Fluoroquinoxalinone derivatives capable of selectively inhibiting PARP1

Abstract

The invention discloses a series of fluorine-substituted heterocyclic compounds, and particularly discloses a compound as shown in a formula (XII) and pharmaceutically acceptable salts thereof. # imgabs0 #

Description

Fluorine-substituted heterocyclic compounds

[0001] This application claims priority to:

[0002] CN202210495459.8, application date: May 7, 2022;

[0003] CN202210693545.X, application date: June 17, 2022;

[0004] CN202211204558.2, application date: September 29, 2022;

[0005] CN202211351783.9, application date: October 31, 2022;

[0006] CN202211408423.8, application date: November 10, 2022;

[0007] CN202211593592.3, application date: December 9, 2022;

[0008] CN202310088997X, application date: February 2, 2023;

[0009] CN2023101237901, application date: February 15, 2023;

[0010] CN2023102323675, application date: March 10, 2023;

[0011] CN2023103034259, application date: March 23, 2023;

[0012] CN2023103408613, application date: March 31, 2023;

[0013] CN202310388951X, application date: April 12, 2023. Technical Field

[0014] The present invention relates to a series of fluorine-substituted heterocyclic compounds, and in particular to compounds represented by formula (XII) and pharmaceutically acceptable salts thereof. Background Art

[0015] Poly-ADP-ribose polymerase (PARP) is a very large protease family that currently consists of 18 members. It plays an important role in many cell cycle processes such as replication, recombination, chromatin remodeling and DNA damage repair.

[0016] Among them, PARP1 and PARP2 have been widely studied for their role in DNA damage repair. PARP1 is activated by DNA damage breaks and plays a role in catalyzing the addition of poly (ADP-ribose) (PAR) chains to target proteins. This post-translational modification, called PARylation, mediates the recruitment of additional DNA repair factors to DNA damage. After completing this recruitment, PARP auto-parylation triggers the release of PARP bound to DNA, allowing the entry of other DNA repair proteins to complete the repair.

[0017] Inhibition of PARP family enzymes has been developed as an anti-tumor strategy to selectively kill cancer cells by inactivating the complementary DNA repair pathway. Many preclinical and clinical studies have shown that tumor cells have BRCA1 or BRCA2 mutations, and BRCA is a key tumor suppressor protein involved in the repair of double-stranded DNA breaks (DSBs) through homologous recombination (HR). These tumors have defective homologous recombination repair (HRD) pathways and rely on the function of PARP enzymes for survival. Tumors with BRCA mutations grow more dependent on the PARP pathway, making them particularly sensitive to PARP1 inhibitors. In addition to being effective against BRCA-mutated cancers, PARP inhibitors have also been clinically verified to show certain efficacy in non-BRCA-mutated tumors that exhibit homologous recombination defects.

[0018] Compared with other clinical PARP1 / 2 inhibitors, highly selective PARP1 inhibitors improve the selectivity of PARP1 over PARP2, reducing the hematologic toxicity associated with PARP2 inhibition, and achieving better clinical efficacy and lower toxicity. Therefore, there is an unmet medical need for effective and safe PARP inhibitors, particularly those that are selective for PARP1.

[0019] PARP1 selective inhibitors can be used alone for solid tumors such as ovarian cancer, breast cancer, triple-negative breast cancer and prostate cancer with BRCA1 / 2 mutations (synthetic lethality mechanism); they can also be used as combination therapy drugs to enhance the effects of anti-cancer drugs with DNA damage mechanisms (such as DNA alkylating agents, topoisomerase inhibitors and platinum chemotherapy drugs) and radiotherapy; they can also be used in combination with other targeted drugs, such as PD-1 inhibitors, androgen receptor (AR) inhibitors, cell cycle checkpoint kinase (Chk) inhibitors, c-Met inhibitors, etc., for the treatment of various BRCA-negative or -positive solid tumors.

[0020] Summary of the Invention

[0021] The present invention provides a compound represented by formula (XII) or a pharmaceutically acceptable salt thereof,

[0022] in,

[0023] X is selected from O and S;

[0024] Structural unit Selected from

[0025] Ring A is selected from phenyl and 6-membered heteroaryl;

[0026] Ring B is selected from 6-membered heteroaryl;

[0027] L is selected from a single bond, and ring C is selected from

[0028] is selected from a single bond or a double bond;

[0029] T2 is selected from C, N and CH;

[0030] T3 and T4 are independently selected from N and CR 10 ;

[0031] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0032] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0033] R3 is selected from H and halogen;

[0034] R9 is absent or selected from H and halogen;

[0035] R4 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0036] R5 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0037] R6 and R7 are independently selected from H, halogen, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0038] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0039] R 10 is selected from H and halogen;

[0040] R 13 and R 14 are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0041] R 15 and R 16 are independently selected from H, D, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0042] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0043] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0044] Each R a are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0045] Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group;

[0046] Each R b are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0047] Each R c are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0048] Each R is independently selected from halogen and D;

[0049] n is selected from 0, 1, 2, 3 and 4;

[0050] The condition is that when the structural unit Selected from L is selected from a single bond, and ring C is selected from When R2 and R4 form a ring or R3 and R5 form a ring;

[0051] The halogen represents F, Cl, Br and I atoms;

[0052] The "hetero" in the 5-6 membered heteroaryl group represents 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -S- and -N-.

[0053] The present invention also provides a compound represented by formula (XII) or a pharmaceutically acceptable salt thereof,

[0054] in,

[0055] X is selected from O, S and N (R d );

[0056] Structural unit Selected from

[0057] Ring A is selected from phenyl and 5-6 membered heteroaryl;

[0058] Ring B is selected from 5-6 membered heteroaryl;

[0059] L is selected from a single bond, and ring C is selected from

[0060] Alternatively, L is selected from N(R 12 ), Ring C is selected from

[0061] is selected from a single bond or a double bond;

[0062] T2 is selected from C, N and CH;

[0063] T3 and T4 are independently selected from N and CR 10 ;

[0064] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0065] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0066] R3 is selected from H and halogen;

[0067] R9 is absent or selected from H and halogen;

[0068] R4 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0069] R5 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0070] R6 and R7 are independently selected from H, halogen, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0071] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0072] R 10 is selected from H and halogen;

[0073] R 12 Selected from H, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0074] R 13 and R 14are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0075] R 15 and R 16 are independently selected from H, D, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0076] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0077] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0078] Each R a are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0079] Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group;

[0080] Each R b are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0081] Each R c are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens;

[0082] R d Selected from C 1-3 Alkoxy, the C 1-3 Alkoxy is optionally substituted with 1, 2 or 3 halogens;

[0083] Each R is independently selected from halogen and D;

[0084] n is selected from 0, 1, 2, 3 and 4;

[0085] The condition is,

[0086] 1) When the structural unit Selected from L is selected from a single bond, and ring C is selected from One of T3 or T4 is selected from N, and the other is selected from CR 10 When R2, R3 and R9 are not H at the same time; or,

[0087] 2) When the structural unit Selected from L is selected from a single bond, and ring C is selected from One of T3 or T4 is selected from N, and the other is selected from CR 10 When R6 and R7 are not H at the same time; or,

[0088] 3) When the structural unit Selected from L is selected from a single bond, and ring C is selected from T3 is selected from N, T4 is selected from CR 10 When R5 is not H; or

[0089] 4) When the structural unit Selected from L is selected from a single bond, and ring C is selected from T4 is selected from N, T3 is selected from CR 10 When R4 is not H; or

[0090] 5) When the structural unit Selected from R 15 Selected from H, R 16 is selected from H, L is selected from a single bond, and ring C is selected from One of T3 or T4 is selected from N, and the other is selected from CR 10 When , Ring B is not pyrrolyl or pyrazolyl; or

[0091] 6) When the structural unit Selected from R 15Selected from H, R 16 is selected from H, L is selected from a single bond, and ring C is selected from One of T3 or T4 is selected from N, and the other is selected from CR 10 When , ring A is not furan.

[0092] The present invention also provides a compound represented by formula (XI) or a pharmaceutically acceptable salt thereof,

[0093] in,

[0094] X is selected from O, S and N (R d );

[0095] Structural unit Selected from

[0096] Ring A is selected from phenyl and 5-6 membered heteroaryl;

[0097] Ring B is selected from 5-6 membered heteroaryl;

[0098] L is selected from a single bond, and ring C is selected from

[0099] Alternatively, L is selected from N(R 12 ), Ring C is selected from

[0100] is selected from a single bond or a double bond;

[0101] T2 is selected from C, N and CH;

[0102] T3 and T4 are independently selected from N and CR 10 ;

[0103] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0104] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0105] R3 is selected from H and halogen;

[0106] R9 is absent or selected from H and halogen;

[0107] R4 is selected from H and halogen;

[0108] R5 is selected from H and halogen;

[0109] R6 and R7 are each independently selected from H and halogen;

[0110] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0111] R 10 is selected from H and halogen;

[0112] R 12 Selected from H, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0113] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0114] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0115] Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group;

[0116] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0117] Each R c are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0118] R d Selected from C 1-3 Alkoxy, the C 1-3 Alkoxy is optionally substituted with 1, 2 or 3 halogens;

[0119] Each R is independently selected from halogen and D;

[0120] n is selected from 0, 1, 2, 3 and 4;

[0121] The condition is,

[0122] 1) When the structural unit Selected from L is selected from a single bond, and ring C is selected from When one of T3 or T4 is selected from N, R2, R3 and R9 are not H at the same time; or

[0123] 2) When the structural unit Selected from L is selected from a single bond, and ring C is selected from When one of T3 or T4 is selected from N, R6 and R7 are not H at the same time; or

[0124] 3) When the structural unit Selected from L is selected from a single bond, and ring C is selected from T3 is selected from N, T4 is selected from CR 10 When R5 is selected from halogen; or

[0125] 4) When the structural unit Selected from L is selected from a single bond, and ring C is selected from T4 is selected from N, T3 is selected from CR 10 When, R4 is selected from halogen.

[0126] The present invention also provides a compound represented by formula (VIII) or a pharmaceutically acceptable salt thereof,

[0127] in,

[0128] X is selected from O, S and N (R d );

[0129] Structural unit Selected from

[0130] Ring A is selected from phenyl and 5-6 membered heteroaryl;

[0131] Ring B is selected from 5-6 membered heteroaryl;

[0132] is selected from a single bond or a double bond;

[0133] T2 is selected from C, N and CH;

[0134] T3 and T4 are independently selected from N and CR 10 ;

[0135] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0136] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0137] R3 is selected from H and halogen;

[0138] R9 is absent or selected from H and halogen;

[0139] R4 is selected from H and halogen;

[0140] R5 is selected from H and halogen;

[0141] R6 and R7 are each independently selected from H and halogen;

[0142] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0143] R 10 is selected from H and halogen;

[0144] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0145] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0146] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0147] Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group;

[0148] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0149] Each R care independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0150] R d Selected from OCH3;

[0151] Each R is independently selected from halogen and D;

[0152] n is selected from 0, 1, 2, 3 and 4;

[0153] The condition is,

[0154] 1) When the structural unit Selected from Structural unit Selected from When one of T3 or T4 is selected from N, R2, R3 and R9 are not H at the same time; or

[0155] 2) When the structural unit Selected from Structural unit Selected from When one of T3 or T4 is selected from N, R6 and R7 are not H at the same time; or

[0156] 3) When the structural unit Selected from Structural unit Selected from T3 is selected from N, T4 is selected from CR 10 When R5 is selected from halogen; or

[0157] 4) When the structural unit Selected from Structural unit Selected from T4 is selected from N, T3 is selected from CR 10 When R4 is selected from halogen; or

[0158] 5) When the structural unit Selected from Structural unit Selected from When either T3 or T4 is selected from N, R6 and R7 are not H at the same time.

[0159] The present invention also provides a compound represented by formula (VIII) or a pharmaceutically acceptable salt thereof,

[0160] in,

[0161] X is selected from O, S and N (Rd );

[0162] Structural unit Selected from

[0163] Ring A is selected from phenyl and 5-6 membered heteroaryl;

[0164] Ring B is selected from 5-6 membered heteroaryl;

[0165] is selected from a single bond or a double bond;

[0166] Structural unit Selected from

[0167] T3 and T4 are independently selected from N and CR 10 ;

[0168] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0169] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0170] R3 and R9 are each independently selected from H and halogen;

[0171] R4 is selected from H and halogen;

[0172] R5 is selected from H and halogen;

[0173] R6 and R7 are each independently selected from H and halogen;

[0174] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0175] R 10 is selected from H and halogen;

[0176] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0177] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0178] T2 is selected from C, N and CH;

[0179] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0180] Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group;

[0181] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0182] Each R c are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0183] R d Selected from OCH3;

[0184] Each R is independently selected from halogen and D;

[0185] n is selected from 0, 1, 2, 3 and 4;

[0186] The condition is,

[0187] 1) When the structural unit Selected from Structural unit Selected from When one of T3 or T4 is selected from N, R2, R3 and R9 are not H at the same time; or

[0188] 2) When the structural unit Selected from Structural unit Selected from When one of T3 or T4 is selected from N, R6 and R7 are not H at the same time; or

[0189] 3) When the structural unit Selected from Structural unit Selected from T3 is selected from N, T4 is selected from CR 10 When R5 is selected from halogen; or

[0190] 4) When the structural unit Selected from Structural unit Selected from T4 is selected from N, T3 is selected from CR 10 When R4 is selected from halogen; or

[0191] 5) When the structural unit Selected from Structural unit Selected from When either T3 or T4 is selected from N, R6 and R7 are not H at the same time.

[0192] The present invention also provides a compound represented by formula (VII-1) or a pharmaceutically acceptable salt thereof,

[0193] in,

[0194] When selected from a double bond, T2 is selected from C, R9 is absent, and R3 is selected from H and halogen;

[0195] When selected from a single bond, T2 is selected from N, CH, R9 and R3 are independently selected from H and halogen;

[0196] X is selected from O, S and N (R d );

[0197] T1 is selected from N;

[0198] Alternatively, T1 and R1 form a ring, so that the structural unit Selected from

[0199] T3 and T4 are independently selected from N and CR 10 ;

[0200] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0201] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0202] R4 is selected from H and halogen;

[0203] R5 is selected from H and halogen;

[0204] R6 and R7 are each independently selected from H and halogen;

[0205] R8 is selected from C 1-3Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0206] R 10 is selected from H and halogen;

[0207] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0208] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0209] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0210] or R on two adjacent atoms a Form double bonds with adjacent atoms;

[0211] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0212] Each R c are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0213] R d Selected from OCH3;

[0214] Each R is independently selected from halogen and D;

[0215] n is selected from 0, 1, 2, 3 and 4;

[0216] The condition is,

[0217] 1) when T1 is selected from N, T2 is selected from N, and one of T3 or T4 is selected from N, R2, R3 and R9 are not H at the same time; or,

[0218] 2) when T1 is selected from N, T2 is selected from N, and one of T3 or T4 is selected from N, R6 and R7 are not H at the same time; or,

[0219] 3) When T1 is selected from N, T2 is selected from N, T3 is selected from N, and T4 is selected from CR 10When R5 is selected from halogen; or

[0220] 4) When T1 is selected from N, T2 is selected from N, T4 is selected from N, and T3 is selected from CR 10 When R4 is selected from halogen; or

[0221] 5) When T1 is selected from N, the structural unit Selected from When either T3 or T4 is selected from N, R6 and R7 are not H at the same time.

[0222] The present invention also provides a compound represented by formula (VI) or a pharmaceutically acceptable salt thereof,

[0223] in,

[0224] When selected from a double bond, T2 is selected from C, R9 is absent, and R3 is selected from H and halogen;

[0225] When selected from a single bond, T2 is selected from N, CH, R9 and R3 are independently selected from H and halogen;

[0226] T1 is selected from N and CR 11 ;

[0227] T3 and T4 are selected from N and CR 10 ;

[0228] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0229] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0230] R4 is selected from H and halogen;

[0231] R5 is selected from H and halogen;

[0232] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0233] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0234] R6 and R7 are each independently selected from H and halogen;

[0235] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0236] R 11 Forming a benzene ring with R1, the structural unit Selected from

[0237] R 10 is selected from H and halogen;

[0238] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0239] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0240] Each R c are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0241] n is selected from 0, 1, 2, 3 and 4;

[0242] Provided that, when T1 is selected from N, T2 is selected from N, and T3 is selected from N,

[0243] 1) R2, R3 and R9 are not H at the same time, or;

[0244] 2) R6 and R7 are not H at the same time, or;

[0245] 3) R5 is selected from halogen.

