hour. TLC analysis indicated the reaction was complete. The reaction was quenched with ice water (50 mL) and extracted with EA (70 mL × 2). The combined organic layers were washed with water (60 mL × 2), dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 20:1 to 15:1) to obtain 260 mg of 5-(4-bromophenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole and 750 mg of 3-(4-bromophenyl)-5-(difluoromethyl)-1-methyl-1H-1,2,4-triazole.

[0983] 5-(4-Bromophenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole: LC-MS (Method A) (ESI+): m / z 288 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.70 (d, J = 6.6, 1.8 Hz, 2H), 7.59 (d, J = 6.6, 1.8 Hz, 2H), 6.73 (t, J = 53.7 Hz, 1H), 4.03 (s, 3H).

[0984] 3-(4-Bromophenyl)-5-(difluoromethyl)-1-methyl-1H-1,2,4-triazole: LC-MS (Method A) (ESI+): m / z 288 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.93 (d, J = 8.7Hz, 2H), 7.58 (d, J = 8.7Hz, 2H), 6.87 (t, J = 53.7Hz, 1H), 4.09 (s, 3H).

[0985] Step 5: Synthesis of 3-(difluoromethyl)-1-methyl-5-(4-vinylphenyl)-1H-1,2,4-triazole:

[0986]

[0987] Under nitrogen protection, a solution of 5-(4-bromophenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole (1.6 g, 5.57 mmol) in toluene (50 mL) was treated once with tributylvinyltinane (2.65 g, 8.36 mmol) and Pd(PPh3)4 (644 mg, 0.56 mmol). The reaction was stirred at 90 °C for 5 h, followed by quenching with water (50 mL) and extraction with EA (50 mL × 2). The combined organic layers were washed with brine (60 mL), dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by rapid silica chromatography (PE:EA = 10:1) to give 1.2 g of the title compound. LC-MS (Method A) (ESI+): m / z 236 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.68 (d, J = 8.1 Hz, 2H), 7.56 (d, J = 8.1 Hz, 2H), 6.74 (t, J = 53. 7Hz, 1H), 6.73 (m, 1H), 5.88 (d, J = 17.4Hz, 1H), 5.40 (d, J = 11.1Hz, 1H), 4.05 (s, 3H).

[0988] Step 6: Synthesis of 5-(difluoromethyl)-1-methyl-3-(4-vinylphenyl)-1H-1,2,4-triazole:

[0989]

[0990] The compound was synthesized according to the procedure in step 5 of general intermediate I-33. LC-MS (Method A) (ESI+): m / z 236 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ8.02 (d, J = 8.1Hz, 2H), 7.51 (d, J = 8.1Hz, 2H), 6.90 (t, J = 53. 7Hz, 1H), 6.78 (m, 1H), 5.83 (d, J = 17.2Hz, 1H), 5.31 (d, J = 10.8Hz, 1H), 4.10 (s, 3H).

[0991] Step 7: Synthesis of 4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)benzaldehyde:

[0992]

[0993] At room temperature, NaIO4 (1.23 g, 5.75 mmol) and OsO4 (4.2 mg, 1 mol%) were added in a single addition to a solution of 3-(difluoromethyl)-1-methyl-5-(4-vinylphenyl)-1H-1,2,4-triazole (450 mg, 1.91 mmol) in THF (15 mL) and water (8 mL). After TLC analysis indicated the completion of the reaction, the reaction mixture was quenched with saturated ammonium chloride solution (20 mL) and extracted with EA (30 mL × 2). The combined organic layers were washed with brine (40 mL), dried over sodium sulfate (20 g), filtered, and concentrated to give 450 mg of the crude title compound. LC-MS (Method A) (ESI+): m / z 238 (M+H) + .

[0994] Step 8: Synthesis of 4-(5-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-3-yl)benzaldehyde:

[0995]

[0996] The compound was synthesized according to the procedure in step 7 of general intermediate I-33. LC-MS (Method A) (ESI+): m / z 238 (M+H) + . 1 H-NMR (300MHz, CDCl3) δ9.99 (s, 1H), 8.17 (d, J = 8.1Hz, 2H), 7.90 (d, J = 8.1Hz, 2H), 6.84 (t, J = 52.5Hz, 1H), 4.06 (s, 3H).

[0997] Step 9: Synthesis of (4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)phenyl)methanol:

[0998]

[0999] NaBH4 (94 mg, 2.47 mmol) was added in a single addition to a solution of 4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)benzaldehyde (450 mg, 1.90 mmol) in THF (15 mL). After TLC analysis indicated the completion of the reaction, the reaction was quenched with ice water (20 mL) and extracted with EA (40 mL × 2). The combined organic layers were washed with brine (40 mL), dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 3:2) to give 340 mg of the title compound. LC-MS (Method A) (ESI+): m / z 240 (M+H) + ; 1H-NMR (300MHz, CDCl3) δ7.71 (d, J = 8.1 Hz, 2H), 7.54 (d, J = 8.1 Hz, 2H), 6.74 (t, J = 53.7 Hz, 1H), 4.81 (d, J = 5.7 Hz, 2H) 4.04 (s, 3H).

[1000] Step 10: Synthesis of (4-(5-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-3-yl)phenyl)methanol:

[1001]

[1002] The compound was synthesized according to the procedure of step 9 of general intermediate I-14. LC-MS (Method A) (ESI+): m / z 240 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ 8.06 (d, J = 8.4 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 7.89 (t, J = 52.5 Hz, 1H), 4.75 (d, J = 6.0 Hz, 2H) 4.13 (s, 3H).

[1003] Step 11: Synthesis of 5-(4-(chloromethyl)phenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole:

[1004]

[1005] SOCl2 (318 mg, 2.68 mmol) was added in a single addition to a solution of (320 mg, 1.34 mmol) of (4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)phenyl)methanol in 15 mL of DCE. The reaction mixture was stirred at 60 °C for 1 h. After TLC analysis indicated completion, the reaction mixture was concentrated to dryness to give 340 mg of the crude title compound. LC-MS (Method A) (ESI+): m / z 258 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.72 (d, J = 8.1 Hz, 2H), 7.57 (d, J = 8.1 Hz, 2H), 6.74 (t, J = 53.7 Hz, 1H), 4.66 (s, 2H), 4.05 (s, 3H).

