USP1 inhibitors

Abstract

The invention provides a novel compound with USP1 inhibitory activity as shown in a formula (II '), a pharmaceutical composition containing the compound, a useful intermediate for preparing the compound and a method for treating USP1 target mediated related diseases by using the compound. # imgabs0 #

Description

USP1 inhibitors

[0001] This application is filed with the China Patent Office on September 2, 2022, with application number CN202211069731.2 and invention name “USP1 inhibitor”; filed with the China Patent Office on October 18, 2022, with application number CN202211272140.5 and invention name “USP1 inhibitor”; filed with the China Patent Office on January 18, 2023, with application number CN202310084635.3 and invention name “USP1 inhibitor”; filed with the China Patent Office on April 23 .... The present invention claims priority from the Chinese patent application filed with the Patent Office of China on the 18th, with application number CN202310416682.3 and invention name “USP1 inhibitor”, the Chinese patent application filed with the Patent Office of China on June 25, 2023, with application number CN202310755391.7 and invention name “USP1 inhibitor”, and the Chinese patent application filed with the Patent Office of China on August 25, 2023, with application number CN202311082622.9 and invention name “USP1 inhibitor”, all of which are incorporated by reference into this application. Technical Field

[0002] The present application belongs to the field of medicinal chemistry and relates to novel compounds with USP1 inhibitory activity, pharmaceutical compositions containing the compounds, useful intermediates for preparing the compounds, and methods for using the compounds of the present application to treat USP1 target-mediated related diseases. Background Art

[0003] Ubiquitin is a small, highly conserved protein composed of 76 amino acids that binds to substrate proteins (including itself) through a three-step enzymatic reaction. The initial covalent attachment occurs primarily between the C-terminal glycine of ubiquitin and the ε-amino group of a lysine residue on the target protein. Additional ubiquitin molecules can be attached to one of the seven internal lysines of ubiquitin, resulting in different ubiquitin chain topologies. The biological consequences of ubiquitination are determined by the length and linkage topology. Similar to other types of post-translational modifications, ubiquitination is a reversible process that is counterregulated by enzymes called deubiquitinases (DUBs), which catalyze the removal of ubiquitin from modified proteins. Importantly, dysfunction of ubiquitin-dependent signaling pathways has been associated with various human diseases, suggesting that inhibition of ubiquitin pathway components is a novel therapeutic target for drug discovery.

[0004] The ubiquitin-proteasome system offers additional opportunities for therapeutic intervention, potentially increasing specificity and improving clinical efficacy. The most obvious targets include enzymes involved in ubiquitin conjugation and deconjugation (i.e., ubiquitin ligases and DUBs), upstream processes of proteasome-mediated protein degradation. Among DUBs, ubiquitin-specific protease 1 (USP1) has emerged as an attractive anticancer target due to its involvement in regulating DNA damage response pathways. USP1 associates with UAF1 (USP1-associated factor 1) to generate the heterodimeric USP1 / UAF1 complex required for deubiquitinase activity. The USP1 / UAF1 complex has been shown to regulate tolerance to DNA cross-linker-induced DNA damage through deubiquitination of PCNA (proliferating cell nuclear antigen) 11 and FANCD2 (Fanconi anemia complementation group D2), proteins that function in translesion synthesis and the Fanconi anemia pathway, respectively.

[0005] Currently, there are many studies based on this mechanism of action, but no USP1 inhibitor drugs have been found on the market. Therefore, there is an urgent need to develop effective USP1 inhibitors for clinical patients.

[0006] Summary of the Invention

[0007] The first aspect of the present application provides a compound represented by formula (II'), its isomers and pharmaceutically acceptable salts thereof,

[0008] in,

[0009] X a C or N;

[0010] Ring A is a benzene ring, a 5-6 membered heteroaryl ring, C 5-6 Cycloalkyl ring or 5-6 membered heterocyclyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4 Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C1-4 alkyl)2P(O)-, Deuterated C 1-4 Alkyl, 4-6 membered heterocycloalkyl; the C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 Alkyl; m is 0, 1, 2, 3 or 4;

[0011] R b For hydrogen, C 1-4 alkyl;

[0012] Ring B is a benzene ring, a 5-10 membered heteroaryl ring, or a 5-10 membered heterocyclyl ring; R c Each independently is halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, deuterated C 1-4 Alkyl; n is 0, 1, 2, 3 or 4; Ring D is a benzene ring, a 5-6 membered heteroaryl ring or a 9-18 membered fused heterocyclic ring; R e Deuterium, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, ring C or halogen, wherein ring C can be 1 R d Substitution; the ring C is a 5-10 membered heteroaryl ring or an 8-10 fused heterocyclic ring containing 1-4 nitrogen atoms; the R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, deuterated C 1-4 Alkyl, C 3-6 Cycloalkyl; l is 1, 2, 3 or 4; p is 1, 2, 3 or 4;

[0013] L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene or chemical bond;

[0014] When ring A is a 5-6 membered heteroaryl ring, the structural unit Not for

[0015] When ring A is a 5-6 membered heterocyclic ring, the structural unit Not for

[0016] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof, characterized in that ring D is

[0017] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof, characterized in that ring D is

[0018] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof, characterized in that ring D is selected from

[0019] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof are characterized in that R e -CF3,

[0020] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof are characterized in that R e It is -OCH3, -F, -D, -CH3.

[0021] In some embodiments of the present application, the compound of formula (II'), its isomers and pharmaceutically acceptable salts thereof are characterized in that R e Selected from -CF3.

[0022] The present application also provides a compound represented by formula (I'), its isomers and pharmaceutically acceptable salts thereof:

[0023] in,

[0024] X a C or N;

[0025] X b 、X c 、X d Each independently represents CH, N or CR', R' represents halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy; Ring A is a benzene ring, a 5-6 membered heteroaryl ring, C 5-6 Cycloalkyl ring or 5-6 membered heterocyclyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C 1-4 alkyl)2P(O)-, The C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 alkyl;

[0026] m is 0, 1, 2, 3 or 4;

[0027] R b For hydrogen, C 1-4 alkyl;

[0028] Ring B is a benzene ring, a 5-10 membered heteroaryl ring, or a 5-10 membered heterocyclyl ring; R c Each independently is halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkyl, C 1-4 Haloalkoxy; n is 0, 1, 2, 3 or 4;

[0029] Ring C is a 5-10 membered heteroaryl ring containing 1-4 nitrogen atoms; R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, deuterated C 1-4 Alkyl; l is 0, 1, 2, 3 or 4;

[0030] L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene or chemical bond.

[0031] When ring A is a 5-6 membered heteroaryl group, the structural unit Not for

[0032] When ring A is a 5-6 membered heterocyclic group, the structural unit Not for

[0033] The present application also provides a compound represented by formula (I'-1), its isomers and pharmaceutically acceptable salts thereof,

[0034] in,

[0035] X b 、X c 、X d Each independently represents CH, N or CR', R' represents halogen, C 1-4 alkoxy;

[0036] Ring A is a benzene ring, pyridine or C 5-6 Cycloalkyl ring; R a Each independently is halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4 Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C 1-4 alkyl)2P(O)-, The C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 Alkyl; m is 0, 1, 2, 3 or 4;

[0037] R b For hydrogen, C 1-4 alkyl;

[0038] Ring B is a 5-6 membered heteroaryl ring or a 5-10 membered heterocyclyl ring; R c Each is independent of C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, halogen; n is 0, 1, 2, 3 or 4;

[0039] Ring C is a 5-6 membered heteroaryl ring containing 1-4 nitrogen atoms; R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy; l is 0, 1, 2, 3 or 4;

[0040] L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene.

[0041] In some embodiments of the present application, the compound represented by formula (I'-1), its isomers and pharmaceutically acceptable salts thereof,

[0042] in,

[0043] X b 、X c 、X d Each independently represents CH, N or CR', R' represents halogen, C 1-4 alkoxy;

[0044] Ring A is a benzene ring, pyridine or C 5-6 Cycloalkyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, cyano, amino, aminosulfonyl, methylsulfonyl, -COOC 1-4 Alkyl, -C 1-4 Alkyl-OH, carbamoyl; m is 0, 1, 2, 3 or 4;

[0045] R b For hydrogen, C 1-4 alkyl;

[0046] Ring B is a 5-6 membered heteroaryl ring; R c Each is independent of C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, halogen; n is 0, 1, 2, 3 or 4;

[0047] Ring C is a 5-6 membered heteroaryl ring containing 1-4 nitrogen atoms; R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy; l is 0, 1, 2, 3 or 4;

[0048] L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene.

[0049] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that ring A is a benzene ring or a 5-6 membered heteroaryl ring.

[0050] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that ring A is

[0051] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring A is

[0052] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring A is

[0053] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring A is

[0054] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring A is selected from

[0055] In some embodiments of the present application, the compounds of the above formulae (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R' is independently -OCH3 or -F.

[0056] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R a Each is independently -F, -OCH3, -CF 3、 -COOCH3, -C(CH3)2-OH, -CHF2, -CN, -Cl, -NH2, CH3-NH-S(O)2-, NH2-S(O)2-.

[0057] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that Ra Each independently represents -F, -OCH3, and -CF3.

[0058] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R a Each independently represents -COOCH3, -C(CH3)2-OH, and -CHF2.

[0059] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R a Each independently is -CH3, -OH, -Br, -CH2CH3, -OCH(CH3)2, D. -CD3,

[0060] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0061] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0062] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0063] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0064] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0065] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from

[0066] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0067] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0068] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0069] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0070] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0071] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0072] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0073] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Where a represents the connection with ring B, and b represents the connection with L1.

[0074] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0075] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from

[0076] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0077] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0078] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0079] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0080] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0081] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0082] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Wherein a represents the connection with ring B, and b represents the connection with L1.

[0083] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0084] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0085] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0086] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0087] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

[0088] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0089]

[0090] Wherein a represents the connection with ring B, and b represents the connection with L1.

[0091] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Wherein a represents the connection with ring B, and b represents the connection with L1.

[0092] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Wherein a represents the connection with ring B, and b represents the connection with L1.

[0093] In some embodiments of the present application, the compounds of formula (II') and (I'), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Wherein a represents the connection with ring B, and b represents the connection with L1.

[0094] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Where a represents the connection with ring B, and b represents the connection with L1.

[0095] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Where a represents the connection with ring B, and b represents the connection with L1.

[0096] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for Where a represents the connection with ring B, and b represents the connection with L1.

[0097] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0098] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0099] In some embodiments of the present application, the compound of formula (I'-1), its isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit Selected from Where a represents the connection with ring B, and b represents the connection with L1.

[0100] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R b For hydrogen.

[0101] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring B is

[0102] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring B is

[0103] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring B is

[0104] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring B is

[0105] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R c Each is independently CH3O-, Cl-, CH(CH3)2-, -OCHF2, -CF3.

[0106] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R c Each is independently -OCH3, -CH(CH3)2, -Cl.

[0107] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R c Each is independent -CH3, -OCD3,.

[0108] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0109] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0110] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0111] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0112] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring C is

[0113] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring C is

[0114] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring C is

[0115] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring C is

[0116] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that ring C is

[0117] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R d Each independently represents -CF3, -CH3, -CH2CH3, -CH(CH3)2, -OCH2CH3, and -CD3.

[0118] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R d Each independently represents -CF3, -CH3, and -CH(CH3)2.

[0119] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R d Each independently represents -CH2CH3.

[0120] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R d Each is independently -Cl,

[0121] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that R d are independently Br and -F.

[0122] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0123] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0124] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0125] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0126] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized by the structural unit for

[0127] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof, are characterized in that L1 is -CH2-, -CH(CH3)-, -C(CH3)2-,

[0128] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that L1 is -CH2-, -CH(CH3)-, or -C(CH3)2-.

[0129] In some embodiments of the present application, the compounds of formula (II'), (I') and (I'-1), their isomers and pharmaceutically acceptable salts thereof are characterized in that L1 is -CH(C2H5)- or a chemical bond.

[0130] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0131] where R a 、R c 、R d 、R e , L1, m are the same as defined above in this application; X is selected from C, N or O.

[0132] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0133] where R a 、R c 、R d , L1, and m are the same as defined above in this application.

[0134] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0135] where R a 、R c 、R d , m are the same as defined above in this application.

[0136] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0137] where R a 、R c 、R d 、R e , m are the same as defined above in this application; X is selected from C, N or O.

[0138] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0139] where R a 、R c 、Rd 、R e , m are the same as defined above in this application.

[0140] The present application also provides the following compounds, their isomers and pharmaceutically acceptable salts thereof:

[0141] The second aspect of the present application provides a method for preparing the compound:

[0142] Preparation method 1

[0143] Among them, ring A, X b 、X c 、Ring C 、 R b 、R c 、R d As defined above in this application;

[0144] Compound 1' is obtained by reacting SM1 and SM2 in the presence of a base and a catalyst, wherein the base is selected from sodium carbonate, potassium carbonate, K3PO4, Na2CO3, CsF, Cs2CO3, t-Bu-Na, etc., and the catalyst is selected from Pd(PPh3)4, Pd(dppf)Cl2, etc.;

[0145] Compound 2' is obtained by reacting compound 1' with 1,2-bis(diphenylphosphino)ethane;

[0146] Compound 3' is obtained by a substitution reaction between compound 2' and SM3 under alkaline conditions, wherein the base is selected from sodium hydride, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, potassium tert-butoxide, etc.;

[0147] Compound 4' is obtained by coupling reaction of compound 3' and SM4 under base and catalyst conditions, wherein the base is selected from sodium carbonate, potassium carbonate, K3PO4, Na2CO3, CsF, Cs2CO3, t-Bu-Na, etc., and the catalyst is selected from Pd(PPh3)4, Pd(dppf)Cl2, etc.

[0148] Preparation method 2

[0149] Among them, X b 、X c , Ring C, R c 、R d , Ra is as defined in the present application above;

[0150] Compound 5' is obtained by reacting SM5 in the presence of a base and a catalyst, wherein the base is selected from sodium carbonate, potassium carbonate, K3PO4, Na2CO3, CsF, Cs2CO3, t-Bu-Na, etc., and the catalyst is selected from Pd(PPh3)4, Pd(dppf)Cl2, etc.;

[0151] Compound 6' is obtained by reacting compound 5' with 1,2-bis(diphenylphosphino)ethane;

[0152] Compound 7' is obtained by a substitution reaction between compound 6' and SM3 under alkaline conditions, wherein the base is selected from sodium hydride, sodium hydroxide, potassium hydroxide, sodium tert-butoxide, potassium tert-butoxide, etc.;

[0153] Compound 8' is obtained by reacting compound 7' in the presence of a base and a catalyst, wherein the base is selected from sodium carbonate, potassium carbonate, K3PO4, Na2CO3, CsF, Cs2CO3, t-Bu-Na, etc., and the catalyst is selected from Pd(PPh3)4, Pd(dppf)Cl2, etc.;

[0154] The third aspect of the present application provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound, its isomer or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0155] A "pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering a biologically active agent to an animal, particularly a mammal, and includes, for example, adjuvants, excipients, or vehicles, such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants, and dispersants, depending on the mode of administration and the nature of the dosage form. Pharmaceutically acceptable carriers are formulated within the purview of those skilled in the art based on a wide range of factors. These include, but are not limited to, the type and nature of the active agent being formulated, the subject to whom the composition containing the agent is to be administered, the intended route of administration of the composition, and the intended therapeutic indication. Pharmaceutically acceptable carriers include both aqueous and non-aqueous media, as well as a variety of solid and semisolid dosage forms. In addition to the active agent, such carriers include a variety of different ingredients and additives, and the inclusion of such additional ingredients in a formulation for various reasons (e.g., to stabilize the active agent, binders, etc.) is well known to those skilled in the art.

[0156] The fourth aspect of the present application provides the use of the above-mentioned compound, its isomer or pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a drug for treating diseases mediated by the USP1 target.

[0157] In some embodiments of the present application, the related diseases mediated by the USP1 target include cellular inflammatory diseases, neurodegenerative diseases, and cancer.

[0158] Technical Effects

[0159] The compound of the present application has obvious USP1 enzyme inhibitory activity and can be used for the treatment of cancer.

[0160] Description and Definition

[0161] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered ambiguous or unclear without a specific definition, but should be understood according to its ordinary meaning.

[0162] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0163] The term "pharmaceutically acceptable salt" refers to derivatives of the compounds of the present invention prepared with relatively nontoxic acids or bases. These salts can be prepared during compound synthesis, isolation, and purification, or by reacting the purified free form of the compound with a suitable acid or base. When the compound contains relatively acidic functional groups, base addition salts are obtained by reaction with alkali metal or alkaline earth metal hydroxides or organic amines. These salts include cations based on alkali and alkaline earth metals, as well as nontoxic ammonium, quaternary ammonium, and amine cations, and also encompass salts of amino acids. When the compound contains relatively basic functional groups, acid addition salts are obtained by reaction with organic or inorganic acids.

[0164] The term "excipient" generally refers to a carrier, diluent and / or vehicle required to formulate an effective pharmaceutical composition.

[0165] The term "effective prophylactic or therapeutic amount" refers to a sufficient amount of a compound of the present application or a pharmaceutically acceptable salt thereof to treat a disorder at a reasonable benefit / risk ratio applicable to any medical treatment and / or prevention. However, it should be recognized that the total daily dosage of the compound represented by Formula II' of the present application or its pharmaceutically acceptable salt and composition must be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific therapeutically effective dosage level must be determined based on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the administration time, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field. For example, it is common practice in the art to start the dose of the compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved.

[0166] Unless otherwise specified, the term "5-10 membered heteroaryl" refers to a monocyclic or bicyclic group consisting of 5-10 ring atoms with a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). 5-6 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or carbon atom. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1, 2, 3-triazolyl, 2H-1, 2, 3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl), pyridyl, pyrimidinyl, benzimidazolyl, etc.

[0167] Unless otherwise specified, the term "fused heterocyclic group" refers to a saturated or partially saturated non-aromatic cyclic group formed by two or more cyclic structures sharing two adjacent atoms, containing at least one ring atom that is a heteroatom; the heteroatom is generally selected from N, O, and S; the ring carbon atoms and heteroatoms in the fused heterocyclic ring may be further oxidized to form a cyclic group containing C(O), NO, SO, or S(O)2 groups, which is also included in the definition of heterocyclic group in this application. The "non-aromatic" mentioned in this definition means that the group does not have aromaticity when it exists independently. The 11-14 membered fused heterocyclic group described in the present application includes “11-14 membered saturated fused heterocyclic group” and “11-14 membered partially saturated fused heterocyclic group”; the fusion mode can be 5-6 membered heterocyclic group and 5-6 membered heterocyclic group, 5-6 membered heterocyclic group and 5-6 membered cycloalkyl group, benzo 5-6 membered heterocyclic group, benzo 5-6 membered saturated heterocyclic group, 5-6 membered heteroaryl and 5-6 membered heterocyclic group, 5-6 membered heteroaryl and 5-6 membered saturated heterocyclic group, benzo 5-6 membered heterocyclic group and 5-6 membered heterocyclic group, 5-6 membered heteroaryl and 5-6 membered heterocyclic group and 5-6 membered heterocyclic group, benzo 5-6 membered cycloalkyl and 5-6 membered heterocyclic group, 5-6 membered heteroaryl and 5-6 membered cycloalkyl and 5-6 membered heterocyclic group; specific examples of the fused heterocyclic group include but are not limited to:

[0168] Unless otherwise specified, the term "heterocyclic group" refers to a substituted or unsubstituted saturated or unsaturated non-aromatic ring and contains 1-3 heteroatoms selected from N, O or S. Any carbon atom group or heteroatom in the heterocyclic group may be oxo-substituted, for example, a carbon atom group forms -C(O)-. The "heterocyclic group" described in this application refers to a non-aromatic cyclic group derived from at least one heteroatom as a ring atom and removing a hydrogen atom; including saturated or partially saturated monocyclic heterocyclic groups; the heterocyclic group is independent of the connection position (i.e., it can be bound through a carbon atom or a heteroatom). Examples of "heterocyclic groups" include but are not limited to

[0169] Unless otherwise specified, "cycloalkyl" refers to a saturated monocyclic or polycyclic hydrocarbon group. 3-12 Cycloalkyl, more preferably C 3-8 Cycloalkyl, more preferably C 5-6 Cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopentyl and cyclohexyl.

[0170] Unless otherwise specified, "cycloalkylene" refers to a divalent group formed by removing a hydrogen atom from a cycloalkyl group.

[0171] Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine or iodine atom.

[0172] Unless otherwise specified, the term “C 1-4 "Alkyl" is used to represent C 1-4 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, and the like.

[0173] Unless otherwise specified, "alkylene" refers to a saturated straight or branched aliphatic hydrocarbon group, which is a residue derived from the removal of two hydrogen atoms from the same carbon atom or two different carbon atoms of a parent alkane, and is a straight or branched group containing 1 to 20 carbon atoms, preferably 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, more preferably 1 to 4 carbon atoms. Alkylene includes, but is not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), and the like.

[0174] Unless otherwise specified, the term “C 1-4 "Haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by a halogen atom. Examples include, but are not limited to, monofluoromethyl, difluoromethyl, trifluoromethyl, trichloromethyl, tribromomethyl, 2,2,2-trifluoroethyl, 2,2,2-trichloroethyl, and the like.

[0175] Unless otherwise specified, the term “C 1-4 "Alkoxy" refers to a C 1-4 Alkyl compounds include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, and tert-butoxy.

[0176] Unless otherwise specified, the term "-C 1-4 Alkyl-OH refers to a C 1-4 Alkyl groups, examples of which include, but are not limited to, -CH2-OH, -(CH2)2-OH, -CH(CH3)2-OH.

[0177] Unless otherwise specified, “C 3-6 "Cycloalkyl" refers to a 3-6 membered monocyclic alkyl group, examples of which include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0178] Unless otherwise specified, the term "5-6 membered heteroaryl containing 1-4 nitrogen atoms" refers to a ring in which the carbon atoms are replaced by 1, 2, 3 or 4 nitrogen atoms. Examples include but are not limited to

[0179] Unless otherwise specified, In the structure It can be a single bond or a double bond. BRIEF DESCRIPTION OF THE DRAWINGS

[0180] The drawings described herein are used to provide further understanding of the present application and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application and do not constitute improper limitations on the present application.

[0181] Figure 1 shows the tumor growth of mice in groups G1 to G4 in Test Example 5 of the present application;

[0182] FIG2 shows the changes in body weight of mice in groups G1 to G4 in Test Example 5 of the present application. DETAILED DESCRIPTION

[0183] To make the objectives, technical solutions, and advantages of this application more clearly understood, the present application is further described in detail below with reference to the accompanying drawings and examples. It is apparent that the described examples are only a portion of the embodiments of this application, rather than all of them. All other embodiments obtained by those skilled in the art based on this application are within the scope of protection of this application.

[0184] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0185] The solvents used in this application are commercially available.

[0186] The structures of the compounds described herein are determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS), or ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker Neo 400M or Bruker Ascend 400 NMR instrument, using deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), deuterated chloroform (CDCl3), and heavy water (D2O) as the internal standard, with tetramethylsilane (TMS) as the internal standard.