[0246] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0247] in,

[0248] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0249] R2 is selected from C 1-3Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0250] R3 is selected from H;

[0251] R4 is selected from halogen;

[0252] R5 is selected from H;

[0253] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0254] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0255] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0256] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens.

[0257] The present invention also provides a compound represented by formula (VII-2) or a pharmaceutically acceptable salt thereof,

[0258] in,

[0259] T3 and T4 are independently selected from N and CR 10 ;

[0260] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0261] R4 is selected from H and halogen;

[0262] R5 is selected from H and halogen;

[0263] R6 and R7 are each independently selected from H and halogen;

[0264] R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently;

[0265] R 10 is selected from H and halogen;

[0266] R 12 Selected from H and C 1-3 alkyl;

[0267] Each R is independently selected from halogen and D.

[0268] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof

[0269] in,

[0270] R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0271] R2 is selected from C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0272] R3 is selected from H;

[0273] R4 is selected from halogen;

[0274] R5 is selected from H;

[0275] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0276] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0277] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0278] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens.

[0279] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0280] in,

[0281] R1 is selected from C1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens;

[0282] R2 is selected from C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0283] R3 is selected from H;

[0284] R4 is selected from halogen;

[0285] R5 is selected from H;

[0286] Alternatively, R2 and R4 form a ring, so that the structural unit Selected from

[0287] Alternatively, R3 and R5 form a ring, so that the structural unit Selected from

[0288] Each R a are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0289] Each R b are independently selected from H, halogen and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens.

[0290] In some embodiments of the present invention, the above X is selected from O, S and N(OCH3), and other variables are as defined in the present invention.

[0291] In some embodiments of the present invention, the above-mentioned X is selected from O and S, and other variables are as defined in the present invention.

[0292] In some embodiments of the present invention, the above-mentioned X is selected from O, and other variables are as defined in the present invention.

[0293] In some embodiments of the present invention, the above R a are independently selected from H, F, Cl, Br, I, CH3, CH2CH3 and CH2CH2CH3, said CH3, CH2CH3 and CH2CH2CH3 are optionally substituted by 1, 2 or 3 halogens, and other variables are as defined herein.

[0294] In some embodiments of the present invention, the above R aare independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0295] In some embodiments of the present invention, the above R a are independently selected from H and CH3, and other variables are as defined in the present invention.

[0296] In some embodiments of the present invention, the two R a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group, making the structural unit Selected from Other variables are as defined in the present invention.

[0297] In some embodiments of the present invention, the two R a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group, making the structural unit Selected from Other variables are as defined in the present invention.

[0298] In some embodiments of the present invention, the two R a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group, making the structural unit Selected from Other variables are as defined in the present invention.

[0299] In some embodiments of the present invention, R on the two adjacent atoms a Together with adjacent atoms, they form double bonds, making the structural unit Selected from Other variables are as defined in the present invention.

[0300] In some embodiments of the present invention, R on the two adjacent atoms a Together with adjacent atoms, they form double bonds, making the structural unit Selected from Other variables are as defined in the present invention.

[0301] In some embodiments of the present invention, the above R b are independently selected from H, F, Cl, Br, I, CH3, CH2CH3 and CH2CH2CH3, said CH3, CH2CH3 and CH2CH2CH3 are optionally substituted by 1, 2 or 3 halogens, and other variables are as defined herein.

[0302] In some embodiments of the present invention, the above R b are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0303] In some embodiments of the present invention, the above Rb are independently selected from H and CH3, and other variables are as defined in the present invention.

[0304] In some embodiments of the present invention, the above R c are independently selected from H, F, Cl, Br, I, CH3, CH2CH3 and CH2CH2CH3, said CH3, CH2CH3 and CH2CH2CH3 are optionally substituted by 1, 2 or 3 halogens, and other variables are as defined herein.

[0305] In some embodiments of the present invention, the above R c are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0306] In some embodiments of the present invention, the above R c are independently selected from H and F, and other variables are as defined in the present invention.

[0307] In some embodiments of the present invention, the above R d is selected from OCH3, and other variables are as defined in the present invention.

[0308] In some embodiments of the present invention, the above T3 is selected from N, CH and CF, and other variables are as defined in the present invention.

[0309] In some embodiments of the present invention, the above T3 is selected from N and CF, and other variables are as defined in the present invention.

[0310] In some embodiments of the present invention, the above T3 is selected from N, and other variables are as defined in the present invention.

[0311] In some embodiments of the present invention, the above T4 is selected from N and CH, and other variables are as defined in the present invention.

[0312] In some embodiments of the present invention, the above R1 is selected from CH3 and CH2CH3, and other variables are as defined in the present invention.

[0313] In some embodiments of the present invention, the above R1 is selected from CH3, and other variables are as defined in the present invention.

[0314] In some embodiments of the present invention, the above R2 is selected from H and CH3, and other variables are as defined in the present invention.

[0315] In some embodiments of the present invention, the above R2 is selected from CH3, and other variables are as defined in the present invention.

[0316] In some embodiments of the present invention, R4, R5, R6 and R7 are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0317] In some embodiments of the present invention, the above R3 is selected from H and F, and the other variables are as defined in the present invention.

[0318] In some embodiments of the present invention, the above R4 is selected from H, F, Cl, CN and CH3, and other variables are as defined in the present invention.

[0319] In some embodiments of the present invention, the above R4 is selected from H, F, Cl and CN, and other variables are as defined in the present invention.

[0320] In some embodiments of the present invention, the above R4 is selected from H, F and CH3, and other variables are as defined in the present invention.

[0321] In some embodiments of the present invention, the above R4 is selected from H and F, and other variables are as defined in the present invention.

[0322] In some embodiments of the present invention, the above R4 is selected from F, and other variables are as defined in the present invention.

[0323] In some embodiments of the present invention, the above R5 is selected from H, F, Cl and CN, and other variables are as defined in the present invention.

[0324] In some embodiments of the present invention, the above R5 is selected from H, F, CN and CH3, and other variables are as defined in the present invention.

[0325] In some embodiments of the present invention, the above R5 is selected from H, F and CH3, and other variables are as defined in the present invention.

[0326] In some embodiments of the present invention, the above R5 is selected from H and F, and the other variables are as defined in the present invention.

[0327] In some embodiments of the present invention, the above R5 is selected from H, and other variables are as defined in the present invention.

[0328] In some embodiments of the present invention, R6 and R7 are independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0329] In some embodiments of the present invention, the above R6 is selected from H, F and CH3, and other variables are as defined in the present invention.

[0330] In some embodiments of the present invention, the above R7 is selected from H, F and CH3, and the other variables are as defined in the present invention.

[0331] In some embodiments of the present invention, the above R6 is selected from H and F, and other variables are as defined in the present invention.

[0332] In some embodiments of the present invention, the above R7 is selected from H and F, and the other variables are as defined in the present invention.

[0333] In some embodiments of the present invention, the above R8 is selected from CH3, CH2CH3, CH2CF3, cyclopropyl and CD3, and other variables are as defined in the present invention.

[0334] In some embodiments of the present invention, the above R8 is selected from CH3, CH2CF3, cyclopropyl and CD3, and other variables are as defined in the present invention.

[0335] In some embodiments of the present invention, the above R8 is selected from CH3, cyclopropyl and CD3, and other variables are as defined in the present invention.

[0336] In some embodiments of the present invention, the above R8 is selected from CH3 and cyclopropyl, and other variables are as defined in the present invention.

[0337] In some embodiments of the present invention, the above R9 is selected from H and F, and the other variables are as defined in the present invention.

[0338] In some embodiments of the present invention, the above R 10 is selected from H and F, and other variables are as defined in the present invention.

[0339] In some embodiments of the present invention, the above R 12 is selected from H and CH3, and other variables are as defined herein.

[0340] In some embodiments of the present invention, the above R 13 is selected from H and CH3, and other variables are as defined herein.

[0341] In some embodiments of the present invention, the above R 14 is selected from H and CH3, and other variables are as defined herein.

[0342] In some embodiments of the present invention, the above R 15 is selected from H, D and CH3, and other variables are as defined in the present invention.

[0343] In some embodiments of the present invention, the above R 16 is selected from H, D and CH3, and other variables are as defined in the present invention.

[0344] In some embodiments of the present invention, Ring A is selected from phenyl and a 6-membered heteroaryl ring, and the other variables are as defined herein.

[0345] In some embodiments of the present invention, Ring A is selected from phenyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furanyl, thienyl, pyridinyl, pyrazinyl and pyrimidinyl, and the other variables are as defined herein.

[0346] In some embodiments of the present invention, Ring A is selected from phenyl, pyridinyl, pyrazinyl and pyrimidinyl, and the other variables are as defined herein.

[0347] In some embodiments of the present invention, Ring B is selected from a 6-membered heteroaryl ring, and the other variables are as defined herein.

[0348] In some embodiments of the present invention, Ring B is selected from pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furanyl, thienyl, pyridinyl, pyrazinyl and pyrimidinyl, and the other variables are as defined herein.

[0349] In some embodiments of the present invention, Ring B is selected from pyridinyl, pyrazinyl and pyrimidinyl, and the other variables are as defined herein.

[0350] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0351] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0352] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0353] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0354] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0355] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0356] In some embodiments of the present invention, the above R 11 Forming a benzene ring with R1, the structural unit Selected from Other variables are as defined in the present invention. In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0357] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0358] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0359] In some embodiments of the present invention, the above R2 and R4 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0360] In some embodiments of the present invention, the above R3 and R5 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0361] In some embodiments of the present invention, the above R3 and R5 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0362] In some embodiments of the present invention, the above R2 and R4 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0363] In some embodiments of the present invention, the above R3 and R5 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0364] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0365] In some embodiments of the present invention, the above R2 and R4 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0366] In some embodiments of the present invention, the above R3 and R5 form a ring, so that the structural unit Selected from Other variables are as defined in the present invention.

[0367] In some embodiments of the present invention, the above L is selected from single bonds, structural units Selected from Other variables are as defined in the present invention.

[0368] In some embodiments of the present invention, the above L is selected from single bonds, structural units Selected from Other variables are as defined in the present invention.

[0369] In some embodiments of the present invention, the above L is selected from single bonds, structural units Selected from Other variables are as defined in the present invention.

[0370] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0371] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0372] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0373] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0374] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0375] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0376] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0377] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0378] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0379] In some embodiments of the present invention, the above structural unit Selected from When R6 and R7 are not H at the same time, other variables are as defined in the present invention.

[0380] In some embodiments of the present invention, the above structural unit Selected from When R6 and R7 are not H at the same time, other variables are as defined in the present invention.

[0381] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

[0382] In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

[0383] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0384] wherein ring A is selected from pyridyl;

[0385] R2, R3, R4, R5, R8, R9, R 13 、R 14 、R 15 、R 16 、R c , T2, n and As defined in the present invention.

[0386] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0387] in, is selected from a single bond or a double bond;

[0388] T2 is selected from C and N;

[0389] R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens;

[0390] R3 is selected from H and halogen;

[0391] R4 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0392] R5 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy;

[0393] R9 is absent or selected from H and halogen;

[0394] R7, R8, R 13 、R 14 、R 15 、R 16 、R c and n are as defined in the present invention.

[0395] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0396] in,

[0397] Ring A is selected from phenyl and 6-membered heteroaryl;

[0398] Ring B is selected from 6-membered heteroaryl;

[0399] R2, R3, R4, R5, R6, R7, R8, R9, R 13 、R 14 、R 15 、R 16 、Rc , X, T2, T3, T4, n and As defined in the present invention.

[0400] In some embodiments of the present invention, the ring A is selected from a pyridine ring, and other variables are as defined in the present invention.

[0401] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0402] Among them, ring A, ring B, R1, R2, R3, R4, R5, R6, R7, R8, R9, X, T3, T4 and structural units As defined in the present invention.

[0403] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0404] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R a 、R b , X, T3 and T4 are as defined in the present invention.

[0405] In some embodiments of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0406] Among them, R1, R2, R3, R4, R5, R6, R7, R8, R9, R a 、R b , X, T3 and T4 are as defined in the present invention;

[0407] The condition is,

[0408] 1) When the compound is selected from formula (VIII-6a), when one of T3 or T4 is selected from N, R2, R3 and R9 are not H at the same time; or,

[0409] 2) When the compound is selected from formula (VIII-6a), when one of T3 or T4 is selected from N, R6 and R7 are not both H; or,

[0410] 3) When the compound is selected from formula (VIII-6a), T3 is selected from N, T4 is selected from CR 10 When R5 is selected from halogen; or

[0411] 4) When the compound is selected from formula (VIII-6a), T4 is selected from N, T3 is selected from CR 10 When R4 is selected from halogen; or

[0412] 5) When the compound is selected from formula (VIII-7a), T3 is selected from N, and R6 and R7 are not H at the same time.

[0413] Some other solutions of the present invention are obtained by any combination of the above variables.

[0414] The present invention also provides the following compounds or pharmaceutically acceptable salts thereof,

[0415] In some embodiments of the present invention, the compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

[0416] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound as defined in the present invention or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier, diluent or excipient.

[0417] The present invention also provides the use of the above compound or its pharmaceutically acceptable salt or the above composition in the preparation of drugs for treating solid tumors.

[0418] In some embodiments of the present invention, the solid tumor refers to solid tumors such as ovarian cancer, breast cancer, prostate cancer and brain glioma.

[0419] The present invention also provides the following synthesis method:

[0420] Method 1

[0421] Method 2

[0422] Method 3

[0423] Method 4

[0424] Method 5

[0425] Method 6

[0426] Method 7

[0427] Method 8

[0428] Method 9

[0429] Method 10

[0430] The present invention also provides the following biological testing method:

[0431] Test method 1: PARP1 enzyme activity test

[0432] 1. Experimental Materials:

[0433] PARP1 chemiluminescence detection kit was purchased from BPS Bioscience; EnVision multi-label analyzer was purchased from PerkinElmer.

[0434] 2. Experimental steps:

[0435] Reagent preparation:

[0436] Preparation of PBST buffer: 1X PBS contains 0.05% Tween-20, that is, add 5 μL 100% Tween-20 to 10 mL PBS

[0437] Preparation of 1X assay buffer: dilute 10X PARP assay buffer 10-fold with double distilled water.

[0438] Compound preparation:

[0439] Compound solution preparation: Dilute the test compound 5-fold in 100% DMSO to the eighth concentration, from 1000 μM to 12.8 nM. Dilute the internal control compound 5-fold in 100% DMSO to the eighth concentration, from 200 μM to 2.56 nM. Then, serially dilute the test compound in 1X assay buffer to a 10% DMSO working solution.

[0440] Experimental methods:

[0441] a) Dilute the histone solution in the kit 5-fold with 1X PBS, dispense 25 μL / well of the dilution into a microplate, and incubate overnight at 4°C.

[0442] b) After incubation, discard the liquid in the wells, take 100 μL / well PBST and wash the plate three times, discarding the remaining liquid in the wells;

[0443] c) Add 100 μL / well of blocking solution to the microplate and incubate at 25°C for 90 minutes. After incubation, discard the liquid in the wells and wash the plate three times with 100 μL / well of PBST, discarding any remaining liquid.

[0444] d) Pipette 12.5 μL / well of substrate mixture solution (1.25 μL 10X PARP assay buffer; 1.25 μL 10X PARP assay mixture; 2.5 μL activated DNA; 7.5 μL double-distilled water) into the microplate.

[0445] e) Pipette 2.5 μL / well of compound working solution into a microplate and set up a duplicate well experiment.

[0446] f) Dilute PARP1 enzyme to 2 ng / μL, add 10 μL / well to the microplate, and incubate the reaction system at 25°C for 60 minutes.

[0447] g) After incubation, discard the liquid in the wells, take 100 μL / well PBST and wash the plate three times, discarding the remaining liquid in the wells;

[0448] h) Dilute Streptavidin-HRP 50-fold with blocking buffer, then dispense 25 μL / well into a microplate and incubate at 25°C for 30 minutes;

[0449] i) After incubation, discard the liquid in the wells, take 100 μL / well PBST and wash the plate three times, discarding the remaining liquid in the wells;

[0450] j) Mix ELISA ECL substrate A and ELISA ECL substrate B at a ratio of 1:1 (v / v), transfer 50 μL / well to a microplate, and read the chemiluminescence value.