[1006] Step 12: Synthesis of 3-(4-(chloromethyl)phenyl)-5-(difluoromethyl)-1-methyl-1H-1,2,4-triazole:

[1007]

[1008] The compound was synthesized according to the procedure of step 11 of general intermediate I-33. LC-MS (Method A) (ESI+): m / z 258 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ8.06 (d, J = 8.1 Hz, 2H), 7.45 (d, J = 8.1 Hz, 2H), 6.89 (t, J = 52.5 Hz, 1H), 4.63 (s, 2H), 4.10 (s, 3H).

[1009] Step 13: Synthesis of 6-chloro-1-(4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-33):

[1010]

[1011] Cs₂CO₃ (474 ​​mg, 1.45 mmol) was added in a single addition to a solution of 5-(4-(chloromethyl)phenyl)-3-(difluoromethyl)-1-methyl-1H-1,2,4-triazole (340 mg, 1.32 mmol) and 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (244 mg, 1.58 mmol) in DMF (15 mL). The reaction was stirred at 80 °C for 1 h, followed by cooling to room temperature. The reaction was then quenched with ice water (40 mL), followed by extraction with EA (50 mL × 2). The combined organic layers were washed with water (40 mL × 3), dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (PE:EA = 1:1 to 1:3) to give 180 mg of the title compound. LC-MS (Method A) (ESI+): m / z 376 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.07 (s, 1H), 8.20 (s, 1H), 7.69 (d, J = 8.4Hz, 2H), 7.52 (d, J = 8.4Hz, 2H), 6.71 (t, J = 53.7Hz, 1H), 5.72 (s, 2H), 4.00 (s, 3H).

[1012] Step 14: Synthesis of 6-chloro-1-(4-(5-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-34):

[1013]

[1014] The compound was synthesized according to the procedure of step 13 of general intermediate I-33. LC-MS (Method A) (ESI+): m / z 376 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.05 (s, 1H), 8.18 (s, 1H), 8.03 (d, J = 8.4Hz, 2H), 7.43 (d, J = 8.4Hz, 2H), 6.86 (t, J = 58.2Hz, 1H), 5.67 (s, 2H), 4.10 (s, 3H).

[1015] Preparation of general intermediate I-35:

[1016]

[1017] Step 1: Synthesis of 5-bromo-1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazole:

[1018]

[1019] Sodium hydride (0.138 g, 3.47 mmol) was added fractionally to an ice-cold solution of 5-bromo-3-(trifluoromethyl)-1H-1,2,4-triazole 1 (0.500 g, 2.31 mmol) in DMF (10 mL). The resulting mixture was stirred for 15 min, followed by the addition of iodoethane (0.360 mL, 0.708 g, 0.462 mmol). The reaction mixture was heated to room temperature and stirred for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (2 × 30 mL). The combined organic layers were washed with water (20 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–5% EA in n-hexane to give the title compound (0.280 g). 1 H-NMR (400MHz, DMSO-d6) δ4.28 (q, J=7.01Hz, 2H), 1.40 (t, J=7.09Hz, 3H).

[1020] Step 2: Synthesis of 4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)benzaldehyde:

[1021]

[1022] In a sealed tube, K3PO4 (0.436 g, 2.07 mmol) and (4-formylphenyl)boronic acid 3 (0.200 g, 1.34 mmol) were added to a stirred solution of 5-bromo-1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazole 2 (0.250 g, 1.03 mmol) in dioxane:H2O (10:4 mL). The resulting mixture was degassed with argon for 10 min, followed by the addition of X-Phos (0.049 g, 0.102 mmol) and X-Phos-Pd-G2 (0.040 g, 0.051 mmol) at room temperature. The reaction mixture was then degassed with argon for another 10 min, followed by heating at 100 °C for 2 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, quenched with water (20 mL), and extracted with EA (3 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography with 0-25% EA elution in hexane to obtain the title compound (0.220 g). 1 H-NMR (400MHz, DMSO-d6) δ 10.13 (s, 1H), 8.11 (d, J = 6.85Hz, 2H), 8.00 (d, J = 7.34Hz, 2H), 4.38 (q, J = 6.85Hz, 2H), 1.44 (t, J = 6.85Hz, 3H).

[1023] Step 3: Synthesis of (4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)phenyl)methanol:

[1024]

[1025] Sodium borohydride (0.056 g, 1.49 mmol) was added fractionally to a stirred solution of 4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)benzaldehyde 4 (0.200 g, 0.743 mmol) in methanol (5 mL). The reaction mixture was heated to room temperature and stirred for 2 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, quenched with water (20 mL), and extracted with EA (3 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography, eluting with 0–40% EA in hexane, to give the title compound (0.150 g). LC-MS (Method B) (ESI+): m / z 271.40 (M+H) + ; 1H-NMR (400MHz, DMSO-d6) δ7.72(d,J=8.31Hz,2H),7.53(d,J=7.82Hz,2H),5.38(t,J= 5.62Hz, 1H), 4.61 (d, J = 5.87Hz, 2H), 4.34 (q, J = 7.34Hz, 2H), 1.42 (t, J = 7.34Hz, 3H).

[1026] Step 4: Synthesis of 6-chloro-1-(4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-35):

[1027]

[1028] At 0 °C, 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.066 g, 0.43 mmol), DEAD (0.163 g, 0.959 mmol), and TPP (0.246 g, 0.959 mmol) were added to a stirred solution of (4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)phenyl)methanol 5 (0.130 g, 0.479 mmol) in THF (5 mL). The reaction mixture was heated to room temperature and then stirred for 6 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–30% EA in hexane as the eluent to give the title compound (0.100 g). 1 H-NMR (400MHz, DMSO-d6) δ9.33 (s, 1H), 8.53 (s, 1H), 7.73 (d, J = 8.31Hz, 2H), 7.4 5(d,J=7.83Hz,2H), 5.77(s,2H), 4.30(q,J=7.17Hz,2H), 1.39(t,J=7.34Hz,3H).