[0187] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1260-6125B single quadrupole mass spectrometer (electrospray ionization as the ion source).

[0188] Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters UPLC H-class SQD mass spectrometer (electrospray ionization as the ion source).

[0189] HPLC analysis was performed using Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high performance liquid chromatography.

[0190] Preparative HPLC was performed using a Waters 2555-2489 (10 μm, ODS 250 cm×5 cm) or a GILSON Trilution LC.

[0191] Chiral HPLC was performed using Waters Acquity UPC2 and a Daicel chi-ring Clpak AD-H column (5 μm, 4.6×250 mm).

[0192] Supercritical fluid chromatography (SFC) was performed using a Waters SFC 80Q.

[0193] The starting materials in the examples of the present application are known and can be purchased on the market, or can be synthesized by or according to methods known in the art.

[0194] Unless otherwise specified, all reactions in this application were carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, the solvent was a dry solvent, and the reaction temperature was expressed in degrees Celsius (°C).

[0195] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any adverse way. The compound of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining the specific embodiments with other chemical synthesis methods, and equivalent replacement modes well known to those skilled in the art, and preferred embodiments include but are not limited to the examples of the present invention. It will be apparent to those skilled in the art that various changes and improvements will be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0196] 1. Preparation method

[0197] As used herein, room temperature refers to about 20-30°C.

[0198] Intermediate INT-1: (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol

[0199] Step A: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (144 g, 0.54 mol) and sodium acetate (80 g, 0.97 mol) in water (320 mL) at room temperature. The solution is then heated to 90°C and stirred for 0.5 hours. The reaction mixture is then cooled to 0°C, and a mixture of methyl 4-formylbenzoate (80 g, 0.49 mol) and 28 wt% ammonia (400 mL) in methanol (120 mL) is slowly added dropwise to the reaction mixture. The reaction mixture is stirred at room temperature for 16 hours.

[0200] After LCMS monitoring indicated the disappearance of the starting material, the reaction mixture was filtered, and the filter cake was rinsed with ethyl acetate (100 mL x 3). The filtrate was then collected and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 104 g of methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0201] MS (ESI) M / Z: 271.0 [M+H] + .

[0202] Step B: Dissolve methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (69 g, 0.25 mol) and cesium carbonate (250 g, 0.76 mol) in acetonitrile (1.2 L) at room temperature and stir for 2 hours. Then, add 2-iodopropane (64.9 g, 0.38 mol) to the solution, heat the reaction to 50°C, and stir for 24 hours. Then, cool the reaction to 0°C and add 2-iodopropane (21.6 g, 0.13 mol). The reaction is then stirred at 50°C for 4 hours.

[0203] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (1.5 L) to quench the mixture. The mixture was extracted with ethyl acetate (500 mL × 3 times), and the organic phases were combined and washed with saturated brine (150 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 33 g of methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0204] MS (ESI) M / Z: 313.2 [M+H] + .

[0205] Step C: Methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (33 g, 0.11 mol) was dissolved in dry tetrahydrofuran (528 mL) at room temperature under nitrogen. A 2.5 M (mol / L) lithium aluminum hydride solution (85 mL, 0.22 mol) was then slowly added dropwise to the solution at 0°C. The reaction was then stirred at room temperature for 1 hour.

[0206] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (1 L) to quench the reaction mixture, filtered, and the filter cake washed with ethyl acetate (100 mL x 3). The filtrate was extracted with ethyl acetate (500 mL x 3), and the organic phases were combined and washed with saturated brine (150 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 28 g of (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol.

[0207] MS (ESI) M / Z: 285.1 [M+H] + .

[0208] 1 H NMR (400MHz, CDCl3): δ7.47-7.32(m,5H),4.72(s,2H),4.59-4.46(m,1H),2.97(br,1H),1.44(d,J=6.8Hz,6H).

[0209] Intermediate INT-2: 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H imidazole

[0210] Step A: Dissolve (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (700 mg, 2.46 mmol) in dichloromethane (12.5 mL) at room temperature. Then, triphenylphosphine (1.29 g, 4.93 mmol), sodium bicarbonate (414 mg, 4.93 mmol), and carbon tetrabromide (1.63 g, 4.93 mmol) were added to the solution in sequence. The reaction was stirred at room temperature for 2 hours.

[0211] After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with dichloromethane (10 mL x 3 times). The organic phases were combined and washed with saturated brine (60 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 820 mg of 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole.

[0212] MS (ESI) M / Z: 347.2 [M+H] + .

[0213] Intermediate INT-3: (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid

[0214] Step A: Dissolve 4-chloro-6-methoxypyrimidine (25 g, 172.94 mmol), cyclopropylboronic acid (25.25 g, 294 mmol), 1,1'-bis(diphenylphosphinoferrocenedichloridopalladium) (6.3 g, 8.68 mmol), potassium phosphate (73.42 g, 345.88 mmol), and silver oxide (20.04 g, 86.47 mmol) in 1,4-dioxane (868 mL) at room temperature under nitrogen. The solution was then heated to 90°C and stirred for 16 hours.

[0215] After LCMS monitoring showed the disappearance of the starting material, the product was filtered while hot and the filtrate was concentrated under reduced pressure. Water (500 mL) was added to the resulting residue to quench the mixture, and the mixture was extracted with dichloromethane (100 mL × 3 times). The organic phases were combined and washed with saturated brine (150 mL × 2 times). The product was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 18.3 g of 4-cyclopropyl-6-methoxypyrimidine.

[0216] MS (ESI) M / Z: 151.2 [M+H] + .

[0217] Step B: Dissolve 4-cyclopropyl-6-methoxypyrimidine (45 g, 299.64 mmol) in ethanol (1.5 L) at -20°C under nitrogen. Then, slowly add liquid bromine (240 g, 1.5 mol) dropwise to the solution. The reaction mixture is stirred at room temperature for 16 hours.

[0218] After LCMS monitoring showed that the starting material disappeared, the reaction solution was concentrated by distillation under reduced pressure. The crude product was slurried with ethyl acetate, filtered, and the filter cake was collected. The filter cake was then added to water (150 mL) and stirred at 0 ° C. Saturated sodium bicarbonate solution was added dropwise until the solution system pH reached 7. Finally, the mixed solution was filtered, the filter cake was rinsed with water (150 mL × 2 times), the filter cake was collected, and concentrated under reduced pressure to obtain 56 g of 5-bromo-4-cyclopropyl-6-methoxypyrimidine.

[0219] MS (ESI) M / Z: 230.1 [M+H] + .

[0220] 1 H NMR (400MHz, DMSO-d6) δ8.53(s,1H),3.99(s,3H),2.50-2.44(m,1H),1.16-1.09(m,2H),1.08-1.02(m,2H).

[0221] Step C: Dissolve 5-bromo-4-cyclopropyl-6-methoxypyrimidine (15.0 g, 65.79 mmol) and triisopropyl borate (16.08 g, 85.53 mmol) in toluene / tetrahydrofuran (150 mL / 45 mL) at room temperature under nitrogen. Cool the reaction mixture to -78°C, stir for 30 minutes, and slowly add 2.5 M n-butyllithium solution (34.2 mL, 85.53 mmol) dropwise. Stir for 30 minutes. Then, warm the reaction mixture to -20°C and continue stirring for 1 hour.

[0222] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (100 mL) to quench the reaction. The mixture was then filtered and the filter cake was rinsed with water (20 mL × 3 times) to obtain 9.7 g (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid.

[0223] MS (ESI) M / Z: 195.0 [M+H] + .

[0224] Intermediate INT-4: 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H imidazole

[0225] Step A: Dissolve methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (5 g, 18.45 mmol) and potassium carbonate (5.1 g, 36.7 mmol) in N,N-dimethylformamide (92 mL) at room temperature. Cool the reaction mixture to 0°C and slowly add iodomethane (3.14 g, 22.14 mmol) dropwise. Stir the reaction mixture at room temperature for 1 hour.

[0226] After LCMS monitoring showed the disappearance of the starting material, water (200 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (60 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3.2 g of methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0227] MS (ESI) M / Z: 285.0 [M+H] + .

[0228] Step B: Dissolve methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (3.2 g, 11.26 mmol) in tetrahydrofuran (28 mL) at room temperature. Then, cool the reaction system to 0°C and slowly add lithium aluminum tetrahydride (2.14 g, 56.32 mmol) dropwise. Stir the reaction system at room temperature for 2 hours.

[0229] After LCMS monitoring showed the disappearance of the starting material, ice water (100 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (60 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2.6 g of (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol.

[0230] MS (ESI) M / Z: 257.0 [M+H] + .

[0231] Step C: At room temperature, under nitrogen, dissolve (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (2.6 g, 10.12 mmol), sodium bicarbonate (1.7 g, 20.24 mmol), and triphenylphosphine (5.3 g, 20.24 mmol) in dichloromethane (51 mL). The reaction mixture was then cooled to 0°C and carbon tetrabromide (6.7 g, 20.24 mmol) was slowly added. The reaction mixture was then allowed to warm to room temperature and stirred for 2 hours.

[0232] After LCMS monitoring showed the disappearance of the starting material, ice water (100 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with dichloromethane (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (80 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2.4 g of (2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole.

[0233] MS (ESI) M / Z: 319.0 [M+H] + .

[0234] Intermediate INT-5: 2-(4-(bromomethyl)phenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole

[0235] Step A: Dissolve methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (700 mg, 2.59 mmol) and sodium hydride (156 mg, 3.89 mmol) in dry N,N-dimethylformamide (13 mL) at 0°C under nitrogen and stir for 30 minutes. Then, slowly add iodoethane (606.6 mg, 3.89 mmol) dropwise to the mixture. Warm the reaction mixture to room temperature and continue stirring for 1 hour.

[0236] After LCMS monitoring showed the disappearance of the starting material, ice water (60 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 300 mg of methyl 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0237] MS (ESI) M / Z: 299.0 [M+H] + .

[0238] Step B: In an ice-water bath under nitrogen, dissolve methyl 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (300 mg, 1.00 mmol) in dry tetrahydrofuran (5 mL). Then, slowly add lithium aluminum hydride (0.8 mL, 2.00 mmol) to the reaction mixture. Warm the reaction mixture to room temperature and stir for 2 hours.

[0239] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction mixture to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (10 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 210 mg of (4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol.

[0240] MS (ESI) M / Z: 271.0 [M+H] + .

[0241] Step C: In an ice-water bath under nitrogen, (4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (200 mg, 0.74 mmol), triphenylphosphine (390 mg, 1.48 mmol), sodium bicarbonate (125 mg, 1.48 mmol), and carbon tetrabromide (390 mg, 1.48 mmol) were dissolved in dichloromethane (3.7 mL). The reaction was then stirred at room temperature for 1 hour.

[0242] After LCMS monitoring showed the disappearance of the starting material, water (50 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 230 mg of 2-(4-(bromomethyl)phenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole.

[0243] MS (ESI) M / Z: 333.0 [M+H] + .

[0244] Intermediate INT-6: 2-chloro-9H-pyrimido[4,5-b]indole

[0245] Step A: Dissolve 1-bromo-2-nitrobenzene (500 mg, 2.49 mmol), (2-chloropyrimidin-5-yl)boronic acid (590 mg, 3.74 mmol), sodium carbonate (792 mg, 7.47 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (305 mg, 0.37 mmol) in 1,4-dioxane / water (11 mL / 1.4 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C under nitrogen for 3 hours.

[0246] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (20 mL) for quenching. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 500 mg of 2-chloro-5-(2-nitrophenyl)pyrimidine.

[0247] MS (ESI) M / Z: 236.0 [M+H] + .

[0248] Step B: 2-Chloro-5-(2-nitrophenyl)pyrimidine (500 mg, 2.13 mmol) and 1,2-bis(diphenylphosphino)ethane (1.06 g, 2.66 mmol) were dissolved in 1,2-dichlorobenzene (7.1 mL) at room temperature and the reaction system was stirred at 160°C for 1 hour.

[0249] After LCMS monitoring showed the disappearance of the starting material, ice water (10 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 200 mg of 2-chloro-9H-pyrimidin[4,5-b]indole.

[0250] MS (ESI) M / Z: 204.0 [M+H] + .

[0251] Intermediate INT-7: 2-chloro-5,6,7,8-tetrahydrocyclopentyl[4,5]pyrrolo[2,3-d]pyrimidine

[0252] Step A: Dissolve 4-amino-5-bromo-2-chloropyrimidine (1.0 g, 4.83 mmol), 1-cyclopentenylboronic acid pinacol ester (1.4 g, 7.25 mmol), potassium phosphate (2.6 g, 12.08 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (389 mg, 0.48 mmol) in 1,4-dioxane / water (15 mL / 3 mL) at room temperature. Evacuate the air and replace it with nitrogen three times. Stir the reaction system at 90°C under nitrogen for 3 hours.

[0253] After TLC monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (20 mL) for quenching. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 600 mg of 2-chloro-5-(cyclopentyl-1-en-1-yl)pyrimidin-4-amine.

[0254] MS (ESI) M / Z: 196.1 [M+H] + .

[0255] 1 H NMR (400MHz, DMSO-d6) δ7.91 (s, 1H), 7.29 (br, 2H), 6.09 (t, J = 2.2Hz, 1H), 2.65-2.57 (m, 2H), 2.49-2.43 (m, 2H), 1.91 (p, J = 7.5Hz, 2H).

[0256] Step B: 2-chloro-5-(cyclopentyl-1-en-1-yl)pyrimidin-4-amine (100 mg, 0.51 mmol) was dissolved in tetrahydrofuran (2 mL) and water (1 mL) at 0°C, and N-bromosuccinimide (100 mg, 0.56 mmol) was added. The mixture was stirred for 1 hour. 2M aqueous sodium hydroxide solution (0.2 mL) was then added and stirring continued for 30 minutes.

[0257] After TLC monitoring showed the disappearance of the starting material, ice water (10 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. Tetrahydrofuran (3 mL) was added to the residue, followed by boron trifluoride etherate (217 mg, 1.53 mmol), and stirred at 60°C for 3 hours.

[0258] After TLC monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding saturated aqueous sodium bicarbonate (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 60 mg of 2-chloro-5,6,7,8-tetrahydrocyclopentyl[4,5]pyrrolo[2,3-d]pyrimidine.

[0259] MS (ESI) M / Z: 194.1 [M+H] + .

[0260] 1H NMR (400MHz, DMSO-d6) δ12.19(s,1H),8.68(s,1H),2.87(t,J=7.1Hz,2H),2.78(t,J=7.1Hz,2H),2.48-2.39(m,2H).

[0261] Intermediate INT-8: 2-(4-(bromomethyl)-2-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)-1H imidazole

[0262] Step A: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (40.2 g, 0.15 mol) and sodium acetate (12.25 g, 0.15 mol) in water (36 mL) at room temperature. The solution was then heated to 90°C and stirred for 0.5 hours. The reaction mixture was then cooled to 0°C and a mixture of methyl 4-formyl-3-methoxybenzoate (10 g, 51.54 mmol), 28 wt% ammonia (45 mL), and methanol (135 mL) was slowly added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 16 hours.

[0263] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered, and the filter cake was rinsed with ethyl acetate / petroleum ether (v / v, 3 / 1). The filtrate was collected and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 10.85 g of methyl 3-methoxy-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0264] MS (ESI) M / Z: 301.0 [M+H] + .

[0265] Step B: Dissolve methyl 3-methoxy-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (6 g, 0.02 mol) and cesium carbonate (19.7 g, 0.06 mol) in acetonitrile (65 mL) at room temperature and stir for 2 hours. Then, add 2-iodopropane (5.1 g, 0.03 mol) to the solution, raise the temperature to 50°C, and stir for 24 hours. Then, cool the reaction system to 0°C, add 2-iodopropane (5.1 g, 0.03 mol), raise the temperature to 50°C, and continue stirring for 4 hours.

[0266] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (300 mL) to quench the mixture. The mixture was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined and washed with saturated brine (50 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 6.9 g of methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxybenzoate.

[0267] MS (ESI) M / Z: 343.0 [M+H] + .

[0268] Step C: Dissolve methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxybenzoate (2.9 g, 8.48 mmol) in dry tetrahydrofuran (30 mL) at 0°C under nitrogen. Then, slowly add lithium aluminum hydride solution (6.8 mL, 17 mmol) dropwise to the solution. The reaction mixture is stirred at room temperature for 1 hour.

[0269] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (100 mL) to quench the reaction mixture, filtered, and the filter cake rinsed with ethyl acetate (50 mL x 3). The filtrate was extracted with ethyl acetate (50 mL x 3), and the organic phases were combined and washed with saturated brine (150 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.95 g of (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxyphenyl)methanol.

[0270] MS (ESI) M / Z: 315.4 [M+H] + .

[0271] Step D: Dissolve (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxyphenyl)methanol (500 mg, 1.59 mmol), sodium bicarbonate (269 mg, 3.2 mmol), and triphenylphosphine (840 mg, 3.2 mmol) in dry dichloromethane (8 mL) at 0°C. Then, carbon tetrabromide (1.06 g, 3.2 mmol) was slowly added to the reaction mixture. The reaction was then stirred at room temperature for 1 hour.

[0272] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 350 mg of 2-(4-(bromomethyl)-2-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole.

[0273] MS (ESI) M / Z: 377.2 [M+H] + .

[0274] Intermediate INT-9: 2-(4-(bromomethyl)-2-fluoro-6-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)-1H imidazole

[0275] Step A: Dissolve 4-bromo-2-fluoro-6-methoxybenzaldehyde (4 g, 17 mmol), triethylamine (12.38 g, 0.12 mol), and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (1.34 g, 18 mmol) in methanol (120 mL) at room temperature. Then, replace the carbon monoxide with air three times under vacuum. The reaction mixture was heated to 90°C and stirred for 24 hours.

[0276] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 1.98 g of methyl 3-fluoro-4-formyl-5-methoxybenzoate.

[0277] MS (ESI) M / Z: 213.0 [M+H] + .

[0278] Step B: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (6.72 g, 25 mmol) and sodium acetate (2.06 g, 25 mmol) in water (7 mL) at room temperature. The solution was then heated to 90°C and stirred for 0.5 hours. The reaction system was then cooled to 0°C, and a mixture of methyl 3-fluoro-4-formyl-5-methoxybenzoate (1.98 g, 9 mmol), 28 wt% aqueous ammonia (17 mL), and methanol (45 mL) was slowly added dropwise to the reaction solution. The mixture was allowed to warm to room temperature and stirred for 16 hours.

[0279] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered, and the filter cake was rinsed with ethyl acetate / petroleum ether (v / v, 3 / 1). The filtrate was collected and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2.06 g of methyl 3-fluoro-5-methoxy-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0280] MS (ESI) M / Z: 319.0 [M+H] + .

[0281] Step C: Dissolve methyl 3-fluoro-5-methoxy-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (2.06 g, 6.5 mmol) and cesium carbonate (6.31 g, 19 mmol) in acetonitrile (25 mL) at room temperature and stir for 2 hours. Then, add 2-iodopropane (1.65 g, 9.7 mmol) to the solution, heat to 50°C, and stir for 24 hours. Then, cool the reaction system to 0°C, add 2-iodopropane (1 g, 5.9 mmol), heat to 50°C, and stir for 4 hours.

[0282] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (50 mL) to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (50 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 1.84 g of methyl 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-methoxybenzoate.

[0283] MS (ESI) M / Z: 361.0 [M+H] + .

[0284] Step D: Methyl 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-methoxybenzoate (1.84 g, 5.1 mmol) was dissolved in dry tetrahydrofuran (25 mL) at 0°C under nitrogen. Lithium aluminum hydride solution (4.2 mL, 0.01 mol) was then slowly added dropwise to the solution. The reaction was then stirred at room temperature for 1 hour.

[0285] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (100 ml) to quench the reaction mixture, filtered, and the filter cake rinsed with ethyl acetate (50 mL x 3 times). The filtrate was extracted with ethyl acetate (50 mL x 3 times), and the organic phases were combined and washed with saturated brine (150 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.38 g of (3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-methoxyphenyl)methanol.

[0286] MS (ESI) M / Z: 333.2 [M+H] + .

[0287] Step E: At 0°C under nitrogen, (3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-methoxyphenyl)methanol (500 mg, 1.5 mmol), sodium bicarbonate (269 mg, 3.2 mmol), and triphenylphosphine (840 mg, 3.2 mmol) were dissolved in dry dichloromethane (8 mL). Carbon tetrabromide (1.06 g, 3.2 mmol) was then slowly added to the reaction mixture. The reaction was then stirred at room temperature for 1 hour.

[0288] After LCMS monitoring showed the disappearance of the starting material, water (50 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 350 mg of 2-(4-(bromomethyl)-2-fluoro-6-methoxyphenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole.

[0289] MS (ESI) M / Z: 395.2 [M+H] + .

[0290] Intermediate INT-10: 4-chloro-1-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole

[0291] Step A: Dissolve 4-chloro-1H-pyrazole (5 g, 0.049 mol), 2-iodopropane (20 g, 0.118 mol), and cesium carbonate (33 g, 0.1 mol) in acetonitrile (40 mL) at room temperature. Stir the reaction system at 80°C for 2 hours.

[0292] After LCMS monitoring showed the disappearance of the starting material, ice water (100 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3.62 g of 4-chloro-1-isopropyl-1H-pyrazole.

[0293] MS (ESI) M / Z: 145.2 [M+H] + .

[0294] Step B: Dissolve 4-chloro-1-isopropyl-1H-pyrazole (2 g, 0.014 mol) in tetrahydrofuran (14 mL) at 0°C under nitrogen. Then, add n-butyllithium (11.2 mL, 0.017 mol) dropwise to the solution, warm to room temperature, and stir for 1 hour. Then, cool the reaction mixture to -78°C and slowly add 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.45 g, 0.018 mol) dropwise. Warm to room temperature and stir for 2 hours.

[0295] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with saturated aqueous ammonium chloride (40 mL). The mixture was extracted with ethyl acetate (30 mL x 3), and the organic phases were combined and washed with saturated brine (20 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2.4 g of 4-chloro-1-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0296] MS (ESI) M / Z: 271.0 [M+H] + .

[0297] Intermediate INT-11: 2-(4-(1-bromopropyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole

[0298] Step A: Methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (300 mg, 0.96 mmol) was dissolved in dry tetrahydrofuran (5 mL) at 0°C under nitrogen. Tetraisopropyl titanate (327 mg, 1.15 mmol) and ethylmagnesium bromide (0.46 mL, 1.15 mmol) were then added to the solution, and the reaction was stirred at room temperature for 16 hours.

[0299] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (30 mL) to quench the reaction, filtered, and the filtrate was extracted with ethyl acetate (20 mL × 3 times). The organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 220 mg of 1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)propan-1-ol.

[0300] MS (ESI) M / Z: 313.2 [M+H] + .