[0451] 3. Experimental Data Processing Method

[0452] The raw data were converted to enzyme activity using the equation (Sample-Min) / (Max-Min) × 100%. The IC50 value was then obtained by four-parameter curve fitting (using the log(inhibitor) vs. response - Variable slope mode in GraphPad Prism).

[0453] Max: Contains 1% DMSO, PARP1 and substrate mixed solution

[0454] Min: Does not contain PARP1 enzyme

[0455] Test method 2: Study on the binding ability of the compounds of the present invention to PARP1 and PARP2

[0456] PARP1 experimental procedures:

[0457] Surface plasmon resonance (SPR) experiments were performed on a Biacore 8K (GE Healthcare) instrument.

[0458] First, biotinylated PARP1 protein (SEQ ID NO: 655-end) was coupled to a streptavidin-coated SA chip (Cytiva, 29699622) at 25°C. The chip surface was activated with 1 mM NaCl / 50 mM NaOH. A 10 μg / mL ligand solution of PARP1 protein was diluted in coupling buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 10 mM MgCl2, 0.05% P20) and passed over the chip surface (injection time 50 s, injection flow rate 5 μL / min) to couple the PARP1 protein to the chip surface. Excess active sites on the chip were blocked with a 50% isopropanol / 1 M NaCl / 50 mM NaOH solution. The final coupling level was 2000–3000 RU (Response Units).

[0459] Small molecule compounds were gradiently diluted with buffer (50mM Tris pH 8.0, 150mM NaCl, 10mM MgCl2, 0.05% Tween 20) to obtain compound solutions of varying concentrations. The solutions were then passed over the protein-coupled chip surface at an injection rate of 50μL / min, an injection time of 60s, and a dissociation time of 20min. The instrument detected the binding and dissociation curves of the protein-small molecule compounds.

[0460] The data of the sample channel and reference channel were analyzed by Biacore 8K evaluation software to generate sensorgrams, and data fitting was performed based on a 1:1 binding mode.

[0461] PARP2 experimental procedures:

[0462] Surface plasmon resonance (SPR) experiments were performed on a Biacore 8K (GE Healthcare) instrument.

[0463] First, biotinylated PARP2 protein (SEQ ID NO: 223-end) was coupled to a streptavidin-coated SA chip (Cytiva, 29699622) at 25°C. The chip surface was activated with 1 mM NaCl / 50 mM NaOH. A 10 μg / mL ligand solution of PARP2 protein was diluted in coupling buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 10 mM MgCl2, 0.05% P20) and passed over the chip surface (injection time 60 s, injection flow rate 10 μL / min) to couple PARP2 protein to the chip surface. Excess active sites on the chip were blocked with a 50% isopropanol / 1 M NaCl / 50 mM NaOH solution. The final coupling level was 3000–4000 RU (Response Units).

[0464] Small molecule compounds were gradiently diluted with buffer (50mM Tris pH 8.0, 150mM NaCl, 10mM MgCl2, 0.05% Tween 20) to obtain compound solutions of varying concentrations. The solutions were passed over the protein-coupled chip surface at an injection rate of 30μL / min, an injection time of 60s, and a dissociation time of 400s. The instrument detected the binding and dissociation curves of the protein-small molecule compounds.

[0465] The data of the sample channel and reference channel were analyzed by Biacore 8K evaluation software to generate sensorgrams, and data fitting was performed based on a 1:1 binding mode.

[0466] Technical Effects

[0467] The compounds of the present invention have excellent binding activity to PARP1, selectively inhibiting PARP1 protein, weaker binding activity to PARP2, and excellent antiproliferative activity against BRCA1-mutated MDA-MB-436 cells and DLD1 (BRCA2 knockout) cells, as well as excellent membrane permeability. The compounds of the present invention exhibit excellent stability in vitro in human and mouse liver microsomes. They also have excellent in vivo metabolic stability, demonstrating excellent oral absorption drug exposure, and oral administration results in excellent brain tissue drug concentrations, a high brain-to-blood ratio, and significant anti-tumor activity.

[0468] Related definitions

[0469] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be construed as indefinite or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0470] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are 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.

[0471] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, which are prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases, having specific substituents. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in a neat solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in a neat solution or a suitable inert solvent. Certain specific compounds of the present invention contain both basic and acidic functional groups and can be converted into either base or acid addition salts.

[0472] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture of the two.

[0473] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0474] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0475] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0476] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0477] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0478] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicate the relative configuration of stereocenters with a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Indicates a straight solid bond or straight dashed key

[0479] Unless otherwise specified, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom on the double bond is connected to two different substituents (in a double bond containing a nitrogen atom, a lone pair of electrons on the nitrogen atom is considered as a substituent to which it is connected), if a wavy line is used between the atom on the double bond and its substituent in the compound, When connected, it represents the (Z) isomer, (E) isomer or a mixture of the two isomers of the compound. For example, the following formula (A) represents that the compound exists in the form of a single isomer of formula (A-1) or formula (A-2) or in the form of a mixture of the two isomers of formula (A-1) and formula (A-2); the following formula (B) represents that the compound exists in the form of a single isomer of formula (B-1) or formula (B-2) or in the form of a mixture of the two isomers of formula (B-1) and formula (B-2). The following formula (C) represents that the compound exists in the form of a single isomer of formula (C-1) or formula (C-2) or in the form of a mixture of the two isomers of formula (C-1) and formula (C-2).

[0480] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and readily interconvert into each other. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0481] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of that isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0482] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0483] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For example, deuterated drugs can be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs have advantages such as reduced toxic side effects, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included within the scope of this invention.

[0484] The terms "optional" or "optionally" mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0485] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be any on a chemically feasible basis.

[0486] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, with each occurrence of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permissible only if such combinations result in stable compounds.

[0487] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0488] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0489] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bond is connected, the number of H atoms at the site will decrease accordingly with the number of connected chemical bonds, and become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy lines For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least In these four connection methods, even if an H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group. It means that R can be connected to either end of the double bond, that is,

[0490] Unless otherwise specified, the terms "two R a Together with the atoms to which they are attached, they form a double bond" means that two R a Together with the single bond between the two carbons, a double bond is formed. For example, the structural unit Two R's a Together with the single bond between the two carbons, a double bond is formed, forming a structural unit

[0491]

[0046] The terms "halo" or "halogen," by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom.

[0492] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1- Examples of 3-alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0493] Unless otherwise specified, the term “C 1-3 "Alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule via an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 、C 2-3 , C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0494] Unless otherwise specified, “C 3-5 "Cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 5 carbon atoms, which is a monocyclic system. 3-5 Cycloalkyl groups include C 3-4 and C 4-5 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent.3-5 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and the like.

[0495] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" are used interchangeably in the present invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 5 to 6 ring atoms, 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 a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. 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, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0496] Unless otherwise specified, the terms "6-membered heteroaromatic ring" and "6-membered heteroaryl" are used interchangeably in the present invention. The term "6-membered heteroaryl" refers to a monocyclic group with a conjugated π electron system consisting of 6 ring atoms, 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). The 5-6 membered heteroaryl group can be attached to the remainder of the molecule through a heteroatom or a carbon atom.

[0497] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 、C 11 , and C12 , also includes any range from n to n+m, such as C 1-12 Including C 1- 3. C 1-6 、C 1-9 、C 3-6 、C 3-9 、C 3-12 、C 6-9 、C 6-12 , and C 9-12 Similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to n+m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

[0498] The compounds 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, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0499] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is performed by collecting diffraction intensity data from a cultured single crystal using a Bruker D8venture diffractometer with CuKα radiation and a φ / ω scanning mode. After collecting the relevant data, the crystal structure can be further analyzed using a direct method (Shelxs97) to confirm the absolute configuration.

[0500] The solvents used in the present invention are commercially available. The present invention uses the following abbreviations: hr represents hours; min represents minutes; DIEA represents N,N-diisopropylethylamine; DDQ represents 2,3-dichloro-5,6-dicyanobenzoquinone; XPHOS-PD-G2 represents chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II); TBSCl represents tert-butyldimethylchlorosilane; DMAP represents 4-dimethylaminopyridine; Cs2CO3 represents carbonic acid Cesium; RuPhos represents 2-dicyclohexylphosphine-2,6-diisopropoxy-1,1-biphenyl; Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium; TBAF represents tetrabutylammonium fluoride; Et3N represents triethylamine; Na2CO3 represents sodium carbonate; NaHCO3 represents sodium bicarbonate; H2O represents water; PE represents petroleum ether; EA represents ethyl acetate; DMF represents N,N-dimethylformamide; MeOH represents methanol; EtOH represents ethanol; THF represents tetrahydrofuran ; DCM represents dichloromethane; NBS represents N-bromosuccinimide; Dess-Martin represents Dess-Martin periodinane, CAS number 87413-09-0; KHMDS represents potassium bis(trimethylsilyl)amide; Pd(OAc)2 represents palladium acetate; PPh3 represents triphenylphosphine; Pd / C represents palladium on carbon; HATU represents 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; (Boc)2O represents di-tert-butyl dicarbonate ; LiAlH4 represents lithium aluminum hydride; N-Boc-piperazine represents 1-tert-butyloxycarbonylpiperazine; Lawesson's reagent represents a compound with CAS number 19172-47-5; DMSO represents dimethyl sulfoxide; NaBH(OAc)3 represents sodium triacetoxyborohydride; KI represents potassium iodide; NH4Cl represents ammonium chloride; HCl represents hydrochloric acid; dioxane represents 1,4-dioxane; Pd(dppf)Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride. BRIEF DESCRIPTION OF THE DRAWINGS

[0501] Figure 1. Tumor growth volume

[0502] Figure 2. Changes in mouse body weight during drug administration DETAILED DESCRIPTION

[0503] The present invention is described in detail below by way of examples, but is not intended to limit the present invention in any way. While the present invention has been described in detail herein, and specific embodiments thereof have been disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0504] Example 1

[0505] Step 1: Synthesis of intermediate 1c

[0506] Compound 1a (9.5 g, 39.92 mmol), DIEA (15.48 g, 119.75 mmol, 20.86 mL), and compound 1b (6.13 g, 43.91 mmol, HCl) were added to DMF (100 mL) and stirred at 25°C for 18 hours. 1000 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined, dried, and concentrated to obtain the crude product. Intermediate 1c was obtained by column chromatography (eluent: PE:EA = 4:1, V / V). MS m / z: 320.8, 322.8 [M+H] + .

[0507] Step 2: Synthesis of intermediate 1d

[0508] Intermediate 1c (7.4 g, 23.05 mmol), NH4Cl (9.86 g, 184.37 mmol), and zinc powder (7.7 g, 117.76 mmol) were added to a mixture of MeOH (100 mL) and H2O (2 mL) at 0°C and stirred for 2 hours. The mixture was filtered, and the methanol was evaporated under reduced pressure. 100 mL of water was added, and the mixture was extracted with ethyl acetate (2 × 30 mL). The organic phases were combined, dried, filtered, and the filtrate was concentrated under reduced pressure to yield Intermediate 1d, which was used directly in the next step. MS m / z: 290.7, 292.7 [M+H] + .

[0509] Step 3: Synthesis of intermediate 1e

[0510] Intermediate 1d (6.7 g, 23.01 mmol) was added to a mixture of dioxane (4 M, 1.81 mL), ethyl acetate (70 mL), and methanol (70 mL) at 0°C and stirred for 2 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure to afford Intermediate 1e (5.5 g, 21.23 mmol). 1 H NMR(400MHz,DMSO-d6)δppm 10.46(s,1H)7.42-7.57(m,1H)7.00(dd,J=8.53,7.28Hz,1H)6.49(d,J=8.03Hz,1H)3.82-3.86(m,1H)1.25(d,J=6.53Hz,3H).

[0511] Step 4: Synthesis of Intermediate 1f

[0512] At 0°C, DDQ (5.26 g, 23.16 mmol) was added portionwise to Intermediate 1e (5.0 g, 19.30 mmol) in DCM (250 mL). The mixture was heated to 25°C and stirred for 2 hours. The solvent was evaporated under reduced pressure, and the reaction was quenched by the addition of 200 mL of saturated aqueous NaHCO₃. The reaction was stirred overnight and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined and concentrated to obtain the desired product. The mixture was purified by column chromatography (eluent: DCM:MeOH = 100:1, V / V) to afford Intermediate 1f. MS m / z: 256.7, 258.7 [M+H] + , 1 H NMR (400MHz, DMSO-d6) δppm 12.61 (br s, 1H) 7.45-7.55 (m, 2H) 2.40 (s, 3H).

[0513] Step 5: Synthesis of Intermediate 1g

[0514] Intermediate 1f (1.0 g, 3.89 mmol), 1-(tributyltin)-methanol (1.50 g, 4.67 mmol), and XPHOS-PD-G2 (153.04 mg, 194.51 μmol) were added to 1,4-dioxane (20 mL) and stirred at 80°C under N2 protection for 4 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (eluent: DCM / MeOH = 20 / 1, V / V) to obtain intermediate 1g. MS m / z: 209.0 [M+1] + .

[0515] Step 6: Synthesis of Intermediate 1h

[0516] Under nitrogen at 0°C, tributylphosphine (388.72 mg, 1.92 mmol, 474.05 μL) was slowly added dropwise to a solution of 1,2-dibromotetrachloroethane (688.23 mg, 2.11 mmol, 253.96 μL) and intermediate 1g (200 mg, 960.67 μmol) in DCM (5 mL). The reaction was stirred at 20°C for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. A mixed solvent (DCM / MeOH = 20 / 1, 5 mL) was added to the crude product and stirred for 30 minutes. The mixture was filtered, and the filter cake was concentrated under reduced pressure to obtain intermediate 1h. 1 H NMR (400MHz, DMSO-d6) δppm 12.55 (s, 1H) 7.47-7.58 (m, 1H) 7.31-7.43 (m, 1H) 4.80-4.90 (m, 2H) 2.42 (s, 3H).

[0517] Step 7: Synthesis of Intermediate 1j

[0518] Triethylamine (1.87 g, 18.49 mmol) was added to a solution of Intermediate 1i (2 g, 9.25 mmol), TBSCl (2.09 g, 13.87 mmol), and DMAP (112.97 mg, 924.74 μmol) in DCM (40 mL). The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to afford the crude product. The crude product was purified by column chromatography (eluent: DCM / MeOH = 20 / 1 to 10 / 1, V / V) to afford Intermediate 1j. 1 H NMR (400MHz, CD3OD) δppm 3.88-4.12(m,2H)3.63(d,J=5.52Hz,2H)3.01(d,J=12.30Hz,1H)2.54-2.94(m,4H)1.48(s,9H)0.95(s,9H)0.12(s,6H).

[0519] Step 8: Synthesis of Intermediate 1k

[0520] Cs2CO3 (197.15 mg, 605.08 μmol), RuPhos (14.12 mg, 30.25 μmol), intermediate 1j (0.1 g, 302.54 μmol), 5-bromo-6-fluoro-pyridine-2-carboxylic acid methyl ester (70.80 mg, 302.54 μmol), and Pd2(dba)3 (55.41 mg, 60.51 μmol) were added to a toluene (4 mL) solution and stirred at 100°C for 24 hours. The reaction solution was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography (eluent: PE:EA = 1:1, V / V) to obtain intermediate 1k. MS m / z: 484.3 [M+H] + .

[0521] Step 9: Synthesis of Intermediate 11

[0522] TBAF (1M THF solution, 258.45 μL, 2.5 eq) and intermediate 1k (50 mg, 103.38 μmol) were added to THF (4 mL) and the reaction was stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain intermediate 1l. MS m / z: 350.2 [M+H] + .

[0523] Step 10: Synthesis of intermediate 1m hydrochloride

[0524] Intermediate 1l (30 mg, 85.87 μmol) was added to EA (2 mL) solution. HCl / EA (4 M, 85.87 μL) was added dropwise to the reaction mixture, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of Intermediate 1m. MS m / z: 250.2 [M+H] + .

[0525] Step 11: Synthesis of Intermediate 1n

[0526] Under nitrogen atmosphere, the hydrochloride salt of intermediate 1m (20 mg) was added to an EtOH (2 mL) solution, and methylamine (2.26 mg, 72.84 μmol) was added to the reaction solution. The reaction was stirred at 20°C for 1 hour. The solution was concentrated under reduced pressure to obtain intermediate 1n. MS m / z: 249.1 [M+H] + .