[1029] The following intermediates are prepared from appropriate heterocyclic compounds and alkylating agents according to the preparation method for I-35:

[1030]

[1031] Preparation of general intermediate I-39:

[1032]

[1033] Step 1: Synthesis of 1-(4-bromophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-ol:

[1034]

[1035] Sodium hydroxide (898 mg, 22.5 mmol) was added in a single addition to a solution of (4-bromophenyl)hydrazine hydrochloride (4.9 g, 22 mmol) in ethanol (60 mL) at room temperature. The resulting mixture was stirred at room temperature for 30 minutes, followed by a single addition of ethyl 4,4,4-trifluoro-3-oxobutyrate (4.9 g, 27 mmol) in ethanol (10 mL) to the reaction mixture. The reaction mixture was heated to reflux and stirred overnight. TLC analysis indicated the reaction was complete. The reaction mixture was then cooled to room temperature and quenched with water (150 mL). The resulting mixture was acidified to pH 5 with dilute hydrochloric acid solution (1 N) and extracted with EA (100 mL × 3). The combined organic layers were dried over sodium sulfate (50 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 30:1 to 6:1) to give 6.2 g of the title compound. LC-MS (Method A) (ESI+): m / z 307 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ7.62-7.74 (m, 4H), 5.84 (s, 1H).

[1036] Step 2: Synthesis of 1-(4-bromophenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole:

[1037]

[1038] Sodium hydride (60% dispersion, 973 mg, 0.024 mmol) was added fractionally to a solution of methyl 1-(4-bromophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-ol (6.2 g, 20 mmol) in DMF (65 mL) over 5 minutes at 0 °C. After stirring the mixture at 0 °C for 30 minutes, methyl iodoform (4.26 g, 0.03 mmol) was added dropwise to the reaction mixture over 2 minutes. The reaction mixture was stirred at room temperature for 3 hours, then quenched with ice water (200 mL) and extracted with EA (100 mL × 3). The organic phase was washed with water (100 mL × 2), dried over sodium sulfate (40 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE to PE:EA = 100:1) to give 4.4 g of the title compound. LC-MS (Method A) (ESI+): m / z 321 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ7.55-7.63(m,4H),5.95(s,1H),4.00(s,3H).

[1039] Step 3: Synthesis of 4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzaldehyde:

[1040]

[1041] At -78°C, n-BuLi (1.5 mL, 3.76 mmol) was added dropwise over 15 minutes to a solution of 1-(4-bromophenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole (1 g, 3.13 mmol) in 10 mL of THF. After stirring at -78°C for 1 hour, anhydrous DMF (0.48 mL, 6.26 mmol) was added dropwise over 5 minutes. Following the addition, the reaction mixture was stirred at -78°C for another 30 minutes, then heated to room temperature. TLC analysis indicated the reaction was complete, and the reaction was quenched with ice water (20 mL). The mixture was extracted with EA (30 mL × 2), dried over sodium sulfate (30 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 30:1 to 20:1) to give 570 mg of the title compound. LC-MS (Method A) (ESI+): m / z 271 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ10.04 (s, 1H), 7.97 (d, J = 6.6Hz, 4H), 3.99 (s, 1H), 4.05 (s, 3H).

[1042] Step 4: Synthesis of (4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol:

[1043]

[1044] At room temperature, NaBH4 (80 mg, 2.11 mmol) was added in a single addition to a solution of 4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzaldehyde (570 mg, 2.11 mmol) in THF (10 mL). The reaction was stirred at room temperature for 1 hour. After TLC analysis indicated completion, the reaction was quenched with water (20 mL) and extracted with EA (20 mL × 2). The combined organic layers were dried over sodium sulfate (20 g), filtered, and concentrated under vacuum to give 620 mg of the title compound, which was used directly in subsequent steps without purification. LC-MS (Method A) (ESI+): m / z 273 (M+H) + ; 1H-NMR (300MHz, CD3OD) δ7.68 (d, J = 8.4 Hz, 2H), 7.45 (d, J = 8.4 Hz, 2H), 5.96 (s, 1H), 4.75 (d, J = 5.1 Hz, 2H), 3.99 (s, 3H), 1.58 (s, 1H).

[1045] Step 5: Synthesis of 1-(4-(chloromethyl)phenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole:

[1046]

[1047] SOCl2 (748 mg, 6.29 mmol) was added in a single addition to a solution of (570 mg, 2.10 mmol) of (4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol in a DCE (20 mL). After addition, the reaction mixture was stirred at 50 °C for 20 min. TLC analysis indicated the reaction mixture was complete. The reaction mixture was then concentrated to dryness under vacuum to give 500 mg of the crude title compound, which was used directly in subsequent steps without purification. LC-MS (Method A) (ESI+): m / z 291 (M+H) + .

[1048] Step 6: Synthesis of 6-chloro-1-(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-39):

[1049]

[1050] To a solution of 1-(4-(chloromethyl)phenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole (520 mg, 1.79 mmol) in DMF (25 mL), 6-chloro-1H-pyrazole[3,4-d]pyrimidine (276 mg, 1.79 mmol) and potassium carbonate (741 mg, 5.37 mmol) were added. After addition, the mixture was stirred at 70 °C for 1 h. TLC analysis indicated the completion of the reaction. The reaction was quenched with water (30 mL) and extracted with EA (30 mL × 2). The combined organic phases were washed with water and brine, dried over sodium sulfate (20 g), filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (PE:EA = 10:1 to 5:1) to give 330 mg of the title compound. LC-MS (Method A) (ESI+): m / z 409 (M+H) + ; 1H-NMR (300MHz, CD3OD) δ9.05 (s, 1H), 8.17 (s, 1H), 7.67 (d, J = 8.4Hz, 2H), 7.47 (d, J = 8.4Hz, 2H), 5.93 (s, 1H), 5.66 (s, 2H), 3.99 (s, 3H).