[0301] Step B: At 0°C under nitrogen, dissolve 1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)propan-1-ol (220 mg, 0.71 mmol), triphenylphosphine (372 mg, 1.42 mmol), and sodium bicarbonate (119 mg, 1.42 mmol) in dichloromethane (4 mL). Subsequently, carbon tetrabromide (471 mg, 1.42 mmol) was slowly added to the reaction mixture. The reaction system was then stirred at room temperature for 30 minutes.

[0302] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with dichloromethane (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 80 mg of 2-(4-(1-bromopropyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole.

[0303] MS (ESI) M / Z: 375.0 [M+H] + .

[0304] Intermediate INT-12: 8-(bromomethyl)-2-(trifluoromethyl)imidazo[2,1-a]isoquinoline

[0305] Step A: Methyl 2-(trifluoromethyl)imidazo[2,1-a]isoquinoline-8-carboxylate (52 mg, 0.18 mmol) was dissolved in tetrahydrofuran (0.9 mL) at room temperature. Lithium aluminum hydride (0.14 mL, 0.35 mmol) was then added to the solution at 0°C. The reaction was then stirred at room temperature for 1 hour.

[0306] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added to ice water (20 mL) to quench the reaction. The mixture was filtered, and the filtrate was extracted with ethyl acetate (10 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 32 mg of (2-(trifluoromethyl)imidazo[2,1-a]isoquinolin-8-yl)methanol.

[0307] MS (ESI) M / Z: 267.2 [M+H] + .

[0308] Step B: Dissolve (2-(trifluoromethyl)imidazo[2,1-a]isoquinolin-8-yl)methanol (53 mg, 0.20 mmol) in dichloromethane (1 mL) at room temperature. Triphenylphosphine (157.4 mg, 0.60 mmol), sodium bicarbonate (33.6 mg, 0.40 mmol), and carbon tetrabromide (199 mg, 0.60 mmol) were then added to the reaction system at 0°C. The reaction was then stirred at room temperature for 1.5 hours.

[0309] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding water (20 mL). The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (30 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 37.8 mg of 8-(bromomethyl)-2-(trifluoromethyl)imidazo[2,1-a]isoquinoline.

[0310] MS (ESI) M / Z: 329.0 [M+H] + .

[0311] Intermediate INT-13: (2-(trifluoromethyl)-6,7-dihydro-5H-imidazo[1,2-a]pyrrolo[2,1-c][1,4]diazepin-9-yl)methanol

[0312] Step A: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (17.5 g, 65.35 mmol) and sodium acetate (5.35 g, 65.35 mmol) in water (19.8 mL) at room temperature and stir at 90°C for 1 hour. Then, cool the reaction mixture to 0°C and add a solution of methyl 5-formyl-1H-pyrrole-2-carboxylate (10 g, 65.35 mmol) in aqueous ammonia / methanol (59.5 mL / 178 mL) and stir at room temperature for 18 hours.

[0313] After LCMS monitoring showed the disappearance of the starting material, the mixed solution was concentrated under reduced pressure, and ice water (200 mL) was added to the resulting concentrate. The mixed solution was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (90 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 5.9 g of methyl 5-(4-(trifluoromethyl)-1H-imidazol-2-yl)-1H-pyrrole-2-carboxylate.

[0314] MS (ESI) M / Z: 260.0 [M+H] + .

[0315] Step B: Dissolve methyl 5-(4-(trifluoromethyl)-1H-imidazol-2-yl)-1H-pyrrole-2-carboxylate (2.6 g, 10.25 mmol) and cesium carbonate (10 g, 30.76 mmol) in acetonitrile (50 mL) at room temperature and stir for 2 hours. Then, add 1,3-diiodopropane (4.5 g, 15.3 mmol) to the reaction mixture, heat to 50°C, and stir for 48 hours.

[0316] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. Ice water (100 mL) was added to the resulting residue to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times). The organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 1.4 g of methyl 2-(trifluoromethyl)-6,7-dihydro-5H-imidazo[1,2-a]pyrrolo[2,1-c][1,4]diazepine-9-carboxylate was obtained.

[0317] MS (ESI) M / Z: 300.1 [M+H] + .

[0318] Step C: Methyl 2-(trifluoromethyl)-6,7-dihydro-5H-imidazo[1,2-a]pyrrolo[2,1-c][1,4]diazepine-9-carboxylate (1.4 g, 4.68 mmol) was dissolved in tetrahydrofuran (23 mL) at room temperature. Lithium aluminum hydride (3.74 mL, 9.36 mmol) was then added to the solution at 0°C. The reaction was then stirred at room temperature for 0.5 hours.

[0319] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added to ice water (40 mL) to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride solution (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 680 mg of (2-(trifluoromethyl)-6,7-dihydro-5H-imidazo[1,2-a]pyrrolo[2,1-c][1,4]diazepin-9-yl)methanol.

[0320] MS (ESI) M / Z: 272.1 [M+H] + .

[0321] Intermediate INT-14: (2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrrolo[2,1-c]pyrazin-8-yl)methanol

[0322] Step A: Dissolve methyl 5-(4-(trifluoromethyl)-1H-imidazol-2-yl)-1H-pyrrole-2-carboxylate (2 g, 7.72 mmol) and cesium carbonate (7.54 g, 23.15 mmol) in acetonitrile (50 mL) at room temperature and stir for 2 hours. Then, add 1,2-dibromoethane (1.45 g, 7.72 mmol). The reaction mixture is stirred at 50°C for 48 hours.

[0323] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. Ice water (100 mL) was added to the resulting residue to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. 860 mg of methyl 2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrrolo[2,1-c]pyrazine-8-carboxylate was obtained.

[0324] MS (ESI) M / Z: 286.2 [M+H] + .

[0325] Step B: Methyl 2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrrolo[2,1-c]pyrazine-8-carboxylate (850 mg, 2.98 mmol) was dissolved in dry tetrahydrofuran (30 mL) at room temperature. Lithium aluminum hydride (2.24 mL, 4.47 mmol) was then added to the solution at 0°C. The reaction was stirred at room temperature for 0.5 hours.

[0326] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added to ice water (40 mL) to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride solution (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 590 mg of (2-(trifluoromethyl)-5,6-dihydroimidazo[1,2-a]pyrrolo[2,1-c]pyrazin-8-yl)methanol.

[0327] MS (ESI) M / Z: 258.2 [M+H] + .

[0328] 1H NMR (400MHz, DMSO-d6): δ7.82(d,J=1.2Hz,1H), 6.50(d,J=3.6Hz,1H), 6.11(d,J=3.6Hz, 1H),5.10(t,J=5.2Hz,1H),4.48(d,J=5.2Hz,2H),4.39-4.33(m,2H),4.32-4.276(m,2H).

[0329] Intermediate INT-15: 7-(Bromomethyl)-2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole

[0330] Step A: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (55.5 g, 205.72 mmol) in water (65 mL) at room temperature. Then, add sodium acetate (16.9 g, 205.72 mmol) to the solution, raise the temperature to 90°C, and stir for 1 hour. Then, cool the solution to 0°C and slowly add methyl 5-bromo-2-formylbenzoate (25 g, 102.86 mmol) dissolved in methanol (685 mL) and aqueous ammonia (150 mL) dropwise. Finally, stir the reaction system at 100°C for 2 hours.

[0331] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure, and water (100 mL) was added to the resulting residue to quench the mixture. The mixture was extracted with ethyl acetate (200 mL x 3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was slurried to yield 41 g of 5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoic acid.

[0332] MS (ESI) M / Z: 334.9 [M+H] + .

[0333] Step B: Dissolve 5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoic acid (36 g, 107.78 mmol) in dry tetrahydrofuran (500 mL) at 0°C. Then, slowly add a solution of borane in tetrahydrofuran (216 mL, 215.27 mmol) dropwise to the solution. The reaction mixture is stirred at room temperature for 2 hours.

[0334] After LCMS monitoring showed the disappearance of the starting material, water (200 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (300 mL x 3 times), and the organic phases were combined and washed with saturated brine (300 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 12.3 g of (5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol.

[0335] MS (ESI) M / Z: 321.0 [M+H] + .

[0336] Step C: Dissolve (5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (12.3 g, 38.44 mmol) in dichloromethane (192 mL) at 0°C. Triethylamine (10.7 mL, 76.88 mmol) and methanesulfonyl chloride (6.6 g, 57.66 mmol) were added sequentially to the solution. The reaction was stirred at room temperature for 1 hour.

[0337] After LCMS monitoring showed the disappearance of the starting material, water (200 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (200 mL × 3 times), and the organic phases were combined and washed with saturated brine (200 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 14.5 g of 5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl methanesulfonate.

[0338] MS (ESI) M / Z: 399.0 [M+H] + .

[0339] Step D: Dissolve 5-bromo-2-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl methanesulfonate (14.5 g, 42.90 mmol) in N,N-dimethylformamide (214 mL) at room temperature. Cesium carbonate (16.8 g, 51.48 mmol) was then added to the solution. The reaction was stirred at 50°C for 1 hour.

[0340] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding water (600 mL). The mixture was extracted with ethyl acetate (200 mL x 3), and the organic phases were combined and washed with saturated brine (200 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.83 g of 7-bromo-2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole and 500 mg of the isomer 7-bromo-3-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole.

[0341] MS (ESI) M / Z: 303.0 [M+H] + .

[0342] 1 H NMR (400MHz, DMSO-d6): δ8.15(d,J=0.8Hz,1H),7.92(d,J=0.8Hz,1H),7.75(d,J=8.0Hz,1H),7.70(dd,J=8.4,1.8Hz,1H),5.17(s,2H).

[0343] Step E: Dissolve 7-bromo-2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole (930 mg, 3.08 mmol) in methanol (35 mL) at room temperature. Then, triethylamine (4.2 mL, 30.8 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (251 mg, 0.31 mmol) were added sequentially. The reaction system was then stirred at 85°C under 40 kg of carbon monoxide pressure for 24 hours.

[0344] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 400 mg of methyl 2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole-7-carboxylate.

[0345] MS (ESI) M / Z: 283.1 [M+H] + .

[0346] Step F: Methyl 2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole-7-carboxylate (400 mg, 1.42 mmol) was dissolved in dry tetrahydrofuran (7 mL) at 0°C. Lithium aluminum hydride (1.13 mL, 2.84 mmol) was then slowly added dropwise to the solution. The reaction was then stirred at room temperature for 1 hour.

[0347] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was poured into ice water (20 mL) for quenching. The mixture was extracted with ethyl acetate (30 mL x 3), and the organic phases were combined and washed with saturated brine (20 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 120 mg of (2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindol-7-yl)methanol.

[0348] MS (ESI) M / Z: 255.1 [M+H] + .

[0349] Step G: Dissolve (2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindol-7-yl)methanol (120 mg, 0.47 mmol) in dichloromethane (2.4 mL) at 0°C. Triphenylphosphine (371 mg, 1.42 mmol), sodium bicarbonate (79 mg, 0.94 mmol), and carbon tetrabromide (471 mg, 1.42 mmol) were added sequentially to the solution. The reaction was stirred at room temperature for 1 hour.

[0350] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding water (10 mL). The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 100 mg of 7-(bromomethyl)-2-(trifluoromethyl)-5H-imidazo[2,1-a]isoindole.

[0351] MS (ESI) M / Z: 317.0 [M+H] + .

[0352] Intermediate INT-16: 9-(chloromethyl)-2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine

[0353] Reaction route:

[0354] Steps:

[0355] Step A: Dissolve 3-bromo-4-formylbenzoic acid (100 g, 436.7 mmol) in water (2.7 L) at room temperature. Add potassium carbonate (90.4 g, 655.1 mmol) and iodomethane (68.2 g, 480.4 mmol) sequentially to the solution. Stir the reaction for 2 hours.

[0356] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was poured into ice water to precipitate a solid, which was filtered and the resulting filter cake was dried to obtain 95 g of methyl 3-bromo-4-formylbenzoate.

[0357] MS (ESI) M / Z: 243.0 [M+H] + .

[0358] Step B: Dissolve 3,3-dibromo-1,1,1-trifluoropropane-2-one (188.7 g, 699.3 mmol) in water (340 mL) at room temperature. Add sodium acetate (57.5 g, 416.1 mmol) to the solution, heat to 90°C, and stir for 1 hour. Then, cool the reaction mixture to 0°C and add a mixture of methyl 3-bromo-4-formylbenzoate (85 g, 349.7 mmol) in methanol / ammonia (1020 mL / 340 mL) dropwise. Finally, stir the reaction mixture at 100°C for 2 hours.

[0359] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure and filtered. The resulting filter cake was slurried with petroleum ether / ethyl acetate to obtain 87 g of methyl 3-bromo-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0360] MS (ESI) M / Z: 349.0 [M+H] + .

[0361] Step C: Methyl 3-bromo-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (96.7 g, 277.0 mmol) was dissolved in N,N-dimethylformamide (185 mL) at 0°C under nitrogen. Potassium carbonate (80.7 g, 583.9 mmol) and 3-bromoprop-1-ene (40.3 g, 333.1 mmol) were then added to the solution. The reaction was stirred at 25°C for 20 hours.

[0362] After LCMS monitoring showed the disappearance of the starting material, water (800 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (300 mL × 3 times), and all organic phases were combined and washed with saturated brine (500 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 83.0 g of methyl 4-(1-allyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-bromobenzoate.

[0363] MS (ESI) M / Z: 389.0 [M+H] + .

[0364] Step D: Methyl 4-(1-allyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-bromobenzoate (107 g, 275.0 mmol) was dissolved in 1,4-dioxane / water (1370 mL / 228 mL) at 0°C under nitrogen. Potassium carbonate (76.2 g, 551.3 mmol), potassium trifluoro(vinyl)borate (96.1 g, 717.4 mmol), 2-dicyclohexylphosphino-2',6'-dimethoxybiphenyl (11.3 g, 27.5 mmol), and palladium acetate (6.2 g, 27.5 mmol) were then added to the solution. The reaction was stirred at 85°C for 4 hours.

[0365] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered, the filter cake was rinsed with ethyl acetate (150 mL x 2), the filtrate was collected, and the mixture was concentrated under reduced pressure. Water (300 mL) was added to the residue and diluted, and the mixture was extracted with ethyl acetate (700 mL x 3 times). All organic phases were combined and washed with saturated brine (400 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 48.4 g of methyl 4-(1-allyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-vinylbenzoate.

[0366] MS (ESI) M / Z: 337.2 [M+H] + .

[0367] Step E: Methyl 4-(1-allyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-vinylbenzoate (61.4 g, 182.6 mmol) was dissolved in dichloromethane (730 mL) at 0°C under nitrogen. (1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(o-isopropoxybenzylidene)ruthenium (8.5 g, 13.7 mmol) was then added to the solution. The reaction was stirred at 25°C for 3 hours.

[0368] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure and filtered. The filter cake was rinsed with dichloromethane, and the solid was collected and dried to obtain 39.0 g of methyl 2-(trifluoromethyl)-5H-benzo[c]imidazo[1,2-a]azepine-9-carboxylate.

[0369] MS (ESI) M / Z: 309.2 [M+H] + .

[0370] Step F: Methyl 2-(trifluoromethyl)-5H-benzo[c]imidazo[1,2-a]azepine-9-carboxylate (39.0 g, 125.8 mmol) and 10% palladium on carbon (8.0 g) were dissolved in methanol (132 mL) at room temperature and the reaction was stirred under a hydrogen atmosphere for 18 hours.

[0371] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered, and the filtrate was collected and concentrated under reduced pressure to obtain 37.2 g of methyl 2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine-9-carboxylate.

[0372] MS (ESI) M / Z: 311.2 [M+H] + .

[0373] Step G: Methyl 2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine-9-carboxylate (37.2 g, 120.0 mmol) was dissolved in tetrahydrofuran (600 mL) at 0°C under nitrogen. A 2.5 M lithium aluminum tetrahydride solution (72 mL) was then slowly added dropwise to the solution. The reaction was then stirred at room temperature for 30 minutes.

[0374] After LCMS monitoring showed the disappearance of the starting material, water (100 mL) was added to the reaction solution to quench the reaction. The mixture was filtered, the filter cake was rinsed with ethyl acetate (100 mL × 2 times), the filtrate was collected, and the layers were separated. The resulting aqueous phase was extracted with ethyl acetate (200 mL × 3 times), and all organic phases were combined and washed with saturated brine (300 mL × 2 times). The organic phase was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by beating to obtain 30 g of (2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine-9-yl)methanol.

[0375] MS (ESI) M / Z: 283.3 [M+H] + .

[0376] Step H: Dissolve (2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine-9-yl)methanol (30.0 g, 106.3 mmol) in 1,2-dichloroethane (530 mL) at 0°C under nitrogen. Then, slowly add thionyl chloride (37.9 g, 319 mmol) dropwise to the solution. The reaction mixture is stirred at 50°C for 20 minutes.

[0377] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure, and the residue was purified by beating to obtain 30.4 g of 9-(chloromethyl)-2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine.

[0378] MS (ESI) M / Z: 301.2 [M+H] + .

[0379] Example 1: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole

[0380] Reaction route:

[0381] Steps:

[0382] Step A: Dissolve 1-bromo-2-nitrobenzene (500 mg, 2.49 mmol), (2-chloropyrimidin-5-yl)boronic acid (590 mg, 3.74 mmol), sodium carbonate (792 mg, 7.47 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (305 mg, 0.37 mmol) in 1,4-dioxane / water (11 mL / 1.4 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C under nitrogen for 3 hours.

[0383] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (20 mL) for quenching. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 500 mg of 2-chloro-5-(2-nitrophenyl)pyrimidine.

[0384] MS (ESI) M / Z: 236.0 [M+H] + .

[0385] Step B: 2-Chloro-5-(2-nitrophenyl)pyrimidine (500 mg, 2.13 mmol) and 1,2-bis(diphenylphosphino)ethane (1.06 g, 2.66 mmol) were dissolved in 1,2-dichlorobenzene (7.1 mL) at room temperature and the reaction system was stirred at 160°C for 1 hour.

[0386] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (10 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 200 mg of 2-chloro-9H-pyrimido[4,5-b]indole.

[0387] MS (ESI) M / Z: 204.0 [M+H] + .

[0388] Step C: Dissolve 2-chloro-9H-pyrimido[4,5-b]indole (200 mg, 0.99 mmol) in tetrahydrofuran (5 mL) at room temperature under nitrogen. Add sodium hydride (47 mg, 1.18 mmol) to the solution at 0°C and stir for 30 minutes. Then, add 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (345 mg, 1.08 mmol). Stir the reaction at room temperature for another 2 hours.

[0389] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (10 mL). The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 300 mg of 2-chloro-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole.

[0390] MS (ESI) M / Z: 442.0 [M+H] + .

[0391] Step D: 2-Chloro-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole (70 mg, 0.16 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (47 mg, 0.24 mmol), sodium carbonate (34 mg, 0.32 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (19 mg, 0.02 mmol) were dissolved in 1,4-dioxane / water (3.2 mL / 0.4 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred in a microwave at 100°C for 3 hours.

[0392] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected and the solvent was removed under reduced pressure to obtain 15.62 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole.

[0393] MS (ESI) M / Z: 556.0 [M+H] + .

[0394] 1 H NMR (400MHz, DMSO-d6): δ9.65(s,1H),8.71(s,1H),8.37(d,J=8.0Hz,1H),7.90(s,1H),7.85(d,J=8.0Hz,1H),7.70-7.56( m,3H),7.54-7.33(m,3H),5.80(s,2H),3.88(s,3H),3.71(s,3H),1.80-1.65(m,1H),1.12-0.96(m,2H),0.91-0.74(m,2H).

[0395] Example 2: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole

[0396] Reaction route:

[0397] Steps:

[0398] Step A: Dissolve (2-chloropyrimidin-5-yl)boronic acid (437 mg, 2.81 mmol) in dry 1,4-dioxane / water (8.4 mL / 0.93 mL) at room temperature under nitrogen. Subsequently, add 1-bromo-2-nitrobenzene (378 mg, 1.87 mmol), sodium carbonate (595 mg, 5.61 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (230 mg, 0.28 mmol) to the above solution. Evacuate the air and replace the nitrogen atmosphere three times. Heat the reaction system to 90°C and stir for 16 hours.

[0399] After LCMS monitoring showed the disappearance of the starting material, water (50 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The resulting residue was purified by silica gel column chromatography to obtain 280 mg of 2-chloro-5-(2-nitrophenyl)pyrimidine.

[0400] MS (ESI) M / Z: 236.0 [M+H] + .

[0401] Step B: Dissolve 2-chloro-5-(2-nitrophenyl)pyrimidine (280 mg, 1.19 mmol) in 1,2-dichlorobenzene (4 mL) at room temperature. Add 1,2-bis(diphenylphosphino)ethane (593 mg, 1.49 mmol). Evacuate the mixture three times to replace the nitrogen atmosphere. Heat the reaction system to 160°C and stir for 2 hours.

[0402] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 120 mg of 2-chloro-9H-pyrimido[4,5-b]indole.

[0403] MS (ESI) M / Z: 204.0 [M+H] + .

[0404] Step C: Dissolve 2-chloro-9H-pyrimido[4,5-b]indole (120 mg, 0.59 mmol) in dry tetrahydrofuran (2 mL) at room temperature under nitrogen. Add sodium hydride (29 mg, 0.71 mmol) to the solution at 0°C and stir for 30 minutes. Then add 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (160 mg, 0.65 mmol), warm to room temperature, and continue stirring for 2 hours.

[0405] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. Ice water (30 mL) was added to the resulting residue to quench the mixture. The mixture was extracted with ethyl acetate (10 mL × 3 times). The organic phases were combined and washed with saturated brine (15 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 60 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole.

[0406] MS (ESI) M / Z: 470.0 [M+H] + .

[0407] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole (60 mg, 0.13 mmol) was dissolved in dry 1,4-dioxane / water (0.9 mL / 0.1 mL) at room temperature under nitrogen. Subsequently, (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (50 mg, 0.27 mmol), cesium carbonate (83 mg, 0.27 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (20 mg, 0.03 mmol) were added sequentially. The atmosphere was evacuated and replaced with nitrogen three times. The reaction system was heated to 100°C in a microwave oven and stirred for 3 hours.

[0408] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by the addition of water (30 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 7.66 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole.

[0409] MS (ESI) M / Z: 583.8 [M+H] + .

[0410] 1 H NMR (400MHz, DMSO-d6): δ9.66(s,1H),8.70(s,1H),8.37(d,J=8.0Hz,1H),8.14(s,1H),7.87(d,J=8.4Hz,1H),7.64(t,J=7.8Hz,1H),7.54- 7.38(m,5H),5.81(s,2H),4.44-4.30(m,1H),3.87(s,3H),1.75-1.69(m,1H),1.35(d,J=6.8Hz,6H),1.10-1.01(m,2H),0.88-0.79(m,2H).

[0411] Example 3: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine

[0412] Reaction route:

[0413] Steps:

[0414] Step A: Dissolve (2-chloropyrimidin-5-yl)boronic acid (1 g, 6.33 mmol), 2-bromo-3-nitropyridine (857 mg, 4.2 mmol), and sodium carbonate (1.34 g, 12.64 mmol) in 1,4-dioxane / water (21 mL / 2.3 mL) at room temperature. Add dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (513 mg, 0.6 mmol) to the solution. Evacuate the air and replace it with nitrogen four times. Heat the reaction system to 100°C and stir for 16 hours.