[0527] Step 12: Synthesis of trifluoroacetate salt of compound 1

[0528] Intermediate 1n (10.50 mg, 36.89 μmol) was added to DMF (2 mL), followed by the addition of Et3N (7.47 mg, 73.78 μmol, 10.27 μL) and the reaction mixture, which was stirred at 25°C for 30 min. KI (2.70 mg, 16.24 μmol, 5.68 μL) and intermediate 1h (10 mg, 36.89 μmol) were then added to the reaction mixture, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was concentrated at 60°C using an oil pump to obtain the crude product. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile percentage: 0%-23% over 8 min) to afford the trifluoroacetate salt of compound 1. MS m / z: 439.2 [M+H] + , 1 H NMR(400MHz,CD3OD)δppm 7.63-7.73(m,2H)7.36-7.50(m,2H)4.44-4.54(m,3H)4.22(dd,J=11.26,7.63Hz,1H)4.13(d, J=11.01Hz,1H)3.51-3.64(m,3H)3.11-3.22(m,2H)2.92(s,3H)2.85-2.91(m,1H)2.56(s,3H).

[0529] Example 2

[0530] Step 1: Synthesis of intermediate 2c

[0531] Compound 2a (0.5 g, 2.14 mmol) and compound 2b (642.89 mg, 3.21 mmol) were added to toluene (10 mL). Cs2CO3 (2.09 g, 6.42 mmol), RuPhos (199.72 mg, 428.00 μmol), and Pd2(dba)3 (195.96 mg, 214.00 μmol) were added to the reaction solution. After nitrogen was purged three times, the reaction was stirred at 100°C for 16 hours. The reaction solution was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (eluent: PE / EA = 4 / 1 to 2 / 1, V / V) to obtain intermediate 2c. MS m / z: 354.2 [M+1] + , 1 H NMR (400MHz, CDCl3) δppm 7.99(d,J=8.03Hz,1H)7.22-7.29(m,1H)4.01(d,J=6.53Hz,1H)4.00-4.24(m,1H)3.98(s,3H)3 .81(d,J=13.30Hz,1H)3.29-3.44(m,2H)3.07-3.25(m,2H)1.50(s,9H)1.08(d,J=6.53Hz,3H).

[0532] Step 2: Synthesis of intermediate 2d

[0533] Intermediate 2c (0.4 g, 1.13 mmol) and methylamine tetrahydrofuran (2 M, 7.27 mL) were added to EtOH (10 mL) and the reaction was stirred at 20°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain crude intermediate 2d, which was used directly in the next step. MS m / z: 353.0 [M+H] + .

[0534] Step 3: Synthesis of intermediate 2e hydrochloride

[0535] Intermediate 2d (0.4 g, 1.14 mmol) was added to EA (5 mL), and ethyl acetate (4 M, 1.42 mL) was added to the reaction mixture. The reaction was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride salt of intermediate 2e, which was used directly in the next step. MS m / z: 253.0 [M+H] + .

[0536] Step 4: Synthesis of compound 2 trifluoroacetate

[0537] Intermediate 2e (21.30 mg) was added to DMF (2 mL), followed by the addition of Et3N (14.93 mg, 147.56 μmol, 20.54 μL). The reaction was stirred at 25°C for 30 min. KI (5.39 mg, 32.49 μmol) and intermediate 1h (20 mg, 73.78 μmol) were then added to the reaction, and the reaction was stirred at 50°C for 2 h. The reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile %: 0%-26% over 8 min) to obtain the trifluoroacetate salt of compound 2. MS m / z: 443.1 [M+H] + , 1 H NMR(400MHz,DMSO-d6)δppm 7.19(d,J=7.28Hz,1H)6.99(s,1H)6.90(d,J=8.78Hz,1H)6.67(d,J=6.78Hz,1H)3.80 (s,2H)2.78(s,1H)2.70(s,3H)2.39-2.48(m,3H)2.14(s,3H)1.76(s,3H)0.32(s,3H).

[0538] Example 3

[0539] Step 1: Synthesis of intermediate 3b

[0540] Compound 3a (1 g, 4.62 mmol) was added to DCM (20 mL), and the reaction mixture was cooled to 0°C. Et3N (935.74 mg, 9.25 mmol, 1.29 mL), TBSCl (1.05 g, 6.94 mmol, 849.87 μL), and DMAP (56.49 mg, 462.37 μmol) were added to the reaction mixture, and the mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (eluent: DCM / MeOH = 20 / 1 to 10 / 1, V / V) to obtain intermediate 3b. 1 H NMR (400MHz, CDCl3) δppm 3.86(s,1H)3.52(s,1H)3.41(dd,J=9.79,7.28Hz,1H)2.92(d,J=11.29Hz,1H)2.63-2.8 4(m,3H)2.48(s,1H)1.90(s,1H)1.28-1.48(m,9H)0.72-0.97(m,9H)-0.08-0.08(m,6H).

[0541] Step 2: Synthesis of intermediate 3c

[0542] Methyl 5-bromo-6-fluoro-pyridine-2-carboxylate (1 g, 4.27 mmol) and intermediate 3b (987.97 mg, 2.99 mmol) were added to toluene (20 mL). RuPhos (398.80 mg, 854.00 μmol), Pd2(dba)3 (391.30 mg, 427.00 μmol), and Cs2CO3 (4.17 g, 12.81 mmol) were added to the reaction solution. The atmosphere was purged with nitrogen three times, and the reaction was stirred at 100°C for 2 hours. The solution was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 4 / 1 to 2 / 1, V / V) to obtain intermediate 3c. MS m / z: 484.3 [M+H] + .

[0543] Step 3: Synthesis of intermediate 3d

[0544] Compound 3c (300 mg, 620.29 μmol) was added to THF (2 mL), and TBAF (1 M THF solution, 1.86 mL) was added to the reaction mixture. The mixture was stirred at 20°C for 2 hours. The reaction mixture was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain crude product 3d, which was used directly in the next step. MS m / z: 350.1 [M+H] + .

[0545] Step 4: Synthesis of intermediate 3e

[0546] Compound 3d (200.00 mg) was added to EtOH (5 mL), and a 40% ethanolic solution of methylamine (1 mL) was added to the reaction mixture. The mixture was stirred at 20°C for 32 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was used directly in the next step to obtain intermediate 3e. MS m / z: 349.1 [M+H] + .

[0547] Step 5: Synthesis of intermediate 3f hydrochloride

[0548] Compound 3e (0.1 g) was added to EA (5 mL), and hydrogen chloride / ethyl acetate (4 M, 358.79 μL) was added to the reaction mixture. The mixture was stirred at 20°C for 8 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride salt of compound 3f, which was used directly in the next step. MS m / z: 249.1 [M+H] + .

[0549] Step 6: Synthesis of trifluoroacetate salt of compound 3

[0550] Compound 3f (10.50 mg, HCl) was added to DMF (2 mL). Et3N (7.47 mg, 73.78 μmol, 10.27 μL) was then added to the reaction mixture, stirred at 25°C for 30 min, and KI (2.70 mg, 16.24 μmol, 5.68 μL) and compound 1h (10 mg, 36.89 μmol) were added to the reaction mixture, stirred at 50°C for 2 hours. The reaction mixture was concentrated at 60°C to afford the crude product. The crude product was purified by preparative HPLC (column: Xtimate C18 150 × 40 mm × 5 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile percentage: 1%-30%, 10 min) to afford the trifluoroacetate salt of compound 3. MS m / z: 439.0 [M+H] + , 1 H NMR(400MHz,CD3OD)δppm 7.66-7.71(m,2H)7.42-7.48(m,1H)7.36-7.42(m,1H)4.50(dd,J=11.19,2.56Hz ,1H)4.43(s,2H)4.22(dd,J=11.13,7.75Hz,1H)4.12(d,J=10.01Hz,1H)3.56(br s,1H)3.51(d,J=9.38Hz,2H)3.14(s,2H)2.92(s,3H)2.85(s,1H)2.56(s,3H).

[0551] Example 4

[0552] Step 1: Synthesis of intermediate 4a

[0553] Toluene (5 mL) was added to a reaction flask containing compound 2a (500 mg, 2.14 mmol) and N-Boc-piperazine (397.93 mg, 2.14 mmol). RuPhos (199.40 mg, 427.31 μmol), cesium carbonate (1.39 g, 4.27 mmol), and Pd2(dba)3 (195.65 mg, 213.65 μmol) were then added sequentially. The mixture was heated to 100°C and stirred under nitrogen for 16 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain the crude product. Intermediate 4a was then isolated and purified by silica gel column chromatography (gradient elution: PE:EA = 100:0 to 80:20) to obtain the product. MS m / z: 340.1 [M+H] + . 1H NMR (400MHz, CDCl3) δppm 7.98-7.98(m,1H)7.91-8.06(m,1H)3.98(s,3H)3.57-3.68(m,4H)3.16-3.31(m,4H)1.51(s,9H).

[0554] Step 2: Synthesis of intermediate 4b

[0555] Compound 4a (200 mg, 589.34 μmol) was dissolved in ethanol (8 mL), and methylamine ethanol solution (1.19 g, 17.68 mmol) was added. The mixture was stirred at 25°C for 16 hours. The reaction solution was concentrated to obtain intermediate 4b, which was used in the next reaction without purification. MS m / z: 339.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 1.42 (s, 9H) 2.93 (d, J = 5.13Hz, 3H) 3.03-3.16 (m, 4H) 3.44-3.63 (m, 4H) 7.24 (dd, J = 10.01, 8.13Hz, 1H) 7.44 (br d, J = 4.13Hz, 1H) 7.93 (dd, J = 8.00, 1.00Hz, 1H).

[0556] Step 3: Synthesis of intermediate 4c

[0557] 4b (50 mg, 147.76 μmol) was dissolved in toluene (2.5 mL), and Lawesson's reagent (71.72 mg, 177.32 μmol) was added. The mixture was stirred at 120°C for 16 hours. The reaction mixture was extracted with water (1 mL) and ethyl acetate (3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by prep-TLC (PE:EA = 5:1) to afford intermediate 4c. MS m / z: 355.1 [M+H] + .

[0558] Step 4: Synthesis of intermediate 4d

[0559] Intermediate 4c (10 mg, 28.21 μmol) was dissolved in DMF (0.5 mL), and sodium hydride (2.82 mg, 70.53 μmol, 60% purity) was added. The mixture was stirred at 0°C for 0.5 hours. Iodomethane (8.01 mg, 56.43 μmol, 3.51 μL) was added, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was quenched with two drops of ammonium chloride solution and extracted with ethyl acetate (5 mL). The organic phase was washed with saturated brine (2 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford Intermediate 4d, which was used directly in the next step without purification. MS m / z: 369.2 [M+H]+ .

[0560] Step 5: Synthesis of intermediate 4e

[0561] Intermediate 4d (5 mg, 13.57 μmol) was dissolved in EtOH (0.5 mL), and triethylamine (6.87 mg, 67.85 μmol, 9.44 μL) and methoxyamine hydrochloride (3.40 mg, 40.71 μmol) were added. The mixture was stirred at 60°C for 1.5 hours. The reaction mixture was extracted with ethyl acetate (5 mL), and the organic phase was washed with water (2 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The mixture was separated and purified by prep-TLC (PE:EA = 1:1). The reaction mixture was filtered and concentrated to yield Intermediate 4e. MS m / z: 368.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δppm 7.48-7.56(m,1H)7.15-7.18(m,1H)3.79(s,3H)3.51-3.56(m,4H)3.00-3.05(m,4H)2.92(s,3H)1.42(s,9H).

[0562] Step 6: Synthesis of intermediate 4f hydrochloride

[0563] Intermediate 4e (10 mg, 27.22 μmol) was dissolved in MeOH (0.5 mL), and hydrogen chloride / dioxane (4 M, 36.74 μL) was added. The mixture was stirred at 25°C for 1.5 hours. LCMS detected the majority of the starting material. An additional 0.2 mL of hydrogen chloride / dioxane (4 M, 36.74 μL) was added, and stirring was continued for 1.5 hours. The reaction solution was concentrated to afford intermediate 4f hydrochloride, which was used directly in the next step without purification.

[0564] Step 7: Synthesis of trifluoroacetate salt of compound 4

[0565] Intermediate 4f (10 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (6.66 mg, 65.84 μmol, 9.16 μL) was added. The mixture was stirred at 25°C for 0.5 h. Intermediate 1h (8.92 mg, 32.92 μmol) and potassium iodide (2.73 mg, 16.46 μmol) were then added, and the mixture was stirred at 50°C for 2 h. The reaction mixture was filtered to obtain the crude product, which was then separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile %: 0%-30%; 8 min) to obtain the trifluoroacetate salt of compound 4. MS m / z: 458.2 [M+H] + . 1H NMR (400MHz, CD3OD) δppm 2.56 (s, 3H) 3.03 (s, 3H) 3.59 (br s, 8H) 3.83-3.94 (m, 3H) 4.64 (s, 2H) 7.46-7.52 (m, 1H) 7.60-7.76 (m, 3H).

[0566] Example 5

[0567] Step 1: Synthesis of intermediate 5a hydrochloride

[0568] Intermediate 4c (19.4 mg, 54.73 μmol) was dissolved in MeOH (1 mL), and hydrogen chloride / dioxane (4 M, 73.89 μL) was added. The mixture was stirred at 25°C for 16 hours. The reaction mixture was concentrated to obtain the hydrochloride salt of intermediate 5a, which was used directly in the next step without purification. MS m / z: 255.0 [M+H] + .

[0569] Step 2: Synthesis of compound 5 trifluoroacetate

[0570] Intermediate 5a (15 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (10.44 mg, 103.17 μmol, 14.36 μL) was added. The mixture was stirred at 25°C for 0.5 hours. Iodomethane (4.28 mg, 25.79 μmol) and intermediate 1h (13.98 mg, 51.58 μmol) were then added and stirred at 50°C for 2 hours. The reaction mixture was filtered to obtain the crude product, which was then purified by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 3%-33% acetonitrile over 8 min) to afford compound 5 trifluoroacetate. MS m / z: 445.1 [M+H] + . 1 H NMR(400MHz,CD3OD)δppm 2.56(s,3H)3.27-3.29(m,3H)3.54(br d,J=3.76Hz,8H)4.58(s,2H)7.47(t,J=7.91Hz,1H)7.59(dd,J=10.16,8.4 1Hz, 1H) 7.70 (d, J = 8.28Hz, 1H) 8.47 (d, J = 8.03Hz, 1H) 10.18-10.34 (m, 1H).

[0571] Example 6

[0572] Step 1: Synthesis of intermediate 6b

[0573] Dioxane (1 mL), water (0.2 mL), intermediate 2a (30 mg, 128.19 μmol), and compound 6a (43.60 mg, 141.01 μmol) were added to a 10 mL thumb flask and stirred. After nitrogen purge, potassium phosphate (54.42 mg, 256.39 μmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (9.38 mg, 12.82 μmol) were added sequentially. The temperature was raised to 80°C and the reaction was allowed to react for 2 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain the crude product. The product was purified by pre-TLC (eluent: PE:EA = 3:1) to obtain intermediate 6b. MS m / z: 337.1 [M+H] + .

[0574] Step 2: Synthesis of intermediate 6c

[0575] Intermediate 6b (37 mg, 110.00 μmol) was dissolved in EtOH (1.5 mL), and ethanolic methylamine (222.81 mg, 3.30 mmol) was added. The mixture was stirred at 25°C for 6 hours. The reaction mixture was directly concentrated to obtain intermediate 6c, which was used in the next step without purification. MS m / z: 336.0 [M+H] + .

[0576] Step 3: Synthesis of intermediate 6d hydrochloride

[0577] Intermediate 6c (17 mg, 50.69 μmol) was dissolved in MeOH (0.5 mL), and hydrogen chloride / dioxane (4 M, 68.43 μL) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was directly concentrated to obtain intermediate 6d hydrochloride, which was used in the next reaction without purification. MS m / z: 236.1 [M+H] + .

[0578] Step 4: Synthesis of compound 6 trifluoroacetate

[0579] Intermediate 6d (10 mg, 36.80 μmol, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (7.45 mg, 73.61 μmol, 10.24 μL) was added. The mixture was stirred at 25°C for 0.5 h. Intermediate 1h (9.98 mg, 36.80 μmol) and potassium iodide (3.05 mg, 18.40 μmol) were then added, and the mixture was stirred at 50°C for 2 h. The reaction mixture was filtered to obtain the crude product, which was then separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min) to afford the trifluoroacetate salt of compound 6. MS m / z: 426.2 [M+H] + .1 H NMR (400MHz, CDCl3) δppm 8.12 (dd, J=7.78, 1.51Hz, 1H) 7.83-7.91 (m, 1H) 7.70-7.79 (m, 1H) 7.57-7.68 (m, 2H) 6.13 (br s,1H)4.53(s,2H)3.74-4.00(m,2H)3.36-3.66(m,2H)3.04-3.09(m,3H)2.91-2.99(m,1H)2.65(s,3H).