[1051] The following intermediates are prepared according to the preparation method of I-39, using appropriate heterocyclic compounds and alkylating agents:

[1052]

[1053]

[1054] Preparation of general intermediate I-45:

[1055]

[1056] Step 1: Synthesis of ethyl 1-(4-bromophenyl)-5-oxo-4,5-dihydro-1H-pyrazole-3-carboxylate:

[1057]

[1058] Potassium carbonate (1.23 g, 8.94 mmol) was added to a solution of (4-bromophenyl)hydrazine hydrochloride (1.0 g, 4.5 mmol) in ethanol (40 mL). After stirring at room temperature for 30 min, diethyl butyrate-2-ynthetic acid (761 mg, 4.47 mmol) in ethanol (10 mL) was added dropwise to the reactants over 5 min. After stirring under reflux for 4 h, the reaction was quenched with water (50 mL) and the pH was adjusted to 5 with dilute hydrochloric acid. The mixture was extracted with EA (40 mL × 3). The combined organic layers were dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 10 / 1 to 5:1) to give 320 mg of the title compound. LC-MS (Method A) (ESI+): m / z 311, 313 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ7.74 (d, J = 9.0 Hz, 2H), 7.71 (d, J = 9.0 Hz, 2H), 6.01 (s, 1H), 4.30-4.38 (q, J = 6.9 Hz, 2H), 1.32-1.38 (t, J = 6.9 Hz, 3H).

[1059] Step 2: Synthesis of ethyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxylate:

[1060]

[1061] DIAD (704 mg, 3.48 mmol) was added dropwise to a mixture of ethyl 1-(4-bromophenyl)-5-oxo-4,5-dihydro-1H-pyrazole-3-carboxylate (720 mg, 2.32 mmol), triphenylphosphine (913 mg, 3.48 mmol), and methanol (96 mg, 3.02 mmol) in toluene (40 mL) over 5 minutes at 0 °C. The mixture was stirred at 0 °C for 30 minutes, then the reaction mixture was slowly heated to room temperature and stirred overnight. After the reaction was indicated by TLC analysis, the mixture was quenched with water (20 mL) and extracted with EA (30 mL × 3). The combined organic layers were dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 20 / 1 to 10 / 1) to give 650 mg of the title compound. LC-MS (Method A) (ESI+): m / z 325, 327 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.65(d,J=8.7Hz,2H),7.56(d,J=8.7Hz,2H),6.22(s ,1H),4.39-4.46(q,J=7.2Hz,2H),3.98(s,3H),1.39-1.44(t,J=7.2Hz,3H).

[1062] Step 3: Synthesis of methyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxamide:

[1063]

[1064] In a sealed tube, ethyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxylate (650 mg, 2.01 mmol) was stirred in a saturated methanol-ammonia solution (30 mL) at 60 °C for 7 hours. After TLC analysis indicated completion, the mixture was quenched with water (30 mL) and extracted with EA (20 mL × 3). The combined organic layers were dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 5 / 1 to 0 / 1) to give 405 mg of the title compound. LC-MS (Method A) (ESI+): m / z 296, 298 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.55-7.65(m,4H),6.82(s,1H),6.26(s,1H),5.40(s,1H),3.99(s,3H).

[1065] Step 4: Synthesis of 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile:

[1066]

[1067] At 0 °C, trifluoroacetic anhydride (1.6 mL) was added dropwise over 5 minutes to a solution of methyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxamide (400 mg, 1.36 mmol) in DCM (40 mL) and pyridine (2 mL). The reaction was stirred at 0 °C for 1 hour. After the reaction was indicated by TLC analysis, the mixture was quenched with saturated NH4Cl aqueous solution (40 mL) and extracted with DCM (40 mL × 3). The organic layer was washed with water (20 mL) and brine (20 mL), dried over sodium sulfate (40 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 20 / 1 to 10 / 1) to give 295 mg of the title compound. LC-MS (Method A) (ESI+): m / z 278, 280 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.59 (s, 4H), 6.06 (s, 1H), 4.00 (s, 3H).

[1068] Step 5: Synthesis of 5-methoxy-1-(4-vinylphenyl)-1H-pyrazole-3-carboxynitrile:

[1069]

[1070] Tributyl(vinyl)stanane (498 mg, 1.57 mmol) and Pd(Ph3)4 (121 mg, 0.11 mmol) were added to a solution of 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile (290 mg, 1.05 mmol) in toluene (8 mL). The reaction was stirred at 90 °C for 6 h. After the reaction was indicated by LC-MS analysis, the mixture was quenched with water (10 mL) and extracted with EA (10 mL × 2). The combined organic layers were dried over sodium sulfate (20 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 20 / 1 to 10 / 1) to give 200 mg of the title compound. LC-MS (Method A) (ESI+): m / z 226 (M+H) + ; 1H-NMR (300MHz, CDCl3) δ7.64(d,J=8.7Hz,2H),7.49(d,J=8.7Hz,2H),6.74(dd,J=17.7Hz, J=10.8Hz,1H),6.06(s,1H),5.80(d,J=17.7Hz,1H),5.33(d,J=10.8Hz,1H),3.99(s,3H).

[1071] Step 6: Synthesis of 1-(4-formylphenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile:

[1072]

[1073] To a solution of 5-methoxy-1-(4-vinylphenyl)-1H-pyrazole-3-carboxynitrile (20 mg, 0.89 mmol) in THF / H₂O (10 mL / 5 mL), NaIO₄ (570 mg, 2.67 mmol) and OsO₄ (22 mg, 0.09 mmol) were added in a single addition. The reaction was stirred at room temperature for 30 min. After the reaction was indicated by TLC analysis, the mixture was quenched with NH₄Cl aqueous solution (10 mL) and extracted with EA (20 mL × 2). The combined organic layers were dried over sodium sulfate (20 g), filtered, and concentrated to give 210 mg of the crude title compound. LC-MS (Method A) (ESI+): m / z 228 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ10.05(s,1H),7.84-8.05(m,4H),6.01(s,1H),3.99(s,3H).