[0415] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (30 mL) to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 600 mg of 2-chloro-5-(3-nitropyridin-2-yl)pyrimidine.

[0416] MS (ESI) M / Z: 237.0 [M+H] + .

[0417] Step B: Dissolve 2-chloro-5-(3-nitropyridin-2-yl)pyrimidine (600 mg, 2.54 mmol) and 1,2-bis(diphenylphosphino)ethane (1.26 g, 3.18 mmol) in 1,2-dichlorobenzene (13 mL) at room temperature. Evacuate the mixture four times to displace nitrogen. Heat the reaction system to 160°C and stir for 2 hours.

[0418] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (30 mL) to quench the mixture. The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (15 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 400 mg of 2-chloro-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0419] MS (ESI) M / Z: 205.2 [M+H] + .

[0420] Step C: 2-Chloro-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine (400 mg, 1.96 mmol), (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (555 mg, 1.96 mmol), and triphenylphosphine (771 mg, 2.94 mmol) were dissolved in dry tetrahydrofuran (10 mL) under nitrogen at room temperature. Diisopropyl azodicarboxylate (594 mg, 2.94 mmol) was then slowly added dropwise to the solution at 0°C. The reaction was stirred at room temperature for 2 hours.

[0421] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by the addition of water (30 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (15 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 250 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0422] MS (ESI) M / Z: 470.8 [M+H] + .

[0423] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine (250 mg, 0.53 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (210 mg, 1.07 mmol), and cesium carbonate (346 mg, 1.06 mmol) were dissolved in 1,4-dioxane / water (3 mL / 0.3 mL) at room temperature. Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (85 mg, 0.11 mmol) was then added to the solution. The air was evacuated under vacuum and replaced with nitrogen four times, and the reaction system was stirred at 100°C for 3 hours under microwave conditions.

[0424] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by the addition of water (30 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 7.35 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[2',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0425] MS (ESI) M / Z: 585.0 [M+H] + .

[0426] 1 H NMR (400MHz, DMSO-d6): δ9.70(s,1H),8.72(s,1H),8.70(dd,J=4.8,1.2Hz,1H),8.37(dd,J=8.4,1.2Hz,1H),8.15(d,J=1.2Hz,1H),7.66(dd,J=8.4,4. 8Hz,1H),7.50(brs,4H),5.85(s,2H),4.45-4.32(m,1H),3.88(s,3H),1.80 -1.70(m,1H),1.35(d,J=6.4Hz,6H),1.11-1.03(m,2H),0.89-0.79(m,2H).

[0427] Example 4: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole

[0428] Reaction route:

[0429] Steps:

[0430] Step A: Dissolve 2-bromo-1-fluoro-3-nitrobenzene (914 mg, 4.15 mmol), (2-chloropyrimidin-5-yl)boronic acid (985 mg, 6.23 mmol), sodium carbonate (1.32 g, 12.45 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (508 mg, 0.62 mmol) in 1,4-dioxane / water (20.7 mL / 2.3 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C for 3 hours.

[0431] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (40 mL) to quench the reaction. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 516 mg of 2-chloro-5-(2-fluoro-6-nitrophenyl)pyrimidine.

[0432] MS (ESI) M / Z: 254.0 [M+H] + .

[0433] Step B: 2-Chloro-5-(2-fluoro-6-nitrophenyl)pyrimidine (400 mg, 1.58 mmol) and 1,2-bis(diphenylphosphino)ethane (786 mg, 1.97 mmol) were dissolved in 1,2-dichlorobenzene (7.9 mL) at room temperature and the reaction was stirred at 160°C for 1 hour.

[0434] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was quenched with ice water (30 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 276 mg of 2-chloro-5-fluoro-9H-pyrimido[4,5-b]indole.

[0435] MS (ESI) M / Z: 222.0 [M+H] + .

[0436] Step C: 2-Chloro-5-fluoro-9H-pyrimido[4,5-b]indole (226 mg, 1.02 mmol) and 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (283 mg, 0.82 mmol) were dissolved in N,N-dimethylformamide (5 mL) at room temperature under nitrogen. Potassium carbonate (281 mg, 2.04 mmol) was then added to the solution. The reaction was stirred at 50°C for 2 hours.

[0437] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (40 mL). The mixture was extracted with ethyl acetate (15 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 220 mg of 2-chloro-5-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole.

[0438] MS (ESI) M / Z: 488.0 [M+H] + .

[0439] Step D: 2-Chloro-5-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole (200 mg, 0.41 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (239 mg, 1.23 mmol), cesium carbonate (200 mg, 0.62 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (65 mg, 0.08 mmol) were dissolved in 1,4-dioxane / water (2 mL / 0.2 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred in a microwave at 100°C for 3 hours.

[0440] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (30 mL). The mixture was extracted with ethyl acetate (30 mL x 3 times), and the organic phases were combined and washed with saturated brine (60 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 16.09 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0441] MS (ESI) M / Z: 602.4 [M+H] + .

[0442] 1H NMR (400MHz, DMSO-d6): δ9.52(s,1H),8.71(s,1H),8.15(d,J=1.2Hz,1H),7.76(d,J=8.0Hz,1H),7.71-7.63(m,1H),7.52-7.47(m,4H),7.31 -7.26(m,1H),5.83(s,2H),4.44-4.33(m,1H),3.88(s,3H),1.78-1.69(m,1H),1.35(d,J=6.4Hz,6H),1.10-1.02(m,2H),0.88-0.80(m,2H).

[0443] Example 5: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole

[0444] Reaction route:

[0445] Steps:

[0446] Step A: Dissolve 1-bromo-4-fluoro-2-nitrobenzene (1.50 g, 6.85 mmol), (2-chloropyrimidin-5-yl)boronic acid (1.62 g, 10.27 mmol), sodium carbonate (2.18 g, 20.55 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (840 mg, 1.03 mmol) in 1,4-dioxane / water (31 mL / 3.5 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C for 3 hours.

[0447] After LCMS monitoring showed the disappearance of the starting material, ice water (60 mL) was added to the reaction mixture to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.1 g of 2-chloro-5-(4-fluoro-2-nitrophenyl)pyrimidine.

[0448] MS (ESI) M / Z: 253.8 [M+H] + .

[0449] Step B: Dissolve 2-chloro-5-(4-fluoro-2-nitrophenyl)pyrimidine (1.1 g, 4.34 mmol) and 1,2-bis(diphenylphosphino)ethane (2.16 g, 5.43 mmol) in 1,2-dichlorobenzene (15 mL) at room temperature. The reaction mixture was stirred at 160°C for 2 hours.

[0450] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was quenched with ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 261 mg of 2-chloro-7-fluoro-9H-pyrimidin[4,5-b]indole.

[0451] MS (ESI) M / Z: 221.9 [M+H] + .

[0452] Step C: 2-Chloro-7-fluoro-9H-pyrimidin[4,5-b]indole (261 mg, 1.18 mmol), 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (490 mg, 1.42 mmol), and potassium carbonate (326 mg, 2.36 mmol) were dissolved in N,N-dimethylformamide (6 mL) at room temperature under nitrogen. The reaction was stirred at 50°C for 2 hours.

[0453] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding water (30 mL). The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (30 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 170 mg of 2-chloro-7-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0454] MS (ESI) M / Z: 487.8 [M+H] + .

[0455] Step D: 2-Chloro-7-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole (130 mg, 0.27 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (156 mg, 0.80 mmol), cesium carbonate (174 mg, 0.53 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (42 mg, 0.05 mmol) were dissolved in 1,4-dioxane / water (1.2 mL / 0.13 mL) at room temperature. The air was evacuated and replaced with nitrogen three times, and the reaction system was stirred in a microwave at 90°C for 3 hours.

[0456] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (30 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 17.11 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-7-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0457] MS (ESI) M / Z: 602.0 [M+H] + .

[0458] 1 H NMR (400MHz, DMSO-d6): δ9.65(s,1H),8.70(s,1H),8.41(dd,J=8.6,5.4Hz,1H),8.15(d,J=1.2Hz,1H),7.85(dd,J=10.0,2.4Hz,1H),7.54-7.46(m,4 H),7.33-7.26(m,1H),5.79(s,2H),4.45-4.31(m,1H),3.87(s,3H),1.75- 1.66(m,1H),1.35(d,J=6.8Hz,6H),1.09-1.00(m,2H),0.88-0.77(m,2H).

[0459] Example 6: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole

[0460] Reaction route:

[0461] Steps:

[0462] Step A: Dissolve 1-bromo-3-fluoro-2-nitrobenzene (1 g, 4.54 mmol), (2-chloropyrimidin-5-yl)boronic acid (1.79 g, 6.82 mmol), sodium carbonate (1.45 g, 13.63 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (498 mg, 0.68 mmol) in 1,4-dioxane / water (19.8 mL / 2.2 mL) at room temperature. Evacuate the mixture three times to displace nitrogen. Stir the reaction system at 90°C for 3 hours.

[0463] After LCMS monitoring showed the disappearance of the starting material, ice water (20 mL) was added to the reaction mixture to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 500 mg of 2-chloro-5-(3-fluoro-2-nitrophenyl)pyrimidine.

[0464] MS (ESI) M / Z: 254.0 [M+H] + .

[0465] Step B: 2-Chloro-5-(3-fluoro-2-nitrophenyl)pyrimidine (500 mg, 1.97 mmol) and 1,2-bis(diphenylphosphino)ethane (979 mg, 2.46 mmol) were dissolved in 1,2-dichlorobenzene (7.1 mL) at room temperature and the reaction was stirred at 160°C for 1 hour.

[0466] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was quenched with ice water (10 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 60 mg of 2-chloro-8-fluoro-9H-pyrimidin[4,5-b]indole.

[0467] MS (ESI) M / Z: 222.0 [M+H] + .

[0468] Step C: 2-Chloro-8-fluoro-9H-pyrimidin[4,5-b]indole (60 mg, 0.27 mmol) and anhydrous potassium carbonate (75 mg, 0.54 mmol) were dissolved in N,N-dimethylformamide (3 mL) at room temperature under nitrogen. 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (113 mg, 0.31 mmol) was then added to the solution. The reaction was stirred at room temperature for 2 hours.

[0469] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (10 mL). The mixture was extracted with ethyl acetate (30 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 90 mg of 2-chloro-8-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0470] MS (ESI) M / Z: 488.3 [M+H] + .

[0471] Step D: 2-Chloro-8-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indole (90 mg, 0.184 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (107.8 mg, 0.55 mmol), cesium carbonate (120 mg, 0.37 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (22 mg, 0.03 mmol) were dissolved in 1,4-dioxane / water (0.8 mL / 0.08 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred in a microwave at 90°C for 2.5 hours.

[0472] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (30 mL). The mixture was extracted with ethyl acetate (30 mL x 3 times), and the organic phases were combined and washed with saturated brine (60 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 4.89 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-fluoro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0473] MS (ESI) M / Z: 602.2 [M+H] + .

[0474] 1 H NMR (400MHz, DMSO-d6): δ9.75(s,1H),8.71(s,1H),8.24(d,J=7.6Hz,1H),8.15(d,J=0.8Hz,1H),7.55-7.46(m,3H),7.45-7.35(m, 3H),5.86(s,2H),4.45-4.32(m,1H),3.87(s,3H),1.77-1.68(m,1H),1.36(d,J=6.8Hz,6H),1.09-1.02(m,2H),0.88-0.81(m,2H).

[0475] Example 7: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-methoxy-9H-pyrimidin[4,5-b]indole

[0476] Reaction route:

[0477] Steps:

[0478] Step A: Dissolve (2-chloropyrimidin-5-yl)boronic acid (1.5 g, 9.49 mmol), 1-bromo-4-methoxy-2-nitrobenzene (1.5 g, 6.49 mmol), and sodium carbonate (2.1 g, 19.81 mmol) in 1,4-dioxane / water (32 mL / 3.5 mL) at room temperature. Add dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium (0.8 g, 0.98 mmol) to the solution, evacuate the air and replace it with nitrogen four times, and stir at 100°C for 16 hours.

[0479] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (30 mL) for quenching. The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (15 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 740 mg of 2-chloro-5-(4-methoxy-2-nitrophenyl)pyrimidine.

[0480] MS (ESI) M / Z: 266.0 [M+H] + .

[0481] Step B: Dissolve 2-chloro-5-(4-methoxy-2-nitrophenyl)pyrimidine (740 mg, 2.79 mmol) and 1,2-bis(diphenylphosphino)ethane (2.22 g, 5.58 mmol) in 1,2-dichlorobenzene (14 mL) at room temperature. Vacuum the reaction mixture four times to displace nitrogen with air. Stir the reaction mixture at 160°C for 2 hours.

[0482] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was quenched by adding ice water (30 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (15 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 440 mg of 2-chloro-7-methoxy-9H-pyrimidin[4,5-b]indole.

[0483] MS (ESI) M / Z: 234.0 [M+H] + .

[0484] Step C: Dissolve 2-chloro-7-methoxy-9H-pyrimidin[4,5-b]indole (440 mg, 1.89 mmol), 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (410 mg, 1.13 mmol), and potassium carbonate (521 mg, 3.78 mmol) in dry N,N-dimethylformamide (10 mL) at room temperature under nitrogen. Stir the reaction at 50°C for 2 hours.

[0485] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (150 mL x 3). The organic phases were combined and washed with saturated brine (25 mL). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 70 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-methoxy-9H-pyrimidin[4,5-b]indole.

[0486] MS (ESI) M / Z: 500.0 [M+H] + .

[0487] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-methoxy-9H-pyrimidin[4,5-b]indole (70 mg, 0.14 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (84 mg, 0.43 mmol), and cesium carbonate (70 mg, 0.21 mmol) were dissolved in 1,4-dioxane / water (1 mL / 0.1 mL) at room temperature. Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (22 mg, 0.03 mmol) was then added to the solution. The atmosphere was evacuated and replaced with nitrogen four times, and the reaction system was stirred in a microwave at 90°C for 3 hours.

[0488] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by the addition of water (30 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. The product was collected and the solvent was removed under reduced pressure to obtain 9.8 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-methoxy-9H-pyrimidin[4,5-b]indole.

[0489] MS (ESI) M / Z: 614.0 [M+H] + .

[0490] 1 H NMR (400MHz, DMSO-d6): δ9.51(s,1H),8.69(s,1H),8.24(d,J=8.4Hz,1H),8.14(s,1H),7.49(brs,4H),7.43(d,J=2.0Hz,1H),7.04(dd,J=8.8,2.4 Hz,1H),5.78(s,2H),4.45-4.32(m,1H),3.90(s,3H),3.86(s,3H),1.74- 1.66(m,1H),1.35(d,J=6.4Hz,6H),1.07-1.00(m,2H),0.85-0.78(m,2H).

[0491] Example 8: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-(trifluoroethyl)-9H-pyrimidin[4,5-b]indole

[0492] Reaction route:

[0493] Steps:

[0494] Step A: Dissolve 1-bromo-2-nitro-4-(trifluoromethyl)benzene (1 g, 3.70 mmol), (2-chloropyrimidin-5-yl)boronic acid (879 mg, 5.60 mmol), sodium carbonate (1.2 g, 11.1 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (406 mg, 0.56 mmol) in 1,4-dioxane / water (18 mL / 2 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C for 16 hours.

[0495] After LCMS monitoring showed the disappearance of the starting material, ice water (20 mL) was added to the reaction mixture to quench the mixture. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 620 mg of 2-chloro-5-(4-trifluoromethyl-2-nitrophenyl)pyrimidine.

[0496] MS (ESI) M / Z: 304.0 [M+H] +.

[0497] Step B: Dissolve 2-chloro-5-(4-trifluoromethyl-2-nitrophenyl)pyrimidine (620 mg, 2.04 mmol) and 1,2-bis(diphenylphosphino)ethane (1.1 g, 2.55 mmol) in 1,2-dichlorobenzene (9.5 mL) at room temperature. The reaction mixture was stirred at 160°C for 2 hours.

[0498] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was quenched with ice water (30 mL). The mixture was extracted with ethyl acetate (20 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 220 mg of 2-chloro-7-(trifluoromethyl)-9H-pyrimidin[4,5-b]indole.

[0499] MS (ESI) M / Z: 272.0 [M+H] + .

[0500] Step C: Dissolve 2-chloro-7-(trifluoromethyl)-9H-pyrimidin[4,5-b]indole (220 mg, 0.81 mmol) and anhydrous potassium carbonate (223.6 mg, 1.62 mmol) in N,N-dimethylformamide (5 mL) at room temperature under nitrogen. Then, add 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (337 mg, 0.97 mmol). The reaction mixture is stirred at 55°C for 2 hours.

[0501] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (10 mL). The mixture was extracted with ethyl acetate (30 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 450 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-(trifluoroethyl)-9H-pyrimidin[4,5-b]indole.

[0502] MS (ESI) M / Z: 538.0 [M+H] + .

[0503] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-(trifluoroethyl)-9H-pyrimido[4,5-b]indole (270 mg, 0.50 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (294 mg, 1.51 mmol), cesium carbonate (327 mg, 1.0 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (60 mg, 0.075 mmol) were dissolved in 1,4-dioxane / water (2.5 mL / 0.25 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred in a microwave at 90°C for 2.5 hours.

[0504] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected and the solvent was removed to obtain 27.13 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7-(trifluoroethyl)-9H-pyrimidin[4,5-b]indole.

[0505] MS (ESI) M / Z: 652.0 [M+H] + .

[0506] 1 H NMR (400MHz, DMSO) δ9.82(s,1H),8.72(s,1H),8.62(d,J=8.0Hz,1H),8.35(s,1H),8.15(d,J=1.2Hz,1H),7.80(d,J=8.0Hz,1H),7.52-7. 46(m,4H),5.93(s,2H),4.42-4.32(m,1H),3.87(s,3H),1.76-1.68(m,1H),1.35(d,J=6.8Hz,6H),1.09-1.03(m,2H),0.86-0.79(m,2H).

[0507] Example 9: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole

[0508] Step A: Dissolve 2-(4-(bromomethyl)phenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (200 mg, 0.60 mmol), 2-chloro-9H-pyrimidin[4,5-b]indole (245 mg, 1.20 mmol), and potassium carbonate (182 mg, 1.20 mmol) in N,N-dimethylformamide (4 mL) at room temperature under nitrogen. The reaction mixture was heated to 50°C and stirred for 2 hours.

[0509] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding water (30 mL). The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 130 mg of 2-chloro-9-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0510] MS (ESI) M / Z: 455.8 [M+H] + .

[0511] Step B: 2-Chloro-9-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole (110 mg, 0.24 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (94 mg, 0.48 mmol), cesium carbonate (158 mg, 0.48 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (38 mg, 0.05 mmol) were dissolved in 1,4-dioxane / water (1.1 mL / 0.12 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred at 90°C in a microwave for 3 hours.

[0512] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixed solution was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). It was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected and the solvent was removed to obtain 55.00 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimidin[4,5-b]indole.

[0513] MS (ESI) M / Z: 570.1 [M+H] + .

[0514] 1H NMR (400MHz, DMSO-d6): δ9.66(s,1H),8.71(s,1H),8.37(d,J=7.6Hz,1H),7.99(d,J= 1.2Hz,1H),7.86(d,J=8.0Hz,1H),7.63(t,J=7.6Hz,1H),7.56(d,J=8.4Hz,2H),7.48 (d,J=8.4Hz,2H),7.44(t,J=8.0Hz,1H),5.80(s,2H),4.01(q,J=7.2Hz,2H),3.87(s, 3H),1.77-1.68(m,1H),1.26(t,J=7.2Hz,3H),1.10-1.02(m,2H),0.89-0.80(m,2H).

[0515] Example 10: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-pyrimidinyl[4,5-b]indole

[0516] Reaction route:

[0517] Steps:

[0518] Step A: Dissolve 4-hydrazinobenzoic acid (5 g, 32.9 mmol) and ethyl 4,4,4-trifluoro-3-oxobutanoate (6 g, 32.9 mmol) in methanol / hydrochloric acid (73 mL / 14.6 mL) at room temperature. Stir the reaction system at room temperature for 3 hours.

[0519] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 8.3 g of methyl 4-(5-hydroxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate.

[0520] MS (ESI) M / Z: 287.0 [M+H] + .

[0521] Step B: Dissolve methyl 4-(5-hydroxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (5 g, 17.5 mmol) in N,N-dimethylformamide (87 mL) at room temperature under nitrogen. Add sodium hydride (1.39 g, 34.96 mmol) to the solution and stir for 30 minutes. Then, slowly add iodoethane (5.4 g, 34.96 mmol) dropwise and stir at room temperature for 1 hour.

[0522] After LCMS monitoring showed the disappearance of the starting material, ice water (250 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (80 mL × 3 times), and the organic phases were combined and washed with saturated brine (250 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3.7 g of methyl 4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate.

[0523] MS (ESI) M / Z: 315.0 [M+H] + .

[0524] Step C: Dissolve methyl 4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (3.7 g, 11.78 mmol) in tetrahydrofuran (59 mL) at 0°C under nitrogen. Then, add lithium aluminum hydride (9.4 mL, 23.56 mmol) and continue stirring for 30 minutes.

[0525] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (150 mL) to quench the reaction. The mixture was filtered and the filtrate was extracted with ethyl acetate (50 mL x 3 times). The organic phases were combined and washed with saturated brine (150 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3 g of (4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol.

[0526] MS (ESI) M / Z: 287.0 [M+H] + .

[0527] Step D: Dissolve (4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol (1.5 g, 5.24 mmol) in dichloromethane (26 mL) at 0°C under nitrogen. Triphenylphosphine (2.7 g, 10.48 mmol), sodium bicarbonate (880 mg, 10.48 mmol), and carbon tetrabromide (3.46 g, 10.48 mmol) were then added to the solution. The reaction was stirred at room temperature for 30 minutes.

[0528] After LCMS monitoring showed the disappearance of the starting material, ice water (150 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with dichloromethane (50 mL x 3 times), and the organic phases were combined and washed with saturated brine (150 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.8 g of 1-(4-(bromomethyl)phenyl)-5-ethoxy-3-(trifluoromethyl)-1H-pyrazole.

[0529] MS (ESI) M / Z: 348.8 [M+H] + .

[0530] Step E: Dissolve 1-(4-(bromomethyl)phenyl)-5-ethoxy-3-(trifluoromethyl)-1H-pyrazole (43.9 mg, 0.13 mmol), 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrimido[4,5-b]indole (40 mg, 0.13 mmol), and potassium carbonate (34.7 mg, 0.26 mmol) in N,N-dimethylformamide (1 mL) at room temperature. Evacuate the air and replace it with nitrogen three times. Stir the reaction at 50°C for 2 hours.

[0531] After LCMS monitoring showed the disappearance of the starting material, water (40 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (25 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected and the solvent was removed to obtain 17.29 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(5-ethoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-9H-pyrimidinyl[4,5-b]indole.