[0580] Example 7

[0581] Step 1: Synthesis of intermediate 7a

[0582] Intermediate 6c (10 mg, 29.82 μmol) was added to a mixture of MeOH (3 mL) and ethyl acetate (1 mL). Palladium on carbon (2 mg, 29.82 μmol, 10%) was added to the reaction solution. The atmosphere was replaced three times with argon and then with hydrogen. The mixture was stirred at 30°C under a 16 psi hydrogen atmosphere for 16 hours. The reaction solution was filtered through celite, and the organic phase was concentrated under reduced pressure to afford Intermediate 7a, which was used directly in the next step. MS m / z: 360.2 [M+23] + .

[0583] Step 2: Synthesis of intermediate 7b hydrochloride

[0584] Intermediate 7a (10 mg, 29.64 μmol) was added to MeOH (0.5 mL). Hydrogen chloride / dioxane (4 M, 37.05 μL) was added to the reaction mixture. The mixture was stirred at 25°C for 2 hours. The reaction mixture was then concentrated under reduced pressure to obtain intermediate 7b hydrochloride, which was used directly in the next step. MS m / z: 238.0 [M+1] + .

[0585] Step 3: Synthesis of compound 7 trifluoroacetate

[0586] Triethylamine (6.94 mg, 68.56 μmol) was added to a solution of intermediate 7b (9.38 mg, 34.28 μmol, crude hydrochloride) in DMF (1 mL). The mixture was stirred at 20°C for 0.5 h. Intermediate 1h (9.29 mg, 34.28 μmol) and potassium iodide (569.03 μg, 3.43 μmol) were added to the reaction mixture, and the mixture was stirred at 50°C for 2 h. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by pre-HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min) to obtain the trifluoroacetate salt of compound 7. MS m / z: 428.3 [M+1] + . 1 H NMR (400MHz, CD3OD) δppm 8.75 (s, 1H) 8.01 (s, 2H) 7.71 (d, J = 8.28Hz, 1H) 7.47 (t, J = 7.65Hz, 1H) 4.58 (s, 2H) 3.72 (br s,2H)3.18-3.30(m,3H)2.95(d,J=5.02Hz,3H)2.56(s,3H)2.15-2.26(m,2H)2.07(d,J=12.80Hz,2H).

[0587] Example 8

[0588] Step 1: Synthesis of intermediate 8b

[0589] Compound 8a (245.05 mg, 976.20 μmol) and N-Boc-piperazine (200 mg, 1.07 mmol) were added to toluene (5 mL). Pd2(dba)3 (8.94 mg, 9.76 μmol), RuPhos (9.11 mg, 19.52 μmol), and Cs2CO3 (636.13 mg, 1.95 mmol) were added to the reaction solution. The atmosphere was purged with nitrogen three times and the reaction was stirred at 100°C for 16 hours. The reaction solution was filtered through celite and the organic phase was concentrated under reduced pressure to obtain a crude product. A mixture of ethyl acetate and n-heptane (ethyl acetate / n-heptane = 5 / 1, 10 mL) was added to the crude product and stirred at 25°C for 0.5 hours. The mixture was then filtered under reduced pressure and the filter cake was concentrated to obtain intermediate 8b. MS m / z: 301.2 [M-56+1] + . 1H NMR (400MHz, DMSO-d6) δppm 7.61 (t, J = 8.19Hz, 1H) 6.93 (t, J = 8.38Hz, 1H) 3.83 (s, 3H) 3.50-3.46 (m, 4H) 3.13-3.23 (m, 4H) 1.43 (s, 9H).

[0590] Step 2: Synthesis of intermediate 8c

[0591] Compound 2 (100 mg, 280.61 μmol) and ethanolic methylamine solution (87.15 mg, 1.29 mmol) were added to EtOH (2 mL) and stirred at 25°C for 16 hours. The reaction temperature was then raised to 50°C and stirred at 50°C for 16 hours. The reaction was concentrated under reduced pressure to obtain a crude product, which was then purified by preparative chromatography (PE / EA = 2 / 1, transfer ratio 0.1) to afford intermediate 8c. MS m / z: 378.2 [M+23] + .

[0592] Step 3: Synthesis of intermediate 8d hydrochloride

[0593] Intermediate 8c (50 mg, 140.69 μmol) was added to MeOH (0.5 mL), and hydrogen chloride / dioxane (4 M, 37.05 μL) was added to the reaction mixture. The reaction mixture was stirred at 25°C for 2 hours. The reaction mixture was directly concentrated under reduced pressure to obtain intermediate 8d hydrochloride, which was used directly in the next step. MS m / z: 256.0 [M+1] + .

[0594] Step 4: Synthesis of trifluoroacetate salt of compound 8

[0595] Triethylamine (10.41 mg, 102.84 μmol) was added to a solution of intermediate 8d (15 mg, 51.42 μmol, crude hydrochloride) in DMF (1 mL). The reaction mixture was stirred at 20°C for 0.5 hours. Intermediate 1h (13.94 mg, 51.42 μmol) and potassium iodide (853.55 μg, 5.14 μmol) were then added to the reaction mixture, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by pre-HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min) to afford the trifluoroacetate salt of compound 8. MS m / z: 468.1 [M+23] + . 1H NMR(400MHz,CD3OD)δppm 8.15(s,1H)7.70(d,J=8.28Hz,1H)7.39-7.57(m,2H)6.89-7.03(m,1H)4.55(s,2H)3.50(s,8H)2.89-2.98(m,3H)2.56(s,3H).

[0596] Example 9

[0597] Step 1: Synthesis of Intermediate 9g Hydrochloride

[0598] Intermediate 4b (50 mg, 147.76 μmol) was dissolved in methanol (0.2 mL) and HCl / dioxane (4 M, 199.48 μL) was added. The mixture was allowed to react at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure without further purification to obtain the hydrochloride salt of intermediate 9g. MS m / z: 238.9 [M+1] + .

[0599] Step 2: Synthesis of intermediate 9b

[0600] Compound 9a (1 g, 4.81 mmol) was added to trifluoroacetic acid (10 mL) at 0°C, followed by the slow addition of potassium nitrate (760 mg, 7.52 mmol) in portions, and then stirred at 20°C for 16 hours. The reaction mixture was poured into 90 mL of saturated sodium bicarbonate solution and extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated sodium chloride (100 mL x 2), filtered, and concentrated. Purification on a silica gel column (EA:PE = 0%-10%) afforded intermediate 9b. 1 H NMR (400MHz, DMSO-d6) δppm 7.68-7.92 (m, 1H), 7.08 (br s, 2H).

[0601] Step 3: Synthesis of intermediate 9c

[0602] Intermediate 9b (100 mg, 395.26 μmol) was dissolved in EtOH (1 mL), cooled to 0°C, and acetic acid (237.36 mg, 3.95 mmol, 226.27 μL) was added. Zinc (240 mg, 3.67 mmol) was then added portionwise and allowed to react for 6 hours. The crude product was filtered and concentrated to afford the product. The crude product was purified using preparative chromatography (PE:EA = 3:1) to afford Intermediate 9c. MS m / z: 224.9, 226.9 [M+1]. + ,[M+3] + . 1 H NMR (400MHz, CDCl3) δppm6.69-6.80 (m, 1H), 3.52 (s, 4H).

[0603] Step 4: Synthesis of a mixture of intermediates 9d1 and 9d2

[0604] Intermediate 9c (25 mg, 112.10 μmol) was added to EtOH (0.32 mL), followed by pyruvic acid (11.85 mg, 134.52 μmol). The mixture was reacted at 100°C for 2 hours, then slowly cooled to 25°C for 14 hours to allow crystallization. The product was filtered and the solid was washed once with 0.5 mL of ethanol. Without further purification, a mixture of intermediates 9d1 and 9d2 was obtained. MS m / z: 274.9 [M+1] + .

[0605] Step 5: Synthesis of a mixture of intermediates 9e1 and 9e2

[0606] A mixture of intermediates 9d1 and 9d2 (95 mg) and XPHOS-PD-G2 (13.59 mg, 17.27 μmol) was dissolved in dioxane (2 mL). 1-(tributyltin)-methanol (133.08 mg, 414.47 μmol) was added, and the mixture was reacted at 80°C for 16 hours after nitrogen substitution. Purification by preparative chromatography (PE:EA = 1:1) afforded a mixture of intermediates 9e1 and 9e2. MS m / z: 227.0 [M+1] + .

[0607] Step 6: Synthesis of a mixture of intermediates 9f1 and 9f2

[0608] Under nitrogen, tributylphosphine (80.50 mg, 397.91 μmol, 98.18 μL) was slowly added dropwise to a solution of intermediates 9f1 and 9f2 (45 mg) and 1,2-dibromotetrachloroethane (142.54 mg, 437.71 μmol) in dichloromethane (1.2 mL) at 0°C. The mixture was reacted at 25°C for 2 hours. Purification by preparative chromatography (dichloromethane:methanol = 15:1) afforded a mixture of intermediates 9f1 and 9f2.

[0609] Step 7: Synthesis of a mixture of trifluoroacetic acid salts of compounds 9-1 and 9-2

[0610] Intermediates 9f1 and 9f2 (43 mg, mixture) were added to DMF (2 mL), followed by triethylamine (29.92 mg, 295.65 μmol). The mixture was stirred at 25°C for 15 minutes. Potassium iodide (11.04 mg, 66.52 μmol) and intermediate 9g (46 mg, 147.82 μmol) were then added to the reaction mixture, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was separated using preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; acetonitrile %: 0%-30%, 8 min). A mixture of trifluoroacetic acid salts of compounds 9-1 and 9-2 was obtained. MS m / z: 447.1 [M+1] + .

[0611] Example 10

[0612] Step 1: Synthesis of intermediate 10c

[0613] Under nitrogen, 10a (50 mg, 231.45 μmol), compound 10b (51.73 mg, 277.74 μmol), Cs2CO3 (150.82 mg, 462.89 μmol), RuPhos (21.60 mg, 46.29 μmol), and Pd2(dba)3 (21.19 mg, 23.14 μmol) were added to toluene (1 mL) and the mixture was heated to 100°C and stirred for 16 hours. The reaction mixture was filtered through celite and washed with DCM (5 mL). The filtrate was concentrated under reduced pressure to obtain the crude product, which was then separated and purified by preparative chromatography (PE:EA = 0:1) to afford intermediate 10c. MS m / z: 322.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δppm 8.06(d,J=2.76Hz,1H),7.82(d,J=8.53Hz,1H),6.93-7.02(m,2H),4.04-4.13(m,1H),3.74-3.80(m,3H),3.53-3. 61(m,1H),3.42-3.47(m,1H),3.32-3.35(m,1H),3.04-3.21(m,1H),2.09-2.20(m,1H),1.74-1.86(m,1H),1.40(br d,J=6.53Hz,9H).

[0614] Step 2: Synthesis of Intermediate 10d

[0615] Intermediate 10c (60 mg, 186.70 μmol) was added to EtOH (1 mL), followed by ethanolic methylamine solution (1.23 g, 11.88 mmol), and stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to afford Intermediate 10d, which was used directly in the next step. MS m / z: 321.2 [M+1] + .

[0616] Step 3: Synthesis of the hydrochloride salt of intermediate 10e

[0617] Intermediate 10d (25 mg, 78.03 μmol) was added to MeOH (0.5 mL), followed by the addition of hydrogen chloride / dioxane (4 M, 105.34 μL), and stirred at 25°C for 2 hr. The mixture was concentrated under reduced pressure to afford the hydrochloride salt of intermediate 10e, which was used directly in the next step. MS m / z: 221.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δ9.35(br s,3H),8.01(d,J=2.51Hz,1H),7.92(dd,J=8.66,4.39Hz,1H),7.21(dd,J=8.41,2.38Hz,1H),4.20-4.30(m,1H),3.4 2-3.51(m,1H),3.24-3.37(m,2H),3.04-3.13(m,1H),2.79(d,J=4.52Hz,3H),2.19-2.31(m,1H),1.86-1.98(m,1H).

[0618] Step 4: Synthesis of compound 10

[0619] Triethylamine (15.77 mg, 155.80 μmol, 21.69 μL) was added to a solution of intermediate 10e (20 mg, hydrochloride) in DMF (1 mL). The reaction was stirred at 20°C for 0.5 hr. Compound 1h (21.12 mg, 77.90 μmol) and potassium iodide (1.29 mg, 7.79 μmol) were added to the reaction mixture, and the temperature was raised to 50°C with stirring for 2 hr. The reaction mixture was washed with water (2 mL) and extracted three times with a mixture of DCM and MeOH (DCM:MeOH = 10:1, 3 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by preparative chromatography (DCM:MeOH = 10:1) to obtain compound 10. MS m / z: 411.1 [M+1] + , 1H NMR(400MHz,DMSO-d6)δppm12.44(s,1H),8.27(br d,J=4.77Hz,1H),7.91(d,J=2.51Hz,1H),7.72(d,J=8.53Hz,1H),7.50(d,J=7.78Hz,1H),7.24- 7.32(m,1H),6.94(dd,J=8.16,2.64Hz,1H),6.65(d,J=6.53Hz,1H),3.94-4.02(m,1H),3.75(br s,2H),3.37-3.42(m,1H),2.86(br t,J=8.66Hz,1H),2.75(d,J=4.77Hz,3H),2.65-2.71(m,1H),2.43-2.47(m,1H),2.41(s,3H),2.20-2.28(m,1H),1.54-1.65(m,1H).

[0620] Example 11

[0621] Step 1: Synthesis of intermediate 11c

[0622] Under nitrogen, 2a (50 mg, 231.45 μmol), compound 10b (51.73 mg, 277.74 μmol), Cs2CO3 (150.82 mg, 462.89 μmol), RuPhos (21.60 mg, 46.29 μmol), and Pd2(dba)3 (21.19 mg, 23.14 μmol) were added to toluene (1 mL) and the mixture was heated to 100°C and stirred for 16 hours. The reaction mixture was filtered through celite, washed with DCM (5 mL), and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Pre-TLC (PE:EA = 0:1) to obtain intermediate 11c. MS m / z: 340.1 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δppm 7.84 (d, J=8.00Hz, 1H), 7.24 (br t, J=9.26Hz, 1H), 6.88 (br d,J=6.38Hz,1H),4.07-4.18(m,1H),3.79(s,3H),3.56-3.64(m,1H),3.38-3.49(m,2H),3.20(br dd,J=11.01,4.75Hz,1H),2.11-2.22(m,1H),1.86-1.96(m,1H),1.40(br d,J=5.38Hz,9H).

[0623] Step 2: Synthesis of intermediate 11d

[0624] Intermediate 11c (60 mg, 186.70 μmol) was added to EtOH (1 mL), followed by ethanolic methylamine solution (1.23 g, 11.88 mmol), and stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to afford Intermediate 11d, which was used directly in the next step. MS m / z: 339.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.21(br d,J=4.63Hz,1H),7.75(d,J=8.25Hz,1H),7.24-7.31(m,1H),6.56(br d,J=6.50Hz,1H),4.04-4.18(m,1H),3.56-3.66(m,1H),3.38-3.47(m,2H),3.16- 3.24(m,1H),2.75(d,J=4.75Hz,3H),2.08-2.24(m,1H),1.84-1.97(m,1H)1.40(br d,J=6.13Hz,9H).

[0625] Step 3: Synthesis of the hydrochloride salt of intermediate 11e

[0626] Intermediate 11d (25 mg, 78.03 μmol) was added to MeOH (0.5 mL), followed by the addition of hydrogen chloride / dioxane (4 M, 105.34 μL), and stirred at 25°C for 4 hours. The mixture was concentrated under reduced pressure to afford the hydrochloride salt of intermediate 11e, which was used directly in the next step. MS m / z: 239.1 [M+1] + .