[1074] Step 7: Synthesis of 1-(4-(hydroxymethyl)phenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile:

[1075]

[1076] NaBH4 (46 mg, 1.2 mmol) was added fractionally to a solution of 1-(4-formylphenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile (210 mg, 0.93 mmol) in THF (30 mL) over 5 minutes. The reaction was stirred at room temperature for 1 hour. After the reaction was indicated by TLC analysis, the mixture was quenched with water (20 mL) and extracted with EA (30 mL × 2). The combined organic layers were dried over sodium sulfate (20 g) and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 5 / 1 to 2 / 1) to give 170 mg of the title compound. LC-MS (Method A) (ESI+): m / z 230 (M+H) + ;1 H-NMR (300MHz, CDCl3) δ7.67 (d, J = 8.7Hz, 2H), 7.47 (d, J = 8.7Hz, 2H), 6.07 (s, 1H), 4.76 (d, J = 6Hz, 2H), 3.99 (s, 3H), 1.75 (t, J = 6Hz, 1H).

[1077] Step 8: Synthesis of 1-(4-(chloromethyl)phenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile:

[1078]

[1079] SOCl2 (176 mg, 1.48 mmol) was added in a single addition to a solution of 1-(4-(hydroxymethyl)phenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile (170 mg, 0.74 mmol) in a DCE (10 mL), and the mixture was stirred at 50 °C for 1 h. After TLC analysis indicated completion of the reaction mixture, the mixture was concentrated to dryness to give 205 mg of the crude title compound. LC-MS (Method A) (ESI+): m / z 248 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.70 (d, J = 8.4Hz, 2H), 7.49 (d, J = 8.4Hz, 2H), 6.07 (s, 1H), 4.62 (s, 2H), 4.00 (s, 3H).

[1080] Step 9: Synthesis of 1-(4-((6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-5-methoxy-1H-pyrazol-3-carboxylonitrile (I-45):

[1081]

[1082] To a solution of 1-(4-(chloromethyl)phenyl)-5-methoxy-1H-pyrazole-3-carboxynitrile (205 mg, 0.83 mmol) in DMF (5 mL), 6-chloro-1H-pyrazole[3,4-d]pyrimidine (153 mg, 1.0 mmol) and potassium carbonate (812 mg, 2.49 mmol) were added in a single addition. Following the addition, the mixture was stirred at 70 °C for 1 h. After TLC analysis indicating the completion of the reaction mixture, the mixture was quenched with water (10 mL) and extracted with EA (30 mL × 2). The combined organic layers were washed with water (20 mL) and brine (10 mL), dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 10 / 1 to 5 / 1) to give 57 mg of the title compound. LC-MS (Method A) (ESI+): m / z 366 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.05 (s, 1H), 8.18 (s, 1H), 7.66 (d, J = 8.4Hz, 2H), 7.48 (d, J = 8.4Hz, 2H), 6.04 (s, 1H), 5.67 (s, 2H), 3.97 (s, 3H).

[1083] Preparation of general intermediate I-46:

[1084]

[1085] Step 1: Synthesis of 4,4,4-trifluoro-3-oxobutyronitrile:

[1086]

[1087] Under an argon atmosphere, ethyl 2,2,2-trifluoroacetate 1 (15.0 g, 106 mmol) and MeCN (12.4 g, 159 mmol) were simultaneously added to a stirred suspension of NaH (10.5 g, 264 mmol) in THF (150 mL) at 0 °C. The resulting mixture was stirred at room temperature for 10 min, then heated to 70 °C and maintained for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (50 mL) and concentrated under reduced pressure to remove the organic phase. The remaining aqueous layer was extracted with diethyl ether (2 × 30 mL). The aqueous layer was then adjusted to pH 2 with concentrated HCl and extracted with diethyl ether (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give crude title compound 2 (18.0 g), which was used in subsequent steps without further purification.

[1088] Step 2: Synthesis of ethyl 4-(5-amino-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[1089]

[1090] At room temperature, methyl 4-hydrazinobenzoate hydrochloride 3 (2.50 g, 12.47 mmol) was added to a stirred solution of 4,4,4-trifluoro-3-oxobutyronitrile 2 (3.39 g, 24.8 mmol) in methanol (30 mL). The reaction mixture was heated to 80 °C and maintained for 16 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 20–25% EA in hexane as eluent to give the title compound (2.00 g). 1 H-NMR (400MHz, DMSO-d6) δ8.09(d,J=8.48Hz,2H),7.79(d,J=8.48Hz,2H),5.98(br s,2H),5.84(s,1H),3.89(s,3H).

[1091] Step 3: Synthesis of methyl 4-(5-acetamido-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[1092]

[1093] To an ice-cold solution of ethyl 4-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate 4 (1.00 g, 3.51 mmol) in pyridine (10 mL), CH3COCl (0.37 mL, 5.3 mmol) was added dropwise. The resulting mixture was stirred for 30 min, and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with saturated CuSO4 solution (30 mL) and extracted with EA (2 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–40% EA in hexane as eluent to give the title compound (0.800 g). LC-MS (Method B) (ESI+): Observation: 328.05 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ8.21 (d, J = 8.48Hz, 2H), 7.59 (d, J = 8.48Hz, 2H), 6.95-7.00 (m, 1H), 3.96 (s, 3H), 2.17 (s, 3H).

[1094] Step 3: Synthesis of methyl 4-(5-(N-methylacetamido)-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[1095]

[1096] At room temperature, potassium carbonate (0.506 g, 3.67 mmol) and methyl 4-(5-acetamido-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate 5 (0.800 g, 2.45 mmol) were added to a stirred solution in CH3CN (15 mL) with stirring. The reaction mixture was then heated in a sealed tube at 85 °C for 16 h. The reaction progress was monitored by TLC. After completion, the reaction mixture was diluted with EA (100 mL) and washed with water (2 × 25 mL), followed by brine (20 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–30% EA in hexane as eluent to give the title compound (0.650 g). LC-MS (Method B) (ESI+): Observation: 342.10 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ8.19 (d, J = 8.48 Hz, 2H), 7.57 (d, J = 8.48 Hz, 2H), 6.62 (s, 1H), 3.96 (s, 3H), 3.15 (s, 3H), 1.85 (s, 3H).