[0532] MS (ESI) M / Z: 586.2 [M+H] + .

[0533] 1 H NMR (400MHz, DMSO-d6): δ9.65(s,1H),8.70(s,1H),8.37(d,J=8.0Hz,1H),7.85(d,J=8.0Hz,1H),7.65-7.58(m,3H),7.49-7.42(m,3H),6.4 2(s,1H),5.78(s,2H),4.25(q,J=6.8Hz,2H),3.87(s,3H),1.77-1.65(m,1H),1.30(t,J=6.8Hz,3H),1.10-1.00(m,2H),0.89-0.76(m,2H).

[0534] Example 11: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4-methyl-9H-pyrimidin[4,5-b]indole

[0535] Reaction route:

[0536] Steps:

[0537] Step A: Dissolve (2-nitrophenyl)boronic acid (1.03 g, 6.15 mmol), 5-bromo-2-chloro-4-methylpyrimidine (850 mg, 4.10 mmol), sodium carbonate (1.3 g, 12.3 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (450 mg, 0.61 mmol) in 1,4-dioxane / water (21 mL / 2.3 mL) at room temperature. Evacuate the air and replace with nitrogen three times. Stir the reaction system at 90°C for 16 hours.

[0538] After LCMS monitoring showed the disappearance of the starting material, ice water (40 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (15 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 600 mg of 2-chloro-4-methyl-5-(2-nitrophenyl)pyrimidine.

[0539] MS (ESI) M / Z: 250.0 [M+H] + .

[0540] Step B: Dissolve 2-chloro-4-methyl-5-(2-nitrophenyl)pyrimidine (600 mg, 2.41 mmol) and 1,2-bis(diphenylphosphino)ethane (1198.8 mg, 3.01 mmol) in 1,2-dichlorobenzene (12 mL) at room temperature. The reaction mixture was stirred at 160°C for 2 hours.

[0541] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (40 mL). The mixture was extracted with ethyl acetate (20 mL x 3), and the organic phases were combined and washed with saturated brine (20 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 320 mg of 2-chloro-4-methyl-9H-pyrimidin[4,5-b]indole.

[0542] MS (ESI) M / Z: 218.0 [M+H] + .

[0543] Step C: Dissolve 2-chloro-4-methyl-9H-pyrimidin[4,5-b]indole (150 mg, 0.69 mmol) and anhydrous potassium carbonate (190.2 mg, 1.38 mmol) in N,N-dimethylformamide (5 mL) at room temperature under nitrogen. Then, add 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (286.7 mg, 0.83 mmol). The reaction mixture is stirred at 55°C for 2 hours.

[0544] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (30 mL). The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 220 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4-methyl-9H-pyrimidin[4,5-b]indole.

[0545] MS (ESI) M / Z: 484.0 [M+H] + .

[0546] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4-methyl-9H-pyrimidin[4,5-b]indole (180 mg, 0.37 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (145 mg, 0.75 mmol), cesium carbonate (243 mg, 0.75 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (59 mg, 0.08 mmol) were dissolved in 1,4-dioxane / water (1.7 mL / 0.20 mL) at room temperature. The air was evacuated and replaced with nitrogen three times. The reaction system was stirred in a microwave at 90°C for 2.5 hours.

[0547] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixed solution was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected and the solvent was removed to obtain 88.80 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4-methyl-9H-pyrimidin[4,5-b]indole.

[0548] MS (ESI) M / Z: 598.2 [M+H] + .

[0549] 1 H NMR (400MHz, DMSO) δ8.70(s,1H),8.27(d,J=7.6Hz,1H),8.14(d,J=0.8Hz,1H),7.88(d,J=8.0Hz,1H),7.63(t,J=7.6Hz,1H),7.52-7.41(m,5H ),5.81(s,2H),4.43-4.30(m,1H),3.87(s,3H),3.03(s,3H),1.74-1.6 3(m,1H),1.35(d,J=6.8Hz,6H),1.09-1.00(m,2H),0.90-0.77(m,2H).

[0550] Example 12: (S)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-9H-pyrimidin[4,5-b]indole and (R)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-9H-pyrimidin[4,5-b]indole

[0551] Prepared with reference to the preparation method of Example 8 and chiral separation (chiral conditions are missing in the synthesis report). The product was separated by chiral supercritical fluid chromatography (SFC) using the following separation conditions: chiral column Daicel IG-3 (25×250 mm, 10 μm); mobile phase: carbon dioxide / methanol [0.2% ammonia (7 M ammonia methanol solution)], to yield 27.65 mg of compound 13-P1 (elution time 1.77 min) and 32.27 mg of compound 13-P2 (elution time 3.27 min).

[0552] Compound 13-P1:

[0553] MS (ESI) M / Z: 598.1 [M+H] + .

[0554] 1H NMR (400MHz, DMSO-d6): δ9.65(s,1H),8.70(s,1H),8.37(d,J=8.0Hz,1H),8.15(d ,J=0.8Hz,1H),7.68(d,J=8.4Hz,1H),7.60-7.47(m,5H),7.41(t,J=7.4Hz,1H),6 .55(q,J=7.2Hz,1H),4.47-4.31(m,1H),3.87(s,3H),2.15(d,J=7.2Hz,3H),1.82 -1.67(m,1H),1.36(dd,J=6.6,1.8Hz,6H),1.11-1.01(m,2H),0.90-0.77(m,2H).

[0555] Compound 13-P2:

[0556] MS (ESI) M / Z: 598.2 [M+H] + .

[0557] 1 H NMR (400MHz, DMSO-d6): δ9.65(s,1H),8.70(s,1H),8.37(d,J=7.6Hz,1H),8.15(d ,J=0.8Hz,1H),7.68(d,J=8.4Hz,1H),7.58-7.48(m,5H),7.41(t,J=7.6Hz,1H),6 .55(q,J=7.2Hz,1H),4.46-4.35(m,1H),3.87(s,3H),2.15(d,J=7.2Hz,3H),1.80 -1.69(m,1H),1.36(dd,J=6.6,1.7Hz,6H),1.09-1.01(m,2H),0.92-0.79(m,2H).

[0558] Example 13: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-imidazolino[2,1-f]purine

[0559] Reaction route

[0560] Steps:

[0561] Step A: Dissolve (4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanamine (2.25 g, 7.96 mmol) and ethyl 2,4-dichloropyrimidine-5-carboxylate (1.76 g, 7.96 mmol) in acetonitrile (40 mL) at 0°C. Then, slowly add N,N-diisopropylethylamine (267 mg, 2.07 mmol) dropwise to the reaction mixture. Stir the reaction mixture for 1 hour.

[0562] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (100 mL) to quench the reaction. The mixed solution was extracted with ethyl acetate (40 mL × 3 times), the organic phases were combined, and the organic phases were washed with saturated brine (20 mL × 2 times). It was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2.4 g of ethyl 2-chloro-4-((4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carboxylate.

[0563] MS (ESI) M / Z: 468.0 [M+H] + .

[0564] 1 H NMR (400MHz, DMSO-d6) δ9.05(t,J=6.2Hz,1H),8.67(s,1H),8.17(d,J=1.2Hz,1H),7.54(d,J=8.0Hz,2H),7.49(d,J=8. 4Hz, 2H), 4.78 (d, J = 6.4Hz, 2H), 4.53-4.42 (m, 1H), 4.34 (q, J = 7.2Hz, 2H), 1.40 (d, J = 6.8Hz, 6H), 1.33 (t, J = 7.2Hz, 3H).

[0565] Step B: Ethyl 2-chloro-4-((4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carboxylate (2.4 g, 4.9 mmol) and lithium hydroxide (413 mg, 9.8 mmol) were dissolved in tetrahydrofuran / water (12.5 mL / 12.5 mL) at 0°C. The reaction was stirred at room temperature for 2 hours.

[0566] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure, and the resulting residue was added to ice water (30 mL) to quench the reaction. The mixture was adjusted to pH 3-4 with 3M dilute hydrochloric acid at 0°C, filtered, and the filter cake was rinsed with water (20 mL × 3 times). The filter cake was collected and concentrated under reduced pressure to obtain 2.2 g of 2-chloro-4-((4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carboxylic acid.

[0567] MS (ESI) M / Z: 440.0 [M+H] + .

[0568] Step C: 2-Chloro-4-((4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)amino)pyrimidine-5-carboxylic acid (2.2 g, 5.01 mmol), diphenylphosphoryl azide (1.38 g, 5.01 mmol), and triethylamine (506 mg, 5.01 mmol) were dissolved in N,N-dimethylacetamide (25 mL) at room temperature. The reaction was then stirred at 115°C for 16 hours.

[0569] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 1.4 g of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,9-dihydro-8H-purin-8-one.

[0570] MS (ESI) M / Z: 437.0 [M+H] + .

[0571] Step D: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-7,9-dihydro-8H-purin-8-one (2.2 g, 5.05 mmol) and triethylamine (1 g, 10.09 mmol) were dissolved in phosphorus oxychloride (25 mL) at 0°C under nitrogen. The reaction was then stirred at 140°C for 24 hours.

[0572] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was quenched with ice water (100 mL). The mixed solution was extracted with ethyl acetate (40 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 70 mg of 2,8-dichloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine.

[0573] MS (ESI) M / Z: 455.0 [M+H]+ .

[0574] Step E: 2,8-dichloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purine (70 mg, 0.15 mmol) was dissolved in ammonia methanol (2 mL, 7 M) at room temperature under nitrogen. The reaction was then stirred at 80°C for 2 hours.

[0575] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 50 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purin-8-amine.

[0576] MS (ESI) M / Z: 436.0 [M+H] + .

[0577] Step F: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-purin-8-amine (25 mg, 0.06 mmol) and 2-chloroacetaldehyde (71 mg, 0.36 mmol) were dissolved in N,N-dimethylacetamide (2 mL) at room temperature under nitrogen. The reaction was stirred at 100°C for 18 hours.

[0578] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 8 mg of 2-chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-imidazolino[2,1-f]purine.

[0579] MS (ESI) M / Z: 460.0 [M+H] + .

[0580] Step G: 2-Chloro-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-imidazolino[2,1-f]purine (8 mg, 0.02 mmol) was dissolved in 1,4-dioxane / water (0.5 mL / 0.05 mL) at room temperature. (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (8 mg, 0.04 mmol), cesium carbonate (7 mg, 0.02 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (3 mg, 0.004 mmol) were then added to the reaction mixture. The atmosphere was evacuated and replaced with nitrogen four times. The reaction system was heated to 90°C in a microwave oven and stirred for 2 hours.

[0581] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected to obtain 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-imidazolino[2,1-f]purine.

[0582] MS (ESI) M / Z: 574.2 [M+H] + .

[0583] 1 H NMR (400MHz, DMSO-d6): δ9.24(s,1H),8.70(s,1H),8.16(d,J=1.4Hz,1H),7.91(d,J=1.7Hz,1H),7.58(d,J=8.4Hz,2H),7.53(d,J=8.3Hz,2H),7. 20(d,J=1.6Hz,1H),5.52(s,2H),4.48-4.35(m,1H),3.86(s,3H),1.79-1 .67(m,1H),1.37(d,J=6.6Hz,7H),1.10-1.01(m,2H),0.89-0.79(m,2H).

[0584] Example 14: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine

[0585] Reaction route

[0586] Steps:

[0587] Step A: Dissolve 4-methoxybenzaldehyde (13 g, 95.59 mmol) and triethylamine (21 g, 210.3 mmol) in dichloromethane (200 mL) at room temperature. Cool the reaction mixture to 0°C and add glycine methyl ester (17 g, 191.2 mmol) and anhydrous sodium sulfate (50 g, 352.1 mmol). Stir the reaction at room temperature for 16 hours.

[0588] After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered and concentrated under reduced pressure to obtain 20 g of methyl 2-((4-methoxybenzylidene)amino)acetate.

[0589] MS (ESI) M / Z: 208.1 [M+H] + .

[0590] Step B: Methyl 2-((4-methoxybenzylidene)amino)acetate (18 g, 86.96 mmol) was dissolved in methanol (200 mL) at 0°C. Then, sodium borohydride (4.96 g, 130.44 mmol) was slowly added. The reaction system was then stirred at room temperature for 2 hours.

[0591] After LCMS monitoring showed the disappearance of the starting material, ice water (500 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (200 mL x 3 times), and the organic phases were combined and washed with saturated brine (100 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 15 g of (4-methoxybenzyl)glycine methyl ester.

[0592] MS (ESI) M / Z: 210.1 [M+H] + .

[0593] Step C: Dissolve (4-methoxybenzyl)glycine methyl ester (15 g, 71.77 mmol), 4-chloro-2-(methylthio)pyrimidine-5-carbaldehyde (13.4 g, 71.77 mmol), and triethylamine (8.7 g, 86.12 mmol) in tetrahydrofuran (360 mL) at room temperature. The reaction was stirred at room temperature for 16 hours.

[0594] After LCMS monitoring showed the disappearance of the starting material, ice water (300 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (100 mL x 3 times), and the organic phases were combined and washed with saturated brine (50 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 22 g of N-(5-formyl-2-(methylthio)pyrimidin-4-yl)-N-(4-methoxybenzyl)glycine methyl ester.

[0595] MS (ESI) M / Z: 362.0 [M+H] + .

[0596] Step D: Under nitrogen at room temperature, dissolve N-(5-formyl-2-(methylthio)pyrimidin-4-yl)-N-(4-methoxybenzyl)glycine methyl ester (16.5 g, 45.71 mmol) in dry toluene (160 mL). Then, slowly add sodium hydride (7.31 g, 182.83 mmol) to the solution in an ice-water bath. The reaction system is then stirred at 70°C for 2 hours.

[0597] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (500 mL). The mixture was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 10 g of methyl 7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylate.

[0598] MS (ESI) M / Z: 344.0 [M+H] + .

[0599] Step E: Methyl 7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylate (10 g, 29.15 mmol) was dissolved in dry dichloromethane (147 mL) at room temperature. Lithium aluminum tetrahydride (1 mol / L) (59.30 mL, 59.30 mmol) was then added to the reaction mixture at -78°C. The reaction was then incubated at -78°C for 30 minutes.

[0600] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (500 mL) to quench the mixture. The mixture was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 10 g of (7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)methanol.

[0601] MS (ESI) M / Z: 316.0 [M+H] + .

[0602] Step F: Dissolve (7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)methanol (10 g, 31.75 mmol) and manganese dioxide (13.65 g, 158.75 mmol) in dichloromethane (635 mL) at room temperature. Evacuate the mixture three times to displace nitrogen. Stir the reaction at room temperature for 2 hours.

[0603] After LCMS monitoring showed the disappearance of the starting material, the mixture was filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 8 g of 7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde.

[0604] MS (ESI) M / Z: 314.0 [M+H] + .

[0605] Step G: Dissolve 7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde (8 g, 25.56 mmol) in N,N-dimethylformamide (128 mL) at room temperature. Then, slowly add N-chlorosuccinimide (5.12 g, 38.34 mmol) to the solution. The reaction mixture is stirred at 50°C for 2 hours.

[0606] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (500 mL). The mixture was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 3.5 g of 5-chloro-7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde.

[0607] MS (ESI) M / Z: 348.0 [M+H] + .

[0608] Step H: Dissolve 5-chloro-7-(4-methoxybenzyl)-2-(methylthio)-7H-pyrrolo[2,3-d]pyrimidine-6-carbaldehyde (3.5 g, 10.09 mmol) and sulfur powder (65 mg, 2.02 mmol) in ammonia / methanol solution (4 mol / L, 34 mL) at room temperature. The reaction system was stirred at 80°C for 6 hours.

[0609] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (40 mL). The mixture was extracted with ethyl acetate (60 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2.9 g of 4-(4-methoxybenzyl)-6-(methylthio)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0610] MS (ESI) M / Z: 343.0 [M+H] + .

[0611] Step I: Dissolve 4-(4-methoxybenzyl)-6-(methylthio)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine (2.9 g, 8.48 mmol) in dry dichloromethane (43 mL) at room temperature. Then, slowly add m-chloroperbenzoic acid (3.22 g, 18.66 mmol) to the reaction mixture at -78°C. The reaction mixture is stirred at room temperature for 4 hours.

[0612] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (100 mL). The mixture was extracted with dichloromethane (60 mL x 3 times), and the organic phases were combined and washed with saturated brine (100 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.8 g of 4-(4-methoxybenzyl)-6-(methylsulfonyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0613] MS (ESI) M / Z: 375.0 [M+H] + .

[0614] Step J: Dissolve 4-(4-methoxybenzyl)-6-(methylsulfonyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine (1.8 g, 4.81 mmol) in dry 1,4-dioxane (16 mL) at room temperature. Ammonia / methanol solution (4 mol / L, 48 mL) was then added to the solution. The reaction was stirred at 80°C for 4 hours.

[0615] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to obtain 840 mg of 4-(4-methoxybenzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidin-6-amine.

[0616] MS (ESI) M / Z: 312.0 [M+H] + .

[0617] Step K: Dissolve 4-(4-methoxybenzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidin-6-amine (840 mg, 2.70 mmol) in dry dichloromethane (68 mL) at room temperature. Chlorotrimethylsilane (910 mg, 8.37 mmol) and tert-butyl nitrite (1.37 g, 13.23 mmol) were then added sequentially to the solution in an ice-water bath. The reaction was stirred at room temperature for 16 hours.

[0618] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (40 mL). The mixture was extracted with dichloromethane (60 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 390 mg of 6-chloro-4-(4-methoxybenzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0619] MS (ESI) M / Z: 331.0 [M+H] + .

[0620] Step L: 6-chloro-4-(4-methoxybenzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine (390 mg, 1.18 mmol) was dissolved in trifluoromethanesulfonic acid (8 mL) at room temperature and the reaction was stirred at 65°C for 2 hours.

[0621] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (10 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 200 mg of crude 6-chloro-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0622] MS (ESI) M / Z: 211.0 [M+H] + .

[0623] Step M: 6-Chloro-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine (200 mg, 0.95 mmol), 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (303 mg, 0.95 mmol), and potassium carbonate (263 mg, 1.90 mmol) were dissolved in N,N-dimethylformamide (5 mL) at room temperature under nitrogen. The reaction was stirred at 50°C for 2 hours.

[0624] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 170 mg of 6-chloro-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0625] MS (ESI) M / Z: 449.0 [M+H] + .

[0626] Step N: 6-Chloro-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine (170 mg, 0.38 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (110 mg, 0.57 mmol), cesium carbonate (186 mg, 0.57 mmol) and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (60 mg, 0.08 mmol) were dissolved in 1,4-dioxane / water (2.7 mL / 0.3 mL) at room temperature. The air was evacuated under vacuum and replaced with nitrogen three times, and the reaction system was reacted at 100°C under microwave conditions for 2 hours.

[0627] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected to obtain 17.95 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4H-isothiazolo[5',4':4,5]pyrrolo[2,3-d]pyrimidine.

[0628] MS (ESI) M / Z: 563.6 [M+H] + .

[0629] 1 H NMR (400MHz, DMSO-d6): δ9.73(s,1H),9.08(s,1H),8.71(s,1H),7.91(d,J=1.2Hz,1H),7.67(d,J=8.4Hz,2H),7.54 (d,J=8.4Hz,2H),5.82(s,2H),3.88(s,3H),3.73(s,3H),1.77-1.68(m,1H),1.10-1.04(m,2H),0.90-0.83(m,2H).

[0630] Example 15: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indole-7-carbonitrile

[0631] Reaction route:

[0632] Steps:

[0633] Step A: Dissolve 5-bromo-2-chloropyrimidin-4-amine (108 g, 518 mmol) in 1,4-dioxane / water (2119 mL / 235 mL) at room temperature under nitrogen. Then, add 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (150 g, 570 mmol), sodium carbonate (110 g, 1036 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (38 g, 46.6 mmol) sequentially to the solution. The reaction mixture is stirred at 90°C for 16 hours.

[0634] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered, the filtrate was collected, and it was concentrated under reduced pressure. The resulting residue was slurried with ethyl acetate / petroleum ether, and the resulting filter cake was slurried again with acetonitrile / water to obtain 94.6 g of a crude product of 4-(4-amino-2-chloropyrimidin-5-yl)-3-chlorobenzonitrile.

[0635] MS (ESI) M / Z: 265.0 [M+H] + .

[0636] Step B: 4-(4-amino-2-chloropyrimidin-5-yl)-3-chlorobenzonitrile (94.6 g, 358 mmol) was dissolved in 1,4-dioxane / water (1394 mL / 232 mL) at room temperature under nitrogen. (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (69.5 g, 358 mmol), cesium carbonate (233 g, 716 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (36.5 g, 46.5 mmol) were then added to the solution. The reaction was stirred at 90°C for 16 hours.

[0637] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered and concentrated under reduced pressure. The resulting residue was slurried with ethyl acetate / petroleum ether, and the filter cake was rinsed with water and dried to obtain 118 g of crude 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrimido[4,5-b]indole-7-carbonitrile.

[0638] MS (ESI) M / Z: 343.2 [M+H] + .

[0639] Step C: Dissolve 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrimido[4,5-b]indole-7-carbonitrile (53.8 g, 157.2 mmol) in N,N-dimethylformamide (436 mL) at room temperature. Cesium carbonate (114 g, 349.2 mmol) and 9-(chloromethyl)-2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine (26.2 g, 87.3 mmol) were then added to the reaction. The reaction was stirred at 45°C for 16 hours.

[0640] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to water (900 mL) to precipitate a solid, which was purified by silica gel column chromatography to give 20.42 g of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indole-7-carbonitrile.

[0641] MS (ESI) M / Z: 607.2 [M+H] + .

[0642] 1 H NMR (400MHz, DMSO-d6) δ9.82 (s, 1H), 8.72 (s, 1H), 8.59 (d, J = 8.4Hz, 1H), 8.50 (s, 1H), 7.9 6(d,J=0.8Hz,1H),7.86(dd,J=8.0,1.2Hz,1H),7.62(d,J=8.0Hz,1H),7.41(d,J=0.8Hz,1 H),7.33(dd,J=8.0,1.6Hz,1H),5.81(s,2H),3.95(t,J=6.8Hz,2H),3.87(s,3H),2.60(t, J=7.0Hz,2H),2.27-2.15(m,2H),1.77-1.67(m,1H),1.10-1.02(m,2H),0.88-0.77(m,2H).

[0643] Example 16: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(5-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-2,3-dihydro-1H-inden-1-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0644] Reaction route:

[0645] Steps:

[0646] Step A: Dissolve methyl 1-oxo-2,3-dihydro-1H-indene-5-carboxylate (1 g, 5.26 mmol) in methanol (27.5 mL) at room temperature. Then, slowly add sodium borohydride (420 mg, 10.53 mmol) to the solution at 0°C. The reaction system is then stirred at room temperature for 1 hour.

[0647] After LCMS monitoring indicated the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (50 mL) to quench the reaction. The mixture was filtered and the filtrate was extracted with ethyl acetate (50 mL x 3). The organic phases were combined and washed with saturated brine (50 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1 g of methyl 1-hydroxy-2,3-dihydro-1H-indene-5-carboxylate.