[0627] Step 4: Synthesis of compound 11

[0628] Triethylamine (11.05 mg, 109.20 μmol, 15.20 μL) was added to the hydrochloride salt of 11e (15 mg) obtained in step 3 in DMF (1 mL). The reaction was stirred at 20°C for 0.5 hr. Compound 1h (14.80 mg, 54.60 μmol) and potassium iodide (906.37 μg, 5.46 μmol) were added to the reaction solution, and the reaction was stirred at 50°C for 2 hr. The reaction solution was washed with water (2 mL) and extracted three times with a mixed solution of DCM and MeOH (DCM:MeOH = 10:1, 3 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative chromatography (DCM:MeOH = 10:1) to obtain compound 11. MS m / z: 429.2 [M+1] + , 1H NMR(400MHz,DMSO-d6)δppm12.43(s,1H),8.15(br d,J=4.63Hz,1H),7.72(d,J=8.00Hz,1H),7.50(d,J=8.25Hz,1H),7.28(t,J=7.63Hz,1H),7.16(dd,J=10.44,8.32Hz,1H),6.41(br d,J=5.88Hz,1H),3.96-4.04(m,1H),3.76(br s,2H),3.29(br s,1H),2.92(br t,J=8.00Hz,1H),2.74(d,J=4.75Hz,3H),2.64-2.70(m,1H),2.53-2.58(m,1H),2.41(s,3H),2.18-2.27(m,1H),1.73-1.81(m,1H).

[0629] Example 12

[0630] Step 1: Synthesis of intermediate 12b

[0631] Under nitrogen, 10a (50 mg, 231.45 μmol), compound 12a (51.73 mg, 277.74 μmol), Cs2CO3 (150.82 mg, 462.89 μmol), RuPhos (21.60 mg, 46.29 μmol), and Pd2(dba)3 (21.19 mg, 23.14 μmol) were added to toluene (1 mL) and stirred at 100°C for 16 hours. The reaction mixture was filtered through celite, washed with DCM (5 mL), and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by Pre-TLC (PE:EA = 0:1) to obtain intermediate 12b. MS m / z: 336.2 [M+1] + .

[0632] Step 2: Synthesis of intermediate 12c

[0633] Intermediate 12b (45 mg, 134.17 μmol) was added to EtOH (1 mL), followed by ethanolic methylamine solution (694.48 mg, 6.71 mmol), and the mixture was stirred at 25°C for 16 hours. The mixture was concentrated under reduced pressure to afford compound 12c. MS m / z: 335.2 [M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 8.34(br d,J=4.75Hz,1H),8.18(d,J=2.88Hz,1H),7.81(d,J=8.88Hz,1H),7.31(dd,J=8.88,3.00Hz,1H),4.61-4.70(m,1H),3.53(br t,J=8.88Hz,1H),3.41-3.46(m,1H),3.30-3.33(m,3H),3.21-3.25(m,1H),2.78(d,J=4.88Hz,3H),1.95-2.11(m,3H),1.41(s,9H).

[0634] Step 3: Synthesis of the hydrochloride salt of intermediate 12d

[0635] Intermediate 12c (44 mg, 131.57 μmol) was added to MeOH (0.5 mL), followed by the addition of hydrogen chloride / dioxane (4 M, 105.34 μL), and stirred at 25°C for 2 hr. The mixture was concentrated under reduced pressure to afford the hydrochloride salt of intermediate 12d, which was used directly in the next step. MS m / z: 235.1 [M+1] + .

[0636] Step 4: Synthesis of compound 12

[0637] Triethylamine (14.95 mg, 147.73 μmol, 20.56 μL) was added to the hydrochloride salt of 12d (20 mg) in DMF (1 mL). The reaction was stirred at 20°C for 0.5 hr. Compound 1h (20.02 mg, 73.87 μmol) and potassium iodide (1.23 mg, 7.39 μmol) were added to the reaction solution, and the reaction was stirred at 50°C for 2 hr. The reaction solution was washed with water (2 mL) and extracted three times with a mixed solution of DCM and MeOH (DCM:MeOH = 10:1, 3 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative chromatography (DCM:MeOH = 10:1) to obtain compound 12. MS m / z: 425.2 [M+1] + , 1H NMR(400MHz,DMSO-d6)δppm 12.45(br s,1H),8.31(br d,J=5.02Hz,1H),8.11(d,J=3.01Hz,1H),7.77(d,J=8.78Hz,1H),7.52(d,J=8.28Hz,1H),7. 30(t,J=7.65Hz,1H),7.22(dd,J=8.78,3.01Hz,1H),4.56-4.67(m,1H),3.75(s,2H),3.32(s, 3H),2.87(td,J=8.34,2.89Hz,1H),2.77(d,J=5.02Hz,3H),2.73(dd,J=10.04,3.26Hz,1H),2 .62(t,J=8.78Hz,1H),2.41(s,3H),2.31-2.37(m,1H),2.17-2.28(m,1H),1.66-1.76(m,1H).

[0638] Example 14

[0639] Step 1: Synthesis of intermediate 14b

[0640] Compound 3a (8 g, 36.99 mmol) was added to DCM (80 mL) and H₂O (80 mL). NaHCO₃ (9.32 g, 110.97 mmol) was added to the reaction solution, and the reaction temperature was cooled to 0°C. Benzyl chloroformate (9.47 g, 55.48 mmol) was added to the reaction solution, and the reaction was stirred at 20°C for 16 hours. 80 mL of dichloromethane was added to the reaction solution, and the layers were separated by extraction. The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure at 50°C to obtain the crude product, which was purified by column chromatography (eluent: PE / EA = 5 / 1 to 2 / 1, v / v) to obtain intermediate 14b.

[0641] Step 2: Synthesis of intermediate 14c

[0642] Intermediate 14b (5 g, 14.27 mmol) was added to DCM (100 mL). Dess-Martin (9.08 g, 21.40 mmol, 6.63 mL) was added portionwise to the reaction mixture at 0°C and stirred for 3 hours. 6 g of calcium hydroxide was added to the reaction mixture, which was stirred at 0°C for 1 hour. The filter cake was washed with dichloromethane (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 50°C to obtain the crude product. The crude product was isolated and purified by flash silica gel column chromatography (eluent: PE:EA = 90:10 to 80:20, v / v) to obtain Intermediate 14c.

[0643] Step 3: Synthesis of Intermediate 14d

[0644] Under nitrogen, KHMDS (1 M, 3.44 mL) was added dropwise to a solution of methyltriphenylphosphonium bromide (1.23 g, 3.44 mmol) in THF (4 mL) at 20°C. The reaction was stirred at 20°C for 1 hour. The temperature was then lowered to -78°C, and a solution of Intermediate 14c (0.4 g, 1.15 mmol) in THF (4 mL) was slowly added dropwise to the reaction mixture with stirring (10 min). The temperature was then raised to 20°C and stirred at this temperature for 1 hour. The reaction mixture was then added with 72 mL of methanol and then 36 mL of saturated potassium sodium tartrate solution. The mixture was extracted with 180 mL of ethyl acetate (3 times). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to afford the crude product. The crude product was purified by silica gel column chromatography (eluent: PE / EA = 90:10 to 80:20, v / v) to afford Intermediate 14d.

[0645] Step 4: Synthesis of intermediate 14e

[0646] Methyl 6-bromo-5-fluoropicolinate (100 mg, 427.31 μmol), intermediate 14d (148.03 mg, 427.31 μmol), Pd(OAc)2 (9.59 mg, 42.73 μmol), PPh3 (22.42 mg, 85.46 μmol), and Na2CO3 (90.58 mg, 854.62 μmol) were added to DMF (5 mL). After nitrogen substitution, the mixture was heated to 130°C and stirred for 6 hours. 5 mL of water was added to the reaction mixture, and the mixture was extracted three times with EA (10 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by thin-layer preparative chromatography (PE:EA = 2:1) to obtain intermediate 14e. MS m / z: 500.2 [M+1] + .

[0647] Step 5: Synthesis of Intermediate 14f

[0648] Intermediate 14e (20 mg, 40.04 μmol) was added to methanol (4 mL), and Pd / C (42.61 mg) was added to the reaction mixture. The reaction atmosphere was replaced with argon three times, then replaced with hydrogen three times. The reaction was stirred at 30°C and 30 psi of hydrogen for 16 hours. The reaction mixture was cooled to 20°C, filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain Intermediate 14f, which was used directly in the next step. MS m / z: 368.2 [M+1] + .

[0649] Step 6: Synthesis of Intermediate 14g

[0650] Potassium carbonate (11.28 mg, 81.65 μmol) was added to a solution of intermediate 14f (15 mg, 40.83 μmol) in DMF (1 mL), and the mixture was heated to 50°C and stirred for 6 hours. 5 mL of water was added to the reaction solution, and the mixture was extracted three times with 10 mL of EA. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by preparative thin-layer chromatography (PE:EA = 1:1) to obtain intermediate 14g. MS m / z: 348.2 [M+1] + ; 1 H NMR (400MHz, CDCl3) δppm 7.82(d,J=8.63Hz,1H),6.94(d,J=8.76Hz,1H),3.98-4.13(m,2H),3.87(s,3H),3.62-3.74(m,1H),3.14(tt,J=10.33,3.42Hz,1H),2.94 -3.06(m,2H),2.84-2.92(m,1H),2.57-2.70(m,1H),2.04(dq,J=13.35,4.39Hz,1H),1.67-1.82(m,1H),1.52-1.63(m,1H),1.42(s,9H).

[0651] Step 7: Synthesis of Intermediate 14h

[0652] Methylamine ethanol solution (0.41 g, 3.96 mmol) was added to a solution of intermediate 14g (20 mg, 57.57 μmol) in EtOH (1 mL) and stirred at 25°C for 16 hours. The mixture was concentrated under reduced pressure to afford intermediate 14h. MS m / z: 347.2 [M+1] + .

[0653] Step 8: Synthesis of the hydrochloride salt of intermediate 14i

[0654] Intermediate 14h (18 mg, 51.96 μmol) was dissolved in MeOH (0.5 mL), and then hydrogen chloride / dioxane (4 M, 64.95 μL) was added and stirred at 25°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 14i. MS m / z: 247.2 [M+1] + .

[0655] Step 9: Synthesis of compound 14

[0656] Triethylamine (70.73 μmol, 9.84 μL) was added to a solution of the hydrochloride salt of intermediate 14i (10 mg) in DMF (1 mL). The mixture was stirred at 20°C for 0.5 hours. Intermediate 1h (9.59 mg, 35.36 μmol) and potassium iodide (587.06 μg, 3.54 μmol) were then added and stirred at 50°C for 2 hours. The reaction solution was filtered, and the residue was separated by preparative HPLC (column: Welch Xtimate C18 100×40 mm×3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min) to obtain the crude product. Methylamine ethanol solution (0.30 g) was added, and the mixture was stirred at 25°C for 24 hours. The mixture was concentrated to obtain compound 14. MS m / z: 437.2 [M+1] + ; 1 H NMR (400MHz, CD3OD) δppm 7.63(d,J=8.53Hz,1H),7.45(d,J=8.53Hz,1H),7.29(t,J=7.65Hz,1H),7.10(d,J=8.78Hz,1H),4.50(br s,1H),3.75(br d,J=11.29Hz,1H),3.67(s,2H),3.03-3.14(m,1H),2.87-2.95(m,2H),2.81(s,3H),2.37-2.43(m,2H) ,2.16-2.27(m,1H),2.09(t,J=7.53Hz,1H),1.88-1.95(m,3H),1.65-1.76(m,1H),1.46-1.54(m,1H).

[0657] Example 15

[0658] Step 1: Synthesis of intermediate 15b

[0659] Compound 15a (0.8 g, 3.96 mmol) and methyl 3-amino-2-fluorobenzoate (803.87 mg, 4.75 mmol) were added to DMF (10 mL). HATU (1.81 g, 4.75 mmol) and triethylamine (1.20 g, 11.88 mmol, 1.65 mL) were added to the reaction solution, and the reaction was stirred at 25°C for 16 hours. The reaction solution was added to 5 mL of water and then extracted with ethyl acetate (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to obtain a crude product. The crude product was purified by thin layer preparative chromatography (PE / EA = 2 / 1, R f =0.14) to give intermediate 15b. MS m / z: 352.7, 354.7 [M+1]+ ; 1 H NMR (400MHz, CDCl3) δppm 8.53-8.62(m,1H)8.43(dd,J=4.75,1.88Hz,2H)8.00(dd,J=7.63,1.88Hz,1H)7.65(td, J=7.41,1.69Hz,1H)7.36(dd,J=7.63,4.75Hz,1H)7.13-7.24(m,1H)3.84-3.91(m,3H).

[0660] Step 2: Synthesis of intermediate 15c

[0661] Intermediate 15b (700 mg, 1.98 mmol) was added to DCM (15 mL). Et3N (5.95 mmol, 827.70 μL), (Boc)2O (519.14 mg, 2.38 mmol), and DMAP (48.43 mg, 396.44 μmol) were added to the reaction mixture, and the reaction was stirred at 25°C for 4 hours. The reaction mixture was added to 5 mL of water and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure at 45°C to obtain the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 5 / 1 to 2 / 1, V / V) to obtain Intermediate 15c. MS m / z: 452.9, 454.9 [M+1] + .

[0662] Step 3: Synthesis of Intermediate 15d

[0663] Intermediate 15c (20 mg, 44.12 μmol) was added to DMF (3 mL). Tributylphosphine (8.93 mg, 44.12 μmol), 1,3-bis(diphenylphosphino)propane (7.28 mg, 17.65 μmol), Pd(OAc)2 (3.96 mg, 17.65 μmol), and potassium carbonate (18.30 mg, 132.37 μmol) were added to the reaction solution, and microwave reaction was performed at 140°C for 10 min. The reaction solution was concentrated under reduced pressure at 60°C to obtain the crude product, which was purified by preparative thin-layer chromatography (DCM / MeOH = 20 / 1) to obtain Intermediate 15d. MS m / z: 272.8 [M+1] + .

[0664] Step 4: Synthesis of intermediate 15e

[0665] Intermediate 15d (20 mg, 73.47 μmol) was added to THF (2 mL). LiAlH4 (2.79 mg, 73.47 μmol) was added to the reaction mixture at 0°C and allowed to react for 30 min. 2 mL of saturated potassium sodium tartrate solution was added to the reaction mixture, followed by extraction with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure at 45°C to afford the crude product. The crude product was purified by preparative thin-layer chromatography (PE / EA = 2 / 1) to afford Intermediate 15e. MS m / z: 244.8 [M+1] + .

[0666] Step 5: Synthesis of Intermediate 15f

[0667] Intermediate 15e (11.5 mg, 47.09 μmol) was added to THF (4 mL). Manganese dioxide (40.94 mg, 470.89 μmol) was added to the reaction mixture at 20°C. The reaction was allowed to proceed for 2 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure at 45°C to afford crude product 15f. MS m / z: 243.0 [M+1] + .

[0668] Step 6: Synthesis of trifluoroacetate salt of compound 15

[0669] Intermediate 9g (12.26 mg, hydrochloride) was added to DMSO (2 mL), and Et3N (9.03 mg, 89.22 μmol) was added to the reaction solution, and the reaction was stirred at 25°C for half an hour. Intermediate 15f (10 mg, 41.29 μmol) was added to the reaction solution, and the pH was adjusted to 6 with acetic acid. The reaction was stirred at 25°C for 2 hours. NaBH(OAc)3 (18.91 mg, 89.22 μmol) was added to the reaction solution, and the reaction was stirred at 25°C for 1 hour. Water (2 mL) was added to the reaction solution, and then extracted with dichloromethane (5 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure at 45°C to obtain the crude product. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 2%-32% acetonitrile, 8 min) to afford the trifluoroacetate salt of compound 15. MS m / z: 487.2 [M+Na] + . 1H NMR(400MHz,DMSO-d6)δppm 12.19(br s,1H),10.03-10.15(m,1H),9.16(d,J=2.40Hz,1H),8.72(d,J=3.20Hz,1H),8.43-8.62(m,2H),7.92(d,J=4.42Hz,1 H),7.71-7.84(m,2H),7.45-7.52(m,1H),4.66(s,2H),3.70-3.81(m,4H),3.19-3.25(m,4H),2.81(d,J=2.40Hz,3H).

[0670] Step 7: Synthesis of Intermediate 15g

[0671] Intermediate 15e (50 mg, 204.73 μmol) was dissolved in DCM (3 mL), and 1,2-dibromotetrachloroethane (146.67 mg, 450.41 μmol) was added. Tributylphosphine (82.84 mg, 409.47 μmol) was added at 0°C, and the mixture was stirred at 25°C for 3 hours. The filter cake was obtained by filtration, which was Intermediate 15g. 1 H NMR(400MHz,DMSO-d6)δppm 12.02(s,1H),9.10(d,J=4.52Hz,1H),8.65(d,J=8.03Hz,1H),8.39-8.52(m,1H),7.69-7.80(m,1H),7.34-7.54(m,1H),4.78-5.01(m,2H).