[1097] Step 5: Synthesis of methyl 4-(5-(methylamino)-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate:

[1098]

[1099] At room temperature, concentrated HCl (7 mL) was added to a stirred solution of methyl 4-(5-(N-methylacetamido)-3-(trifluoromethyl)-1H-pyrazole-1-yl)benzoate 6 (0.650 g, 1.91 mmol) in methanol (10 mL). The reaction mixture was then heated at 90 °C for 4 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with EA (3 × 25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–30% EA in hexane as eluent to give the title compound (0.500 g). LC-MS (Method C) (ESI+): m / z 299.98 (M+H) + ; 1 H-NMR (400MHz, CDCl3) δ8.16(d,J=8.48Hz,2H),7.68(d,J=8.48Hz,2H),5.80(s,1H),3.95(s,3H),2.89(s,3H).

[1100] Step 6: Synthesis of (4-(5-(methylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol:

[1101]

[1102] At 0 °C, methyl 4-(5-(methylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate 7 (0.500 g, 1.67 mmol) in a stirred solution of THF (15 mL) was added to DIBAL-H (1 M solution in toluene, 5.00 mL, 5.02 mmol). The resulting mixture was stirred for 30 min, and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with an aqueous solution of NH4Cl (10 mL). The resulting solution was diluted with EA (20 mL) and filtered through a diatomaceous earth mat. The aqueous layer was separated and re-extracted with EA (3 × 15 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 50–80% EA in hexane as the eluent to give the title compound (0.400 g). 1 H-NMR (400MHz, DMSO-d6) δ7.42-7.51 (m, 4H), 5.89 (s, 1H), 5.83 (d, J = 4.40Hz ,1H),5.33(t,J=5.62Hz,1H),4.56(d,J=5.87Hz,2H),2.69(d,J=4.89Hz,3H).

[1103] Step 7: Synthesis of 1-(4-((6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-N-methyl-3-(trifluoromethyl)-1H-pyrazol-5-amine (I-46):

[1104]

[1105] To a stirred solution of (4-(5-(methylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol 8 (0.200 g, 0.738 mmol) in THF (10 mL), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.102 g, 0.664 mmol), DIAD (0.336 g, 1.48 mmol), and TPP (0.382 g, 1.48 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour, and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with water (20 mL) and extracted with EA (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–40% EA in hexane as the eluent to give the title compound (0.160 g). LC-MS (Method C) (ESI+): m / z 407.86 (M+H) + .

[1106] Preparation of general intermediate I-47:

[1107]

[1108] Step 1: Synthesis of ethyl 4-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorobenzoate:

[1109]

[1110] At room temperature, ethyl 3-fluoro-4-hydrazinobenzoate hydrochloride 2 (0.731 g, 5.341 mmol) was added to a stirred solution of 4,4,4-trifluoro-3-oxobutyronitrile 1 (0.500 g, 2.136 mmol) in methanol (20 mL). The reaction mixture was then heated to 80 °C and maintained for 16 hours. The reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 15–25% EA in hexane as eluent to give the title compound (0.070 g). 1 H-NMR (400MHz, DMSO-d6) δ7.92(d,J=9.29Hz,2H),7.72(t,J=7.83Hz,1H),5.91(s,2H),5.75(s,1H),4.37(q,J=6.85Hz,2H),1.35(t,J=7.09Hz,3H).

[1111] Step 2: Synthesis of ethyl 4-(5-(dimethylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorobenzoate:

[1112]

[1113] At 0 °C, a dispersion of 60% sodium hydride in oil (0.063 g, 1.577 mmol) and methyl iodide (0.145 mL, 2.365 mmol) were added to a stirred solution of ethyl 4-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorobenzoate 3 (0.250 g, 0.788 mmol) in DMF (10 mL). The resulting mixture was stirred for 16 h, and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with ice-cold water (10 mL) and extracted with EA (2 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography using 0–10% EA in hexane as the eluent to give the title compound (0.070 g). 1 H-NMR (400MHz, DMSO-d6) δ7.94-8.00(m,2H),7.79-7.85(m,1H),6.40(s,1H),4.37(q,J=6.85Hz,2H),2.56(s,6H),1.35(t,J=7.09Hz,3H).

[1114] Step 3: Synthesis of (4-(5-(dimethylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorophenyl)methanol:

[1115]

[1116] Ethyl 4-(5-(dimethylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorobenzoate 4 (0.150 g, 0.434 mmol) was added fractionally to an ice-cold solution of 4-(5-(dimethylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorobenzoate in 10 mL of methanol. After the addition was complete, the reaction mixture was stirred for 5 hours and the reaction progress was monitored by TLC. After completion, the reaction mixture was quenched with 10 mL of aqueous NH4Cl solution and extracted with EA (3 × 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude compound was purified by silica gel chromatography, eluting with 40–50% EA in hexane, to give the title compound (0.090 g). LC-MS (Method B) (ESI+): m / z 304.00 (M+H) + ; 1H-NMR (400MHz, DMSO-d6) δ7.56(t,J=7.98Hz,1H),7.38(d,J=11.47Hz,1H),7.32(d,J= 7.98Hz, 1H), 6.35 (s, 1H), 5.49 (t, J = 5.73Hz, 1H), 4.59 (d, J = 5.49Hz, 2H), 2.54 (s, 6H).