[0648] MS (ESI) M / Z: 193.1 [M+H] + .

[0649] Step B: Methyl 1-hydroxy-2,3-dihydro-1H-indene-5-carboxylate (710 mg, 3.68 mmol), 3,4-dihydro-2H-pyran (621.25 mg, 7.40 mmol), and pyridinium p-toluenesulfonate (95 mg, 0.37 mmol) were dissolved in dichloromethane (18.50 mL) at room temperature under nitrogen. The reaction was stirred at 60°C for 1 hour.

[0650] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 850 mg of methyl 1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-indene-5-carboxylate.

[0651] MS (ESI) M / Z: 299.2 [M+23] + .

[0652] Step C: Methyl 1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-indene-5-carboxylate (850 mg, 2.90 mmol) was dissolved in tetrahydrofuran (15.50 mL) at 0°C under nitrogen. Lithium aluminum hydride (2.5 mL, 6.18 mmol) was then slowly added to the solution. The reaction was then stirred at room temperature for 1 hour.

[0653] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (100 mL) to quench the reaction. The mixture was filtered, and the filtrate was extracted with ethyl acetate (50 mL x 3 times). The organic phases were combined and washed with saturated brine (100 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 690 mg of (1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)methanol.

[0654] MS (ESI) M / Z: 271.2 [M+H] + .

[0655] Step D: (1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)methanol (690 mg, 2.78 mmol) and manganese dioxide (2.42 g, 27.82 mmol) were dissolved in dichloromethane (14 mL) at room temperature under nitrogen. The reaction was stirred at 60°C for 1 hour.

[0656] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered while hot, and the filtrate was concentrated under reduced pressure to obtain 690 mg of 1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-indene-5-carbaldehyde.

[0657] MS (ESI) M / Z: 247.2 [M+H] + .

[0658] Step E: Dissolve 3,3-dibromo-1,1,1-trifluoropropan-2-one (833.05 mg, 3.09 mmol) and sodium acetate (460 mg, 5.61 mmol) in water (14 mL) at room temperature, raise the temperature to 90°C, and stir for 0.5 hours. Then, cool the temperature to 0°C and add a mixture of 1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-indene-5-carbaldehyde (690 mg, 2.80 mmol) in aqueous ammonia / methanol (49 mL / 15 mL). The reaction mixture is then stirred at room temperature for 16 hours.

[0659] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (200 mL) to quench the mixture. The mixture was extracted with ethyl acetate (100 mL × 3 times), and the organic phases were combined and washed with saturated brine (100 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 590 mg of 2-(1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)-4-(trifluoromethyl)-1H-imidazole.

[0660] MS (ESI) M / Z: 353.2 [M+H] + .

[0661] Step F: 2-(1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)-4-(trifluoromethyl)-1H-imidazole (590 mg, 1.67 mmol) and potassium carbonate (461.30 mg, 3.34 mmol) were dissolved in N,N-dimethylformamide (8.40 mL) at room temperature. Methyl iodide (617.08 mg, 4.35 mmol) was then added to the solution at 0°C. The reaction was stirred at room temperature for 4 hours.

[0662] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was slowly added dropwise to ice water (100 mL) to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (50 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 400 mg of 1-methyl-2-(1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)-4-(trifluoromethyl)-1H-imidazole.

[0663] MS (ESI) M / Z: 367.2 [M+H] + .

[0664] Step G: At room temperature, under nitrogen protection, 1-methyl-2-(1-((tetrahydro-2H-pyran-2-yl)oxy)-2,3-dihydro-1H-inden-5-yl)-4-(trifluoromethyl)-1H-imidazole (400 mg, 1.09 mmol) and p-toluenesulfonic acid (41.53 mg, 0.22 mmol) were dissolved in methanol (6 mL), the temperature was raised to 60°C, and stirred for 2 hours.

[0665] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure and the resulting residue was purified by silica gel column chromatography to give 180 mg of 5-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-2,3-dihydro-1H-inden-1-ol.

[0666] MS (ESI) M / Z: 283.1 [M+H] + .

[0667] Step H: 5-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-2,3-dihydro-1H-inden-1-ol (80 mg, 0.28 mmol), 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (90.21 mg, 0.28 mmol), and triphenylphosphine (111.49 mg, 0.43 mmol) were dissolved in anhydrous tetrahydrofuran (1.5 mL) at room temperature under nitrogen. Diisopropyl azodicarboxylate (85.96 mg, 0.43 mmol) was then slowly added dropwise to the solution at 0°C. The reaction was stirred at room temperature for 1 hour.

[0668] After LCMS monitoring showed the disappearance of the starting material, water (40 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (10 mL × 3 times), the organic phases were combined, and the organic phases were washed with saturated brine (25 mL × 2 times). It was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. 28.22 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(5-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-2,3-dihydro-1H-inden-1-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine was obtained.

[0669] MS (ESI) M / Z: 583.3 [M+H] + .

[0670] 1 H NMR (400MHz, DMSO-d6): δ9.82(s,1H),8.71(s,1H),8.58(d,J=5.2Hz,1H),8.53-8.22( m,2H),7.94(s,1H),7.80(s,1H),7.48(d,J=8.0Hz,1H),7.03(d,J=7.6Hz,1H),6.92(t, J=8.4Hz,1H),3.86(s,3H),3.79(s,3H),3.47-3.36(m,1H),3.27-3.14(m,1H),2.86-2. 75(m,1H),2.71-2.57(m,1H),1.80-1.68(m,1H),1.12-1.00(m,2H),0.93-0.80(m,2H).

[0671] Example 17: 9-((2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrido[4′,3′:4,5]pyrrolo[2,3-d]pyrimidin-9-yl)methyl)-2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine

[0672] Reaction route

[0673] Steps:

[0674] Step A: Dissolve (2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)methanol (70 mg, 0.25 mmol), 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (79 mg, 0.25 mmol), and triphenylphosphine (97 mg, 0.37 mmol) in dry tetrahydrofuran (1.5 mL) at room temperature under nitrogen. Diisopropyl azodicarboxylate (75 mg, 0.37 mmol) was then slowly added dropwise to the solution in an ice-water bath. The reaction was stirred at room temperature for 1 hour.

[0675] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched with ice water (20 mL). The mixture was extracted with ethyl acetate (30 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to obtain 48.59 mg of 9-((2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidin-9-yl)methyl)-2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine.

[0676] MS (ESI) M / Z: 585.2 [M+H] + .

[0677] 1H NMR (400MHz, DMSO-d6) δ9.81 (s, 1H), 9.27 (s, 1H), 8.73 (s, 1H), 8.63 (d, J = 5. 2Hz,1H),8.34(d,J=5.2Hz,1H),8.26(d,J=8.0Hz,1H),7.94(d,J=0.8Hz,1H), 7.20(dd,J=8.4,1.6Hz,1H),7.04(d,J=1.6Hz,1H),5.80(s,2H),4.47-4.38( m,4H),3.89(s,3H),1.79-1.70(m,1H),1.13-1.04(m,2H),0.93-0.85(m,2H).

[0678] Example 18: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)methyl)-9H-pyrimido[4,5-b]indole-7-carbonitrile

[0679] Reaction route

[0680] Steps:

[0681] Step A: Dissolve methyl 3-hydroxy-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (1.13 g, 3.95 mmol) and cesium carbonate (3.9 g, 11.85 mmol) in acetonitrile (20 mL) at room temperature and stir for 1 hour. Then, add 1,2-dibromoethane (3.7 g, 19.76 mmol) to the reaction mixture, heat the temperature to 60°C, and continue stirring for 12 hours.

[0682] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered, the filter cake was washed with ethyl acetate, and the combined filtrates were concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 453 mg of methyl 2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine-9-carboxylate.

[0683] MS (ESI) M / Z: 313.2 [M+H] + .

[0684] Step B: Dissolve methyl 2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine-9-carboxylate (453 mg, 1.45 mmol) in tetrahydrofuran (7.3 mL) at 0°C under nitrogen. Then, slowly add lithium aluminum tetrahydride (1.2 mL, 2.9 mmol) dropwise to the solution, warm to room temperature, and continue stirring for 30 minutes.

[0685] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction mixture to quench the mixture, followed by filtration. The filter cake was washed with ethyl acetate (20 mL x 2). The filtrate was collected, the layers separated, and the aqueous phase was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and washed with saturated brine (40 mL x 2). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 200 mg of (2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepine-9-yl)methanol.

[0686] MS (ESI) M / Z: 285.3 [M+H] + .

[0687] Step C: At 0°C under nitrogen, (2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)methanol (50 mg, 0.18 mmol), triphenylphosphine (92 mg, 0.35 mmol), and 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrimido[4,5-b]indole-7-carbonitrile (90 mg, 0.26 mmol) were dissolved in dry tetrahydrofuran (4.9 mL). Diisopropyl azodicarboxylate (71 mg, 0.35 mmol) was then slowly added dropwise to the solution. The reaction was then stirred at room temperature for 1 hour.

[0688] After LCMS monitoring showed the disappearance of the starting material, ice water (50 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (30 mL × 3 times), and the organic phases were combined, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography to obtain 38.75 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-5,6-dihydrobenzo[f]imidazo[1,2-d][1,4]oxazepin-9-yl)methyl)-9H-pyrimido[4,5-b]indole-7-carbonitrile.

[0689] MS (ESI) M / Z: 609.2 [M+H] + .

[0690] 1H NMR(400MHz,DMSO-d6)δ9.81(s,1H),8.72(s,1H),8.57(d,J=8.0Hz,1H),8.52(s,1H), 8.25(d,J=8.4Hz,1H),7.94(s,1H),7.85(d,J=8.0Hz,1H),7.20(dd,J=8.4,1.6Hz,1H), 7.03(s,1H),5.76(s,2H),4.42(s,4H),3.88(s,3H),1.79-1.70(m,1H),1.12-1.04(m,2H),0.92-0.82(m,2H).

[0691] Example 19: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0692] Reaction route

[0693] Steps:

[0694] Step A: Methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (2 g, 7.4 mmol) was dissolved in N,N-dimethylformamide (37 mL) at room temperature. The solution was then cooled to 0°C and potassium carbonate (2.05 g, 14.8 mmol) and iodomethane-D3 (1.29 g, 8.89 mmol) were added sequentially. The reaction was then stirred at room temperature for 1 hour.

[0695] After LCMS monitoring showed the disappearance of the starting material, water (200 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (80 mL × 3 times), and the organic phases were combined and washed with saturated brine (80 mL × 2 times). The combined organic phases were purified by silica gel column chromatography to obtain 1.6 g of methyl 4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate.

[0696] MS (ESI) M / Z: 288.3 [M+H] + .

[0697] Step B: Methyl 4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (1.6 g, 5.57 mmol) was dissolved in tetrahydrofuran (28 mL) at 0°C under nitrogen. A 2M lithium aluminum tetrahydride solution (4.8 mL) was then slowly added dropwise to the solution. The reaction was then stirred at room temperature for 30 minutes.

[0698] After LCMS monitoring showed the disappearance of the starting material, water (100 mL) was added to the reaction solution to quench the reaction. The mixture was filtered, the filter residue was rinsed with ethyl acetate (70 mL × 2 times), the filtrate was collected, and the layers were separated. The aqueous phase was extracted with ethyl acetate (60 mL × 3 times), all organic phases were combined, and the organic phase was washed with saturated brine (50 mL × 2 times). It was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 1.34 g (4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol.

[0699] MS (ESI) M / Z: 260.2 [M+H] + .

[0700] Step C: Dissolve (4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (300 mg, 1.16 mmol) in dichloromethane (5.8 mL) at room temperature. The solution was then cooled to 0°C and triphenylphosphine (607 mg, 2.32 mmol), sodium bicarbonate (187 mg, 1.16 mmol), and carbon tetrabromide (767 mg, 2.32 mmol) were added sequentially. The reaction was then stirred at room temperature for 2 hours.

[0701] After LCMS monitoring showed the disappearance of the starting material, water (30 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with dichloromethane (20 mL × 3 times), and the organic phases were combined and washed with saturated brine (30 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 210 mg of 2-(4-(bromomethyl)phenyl)-1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazole.

[0702] MS (ESI) M / Z: 322.0 [M+H] + .

[0703] Step D: 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (74 mg, 0.22 mmol) was dissolved in N,N-dimethylformamide (1.2 mL) at 0°C under nitrogen. Sodium hydride (37 mg, 0.88 mmol) was then added to the solution and stirred for 20 minutes. 2-(4-(bromomethyl)phenyl)-1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazole (70 mg, 0.22 mmol) was then added, and the mixture was allowed to warm to room temperature and stirred for 30 minutes.

[0704] After LCMS monitoring showed the disappearance of the starting material, water (30 mL) was added to the reaction solution to quench the reaction. The mixed solution was washed with ethyl acetate (10 mL × 2 times), the organic phases were combined, and the organic phases were washed with saturated brine (20 mL × 2 times). It was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. 25.35 mg of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-(methyl-d3)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine was obtained.

[0705] MS (ESI) M / Z: 560.3 [M+H] + .

[0706] 1 H NMR (400MHz, DMSO-d6) δ9.82(s,1H),9.30(s,1H),8.73(s,1H),8.64(d,J=5.2Hz,1H),8.35(dd,J=5.2,0.8Hz,1H),7.90(d,J=1.2Hz, 1H),7.67(d,J=8.4Hz,2H),7.53(d,J=8.0Hz,2H),5.88(s,2H),3.89(s,3H),1.79-1.70(m,1H),1.12-1.04(m,2H),0.92-0.82(m,2H).

[0707] Example 20: 2-(2-isopropylphenyl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0708] Reaction route

[0709] Steps:

[0710] Step A: Dissolve (2-isopropylphenyl)boronic acid (170 mg, 1.04 mmol), 2-chloro-5-(3-chloropyridin-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (400 mg, 1.04 mmol), cesium carbonate (676 mg, 2.07 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (163 mg, 0.21 mmol) in 1,4-dioxane / water (4.40 mL / 0.74 mL) at room temperature. The air was evacuated and replaced with nitrogen three times, and the reaction system was stirred in a microwave at 90°C for 8 hours.

[0711] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (20 mL). The mixture was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 85 mg of 9-(2,4-dimethoxybenzyl)-2-(2-isopropylphenyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0712] MS (ESI) M / Z: 439.2 [M+H] + .

[0713] Step B: 9-(2,4-dimethoxybenzyl)-2-(2-isopropylphenyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (80 mg, 0.21 mmol) was dissolved in trifluoroacetic acid (3 mL) at room temperature and the reaction was stirred for 12 hours.

[0714] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 76 mg of 2-(2-isopropylphenyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0715] MS (ESI) M / Z: 289.1 [M+H] + .

[0716] Step C: Dissolve 2-(2-isopropylphenyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (70 mg, 0.24 mmol) in dry N,N-dimethylformamide (1.5 mL) at 0°C under nitrogen. Add sodium hydride (58 mg, 0.97 mmol) and stir for 20 minutes. Then add 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (77 mg, 0.24 mmol), warm to room temperature, and continue stirring for 10 minutes.

[0717] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected to obtain 11.73 mg of 2-(2-isopropylphenyl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0718] MS (ESI) M / Z: 527.2 [M+H] + .

[0719] 1 H NMR (400MHz, DMSO-d6) δ9.79(s,1H),9.22(s,1H),8.62(d,J=5.2Hz,1H),8.33(d,J=5.2Hz,1H),7.90(s,1H),7.73(d,J=8.0Hz,1H) ,7.67(d,J=8.4Hz,2H),7.54-7.43(m,4H),7.37-7.30(m,1H),5.89(s,2H),3.72(s,3H),3.67-3.55(m,1H),1.17(d,J=6.8Hz,6H).

[0720] Example 21: 2-(2-isopropylpyridin-3-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0721] Reaction route

[0722] Steps:

[0723] Step A: (2-isopropylpyridin-3-yl)boronic acid (500 mg, 2.02 mmol), 2-chloro-5-(3-chloropyridin-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (790 mg, 2.02 mmol), cesium carbonate (1.32 g, 04.05 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (320 mg, 0.41 mmol) were dissolved in 1,4-dioxane / water (8.70 mL / 1.45 mL) at room temperature. The air was evacuated and replaced with nitrogen three times, and the reaction system was stirred in a microwave at 90°C for 16 hours.

[0724] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (20 mL). The mixture was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 235 mg of 9-(2,4-dimethoxybenzyl)-2-(2-isopropylpyridin-3-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0725] MS (ESI) M / Z: 440.2 [M+H] + .

[0726] Step B: 9-(2,4-Dimethoxybenzyl)-2-(2-isopropylpyridin-3-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (210 mg, 0.48 mmol) was dissolved in trifluoroacetic acid (4 mL) at room temperature and the reaction was stirred for 16 hours.

[0727] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 167 mg of 2-(2-isopropylpyridin-3-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0728] MS (ESI) M / Z: 291.2 [M+H] + .

[0729] Step C: Dissolve 2-(2-isopropylpyridin-3-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (157 mg, 0.54 mmol) in dry N,N-dimethylformamide (2.7 mL) at 0°C under nitrogen. Add sodium hydride (87 mg, 2.17 mmol) to the solution and stir for 20 minutes. Then add 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (172 mg, 0.54 mmol), warm to room temperature, and continue stirring for 10 minutes.

[0730] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. The product was collected to obtain 62.29 mg of 2-(2-isopropylpyridin-3-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0731] MS (ESI) M / Z: 528.2 [M+H] + .

[0732] 1 H NMR(400MHz,DMSO-d6)δ9.82(s,1H),9.24(s,1H),8.68(dd,J=4.6,1.8Hz,1H),8 .63(d,J=5.2Hz,1H),8.35(dd,J=5.2,0.4Hz,1H),8.19(dd,J=7.6,1.8Hz,1H),7. 90(d,J=1.2Hz,1H),7.68(d,J=8.4Hz,2H),7.48(d,J=8.0Hz,2H),7.40(dd,J=8.0 ,4.8Hz,1H),5.91(s,2H),3.86-3.75(m,1H),3.73(s,3H),1.22(d,J=6.4Hz,6H).

[0733] Example 22: 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0734] Reaction route

[0735] Steps:

[0736] Step A: Dissolve 4-methoxy-1H-pyrazole (6.0 g, 61.2 mmol) and potassium carbonate (33.8 g, 244.8 mmol) in N,N-dimethylformamide (250 ml) at room temperature and stir for 30 minutes. Then, add cyclobutyl bromide (24.6 g, 183.6 mmol) dropwise to the solution, heat the temperature to 70°C, and stir for 24 hours.

[0737] After LCMS monitoring showed the disappearance of the starting material, ice water (800 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (250 mL x 3 times), and the organic phases were combined and washed with saturated brine (250 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 600 mg of 1-cyclobutyl-4-methoxy-1H-pyrazole.

[0738] MS (ESI) M / Z: 153.2 [M+H] + .

[0739] Step B: Dissolve 1-cyclobutyl-4-methoxy-1H-pyrazole (580 mg, 3.81 mmol) in dry tetrahydrofuran (40 mL) at -78°C under nitrogen. Slowly add 2.5 M n-butyllithium solution (2.0 mL, 5.0 mmol) dropwise to the solution and stir for 1 hour. Then, add 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (922 mg, 4.95 mmol) dropwise. Warm the mixture to room temperature and continue stirring for 2 hours.

[0740] After LCMS monitoring showed the disappearance of the starting material, ice water (20 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (10 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 600 mg of (1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)boronic acid.

[0741] MS (ESI) M / Z: 197.2 [M+H] + .

[0742] Step C: (1-Cyclobutyl-4-methoxy-1H-pyrazol-5-yl)boronic acid (84 mg, 0.43 mmol), 2-chloro-5-(3-chloropyridin-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (168 mg, 0.43 mmol), cesium carbonate (281 mg, 0.86 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (84 mg, 0.08 mmol) were dissolved in 1,4-dioxane / water (1.85 mL / 0.31 mL) at room temperature. The atmosphere was replaced with nitrogen three times under vacuum, and the reaction system was stirred at 90°C in a microwave for 12 hours.

[0743] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (20 mL). The mixture was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 115 mg of 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9-(2,4-dimethoxybenzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0744] MS (ESI) M / Z: 471.2 [M+H] + .

[0745] Step D: Dissolve 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9-(2,4-dimethoxybenzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (110 mg, 0.23 mmol) in trifluoroacetic acid (3 mL) at room temperature, bring to room temperature, and stir for 12 hours.

[0746] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 100 mg of 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0747] MS (ESI) M / Z: 321.2 [M+H] + .

[0748] Step E: Dissolve 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine (100 mg, 0.31 mmol) in dry N,N-dimethylformamide (1.6 mL) at 0°C under nitrogen. Add sodium hydride (50 mg, 1.25 mmol) to the solution and stir for 20 minutes. Then, add 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (99 mg, 0.31 mmol), warm to room temperature, and continue stirring for 10 minutes.

[0749] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography to obtain 15.44 mg of 2-(1-cyclobutyl-4-methoxy-1H-pyrazol-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrido[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0750] MS (ESI) M / Z: 559.2 [M+H] + .

[0751] 1 H NMR (400MHz, DMSO-d6) δ9.75(s,1H),9.27(s,1H),8.61(d,J=5.2Hz,1H),8.29(d,J=5.2Hz,1H),7.90(d,J=0.8Hz,1H),7.69(d,J=8.4Hz,2H),7.60 (s,1H),7.57(d,J=8.4Hz,2H),5.89(s,2H),5.68-5.56(m,1H),3.84(s,3 H),3.72(s,3H),2.61-2.53(m,2H),2.31-2.19(m,2H),1.79-1.54(m,2H).

[0752] Example 23: N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indol-7-yl)-N-methylmethanesulfonamide

[0753] Reaction route

[0754] Steps:

[0755] Step A: Dissolve N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indol-7-yl)methanesulfonamide (80 mg, 0.12 mmol) in N,N-dimethylformamide (1.2 mL) at 0°C under nitrogen. Sodium hydride (8.8 mg, 0.22 mmol) was then added to the solution, and the mixture was stirred for 0.5 hours. Methyl iodide (31 mg, 0.22 mmol) was then slowly added dropwise, and the mixture was allowed to warm to room temperature and stirred for 10 minutes.

[0756] After LCMS monitoring showed the disappearance of the starting material, water (20 mL) was added to the reaction solution to quench the reaction. The mixed solution was extracted with ethyl acetate (10 mL × 3 times), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. 42.99 mg of N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-9H-pyrimido[4,5-b]indol-7-yl)-N-methylmethanesulfonamide was obtained.

[0757] MS (ESI) M / Z: 663.2 [M+H] + .

[0758] 1 H NMR (400MHz, DMSO-d6) δ9.65(s,1H),8.71(s,1H),8.38(d,J=8.4Hz,1H),7.95(d,J=2.0Hz,1H),7.90(d,J=1.2Hz,1H),7.64(d,J=8.4Hz,2H), 7.54-7.45(m,3H),5.80(s,2H),3.88(s,3H),3.71(s,3H),3.36(s,3H) ,3.00(s,3H),1.76-1.67(m,1H),1.10-1.01(m,2H),0.89-0.81(m,2H).