[0672] Step 8: Synthesis of compound 15

[0673] Intermediate 9g (38.02mg, hydrochloride) was dissolved in DMF (1mL), and triethylamine (32.95mg, 325.61μmol) was added. After stirring at 25°C for 0.5 hours, intermediate 15g (50mg, 162.80μmol) and KI (2.70mg, 16.28μmol) were added, and the mixture was stirred at 50°C for 3 hours. Water (2mL) was added to the reaction solution and stirred for 10 minutes. A solid precipitated, which was filtered to obtain the filter cake, which was compound 15. MS m / z: 465.1[M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=4.63,1.75Hz,1H),8.65(dd,J=7.94,1.81Hz,1H ),8.39-8.49(m,2H),7.84(dd,J=8.13,1.25Hz,1H),7.72(dd,J=8.00,4.5 0Hz,1H),7.57(dd,J=10.76,8.13Hz,1H),7.37(dd,J=7.94,6.44Hz,1H),3 .75(s,2H),3.15-3.24(m,4H),2.76(d,J=4.75Hz,3H),2.61-2.65(m,4H).

[0674] Example 16

[0675] Step 1: Synthesis of intermediate 16b

[0676] Compound 1i (5 g, 23.12 mmol) was added to H₂O (40 mL) and DCM (40 mL). NaHCO₃ (5.83 g, 69.36 mmol) was added to the reaction mixture. The reaction temperature was lowered to 0°C. Benzyl chloroformate (5.92 g, 34.68 mmol, 4.95 mL) was added to the reaction mixture, and the mixture was stirred at 20°C for 16 hours. 40 mL of dichloromethane was added to the reaction mixture. The layers were separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 50°C to obtain the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 5 / 1 to 2 / 1, v / v) to obtain intermediate 16b.

[0677] Step 2: Synthesis of intermediate 16c

[0678] Intermediate 16b (8 g, 22.83 mmol) was added to DCM (120 mL). Dess-Martin (14.52 g, 34.25 mmol, 10.61 mL) was added portionwise to the reaction mixture at 0°C and stirred for 3 hours. 10 g of calcium hydroxide was added to the reaction mixture, which was stirred at 0°C for 1 hour. The mixture was filtered, and the filter cake was washed with dichloromethane (60 mL x 3). The filtrate was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 50°C to obtain the crude product. The crude product was purified by column chromatography (eluent: ethyl acetate / n-heptane = 5 / 1 to 2 / 1, v / v) to obtain Intermediate 16c.

[0679] Step 3: Synthesis of Intermediate 16d

[0680] Under nitrogen, KHMDS (1 M, 51.67 mL) was added dropwise to a solution of methyltriphenylphosphonium bromide (18.46 g, 51.67 mmol) in THF (50 mL) at 20°C. The reaction was stirred at 20°C for 1 hour. The temperature was then lowered to -70°C, and a solution of Intermediate 16c (6 g, 17.22 mmol) in THF (50 mL) was slowly added dropwise to the reaction mixture with stirring (10 min). The temperature was then raised to 20°C and stirred at this temperature for 3 hours. To the reaction mixture were added 50 mL of methanol, followed by 50 mL of saturated potassium sodium tartrate solution. The mixture was extracted with 100 mL of ethyl acetate (3 times). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to afford the crude product, which was then purified by column chromatography (eluent: PE / EA = 5 / 1 to 1 / 1, v / v) to afford Intermediate 16d. 1 H NMR(400MHz, CDCl3)δppm 7.14-7.28(m,5H)5.61(ddd,J=17.39,10.76,4.50Hz,1H)5.02-5.11(m,2H)5.00(d,J=2.63Hz,2H)4.58(br s,1H)3.67-3.96(m,3H)2.58-3.04(m,3H)1.30(s,9H).

[0681] Step 4: Synthesis of intermediate 16e

[0682] Intermediate 16d (500.00 mg, 1.44 mmol), methyl 6-bromo-5-fluoropicolinate (337.77 mg, 1.44 mmol), PPh3 (75.71 mg, 288.67 μmol), Na2CO3 (458.93 mg, 4.33 mmol), and Pd(OAc)2 (32.40 mg, 144.33 μmol) were added to DMF (5 mL). The atmosphere was purged with nitrogen three times and stirred at 130°C for 12 hours. 10 mL of water was added to the reaction mixture, which was then extracted with ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 50°C to afford the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 4 / 1 to 2 / 1, v / v) to afford Intermediate 16e. MS m / z: 522.1 [M+23] + .

[0683] Step 5: Synthesis of Intermediate 16f

[0684] Intermediate 16e (0.2 g, 400.38 μmol) was added to a hydrogenation flask containing MeOH (10 mL). The atmosphere was replaced with argon once. Pd / C (200 mg) was added to the reaction flask and the atmosphere was replaced with hydrogen three times. The reaction was stirred at 30°C under 30 psi of hydrogen for 16 hours. The reaction solution was filtered through celite under reduced pressure, and the filtrate was concentrated under reduced pressure at 40°C to obtain intermediate 16f. MS m / z: 368.1 [M+1] + .

[0685] Step 6: Synthesis of Intermediate 16g

[0686] Intermediate 16f (100 mg, 272.17 μmol) was added to DMF (4 mL), and then potassium carbonate (75.23 mg, 544.34 μmol) was added to the reaction solution. The reaction was stirred at 50°C for 2 hours. The reaction solution was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure to obtain intermediate 16g.

[0687] MS m / z:348.2[M+1] + .

[0688] Step 7: Synthesis of Intermediate 16h

[0689] Intermediate 16g (80mg, 230.28μmol) was added to ethanol (2mL), and then methylamine ethanol solution (2.53g, 24.44mmol) was added to the reaction solution. The reaction was stirred at 30°C for 16 hours, and the reaction solution was concentrated under reduced pressure to obtain intermediate 16h. MS m / z: 347.1[M+1] +

[0690] Step 8: Synthesis of the hydrochloride salt of intermediate 16i

[0691] Intermediate 16h (40 mg, 115.47 μmol) was added to MeOH (2 mL), and then hydrogen chloride / dioxane (4 M, 144.33 μL) was added to the reaction mixture. The mixture was stirred at 30°C for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 16i. MS m / z: 269.1 [M+23] + .

[0692] Step 9: Synthesis of trifluoroacetate salt of compound 16

[0693] Et3N (10.74 mg, 106.09 μmol, 14.77 μL) was added to the hydrochloride salt of intermediate 16i (15 mg) in DMF (1 mL). The reaction was stirred at 20°C for half an hour. Intermediate 1h (14.38 mg, 53.05 μmol) and potassium iodide (880.58 μg, 5.30 μmol) were added to the reaction mixture, and the reaction was stirred at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was purified by preparative HPLC (column: Welch Xtimate C18 100×40 mm×3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 24%-54% acetonitrile) to obtain the trifluoroacetate salt of compound 16. MS m / z: 437.2 [M+1] + ; 1 H NMR (400MHz, CD3OD) δppm 7.82 (d, J = 8.53Hz, 1H) 7.71 (d, J = 8.28Hz, 1H) 7.44-7.51 (m, 1H) 7.37 (d, J = 8.78Hz, 1H) 4.59 (s, 2H) 4.23 (br d,J=14.56Hz,1H)3.61-3.75(m,2H)3.38-3.54(m,1H)3.08-3.23(m,3H)2.9 9-3.06(m,2H)2.94(s,3H)2.56(s,3H)2.14-2.24(m,1H)1.85-1.98(m,1H).

[0694] Example 17

[0695] Step 1: Synthesis of intermediate 17b

[0696] Triethylamine (19.46 g, 192.30 mmol) was added dropwise to a solution of compound 17a (10 g, 48.08 mmol) in DCM (50 mL) at 0°C. After stirring for 30 min, acetyl chloride (6.04 g, 76.92 mmol, 5.47 mL) was slowly added dropwise. The temperature was then raised to 25°C and the reaction mixture was allowed to react for 3 hours. The reaction solution was added to 100 mL of water and extracted with 40 mL of dichloromethane (5 × 40 mL). The organic phases were combined and washed with 100 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The product was then purified by column chromatography (eluent: EA / PE = 0%-10%, v / v) to afford intermediate 17b. MS m / z: 249.9, 251.9 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm 10.41 (s, 1H) 7.43-7.45 (m, 2H) 2.07 (s, 3H).

[0697] Step 2: Synthesis of intermediate 17c

[0698] Fuming nitric acid (71.99 mmol, 3.24 mL) was added to a three-necked flask, cooled to 0°C, and intermediate 17b was added portionwise. Sulfuric acid (334.75 mmol, 17.84 mL) was then slowly added dropwise, maintaining the temperature at 0°C over 1 hour. The reaction mixture was then heated to 25°C and allowed to react for 1 hour. The reaction mixture was poured into 100 mL of 3M sodium hydroxide solution at 0°C and the pH was adjusted to approximately 7. Extraction was then performed with 30 mL of ethyl acetate (5 times). The organic phases were combined and washed with saturated brine (2 times with 100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield intermediate 17c. 1 H NMR (400MHz, DMSO-d6) δppm 10.55 (s, 1H) 7.50-7.53 (m, 1H) 2.08 (s, 3H).

[0699] Step 3: Synthesis of Intermediate 17d

[0700] Intermediate 17c (9.50 g, 32.20 mmol) was dissolved in sulfuric acid (90 mL) and reacted at 50°C for 2 hours. The reaction solution was poured into 100 mL of ice water, adjusted to pH 7-8 with 3M NaOH solution, and filtered. The filter cake was washed with 10 mL of water and dried in vacuo to obtain Intermediate 17d. 1 H NMR (400MHz, DMSO-d6) δppm 7.45 (br, s, 2H) 6.74-6.77 (m, 1H).

[0701] Step 4: Synthesis of Intermediate 17e

[0702] Intermediate 17d (6.2 g, 24.51 mmol) was dissolved in EtOH (70 mL), cooled to 0°C, and acetic acid (14.72 g, 245.06 mmol, 14.03 mL) was added. Zinc (14.88 g, 227.56 mmol) was then added portionwise and allowed to react for 1 hour. The reaction mixture was poured into 500 mL of water and extracted with 80 mL of ethyl acetate (5 times). The organic phase was washed with 300 mL of saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to yield Intermediate 17e. 1 H NMR(400MHz, DMSO-d6)δppm 6.37-6.40(m,1H)5.21(br,s,2H)4.65(br,s,2H).

[0703] Step 5: Synthesis of Intermediate 17f

[0704] Intermediate 17e (3.15 g, 14.12 mmol) was added to EtOH (30 mL), followed by the dropwise addition of pyruvic acid (1.49 g, 16.95 mmol, 1.19 mL). The mixture was reacted at 100°C for 2 hours. The reaction solution was concentrated under reduced pressure, and 10 mL of ethanol was added, stirred for 20 minutes, and filtered to obtain the filter cake to obtain the intermediate 17f. MS m / z: 274.9, 276.9 [M+1] + .

[0705] Step 6: Synthesis of Intermediate 17g

[0706] A mixture of Intermediate 17f (3.05 g, 11.09 mmol) and 1-(tributyltin)methanol (4.27 g, 13.31 mmol) was dissolved in dioxane (30 mL). Chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (436.24 mg, 554.45 μmol) was then added. After nitrogen substitution, the mixture was reacted at 110°C for 3 hours. The reaction solution was concentrated under reduced pressure and purified on a flash silica gel column (EA / PE = 0%-30%, MeOH / DCM = 0%-5%) to afford the crude product. This product was then separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: water (trifluoroacetic acid)-acetonitrile; gradient: 0%-26% acetonitrile, 8 min) to afford Intermediate 17g. MS m / z:227.1[M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm 12.51 (s, 1H) 7.35-7.37 (m, 1H) 5.25 (s, 1H) 4.57 (s, 2H) 2.41 (s, 3H).

[0707] Step 7: Synthesis of Intermediate 17h

[0708] Dess-Martin (84.39 mg, 198.96 μmol) was added to a solution of Intermediate 17g (30 mg, 132.64 μmol) in DCM (2 mL) at 25°C and stirred for 2 hours. 0.057 g of calcium hydroxide was added to the reaction mixture, which was stirred at 25°C for 1 hour. The mixture was filtered, and the filter cake was washed with dichloromethane (10 mL x 3). The filtrate was concentrated under reduced pressure at 50°C to afford the crude product. The crude product was purified by preparative thin-layer chromatography (PE:EA = 1:1) to afford Intermediate 17h. 1 H NMR (400MHz, DMSO-d6) δppm 12.72 (s, 1H), 10.27 (s, 1H), 7.53-7.63 (m, 1H), 2.47 (s, 3H).

[0709] Step 8: Synthesis of compound 17

[0710] Intermediate 9g (21.86 mg, hydrochloride) was dissolved in DMSO (1 mL). Triethylamine (13.54 mg, 133.83 μmol) was added to the reaction solution with stirring, and the mixture was stirred at 25°C for 30 min. Intermediate 17h (15 mg, 66.92 μmol) was added to the reaction solution, and the pH was adjusted to 6-7 with acetic acid. The mixture was stirred at 25°C for 2 hours. NaBH(OAc)3 (28.36 mg, 133.83 μmol) was added to the reaction solution, and the mixture was stirred at 25°C for 16 hours. 2 mL of water was added to the reaction solution, and the mixture was extracted three times with EA (5 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min). The obtained product was basified with triethylamine (3 μL) and then purified by preparative thin-layer chromatography (100% EA) to obtain compound 17. MS m / z: 447.1 [M+1] + ; 1 H NMR (400MHz, CDCl3) δppm 7.88(dd,J=8.16,1.13Hz,1H),7.49(dd,J=10.29,8.03Hz,1H),7.37(dd,J=9.79,1.76 Hz,1H),3.87(s,2H),3.22-3.25(m,4H),2.91(s,3H),2.71-2.77(m,4H),2.53(s,3H).

[0711] Example 18

[0712] Step 1: Synthesis of compound 18

[0713] Intermediate 14i (19.87 mg, hydrochloride) was dissolved in DMSO (1 mL), and triethylamine (133.83 μmol, 18.63 μL) was added to the reaction solution with stirring, and the mixture was stirred at 25°C for 30 min. Intermediate 17h (15.00 mg, 66.92 μmol) was added to the reaction solution, and the pH was adjusted to 6-7 with acetic acid. The mixture was stirred at 25°C for 2 h. NaBH(OAc)3 (28.36 mg, 133.83 μmol) was added to the reaction solution, and the mixture was stirred at 25°C for 16 h. 2 mL of water was added to the reaction mixture, and the mixture was extracted with EA (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile, 8 min) to obtain the crude product. 3 μL of triethylamine was added to the crude product, and the product was purified by preparative thin-layer chromatography (100% EA) to obtain compound 18. MS m / z: 455.2 [M+1] + ; 1 H NMR (400MHz, CD3OD) δppm 7.64 (d, J=8.53Hz, 1H), 7.06-7.16 (m, 2H), 3.76 (br d,J=12.55Hz,1H),3.64(s,2H),3.06-3.15(m,1H),2.83-2.95(m,4H),2.81-2.83(m,3H),2.43( s,3H),2.23(td,J=11.54,3.26Hz,1H),1.92-2.00(m,2H),1.64-1.78(m,1H),1.22-1.27(m,1H).

[0714] Example 19

[0715] Step 1: Synthesis of trifluoroacetate salt of compound 19

[0716] Intermediate 16i (12.5 mg, hydrochloride) was dissolved in DMSO (1 mL), and triethylamine (74.58 μmol, 10.38 μL) was added. The mixture was stirred at 20°C for 0.5 h, followed by the addition of 17h (8.36 mg, 37.29 μmol). The pH was adjusted to 6-7 with acetic acid, and the mixture was stirred at 20°C for 2 h. NaBH(OAc)3 (15.81 mg, 74.58 μmol) was then added, and the mixture was stirred for 16 h. The reaction mixture was extracted with 3 mL of water and 3 × 3 mL of ethyl acetate. The organic phases were combined, washed with 3 mL of saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain the crude product. The crude product was then separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: water (trifluoroacetic acid)-acetonitrile; gradient: 0%-30% acetonitrile, 8 min) to afford the trifluoroacetate salt of compound 19. MS m / z:455.1[M+1] + ; 1 H NMR (400MHz, CD3OD) δppm 1.63-1.77(m,1H)1.93-2.05(m,2H)2.18-2.31(m,1H)2.42(s,3H)2.81(s,3H)2.82-2.86(m,2H)2.93(br t,J=10.42Hz,2H)3.07(s,2H)3.72-3.81(m,3H)7.09(d,J=8.78Hz,1H)7.27(dd,J=9.54,1.76Hz,1H)7.59-7.68(m,1H).