[1117] Step 4: Synthesis of 1-(4-((6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)-2-fluorophenyl)-N,N-dimethyl-3-(trifluoromethyl)-1H-pyrazol-5-amine (I-47):

[1118]

[1119] At room temperature, 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (0.036 g, 0.237 mmol), DTAD (0.119 g, 0.528 mmol), and TPP (0.136 g, 0.528 mmol) were added to a stirred solution of (4-(5-(dimethylamino)-3-(trifluoromethyl)-1H-pyrazol-1-yl)-3-fluorophenyl)-methanol 5 (0.080 g, 0.264 mmol) in THF (5 mL). The resulting mixture was stirred for 1 hour, and the reaction progress was monitored by TLC. After completion, the reaction mixture was concentrated under reduced pressure. The crude compound was purified by silica gel chromatography, eluting with 0–60% EA in hexane, to give the title compound (0.060 g). LC-MS (Method B) (ESI+): m / z 440.00 (M+H) + ; 1 H-NMR (400MHz, DMSO-d6) δ9.33 (s, 1H), 8.54 (s, 1H), 7.60 (t, J = 8.07Hz, 1H), 7.38 ( d,J=10.76Hz,1H),7.21(d,J=8.31Hz,1H),6.35(s,1H),5.77(s,2H),2.52(s,6H).

[1120] The following intermediates are prepared from appropriate heterocyclic compounds and alkylating agents according to the preparation method for I-47:

[1121]

[1122] Preparation of general intermediate I-50:

[1123]

[1124] Step 1: Synthesis of 1-(4-bromophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-thiol:

[1125]

[1126] A solution of 1-(4-bromophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-ol (900 mg, 2.94 mmol) in toluene (40 mL) was added with Lawstmann's reagent (2.38 g, 5.88 mmol). The reaction was then stirred under reflux for 4 hours. After the reaction was indicated by TLC analysis, the suspension was filtered, and the filtrate was concentrated to dryness to give 2.6 g of the crude title compound. LC-MS (Method A) (ESI+): m / z 323, 325 (M+H) + .

[1127] Step 2: Synthesis of 1-(4-bromophenyl)-5-(methylthio)-3-(trifluoromethyl)-1H-pyrazole:

[1128]

[1129] At 0 °C, NaH (388 mg, 9.69 mmol) was added fractionally over 5 minutes to a solution of crude methyl 1-(4-bromophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-thiol (2.6 g, 8.1 mol) in DMF (20 mL). The mixture was stirred at 0 °C for 30 minutes, followed by dropwise addition of MeI (1.72 g, 12.1 mmol) over 2 minutes. The reaction mixture was stirred at 0 °C and then allowed to warm to room temperature over 3 hours. After TLC analysis indicated the completion of the reaction, the mixture was quenched with ice water (20 mL) and extracted with EA (20 mL × 3). The organic layer was washed with water (20 mL × 2) and brine (20 mL), dried over sodium sulfate (20 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 50 / 0 to 50 / 1) to give 650 mg of the title compound. LC-MS (Method A) (ESI+): m / z 337,339 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.63 (d, J = 6.9 Hz, 2H), 7.48 (d, J = 6.9 Hz, 2H), 6.57 (s, 1H), 2.44 (s, 3H).

[1130] Step 3: Synthesis of 4-(5-(methylthio)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzaldehyde:

[1131]

[1132] At -78°C, n-BuLi (0.98 mL, 2.46 mmol) was added dropwise over 5 minutes to a solution of 1-(4-bromophenyl)-5-(methylthio)-3-(trifluoromethyl)-1H-pyrazole 3 (550 mg, 1.64 mmol) in anhydrous THF (10 mL). After stirring at -78°C for 1 hour, DMF (240 mg, 3.28 mmol) was added dropwise over 5 minutes, and the mixture was slowly heated to room temperature over 2 hours. The reaction was indicated by TLC analysis, and the mixture was quenched with ice water (20 mL). The mixture was extracted with EA (30 mL × 2). The organic layer was dried over sodium sulfate (30 g), filtered, and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 100 / 1 to 50 / 1) to give 100 mg of the title compound. LC-MS (Method A) (ESI+): m / z 287 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ10.08 (s, 1H), 8.02 (d, J = 8.7Hz, 2H), 7.84 (d, J = 8.7Hz, 2H), 6.61 (s, 1H), 2.48 (s, 3H).

[1133] Step 4: Synthesis of (4-(5-(methylthio)-3-(trifluoromethyl)-1H-pyrazole-1-yl)phenyl)methanol:

[1134]

[1135] Prepared according to the reduction procedure of I-8. LC-MS (Method A) (ESI+): m / z 289 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.57 (d, J = 8.4Hz, 2H), 7.49 (d, J = 8.4Hz, 2H), 6.57 (s.1H), 4.78 (s, 2H), 2.43 (s, 3H), 1.83 (br s, 1H).

[1136] Step 5: Synthesis of 6-chloro-1-(4-(5-(methylthio)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine (I-50):

[1137]

[1138] DIAD (136 mg, 0.68 mmol) was added dropwise over 5 minutes to a suspension of 5 (130 mg, 0.45 mol), 6-chloro-1H-pyrazolo[3,4-d]pyrimidine (90 mg, 0.59 mmol), and PS-TPP (300 mg, 0.9 mmol) in anhydrous THF (10 mL). The reaction was stirred in an ice-water bath for 30 minutes, then slowly heated to room temperature overnight. After TLC analysis indicated the completion of the reaction, the reaction was quenched with water (20 mL) and extracted with EA (20 mL × 2). The combined organic layers were dried and concentrated. The residue was purified by silica gel chromatography (eluent: PE / EA = 10 / 1 to 1 / 0) to give 160 mg of crude product. The crude product was further purified by preparative HPLC (Method A) to give 30 mg of the title compound. LC-MS (Method A) (ESI+): m / z 425 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ9.06(s.1H),8.19(s,1H),7.59-7.45(m,4H),6.55(s.1H),5.70(s,2H),2.42(s,3H).