[0759] Example 24: N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indol-7-yl)methanesulfonamide

[0760] Reaction route

[0761] Steps:

[0762] Step A: Dissolve N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9H-pyrimido[4,5-b]indol-7-yl)-N-((2-(trimethylsilyl)ethoxy)methyl)methanesulfonamide (366 mg, 0.68 mmol) in N,N-dimethylformamide (6.8 mL) at room temperature under nitrogen. Then, cool the solution to 0°C and slowly add sodium hydride (48.8 mg, 1.22 mmol) and stir for 0.5 hours. Then, add 9-(bromomethyl)-2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepine (233 mg, 0.68 mmol), warm to room temperature, and stir for 10 minutes.

[0763] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by adding water (20 mL). The mixture was extracted with ethyl acetate (10 mL x 3), then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 350 mg of N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indol-7-yl)-N-(2-(trimethylsilyl)ethoxy)methylmethyl)methanesulfonamide.

[0764] MS (ESI) M / Z: 805.2 [M+H] + .

[0765] Step B: Dissolve N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indol-7-yl)-N-(2-(trimethylsilyl)ethoxy)methylmethyl)methanesulfonamide (350 mg, 0.43 mmol) in dichloromethane (4 mL) at room temperature. Then, cool the solution to 0°C, slowly add trifluoroacetic acid (4 mL), warm to room temperature, and stir for 1 hour.

[0766] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was quenched by the addition of water (30 mL). The mixture was extracted with ethyl acetate (15 mL x 3 times), and the organic phases were combined and washed sequentially with saturated sodium bicarbonate solution (15 mL) and saturated brine (30 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified with dichloromethane / ethyl acetate to yield 168.30 mg of N-(2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-9-((2-(trifluoromethyl)-6,7-dihydro-5H-benzo[c]imidazo[1,2-a]azepin-9-yl)methyl)-9H-pyrimido[4,5-b]indol-7-yl)methanesulfonamide.

[0767] MS (ESI) M / Z: 675.2 [M+H] + .

[0768] 1 H NMR(400MHz,DMSO-d6)δ10.12(s,1H),9.56(s,1H),8.71(s,1H),8.28(d,J=8.4Hz,1H), 7.96(d,J=1.2Hz,1H),7.61(d,J=8.0Hz,1H),7.58(d,J=1.6Hz,1H),7.39(s,1H),7.27- 7.23(m,2H),5.70(s,2H),3.95(t,J=6.8Hz,2H),3.87(s,3H),3.04(s,3H),2.61(t,J=7 .0Hz,2H),2.28-2.15(m,2H),1.77-1.67(m,1H),1.10-1.02(m,2H),0.90-0.79(m,2H).

[0769] Examples 25-140

[0770] The target compounds shown in Table 1 below were prepared by referring to the synthesis method of Example 8 above:

[0771] Table 1

[0772] Example 141: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0773] Reaction route

[0774] Steps:

[0775] Step A: Dissolve 3-chloro-1H-pyrazole (5 g, 49.0 mmol) in N,N-dimethylformamide (50 mL) at 0°C. Slowly add N-iodosuccinimide (14.3 g, 63.7 mmol) to the solution. Stir the reaction at room temperature for 2 hours.

[0776] After LCMS monitoring showed the disappearance of the starting material, ice water (200 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (35 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 11 g of 3-chloro-4-iodo-1H-pyrazole.

[0777] MS (ESI) M / Z: 229.0 [M+H] + .

[0778] Step B: Dissolve 3-chloro-4-iodo-1H-pyrazole (3.5 g, 15.35 mmol) in dry tetrahydrofuran (77 mL) at 0°C under nitrogen. Slowly add sodium hydride (60%) (1.2 g, 30.70 mmol) to the solution and stir for 30 minutes. Then, slowly add iodomethane (4.3 g, 30.70 mmol) dropwise, warm to room temperature, and stir for 1 hour.

[0779] After LCMS monitoring showed the disappearance of the starting material, ice water (150 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2 g of 3-chloro-4-iodo-1-methyl-1H-pyrazole.

[0780] MS (ESI) M / Z: 243.0 [M+H] + .

[0781] 1 H NMR (400MHz, DMSO-d6) δ7.93 (s, 1H), 3.81 (s, 3H).

[0782] Step C: Dissolve 3-chloro-4-iodo-1-methyl-1H-pyrazole (1.6 g, 6.61 mmol) in dry tetrahydrofuran (33 mL) at room temperature. Slowly add 2M isopropylmagnesium chloride (10.5 mL, 21.02 mmol) dropwise to the solution and stir for 1 hour. Then, add 2-methoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.43 mg, 28.03 mmol) and continue stirring for 1 hour.

[0783] After LCMS monitoring showed the disappearance of the starting material, water (150 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 2 g of 3-chloro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)-1H-pyrazole.

[0784] MS (ESI) M / Z: 243.2 [M+H] + .

[0785] Step D: 3-Chloro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-yl)-1H-pyrazole (2 g, 8.23 ​​mmol), 5-bromo-2-chloro-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (1.5 g, 4.12 mmol), and sodium carbonate (0.87 g, 8.23 ​​mmol) were dissolved in 1,4-dioxane / water (20 mL / 2.2 mL) at room temperature. [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (0.33 g, 0.41 mmol) was then added to the solution. The atmosphere was evacuated and replaced with nitrogen three times under vacuum. The reaction system was stirred at 90°C for 16 hours.

[0786] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride aqueous solution (20 mL × 2 times). The organic phases were then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 780 mg of 2-chloro-5-(3-chloro-1-methyl-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine.

[0787] MS (ESI) M / Z: 394.2 [M+H] + .

[0788] Step E: 2-chloro-5-(3-chloro-1-methyl-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (780 mg, 1.98 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (386 mg, 1.98 mmol), and cesium carbonate (1.29 g, 3.96 mmol) were dissolved in 1,4-dioxane / water (10 mL / 1.7 mL) at room temperature. Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (312 mg, 0.40 mmol) was then added to the solution. The atmosphere was evacuated and replaced with nitrogen three times. The reaction system was stirred at 90°C in a microwave oven for 3 hours.

[0789] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride aqueous solution (20 mL × 2 times). The product was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 650 mg of 5-(3-chloro-1-methyl-1H-pyrazol-4-yl)-4'-cyclopropyl-N-(2,4-dimethoxybenzyl)-6'-methoxy-[2,5'-bipyrimidin]-4-amine.

[0790] MS (ESI) M / Z: 508.2 [M+H] + .

[0791] Step F: 5-(3-chloro-1-methyl-1H-pyrazol-4-yl)-4'-cyclopropyl-N-(2,4-dimethoxybenzyl)-6'-methoxy-[2,5'-bipyrimidin]-4-amine (650 mg, 1.28 mmol), N,N'-dimethylethylenediamine (68 mg, 0.77 mmol), and potassium carbonate (353 mg, 2.56 mmol) were dissolved in acetonitrile (6.4 mL) at room temperature. Cuprous iodide (73 mg, 0.38 mmol) was then added, and the atmosphere was evacuated and replaced with nitrogen three times. The reaction system was stirred at 120°C for 16 hours.

[0792] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 180 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0793] MS (ESI) M / Z: 472.2 [M+H] + .

[0794] Step G: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (180 mg, 0.38 mmol) was dissolved in trifluoroacetic acid (8 mL) at room temperature under nitrogen. The reaction was then stirred at 85°C for 16 hours.

[0795] After LCMS monitoring showed the disappearance of the starting material, the mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 120 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0796] MS (ESI) M / Z: 322.2 [M+H] + .

[0797] Step H: Dissolve 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (100 mg, 0.31 mmol) in dry N,N-dimethylformamide (1.5 mL) at 0°C under nitrogen. Slowly add sodium hydride (60%) (50 mg, 1.24 mmol) to the solution and stir for 0.5 hour. Then, add 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (162 mg, 0.47 mmol), raise the temperature to 50°C, and continue stirring for 16 hours.

[0798] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography. 25.98 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-methyl-2,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (Compound 88) was obtained.

[0799] MS (ESI) M / Z: 588.3 [M+H] + .

[0800] 1 H NMR (400MHz, DMSO-d6) δ9.14(s,1H),8.68(s,1H),8.26(s,1H),8.16(d,J=1.2Hz,1H),7.51(d,J=8.4Hz,2H),7.46(d,J=8.4Hz,2H),5.5 1(s,2H),4.45-4.36(m,1H),4.05(s,3H),3.85(s,3H),1.76-1.67(m,1H),1.37(d,J=6.4Hz,6H),1.08-1.00(m,2H),0.87-0.80(m,2H).

[0801] Example 142: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0802] Reaction route:

[0803] Steps:

[0804] Step A: Dissolve 3-chloro-1H-pyrazole (5 g, 49.02 mmol) in N,N-dimethylformamide (50 mL) at 0°C. Then, slowly add N-iodosuccinimide (14.3 g, 63.73 mmol) to the solution. The reaction mixture is stirred at room temperature for 2 hours.

[0805] After LCMS monitoring showed the disappearance of the starting material, ice water (200 mL) was added to the reaction solution to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (35 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 11 g of 3-chloro-4-iodo-1H-pyrazole.

[0806] MS (ESI) M / Z: 229.0 [M+H] + .

[0807] Step B: Dissolve 3-chloro-4-iodo-1H-pyrazole (4 g, 17.54 mmol) in N,N-dimethylformamide (88 mL) at room temperature under nitrogen. Add sodium hydride (1.05 g, 26.32 mmol) to the solution in an ice-water bath and stir for 30 minutes. Then, slowly add 2-(trimethylsilyl)ethoxymethyl chloride (4.4 g, 26.32 mmol), warm to room temperature, and stir for 1 hour.

[0808] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (500 mL) to quench the reaction. The mixed solution was extracted with ethyl acetate (80 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 5 g of 3-chloro-4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole.

[0809] MS (ESI) M / Z: 359.0 [M+H] + .

[0810] Step C: Dissolve 3-chloro-4-iodo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (5 g, 13.97 mmol) in dry tetrahydrofuran (70 mL) at room temperature. Add 2M isopropylmagnesium chloride (22.5 mL, 44.70 mmol) to the solution and stir for 1 hour. Then, slowly add 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (9.5 g, 60.07 mmol) dropwise and continue stirring for 1 hour.

[0811] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was added to ice water (100 mL) to quench the reaction. The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated brine (60 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 4 g of 3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole.

[0812] MS (ESI) M / Z: 359.2 [M+H] + .

[0813] Step D: 3-Chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (4 g, 11.17 mmol), (5-bromo-2-chloro-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (2 g, 5.59 mmol), sodium carbonate (1.18 g, 11.17 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (915 mg, 1.12 mmol) were dissolved in 1,4-dioxane / water (54 mL / 6 mL) at room temperature. The air was evacuated and replaced with nitrogen three times, and the reaction system was stirred at 100°C for 2 hours.

[0814] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 3 g of 2-chloro-5-(3-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine.

[0815] MS (ESI) M / Z: 510.2 [M+H] + .

[0816] Step E: 2-Chloro-5-(3-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (3 g, 5.89 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (2.3 g, 11.78 mmol), cesium carbonate (2.9 g, 8.84 mmol), and chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (2.3 g, 2.95 mmol) were dissolved in 1,4-dioxane / water (54 mL / 9 mL) at room temperature. The air was evacuated under vacuum three times to replace the nitrogen atmosphere, and the reaction was stirred at 130°C for 16 hours.

[0817] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide 240 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0818] MS (ESI) M / Z: 588.2 [M+H] + .

[0819] Step F: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (200 mg, 0.34 mmol) was dissolved in trifluoroacetic acid (3 mL) at room temperature. The reaction was stirred at 85°C for 5 hours.

[0820] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 70 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0821] MS (ESI) M / Z: 308.2 [M+H] + .

[0822] Step G: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (25 mg, 0.08 mmol), 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (28 mg, 0.08 mmol), and potassium carbonate (23 mg, 0.16 mmol) were dissolved in N,N-dimethylformamide (1 mL) at room temperature. The reaction was stirred at 50°C for 2 hours.

[0823] After LCMS monitoring showed the disappearance of the starting material, ice water (30 mL) was added to the reaction solution to quench the mixture. The mixture was extracted with ethyl acetate (10 mL × 3 times), and the organic phases were combined and washed with saturated brine (20 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative high-performance liquid chromatography to obtain 3.60 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0824] MS (ESI) M / Z: 574.2 [M+H] + .

[0825] 1 H NMR(400MHz,DMSO-d6)δ13.17(s,1H),9.13(s,1H),8.68(s,1H),8.30(s,1H),8.15(s,1H),7.53-7.46(m,4H),5.53(s ,2H),4.45-4.34(m,1H),3.86(s,3H),1.76-1.66(m,1H),1.36(d,J=6.8Hz,6H),1.07-1.00(m,2H),0.91-0.80(m,2H).

[0826] Example 143: 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine

[0827] Reaction route

[0828] Steps:

[0829] Step A: Dissolve 3-chloro-4-iodo-1H-pyrazole (112 g, 491 mmol) in dry tetrahydrofuran (1.4 L) at 0°C under nitrogen. Slowly add sodium hydride (42 g, 1.05 mol) to the solution and stir for 30 minutes. Then, add iodomethane (104 g, 736 mmol) dropwise. Warm the mixture to room temperature and continue stirring for 4 hours.

[0830] After LCMS monitoring showed the disappearance of the starting material, ice water (5 L) was added to the reaction solution to quench it. The mixture was extracted with ethyl acetate (1000 mL × 3 times), and the organic phases were combined and washed with saturated brine (600 mL × 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 23 g of 5-chloro-4-iodo-1-methyl-1H-pyrazole.

[0831] MS (ESI) M / Z: 243.0 [M+H] + .

[0832] 1 H NMR (400MHz, DMSO-d6) δ7.64 (s, 1H), 3.87 (s, 3H).

[0833] Step B: Dissolve 5-chloro-4-iodo-1-methyl-1H-pyrazole (3 g, 12.39 mmol) in dry tetrahydrofuran (62 mL) at room temperature. Add 2M isopropylmagnesium chloride solution (19.7 mL, 39.4 mmol) dropwise to the solution and stir for 1 hour. Then, add 2-methoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (6.72 g, 52.5 mmol) and continue stirring for 1 hour.

[0834] After LCMS monitoring showed the disappearance of the starting material, water (150 mL) was added to the reaction mixture to quench the reaction. The mixture was extracted with ethyl acetate (50 mL x 3 times), and the organic phases were combined and washed with saturated brine (20 mL x 2 times). The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 2.1 g of 5-chloro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborin-2-)-1H-pyrazole.

[0835] MS (ESI) M / Z: 243.2 [M+H] + .

[0836] Step C: Dissolve 5-chloro-1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborol-2-)-1H-pyrazole (2 g, 8.23 ​​mmol), 5-bromo-2-chloro-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (1.5 g, 4.12 mmol), and sodium carbonate (0.87 g, 8.23 ​​mmol) in 1,4-dioxane / water (20 mL / 2.2 mL) at room temperature. Then, add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (0.33 g, 0.41 mmol) to the solution. Evacuate the air and replace it with nitrogen three times. Stir the reaction system at 90°C for 16 hours.

[0837] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (20 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride solution (20 mL × 2 times). The organic phases were then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 1.8 g of 2-chloro-5-(5-chloro-1-methyl-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine.

[0838] MS (ESI) M / Z: 394.2 [M+H] + .

[0839] Step D: 2-Chloro-5-(5-chloro-1-methyl-1H-pyrazol-4-yl)-N-(2,4-dimethoxybenzyl)pyrimidin-4-amine (1.8 g, 4.57 mmol), (4-cyclopropyl-6-methoxypyrimidin-5-yl)boronic acid (891 mg, 4.57 mmol), and cesium carbonate (3 g, 9.14 mmol) were dissolved in 1,4-dioxane / water (23 mL / 2.3 mL) at room temperature. Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (644 mg, 0.91 mmol) was then added to the solution. The atmosphere was evacuated and replaced with nitrogen three times under vacuum. The reaction system was stirred at 90°C in a microwave oven for 16 hours.

[0840] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (150 mL). The mixture was extracted with ethyl acetate (50 mL × 3 times), and the organic phases were combined and washed with saturated sodium chloride aqueous solution (20 mL × 2 times). The product was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to yield 800 mg of 5-(5-chloro-1-methyl-1H-pyrazol-4-yl)-4'-cyclopropyl-N-(2,4-dimethoxybenzyl)-6'-methoxy-[2,5'-bipyrimidin]-4-amine.

[0841] MS (ESI) M / Z: 508.8 [M+H] + .

[0842] Step E: 5-(5-chloro-1-methyl-1H-pyrazol-4-yl)-4'-cyclopropyl-N-(2,4-dimethoxybenzyl)-6'-methoxy-[2,5'-bipyrimidin]-4-amine (800 mg, 1.58 mmol), N,N'-dimethylethylenediamine (278 mg, 3.16 mmol), and potassium carbonate (404 mg, 3.16 mmol) were dissolved in N,N-dimethylformamide (8 mL) at room temperature. Cuprous iodide (300 mg, 1.58 mmol) was then added to the solution. The atmosphere was evacuated and replaced with nitrogen three times under vacuum. The reaction system was stirred at 120°C for 16 hours.

[0843] After LCMS monitoring showed the disappearance of the starting material, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 520 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0844] MS (ESI) M / Z: 472.2 [M+H] + .

[0845] Step F: Dissolve 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(2,4-dimethoxybenzyl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (520 mg, 1.1 mmol) in trifluoroacetic acid (6 mL) at room temperature under nitrogen. The reaction was stirred at 85°C for 8 hours.

[0846] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to obtain 250 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0847] MS (ESI) M / Z: 322.2 [M+H] + .

[0848] Step G: Dissolve 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine (50 mg, 0.16 mmol) in N,N-dimethylformamide (1.0 mL) at 0°C under nitrogen. Add sodium hydride (25 mg, 0.62 mmol) and stir for 20 minutes. Then, add 2-(4-(bromomethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (60 mg, 0.17 mmol), warm to room temperature, and continue stirring for 1 hour.

[0849] After LCMS monitoring showed the disappearance of the starting material, the reaction solution was quenched by adding ice water (50 mL). The mixture was extracted with ethyl acetate (15 mL x 3), and the organic phases were combined and washed with saturated brine (20 mL x 2). The product was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative HPLC. Purification conditions were as follows: preparative column: YMC-Actus Triart C18 20.0 mm x 150 mm; mobile phase: water (containing 0.1% ammonium bicarbonate) and acetonitrile. This yielded 22.69 mg of 6-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-8-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-1-methyl-1,8-dihydropyrazolo[4',3':4,5]pyrrolo[2,3-d]pyrimidine.

[0850] MS (ESI) M / Z: 588.2 [M+H] + .

[0851] 1H NMR (400MHz, DMSO-d6) δ9.22 (s, 1H), 8.67 (s, 1H), 8.16 (d, J = 0.8Hz, 1H), 7.95 (s, 1H),7.54(d,J=8.0Hz,2H),7.35(d,J=8.0Hz,2H),5.85(s,2H),4.49-4.33(m,1H),3.97(s,3H) ,3.85(s,3H),1.74-1.65(m,1H),1.37(d,J=6.8Hz,6H),1.06-0.99(m,2H),0.86-0.78(m,2H).

[0852] Examples 144-166

[0853] The target compounds shown in Table 2 below were prepared by referring to the synthesis method of Example 142:

[0854] Table 2

[0855] 2. Biological Activity Test

[0856] As used herein, room temperature refers to about 20-30°C.

[0857] Test Example 1 USP1 enzyme activity test method

[0858] 1. Experimental plan:

[0859] USP1 enzyme activity assay was used to screen USP1i compounds.

[0860] 1.1 Experimental Materials: Recombinant human His6-USP1 / His6-UAF1 complex protein (R&D, catalog number E-568-050); Ubiquitin Rhodamine 110 (Ub-Rho) (R&D, catalog number U-555-050); Fluorescence 384-well plate (Perkin Elmer, catalog number 6007279).

[0861] 1.2 Test sample: The compound in Table 3 of this application, whose structural formula and preparation method are shown in the above examples.

[0862] 1.3 Experimental process:

[0863] (1) Prepare 1× assay buffer (modified Tris buffer, components: 50 mM Tris-HCl (pH 7.8) (Sigma, Catalog No.: T2569-1L), 0.01% Tween-20 (Sigma, Catalog No.: P2287-100ML), 1 mM DTT (Sigma, Catalog No.: D0632-10G), 0.01% BSA (Sigma, Catalog No.: B2064-100G), 0.5 mM EDTA (Invitrogen, Catalog No.: 15575020)).

[0864] (2) Compound dilution: Use dimethyl sulfoxide (DMSO, 100% purity) to prepare a 10 mM (mol / L) solution of the test compound; dilute the test compound solution 3-fold to 10 concentrations, with the highest concentration being 10 mM; transfer the diluted test compound solution to a fluorescent 384-well plate using an Echo acoustic pipetting system, with two replicate wells set for each concentration, and the final DMSO concentration is 1 vol%; the final concentrations of the test compound solution are 10000 nM, 3333 nM, 1111 nM, 370 nM, 123 nM, 41 nM, 13.7 nM, 4.6 nM, 1.5 nM, and 0.5 nM.

[0865] (3) Prepare enzyme solution: Prepare enzyme solution in 1× assay buffer.

[0866] (4) Prepare substrate solution: Add ubiquitin rhodamine 110 (Ub-Rho) to 1× detection buffer to form a substrate solution.

[0867] (5) Transfer 10 μL of the enzyme solution prepared in step (3) to a fluorescent 384-well plate.

[0868] (6) Incubate at room temperature for 1 hour.

[0869] (7) Add 10 μL of the substrate solution prepared in step (4) to each well to start the reaction. The final reaction system is 200 nL of the test compound + 10 uL of the enzyme solution + 10 uL of the substrate solution. The final concentration of the enzyme is 0.05 nM, and the final concentration of the substrate solution is 300 nM. Centrifuge for 30 s and shake for 30 s.

[0870] (8) The plate was read on a SpectraMax Paradigm multifunctional microplate reader for 30 minutes with an excitation wavelength of 480 nm and an emission wavelength of 540 nm.

[0871] (9) Collect data on SpectraMax Paradigm.

[0872] (10) Curve fitting:

[0873] The data were fitted in Excel using equation (I) to obtain inhibition values;

[0874] Equation (I): Inhibition rate % = (maximum signal value - target signal value) / (maximum signal value - minimum signal value) × 100;

[0875] Among them, the maximum signal value represents the luminescent signal intensity of the positive control well without the compound of the present application; the minimum signal value represents the luminescent signal intensity of the negative control well without the enzyme; and the target signal value represents the luminescent signal intensity of the test compound.

[0876] Fit the data in XL-Fit using equation (II) to obtain IC 50 value;

[0877] Equation (II): Y = Bottom + (Top - Bottom) / (1 + (IC 50 / X)×HillSlope)

[0878] Where Y is the inhibition percentage, X is the compound concentration; Bottom is the lowest inhibition rate; Top is the highest inhibition rate; HillSlope is the slope.

[0879] 2. Experimental results:

[0880] The results show that the compounds of the present invention have a good inhibitory effect on USP1, and their IC 50 The values ​​are generally lower than 1000 nanomolar (nM); IC values ​​of some compounds in the examples of this application are 50 The value is lower than 100nM, and the IC of the more preferred compounds of the present invention is 50 The inhibition results of some compounds of the present invention on USP1 are shown in Table 3.

[0881] Table 3 Enzyme inhibition results

[0882] 3. Conclusion:

[0883] It can be seen from the above experimental results that the compounds prepared in the examples of the present application have a good inhibitory effect on USP1 and are effective USP1 inhibitors.

[0884] Test Example 2 USP1CTG Test Method

[0885] 1. Experimental Plan

[0886] The in vitro biological activity of USP1i compounds was verified using the NCI-H1693 cell killing assay.

[0887] 1.1 Experimental Materials:

[0888] NCI-H1693 cells were purchased from ATCC (Cat. No. CRL-5866) and cultured in a 37°C, 5% CO2 (containing 5% CO2 and 95% air) cell culture incubator; 1640 complete medium: 94wt% RPMI-1640 liquid medium (Gibco Cat. No. 11875-093), 5wt% FBS (Gibco Cat. No. 10099-141), 1wt% Pen Strep (Gibco Cat. No. 15070-063); fluorescent 384-well plates (Perkin Elmer, Cat. No. 6007279); trypsin (Gibco Cat. No. 25200056); CTG buffer ( 2.0 Cell Viability Assay, Promega, Catalog No.: G9241).

[0889] 1.2 Test sample:

[0890] The compounds in Table 4 of this application; the structural formulas and preparation methods of compounds 58 and 88 are shown in the above examples;

[0891] The structural formula of comparative compound 1 is:

[0892] The structural formula of comparative compound 2 is:

[0893] 1.3 Experimental process:

[0894] (1) 75cm 2 Disintegrate NCI-H1693 cells in a culture flask with 2 mL of trypsin for 2-3 minutes, then neutralize with 2 mL of 1640 complete medium. Centrifuge at 1200 rpm for 5 minutes. Remove the supernatant and retain the cell pellet, then resuspend it in 4 mL of 1640 complete medium. Count 500 μL of the cell suspension using a Vi-CELL-XR cell counter.

[0895] (2) Using a Multidrop instrument, 500 NCI-H1693 cells were seeded per well in a fluorescent 384-well plate (each well contained 40 μL of 1640 complete medium), and drugs were added after 24 hours.

[0896] (3) Using an ultra-micropipette, the test sample (concentration: 3.33 mM, dissolved in DMSO) was diluted with 10 uM as the highest starting concentration, and then diluted with 9 gradients (10000 nM, 3333 nM, 1111 nM, 370 nM, 123 nM, 41 nM, 13.7 nM, 4.6 nM, 1.5 nM) in a 1:3 ratio, and the drug was added. Two replicate wells were set for each concentration; 9 wells were set for positive control and blank control, with the positive control being the replicate wells of 10 μM positive compound and the blank control being the replicate wells of DMSO.

[0897] (4) After drug addition, the cells were placed in a 37°C, 5% CO2 (containing 5% CO2 by volume and the rest being air) incubator and cultured for 6 days. After 6 days, 25 μL of CTG buffer was added to each well for CTG detection, and the plate was read and analyzed using a microplate reader.

[0898] (5) Analyze the CTG readings:

[0899] The average inhibition rate of the positive control replicates was set as 100%;

[0900] The average value of negative control replicates was set as the relative inhibition rate of 0%.

[0901] The CTG readings were converted into relative inhibition rates, and the inhibition rates (inhibition %) of the cells at various compound concentrations were calculated according to the following formula.

[0902] Inhibition%=(bx) / (ba×100%;

[0903] a=CTG value (highest concentration well)

[0904] b=CTG value (blank control well)

[0905] x=CTG value(x nM)

[0906] (6) Using GraphPad PRISM 8 to analyze IC 50 Perform calculations.

[0907] The concentrations and inhibition rates corresponding to 10000 nM, 3333 nM, 1111 nM, 370 nM, 123 nM, 41 nM, 13.7 nM, 4.6 nM, and 1.5 nM were statistically analyzed and calculated using Log10 (compound concentration).

[0908] Input the data into GraphPad PRISM 8, select "Analysis", select "Nonlinear regression (curve fit)", select "Log (inhibitor) vs. response-Variable slope", select the calculation formula, and calculate it according to the following formula: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)×HillSlope))

[0909] X:Log10(compound concentration)

[0910] Y: corresponding response

[0911] Top and Bottom: corresponding responses with the same units as Y

[0912] Notice:

[0913] - If X is not already the logarithm of dose, go back and transform the data.

[0914] - If any base response is subtracted, clamp Bottom to a constant value of 0.0.

[0915] Fit the data to obtain IC 50 value.

[0916] Adjust the fitting conditions according to the specific data conditions. Make appropriate constraint adjustments to Bottom, Top, and HillSlope to achieve the curve that best fits the actual situation.

[0917] 2. Experimental results:

[0918] The inhibition results of some of the compounds in the examples of this application on NCI-H1693 cells are shown in Table 4.

[0919] Table 4 NCI-H1693 cell inhibition results

[0920] 3. Conclusion:

[0921] It can be seen from the above experimental results that the compound of the present application has obvious cell inhibitory activity.

[0922] Test Example 3 Pharmacokinetic Determination of the Compound of the Application in Male CD1 Mice

[0923] 1. Experimental plan:

[0924] Male CD1 mice were used as test animals to study the pharmacokinetic characteristics of the compound of the present application in mouse plasma after intravenous and oral administration. Plasma samples were collected at specific time points, and the compound concentration in plasma was detected by LC-MS / MS. Drug metabolism and pharmacokinetic (DMPK) parameters were calculated to evaluate the pharmacokinetic characteristics of the compound of the present application in mouse plasma.

[0925] 1.1 Experimental drugs:

[0926] The present application lists 5 compounds; the structural formula and preparation method of compound 88 are shown in the above examples; the structural formula of comparative compound 1 is:

[0927] 1.2 Experimental Animals

[0928] Male CD1 mice, weighing 20-30 g, were purchased from Vital River.

[0929] 1.3 Administration:

[0930] Dosing information for the compound of this application: 3 mice each were administered in the IV group (intravenous injection) and the PO group (oral administration). The IV dose was 1 mg / kg in a 5 mL / kg volume, and the PO group was 10 mg / kg in a 10 mL / kg volume. The dosing vehicle consisted of 5 vol% DMSO, 10 vol% Solutol (Kolliphor HS 15), and 85 vol% saline.

[0931] 1.4 Sample collection:

[0932] After administration to mice, 0.02-0.03 mL of blood was collected from the saphenous vein at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours for the IV group and at 0.25, 0.5, 1, 2, 4, 8, and 24 hours for the PO group. The blood was placed in EDTA-K2 tubes and centrifuged at 4000 g for 5 minutes at 4°C to separate plasma, which was then stored at -80°C until sample analysis.

[0933] 1.5 Sample processing:

[0934] Mouse plasma sample processing:

[0935] 1) 10 μL of plasma sample was added to 200 μL of acetonitrile for precipitation, vortexed for 30 seconds, and centrifuged at 3900 rpm at 4°C for 15 minutes.

[0936] 2) The supernatant after treatment was diluted 3-fold with water, and the concentration of the test compound was analyzed by LC / MS / MS.

[0937] 2. Experimental results:

[0938] Table 5

[0939] 3. Conclusion:

[0940] It can be seen from the above experimental results that the compound of the present application has obvious pharmacokinetic advantages compared with the control compound.

[0941] Test Example 4 Pharmacokinetics of the Compound of the Application in Male Beagle Dogs

[0942] 1. Experimental plan:

[0943] Male Beagle dogs were used as test animals to study the pharmacokinetic characteristics of the compound of the present application in dog plasma after intravenous and oral administration. Plasma samples were collected at specific time points, and the compound concentration in plasma was detected by LC-MS / MS. DMPK parameters were calculated and the pharmacokinetic characteristics of the compound of the present application in dog plasma were evaluated.

[0944] 1.1 Experimental drugs:

[0945] The present application lists 6 compounds; among them, the structural formula and preparation method of compound 58 and compound 88 are shown in the above examples; the structural formula of KSQ-4279 is:

[0946] 1.2 Experimental Animals

[0947] Male Beagle dogs, weighing 9-13kg, supplied by Beijing Mas Biotechnology Co., Ltd.

[0948] 1.3 Administration:

[0949] Dosing information for the compound of this application: 3 dogs each received IV injection (0.2 mg / kg in a 0.5 mL / kg volume) and PO administration (3 mg / kg in a 3 mL / kg volume). The dosing vehicle consisted of 5 vol% DMSO, 10 vol% Solutol (Kolliphor HS 15), and 85 vol% saline.

[0950] 1.4 Sample collection:

[0951] After administration to the dogs, 0.5 mL of blood was collected from the cephalic vein at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours for the IV group and at 0.25, 0.5, 1, 2, 4, 8, and 24 hours for the PO group. The blood was placed in EDTA-K2 tubes and centrifuged at 2000 g for 10 minutes at 2-8°C to separate plasma. The plasma was then stored at -80°C until sample analysis.

[0952] 1.5 Sample processing:

[0953] Canine plasma sample processing:

[0954] 1) 50 μL of plasma sample was added to 200 μL of acetonitrile for precipitation, vortexed for 30 seconds, and centrifuged at 3900 rpm at 4°C for 15 minutes.

[0955] 2) The supernatant after treatment was diluted 3-fold with water, and the concentration of the test compound was analyzed by LC / MS / MS.

[0956] 2. Experimental results:

[0957] Table 6

[0958] Note: In Table 6, “CL” is total clearance; “AUC last ” is the area under the plasma drug concentration-time curve from time 0 to the final quantifiable time point.

[0959] 3. Conclusion:

[0960] It can be seen from the above experimental results that the compounds of the present application have obvious advantages in clearance level and drug exposure compared with the control compounds.

[0961] Test Example 5: OV0589 transplant tumor model (PDX) efficacy experiment

[0962] 1. Experimental plan:

[0963] Female BALB / c nude mice were used as test animals. After oral administration of the compounds of the present application, tumors were measured and weighed regularly to investigate tumor growth inhibition and tolerance in different dosing groups. Plasma and tumor samples were collected at specific time points after the study for pharmacokinetic / pharmacodynamic (PK / PD) analysis.

[0964] 1.1 Experimental drugs:

[0965] Compound 58, whose structural formula and preparation method are shown in the above examples;

[0966] Compound KSQ4279 has the structural formula:

[0967] 1.2 Experimental Materials:

[0968] Tumor-bearing mouse tumor (R10P8) was purchased from Sino-US Crown Biotechnology (Beijing) Co., Ltd.; Ubiquityl-PCNA (Lys164) (D5C7P) mAb antibody was purchased from Cell Signaling, catalog number: 13439S; PCNA (PC10) antibody was purchased from Santa Cruz, catalog number: sc-56.

[0969] 1.3 Experimental Animals

[0970] Female BALB / c nude mice, weighing 21-25 g, supplier: Beijing Ankai Yibo Biotechnology Co., Ltd.

[0971] 2. Experimental steps:

[0972] Under the conditions of temperature 20-26℃, humidity 30-70%, and alternating light and day, the tumor (R10P8) of the tumor-bearing mice grew to a diameter of about 1 cm (the tumor size reached 50-800 mm). 3 ) when the tumor was collected and cut into pieces of about 2 to 3 mm 3 The tumor mass (R10P8) was inoculated subcutaneously in the right anterior scapula of female BALB / c nude mice. The tumor growth in the mice was regularly observed. 35 days after inoculation, when the tumor (R10P9) grew to an average volume of approximately 150 mm 3 (Inclusion range 86.54mm 3 ~262.96mm 3 ) According to the tumor size and mouse body weight, the mice were randomly divided into groups using StudyDirector™ (version 3.1.399.19, supplier: Studylog System, Inc., S. San Francisco, CA, USA), and the drugs were administered on the next day after grouping.

[0973] Twelve mice were randomly divided into four groups, G1 to G4. Group G1 served as the vehicle control group, with the vehicle consisting of 10 vol% DMSO + 20 vol% Solutol + 70 vol% Water + 5 vol% DMSO + 10 vol% Solutol + 85 vol% Saline. Group G2 received oral administration of KSQ4279 at a dose of 100 mg / kg (mpk) in a vehicle consisting of 5 vol% DMSO + 10 vol% Solutol + 85 vol% Saline. Group G3 received oral administration of compound 58 at a dose of 30 mg / kg in a vehicle consisting of 10 vol% DMSO + 20 vol% Solutol + 70 vol% Water. Group G4 received oral administration of compound 58 at a dose of 100 mg / kg in a vehicle consisting of 10 vol% DMSO + 20 vol% Solutol + 70 vol% Water.

[0974] The growth of tumors in mice was observed regularly, and the tumor volume was measured (see Figure 1 for the results). The changes in the weight of mice were also observed regularly (see Figure 2 for the results). After 36 days of administration, plasma and tumor samples were collected for PK / PD analysis.

[0975] PK assay The pharmacokinetic assay in mice was performed as described in the previous test example 3.

[0976] PD detection: Tumor samples were collected 8 hours after the last administration and divided into tumor samples weighing 50-100 mg. 1× tissue lysis buffer was prepared using ddH2O, and then 20uL of tissue lysis buffer (Cell Signaling Cell Lysis Buffer (10×) Catalog Number: #9803) was added per mg of sample. The tumor samples were ground using a tissue grinder. The ground tumor sample lysate was placed on ice for 30 minutes. Subsequently, centrifuged at 12000rpm and 4°C for 15 minutes. The sample supernatant was retained. The PD marker (Ubiquityl-PCNA / PCNA) in the sample was detected using a protein blotting assay.

[0977] 3. Experimental results:

[0978] Figure 1 shows the tumor growth of mice in groups G1 to G4. As can be seen from Figure 1, compound 58 (at a dose of 100 mg / kg) had the best inhibitory effect on tumor growth, followed by compound 58 (at a dose of 30 mg / kg). KSQ4279 (at a dose of 100 mg / kg) had a significantly less inhibitory effect on tumor growth than compound 58 (at a dose of 100 mg / kg) and compound 58 (at a dose of 30 mg / kg). Figure 2 shows the weight changes of mice in groups G1 to G4. As can be seen from Figure 2, the weight changes of mice in groups G1-G4 were all within the normal range and were well tolerated.

[0979] Table 7 shows the statistical results of tumor growth inhibition rates in mice in groups G2 to G4. As can be seen from Table 7, compound 58 (dosage of 100 mg / kg) has the best inhibitory effect on tumor growth, followed by compound 58 (dosage of 30 mg / kg). The inhibitory effect of KSQ4279 (dosage of 100 mg / kg) on ​​tumor growth is significantly inferior to that of compound 58 (dosage of 100 mg / kg) and compound 58 (dosage of 30 mg / kg).

[0980] Table 7 Statistical results of tumor growth inhibition rate Note: "TGI%" is tumor growth inhibition rate; TGI% = [1-ΔT / C] × 100%, ΔT / C = (mean(T)-mean(T0)) / (mean(C)-mean(C0)), T and C are the mean tumor volumes of the drug-treated group and the vehicle control group on day 38, respectively, and T0 and C0 are the mean tumor volumes of the drug-treated group and the vehicle control group on day 0, respectively.

[0981] 4. Conclusion:

[0982] From the above data, it can be seen that compared with the control compound, the compound of the present application has obvious advantages in inhibiting tumor growth, and the dosage is significantly lower than that of the control compound.

Claims

1. A compound represented by formula (II'), its isomers and pharmaceutically acceptable salts thereof, in, X a C or N; Ring A is a benzene ring, a 5-6 membered heteroaryl ring, C 5-6 Cycloalkyl ring or 5-6 membered heterocyclyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4 Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C 1-4 alkyl)2P(O)-, Deuterated C 1-4 Alkyl, 4-6 membered heterocycloalkyl; the C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 Alkyl; m is 0, 1, 2, 3 or 4; R b For hydrogen, C 1-4 alkyl; Ring B is a benzene ring, a 5-10 membered heteroaryl ring, or a 5-10 membered heterocyclyl ring; R c Each independently is halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, deuterated C 1-4 Alkyl; n is 0, 1, 2, 3 or 4; Ring D is a benzene ring, a 5-6 membered heteroaryl ring or a 9-18 membered fused heterocyclic ring; R e Deuterium, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, ring C or halogen, wherein ring C can be 1 R d Substitution; the ring C is a 5-10 membered heteroaryl ring or an 8-10 fused heterocyclic ring containing 1-4 nitrogen atoms; the R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, deuterated C 1-4 Alkyl, C 3-6 Cycloalkyl; l is 1, 2, 3 or 4; p is 1, 2, 3 or 4; L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene or chemical bond; When ring A is a 5-6 membered heteroaryl ring, the structural unit Not for When ring A is a 5-6 membered heterocyclic ring, the structural unit Not for 2. The compound according to claim 1, its isomers and pharmaceutically acceptable salts thereof, having the structure represented by the following formula (I'): in, X a C or N; X b 、X c 、X d Each independently represents CH, N or CR', R' represents halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy; Ring A is a benzene ring, a 5-6 membered heteroaryl ring, C 5-6 Cycloalkyl ring or 5-6 membered heterocyclyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4 Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C 1-4 alkyl)2P(O)-, The C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 alkyl; m is 0, 1, 2, 3 or 4; R b For hydrogen, C 1-4 alkyl; Ring B is a benzene ring, a 5-10 membered heteroaryl ring, or a 5-10 membered heterocyclyl ring; R c Each independently is halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkyl, C 1-4 Haloalkoxy; n is 0, 1, 2, 3 or 4; Ring C is a 5-10 membered heteroaryl ring containing 1-4 nitrogen atoms; R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, deuterated C 1-4 Alkyl; l is 0, 1, 2, 3 or 4; L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene or chemical bond.

3. The compound according to claim 1, its isomers and pharmaceutically acceptable salts thereof, having the structure represented by the following formula (I'-1): in, X b 、X c 、X d Each independently represents CH, N or CR', R' represents halogen, C 1-4 alkoxy; Ring A is a benzene ring, pyridine or C 5-6 Cycloalkyl ring; R a Each independently represents deuterium, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, oxo, cyano, amino, hydroxy, aminosulfonyl, C 1-4 Alkylsulfonyl, carbamoyl, C 1-4 Alkylamino, C 3-6 Cycloalkyl, C 1-4 Alkylsulfonylamino, dimethylphosphonyl, -C 1-4 Alkyl-OH, -COOC 1-4 Alkyl, C 1-4 Alkyl-SO2-NR f -、HO-C 1-4 Alkyl-SO2-NR f -, 3-6 membered heterocyclic group, -C 1-4 Haloalkyl-OH, (C 1-4 alkyl)2P(O)-, The C 1-4 Alkyl, C 1-4 The C atoms in the haloalkyl group can be arbitrarily replaced by N or O; R f C 3-6 Cycloalkyl, C 1-4 Alkyl; m is 0, 1, 2, 3 or 4; R b For hydrogen, C 1-4 alkyl; Ring B is a 5-6 membered heteroaryl ring or a 5-10 membered heterocyclyl ring; R c Each is independent of C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, halogen; n is 0, 1, 2, 3 or 4; Ring C is a 5-6 membered heteroaryl ring containing 1-4 nitrogen atoms; R d Each is independent of C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy; l is 0, 1, 2, 3 or 4; L1 is C 1-4 Alkylene, C 3-6 Cycloalkylene.

4. The compound according to claim 1, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Ring D is 5. The compound according to claim 1, its isomers and pharmaceutically acceptable salts thereof, characterized in that: R e -CF3, -OCH3, -F, -D, -CH3.

6. The compound according to any one of claims 1 to 5, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Ring A is 7. The compound according to any one of claims 1 to 6, its isomers and pharmaceutically acceptable salts thereof, characterized in that: R a -F, -OCH3, -CF3, -COOCH3, -C(CH3)2-OH, -CHF2, -CN, -Cl, -NH2, CH3-NH-S(O)2-, NH2-S(O)2-, -CH3, -OH, -Br, -CH2CH3, CH(CH3)2, -OCH(CH3)2, D, -CD3, 8. The compound according to any one of claims 1 to 7, its isomers and pharmaceutically acceptable salts thereof, characterized in that Structural unit or structural unit for 9. The compound according to any one of claims 1 to 8, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Structural unit or structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

10. The compound according to any one of claims 1 to 9, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Structural unit or structural unit for Wherein a represents the connection with ring B, and b represents the connection with L1.

11. The compound according to any one of claims 1 to 10, its isomers and pharmaceutically acceptable salts thereof, characterized in that: R b For hydrogen.

12. The compound according to any one of claims 1 to 11, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Ring B is 13. The compound according to any one of claims 1 to 12, its isomers and pharmaceutically acceptable salts thereof, characterized in that: R c is -OCH3, -CH(CH3)2, -Cl, -OCHF2, -CF3, -CH3, -OCD3.

14. The compound according to any one of claims 1 to 13, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Structural unit for 15. The compound according to any one of claims 1 to 14, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Ring C is 16. The compound according to any one of claims 1 to 15, its isomers and pharmaceutically acceptable salts thereof, characterized in that: R d is -CF3, -CH3, -CH2CH3, -CH(CH3)2, -OCH2CH3, -CD3, -Cl, Br, -F.

17. The compound according to any one of claims 1 to 16, its isomers and pharmaceutically acceptable salts thereof, characterized in that: Structural unit for 18. The compound according to any one of claims 1 to 17, its isomers and pharmaceutically acceptable salts thereof, characterized in that: L1 is -CH2-, -CH(CH3)-, -C(CH3)2-, -CH(C2H5)-, Or a chemical bond.

19. The compound according to any one of claims 1 to 18, its isomers and pharmaceutically acceptable salts thereof, characterized in that: The compound, its isomer and pharmaceutically acceptable salt thereof are selected from the structures shown below, where R a 、R c 、R d 、R e , L1, m are the same as defined above; X is selected from C, N or O.

20. The compound according to any one of claims 1 to 18, its isomers and pharmaceutically acceptable salts thereof, characterized in that: The compound, its isomer and pharmaceutically acceptable salt thereof are selected from the structures shown below, where R a 、R c 、R d 、R e , m are as defined above; X is selected from C, N or O.

21. The compound according to any one of claims 1 to 20, its isomers and pharmaceutically acceptable salts thereof, selected from the following structures:

22. A pharmaceutical composition comprising the compound according to any one of claims 1 to 21, its isomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

23. Use of the compound according to any one of claims 1 to 21, its isomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 22 in the preparation of a drug for treating diseases mediated by USP1.

24. The use according to claim 23, wherein the related diseases mediated by the USP1 target include cellular inflammatory diseases, neurodegenerative diseases, and cancer.

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