[1139] Preparation of general intermediate I-51

[1140]

[1141] Step 1: Synthesis of methyl 4-(5-methoxy-1-methyl-1H-1,2,4-triazol-3-yl)benzoate:

[1142]

[1143] Under nitrogen atmosphere, Pd2(dba)3 (238.45 mg, 0.26 mmol) was added to a mixture of 3-bromo-5-methoxy-1-methyl-1H-1,2,4-triazole (500 mg, 2.60 mmol), methyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)benzoate (1.02 g, 3.91 mmol), tricyclohexylphosphine (219.07 mg, 0.78 mmol), K3PO4-3H2O (2.08 g, 7.81 mmol) in dioxane (26 mL) and H2O (1.3 mL). The resulting mixture was stirred at 90 °C for 1 hour. TLC analysis indicated the completion of the reaction, which was then quenched with water (30 mL) and extracted with EA (25 mL × 3). The combined organic layers were dried over anhydrous Na2SO4 (30 g), filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel chromatography (PE:EA = 10:1) to obtain 650 mg of the title compound.1 H-NMR (300MHz, CDCl3) δ8.08 (s, .4H), 4.17 (s, 3H), 3.93 (s, 3H), 3.68 (s, 3H).

[1144] Step 2: Synthesis of (4-(5-methoxy-1-methyl-1H-1,2,4-triazol-3-yl)phenyl)methanol:

[1145]

[1146] At 0 °C, DIBAL-H (1.5 M, 7.9 mL, 11.9 mmol) was added dropwise over 15 minutes to a solution of methyl 4-(5-methoxy-1-methyl-1H-1,2,4-triazol-3-yl)benzoate (650 mg, 2.63 mmol) in THF (26 mL). The resulting mixture was stirred at 0 °C for 1 hour, followed by slow dilution with saturated NH4Cl aqueous solution (10 mL). The resulting suspension was stirred for another 30 minutes in an ice-water bath, followed by filtration to remove the solids. The filter cake was washed with EA (30 mL × 4), and the filtrate was washed with brine (50 mL). After separation, the organic layer was dried over anhydrous Na2SO4 (30 g), filtered, and concentrated to obtain a crude residue, which was purified by silica gel chromatography (PE:EA = 3:1 to 2:1) to give 500 mg of ...

Claims

1. A compound, said compound being selected from the group consisting of: 6-(3-methoxypyridin-4-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-(methylsulfonyl)-phenyl)-1H-pyrazolo[3,4-d]pyrimidine; 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-methyl-6-(methylsulfonyl)phenyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-(ethylsulfonyl)phenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 1-(4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)benzyl)-6-(2-isopropylpyridin-3-yl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-methoxy-4-methylpyridin-3-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-methoxy-6-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(2-isopropylpyridin-3-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2,6-dimethoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-methoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-methoxy-6-methylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 2-Methoxy-3-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)isonicotinonitrile; 2-(2-Isopropylphenyl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(2-isopropylpyridin-3-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(2-methyl-6-(methylthio)phenyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(2-Cyclopropyl-6-methoxyphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-1-(4-(1-(1-methylazacyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-4-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(3-fluoro-2-isopropylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-3-methyl-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(2-isopropylphenyl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-(tert-butyl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-(2-fluoropropane-2-yl)-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin; (R)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropyl-6-ethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-isopropoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-Cyclopropyl-N,N-Dimethyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine; 6-(4,6-dimethoxypyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-isopropyl-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methyl-6-(methylthio)pyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin; 6-(1-Isopropyl-4-methyl-1H-pyrazol-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7H-pyrrolo[2,3-d]pyrimidinyl; 5-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-3-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3H-[1,2,3]triazolo[4,5-d]pyrimidinyl; 6-(4,6-diethoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-5-(trifluoromethyl)-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(1-Cyclopropyl-4-methoxy-1H-pyrazol-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-((6-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)pyridin-3-yl)methyl)-1H-pyrazolo[3,4-d]pyrimidin; (R)-6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin; (S)-6-(4-cyclobutyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin; (R)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin; (S)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-1H-pyrazolo[3,4-d]pyrimidin; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-((6-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)pyridin-3-yl)methyl)-1H-pyrazolo[3,4-d]pyrimidin; 1-(4-((6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-5-methyl-1H-pyrazol-3-carboxylonitrile; 6-(4-cyclopropyl-1-ethyl-1H-pyrazol-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropoxy-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclobutoxy-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-(difluoromethoxy)pyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxy-2-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(oxecyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-(tert-butoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-(2-fluoroethoxy)pyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 1-(4-(1-cyclobutyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4,6-dimethoxypyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 1-(4-(1-cyclopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(2,4-Dimethoxy-6-methylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-(2-methoxyethoxy)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-(2-methoxyethoxy)pyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; N,N,6-Trimethyl-5-(1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-(1-methylazacyclobutane-3-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-methoxy-6-methylpyrimidin-5-yl)-1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 3-(1-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyridinecarboxylonitrile; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(3-(difluoromethyl)-5-methyl-1H-pyrazol-1-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-Cyclopropyl-N-methyl-5-(1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin-6-yl)pyrimidin-4-amine; 1-(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6-(4-methoxy-6-methylpyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidine; 6-(4-cyclopropyl-6-((tetrahydrofuran-3-yl)oxy)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-(benzoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(3-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-5-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-(benzoxy)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-methoxy-1-methyl-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 2-(2-(4-((6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl)methyl)phenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)-N,N-dimethylacetamide; 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(5-(difluoromethyl)-1-methyl-1H-1,2,4-triazol-3-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidinyl; 6-(4-cyclopropyl-6-(pyrrolidone-1-yl)pyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidin; and 6-(4-(azacyclobutan-1-yl)-6-cyclopropylpyrimidin-5-yl)-1-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine, Or its pharmaceutically acceptable salt.

2. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol)-2-yl)benzyl)-1H-pyrazolo[3,4-d]pyrimidine.

3. A pharmaceutical composition comprising the compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

4. Use of a compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the compound or a pharmaceutically acceptable salt thereof is used to manufacture an agent for treating cancer, wherein the cancer is selected from the group consisting of: BRCA1-mutated breast cancer, BRCA1-mutated ovarian cancer, BRCA2-mutated lung cancer, and BRCA2-mutated colon cancer.

5. A kit comprising a compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 3, and instructions for administering the compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, to a patient suffering from cancer. The cancers mentioned are selected from the group consisting of: BRCA1-mutated breast cancer, BRCA1-mutated ovarian cancer, BRCA2-mutated lung cancer, and BRCA2-mutated colon cancer.

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