[0717] Example 20

[0718] Step 1: Synthesis of intermediate 20b

[0719] Triethylamine (7.78 g, 76.92 mmol, 10.71 mL) was added dropwise to a solution of compound 20a (4 g, 19.23 mmol) in DCM (20 mL) at 0°C. After stirring for 0.5 hours, acetyl chloride (2.42 g, 30.77 mmol, 2.19 mL) was slowly added dropwise. The temperature was then raised to 18°C ​​and the reaction was allowed to react for 3 hours. The reaction solution was added dropwise to 100 mL of water and extracted with 20 mL of dichloromethane (5 times). The organic phase was washed with 100 mL of saturated brine and dried over anhydrous sodium sulfate. The crude product was concentrated under reduced pressure to obtain the product, which was then purified on a flash silica gel column (EA / PE = 0%-10%) to obtain intermediate 20b. MS m / z: 250.0 [M+1] + ; 1H NMR (400MHz, DMSO-d6) δppm 10.03 (br s, 1H) 8.02-8.17 (m, 1H) 7.77 (dd, J = 10.16, 6.40Hz, 1H) 2.12 (s, 3H).

[0720] Step 2: Synthesis of intermediate 20c

[0721] Intermediate 20b (2.52 g, 10.08 mmol) was suspended in nitric acid (1.27 g, 20.16 mmol, 907.24 μL) at 0°C. Sulfuric acid (9.19 g, 93.73 mmol, 5.00 mL) was then slowly added dropwise. The mixture was reacted at 25°C for 1 hour. The reaction solution was added dropwise to water, resulting in the precipitation of a solid. The pH was adjusted to 6-7 with 2M NaOH solution, and the mixture was filtered and the filter cake dried to afford Intermediate 20c. 1 H NMR (400MHz, DMSO-d6) δppm10.48(s,1H)8.21-8.38(m,1H)2.07(s,3H).

[0722] Step 3: Synthesis of intermediate 20d

[0723] Intermediate 20c (2.28 g, 7.73 mmol) was dissolved in sulfuric acid (22 mL) and stirred at 50°C for 2 hours. The reaction solution was added to 400 mL of water and the pH was adjusted to 6-7 with 2M NaOH solution. A solid precipitated and was filtered to obtain a filter cake, which was then dried in vacuo to afford Intermediate 20d. 1 H NMR (400MHz, DMSO-d6) δppm 7.76-7.89 (m, 1H) 7.09 (s, 2H).

[0724] Step 4: Synthesis of intermediate 20e

[0725] Intermediate 20d (1.49 g, 5.89 mmol) was dissolved in EtOH (15 mL). Zinc powder (3.66 g, 55.95 mmol) and acetic acid (3.54 g, 58.89 mmol, 3.37 mL) were added portionwise at 0°C and stirred at 40°C for 1 hour. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by column chromatography (eluent: PE:EA = 100:0 to 90:10, V / V) to obtain Intermediate 20e. MS m / z: 223.0 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm 6.65 (dd, J = 10.04, 6.02Hz, 1H) 5.06 (s, 2H) 4.90 (s, 2H).

[0726] Step 5: Synthesis of Intermediate 20f

[0727] Intermediate 20e (1.14 g, 5.11 mmol) was dissolved in EtOH (12 mL), followed by the addition of pyruvic acid (540.17 mg, 6.13 mmol, 432.14 μL). The mixture was stirred at 100°C for 2 hours and then slowly cooled to 25°C for 14 hours to allow crystallization. The reaction mixture was filtered to obtain the filter cake, which was Intermediate 20f (mixture). MS m / z: 274.9 [M+1] + .

[0728] Step 6: Synthesis of Intermediate 20g

[0729] Under nitrogen, intermediate 20f (500 mg, 1.82 mmol) was dissolved in dioxane (5 mL), and 1-(tributyltin)-methanol (700.42 mg, 2.18 mmol) and chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (71.51 mg, 90.89 μmol) were added, and the mixture was stirred at 110 °C for 4 h. The reaction solution was concentrated under reduced pressure and separated and purified by column chromatography (eluent: PE:EA = 90:10 to 50:50, V / V) to obtain a crude product. The crude product was separated by preparative HPLC (column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-26% acetonitrile) to obtain 20g of the intermediate; MS m / z: 226.8 [M+1] + ; 1 H NMR (400MHz, DMSO-d6) δppm 12.58 (br s, 1H) 6.99-7.25 (m, 1H) 4.62 (br d, J = 3.51Hz, 2H) 2.43 (br s, 3H).

[0730] Step 7: Synthesis of Intermediate 20h

[0731] Intermediate 20g (100mg, 442.13μmol) was dissolved in DCM (2mL), and Dess-Martin (281.29mg, 663.19μmol, 205.47μL) was added. The mixture was stirred at 20°C for 2 hours. 0.1899g of calcium hydroxide was added to the reaction mixture and stirred at 20°C for 1 hour. The crude product was filtered and purified by preparative thin-layer chromatography (DCM:MeOH = 20:1) to afford Intermediate 20h. 1 H NMR (400MHz, DMSO-d6) δppm 12.96 (br, s, 1H), 10.26 (s, 1H), 7.40-7.44 (m, 1H), 2.49 (s, 3H).

[0732] Step 8: Synthesis of compound 20

[0733] Intermediate 14i (13.25 mg, hydrochloride) was dissolved in DMSO (1 mL). Triethylamine (9.03 mg, 89.22 μmol, 12.42 μL) was added to the reaction solution with stirring, and the mixture was stirred at 25°C for 30 min. Intermediate 20h (10 mg, 44.61 μmol) was added to the reaction solution, and the pH was adjusted to 6-7 with acetic acid. The mixture was stirred at 25°C for 2 h. NaBH(OAc)3 (18.91 mg, 89.22 μmol) was added to the reaction solution, and the mixture was stirred at 25°C for 16 h. 2 mL of water was added to the reaction mixture, and the mixture was extracted with EA (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain the crude product. 3 μL of triethylamine was added to the crude product, and the product was purified by preparative thin-layer chromatography (100% EA) to obtain compound 20. MS m / z: 455.2 [M+1] + ; 1 H NMR(400MHz,CD3OD)δppm 7.75(d,J=8.78Hz,1H),7.38(dd,J=9.41,1.63Hz,1H),7.21(d,J=8.78Hz,1H),4.59(s,1H),3.83-3.90(m,3H),3.14-3.23(m,1H),3.01-3. 11(m,2H),2.94-2.99(m,1H),2.92-2.94(m,3H),2.85-2.92(m,1H),2 .54(s,3H),2.31-2.41(m,1H),1.93-2.23(m,2H),1.75-1.88(m,1H).

[0734] Example 21

[0735] Step 1: Synthesis of compound 21

[0736] Intermediate 16i (15 mg, hydrochloride) was dissolved in DMSO (1 mL), and triethylamine (9.06 mg, 89.50 μmol, 12.46 μL) was added. The mixture was stirred at 20°C for 0.5 h, followed by the addition of intermediate 20h (12.54 mg, 44.75 μmol). The pH was adjusted to 6-7 with acetic acid, and the mixture was stirred at 20°C for 2 h. NaBH(OAc)3 (18.97 mg, 89.50 μmol) was then added, and the mixture was stirred for 16 h. The reaction mixture was extracted with 3 mL of water and 3 × 3 mL of ethyl acetate. The combined organic phases were washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate, and filtered to obtain the crude product. The crude product was then separated by preparative HPLC (column: Welch Xtimate C18, 100 × 40 mm × 3 μm; mobile phase: water (trifluoroacetic acid)-acetonitrile; gradient: 0% to 30% acetonitrile, 8 min). The crude product was added with 3 μL triethylamine and separated and purified by thin layer preparative chromatography (EA=100%) to obtain compound 21. MS m / z: 455.0 [M+1] + ; 1 H NMR (400MHz, CD3OD) δppm 7.58-7.69 (m, 1H) 7.10 (d, J = 8.78Hz, 2H) 3.76 (br d,J=11.80Hz,1H)3.64(d,J=1.51Hz,2H)3.04-3.17(m,1H)2.82-2.96(m,5H)2 .82(s,3H)2.43(s,3H)2.15-2.30(m,1H)1.88-2.01(m,2H)1.61-1.79(m,1H).

[0737] Example 22

[0738] Step 1: Synthesis of trifluoroacetate salt of compound 22

[0739] Intermediate 9g (7.29mg, hydrochloride) was dissolved in DMSO (1mL), and triethylamine (44.61μmol, 6.21μL) was added. The mixture was stirred at 20°C for 0.5 hours. Intermediate 20h (5mg, 22.31μmol) was then added, the pH was adjusted to 6-7 with acetic acid, and the mixture was stirred for 2 hours. NaBH(OAc)3 (9.45mg, 44.61μmol) was added and stirred for 4 hours. The reaction mixture was extracted with 1mL of water and 3×1mL of ethyl acetate. The organic phase was washed with 10mL of saturated brine, dried, and filtered to obtain the crude product. The crude product was separated by preparative HPLC (column: Welch Xtimate C18 100×40mm×3μm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 1%-31% acetonitrile, 8min) to obtain the trifluoroacetate salt of compound 22. MS m / z: 447.1[M+1]+ ; 1 H NMR(400MHz,CD3OD)δppm 7.92-7.99(m,1H)7.54-7.68(m,1H)7.27(dd,J=10.16,5.40Hz,1H)4.22- 4.39(m,2H)3.42-3.51(m,4H)3.23-3.31(m,4H)2.93(s,3H)2.57(s,3H).

[0740] Example 23

[0741] Step 1: Synthesis of intermediate 23a

[0742] Compound 17g (100.00 mg, 442.13 μmol) and 1,2-dibromo-1,1,2,2-tetrachloroethane (316.74 mg, 972.68 μmol) were dissolved in DCM (5 mL). Tri-n-butylphosphine (178.90 mg, 884.25 μmol) was added at 0°C. After addition, the temperature was raised to 25°C and stirred for 12 hours. The reaction mixture was filtered to obtain a filter cake, Intermediate 23a. 1 H NMR (400MHz, DMSO-d6) δppm 12.63 (br s, 1H), 7.33-7.72 (m, 1H), 4.71-4.94 (m, 2H), 2.43 (s, 3H).

[0743] Step 2: Synthesis of compound 23

[0744] Triethylamine (21.00 mg, 207.56 μmol) was added to a solution of intermediate 6d (25.64 mg, hydrochloride) in DMF (2 mL) and stirred at 25°C for 30 minutes. Intermediate 23a (30 mg, 103.78 μmol) and KI (1.72 mg, 10.38 μmol) were then added, and the reaction was stirred at 50°C for 2 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product. The crude product was separated and purified by preparative HPLC (column: Welch Xtimate C18 100×40 mm×3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain a crude product. Triethylamine (20 μL) was added to the crude product, and the product was separated and purified by preparative chromatography (developing ratio: DCM:MeOH = 10:1) to obtain compound 23. MS m / z: 444.1 [M+1] + ; 1H NMR(400MHz,DMSO-d6)δppm 12.54(br s,1H),8.63(br d,J=5.00Hz,1H),8.05(dd,J=9.82,7.82Hz,1H),7.91(dd,J=7.69,1.69Hz,1H),7.46(d,J=8.88Hz,1H),6 .14-6.27(m,1H),3.80(s,2H),3.13-3.26(m,3H),2.79(d,J=4.88Hz,3H),2.60-2.76(m,3H),2.44(s,3H).

[0745] Example 24

[0746] Step 1: Synthesis of intermediate 24a

[0747] Compound 20g (100.00 mg, 442.13 μmol) and 1,2-dibromo-1,1,2,2-tetrachloroethane (316.74 mg, 972.68 μmol) were dissolved in DCM (5 mL). Tri-n-butylphosphine (178.90 mg, 884.25 μmol) was added at 0°C. After addition, the mixture was heated to 25°C and stirred for 8 hours. The reaction mixture was filtered to obtain a filter cake, Intermediate 24a. 1 H NMR (400MHz, DMSO-d6) δppm 12.72 (br s, 1H), 7.32 (dd, J = 10.54, 5.77Hz, 1H), 4.76 (s, 2H), 2.44 (s, 3H).

[0748] Step 2: Synthesis of compound 24

[0749] Triethylamine (21.00 mg, 207.56 μmol) was added to a solution of intermediate 6d (25.64 mg, hydrochloride) in DMF (2 mL) and stirred at 25°C for 30 min. Intermediate 24a (30 mg, 103.78 μmol) and KI (1.72 mg, 10.38 μmol) were then added, and the reaction was stirred at 50°C for 2 hr. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product. The crude product was separated and purified by preparative HPLC (column: Welch Xtimate C18 100×40 mm×3 μm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain a crude product. Triethylamine (20 μL) was added to the crude product, and the product was separated and purified by preparative chromatography (developing ratio: DCM:MeOH = 10:1) to obtain compound 24. MS m / z: 444.1 [M+1] + ; 1H NMR (400MHz, DMSO-d6) δppm 12.63 (br s, 1H), 8.64 (br d, J=4.63Hz, 1H), 8.01-8.17 (m, 1H), 7.92 (br d, J=7.38Hz, 1H), 7.07-7.28 (m, 1H), 6.25 (br s,1H),3.75(s,2H),3.18-3.25(m,3H),2.80(br d,J=4.63Hz,3H),2.65-2.76(m,3H),2.44(s,3H).

[0750] Example 25

[0751] Step 1: Synthesis of intermediate 25a

[0752] Compound 1a (6.1 g, 25.63 mmol), methyl aminoacetate (3.54 g, 28.19 mmol, HCl), and N,N-diisopropylethylamine (9.94 g, 76.90 mmol) were added to DMF (60 mL) and reacted at 25°C for 18 hours. The reaction solution was poured into 300 mL of water and extracted with ethyl acetate (100 mL x 5). The organic phase was washed with 200 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified on a flash silica gel column (EA / PE = 0%-10%) to obtain Intermediate 25a. 1 H NMR (400MHz, DMSO-d6) δppm 7.54-7.68 (m, 2H), 6.70 (d, J = 7.8Hz, 1H), 4.20 (d, J = 4.2Hz, 2H), 3.68 (s, 3H).

[0753] Step 2: Synthesis of intermediate 25b

[0754] Intermediate 25a (1.82 g, 5.93 mmol) and NH4Cl (2.54 g, 47.42 mmol) were added to a mixture of MeOH (26 mL) and H2O (0.468 mL), followed by Zn (1.98 g, 30.29 mmol). The mixture was allowed to react at 0°C for 2 hours. The mixture was filtered, and 50 mL of water was added to the filtrate. The mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to yield Intermediate 25b. MS m / z: 276.9, 278.9 [M+1] + .

[0755] Step 3: Synthesis of intermediate 25c

[0756] Intermediate 25b (1.35 g, 4.87 mmol) was added to a mixture of hydrogen chloride / dioxane (4 M, 15.64 mL), EA (13.5 mL), and MeOH (13.5 mL) at 0°C and allowed to react for 2 hours. The reaction mixture was then concentrated under reduced pressure to afford Intermediate 25c. MS m / z: 245.0, 247.0 [M+1] + .

[0757] Step 4: Synthesis of intermediate 25d

[0758] At 0°C, DDQ (1.56 g, 6.86 mmol) was added to a solution of intermediate 25c (1.4 g, 5.71 mmol) in DCM (14 mL), and the temperature was raised to 25°C for 4 hours. The reaction solution was concentrated under reduced pressure, and 70 ml of saturated sodium bicarbonate solution was slowly added and stirred overnight. The mixture was extracted with ethyl acetate (3 × 50 mL), and the combined organic phases were washed with 70 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash silica gel column chromatography (elution ratio: MeOH / DCM = 0%-5%) to obtain intermediate 25d. MS m / z: 243.0, 245.0 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 12.75 (br s, 1H), 8.24 (s, 1H), 7.53-7.60 (m, 2H).

[0759] Step 5: Synthesis of intermediate 25e

[0760] Intermediate 25d (660 mg, 2.72 mmol) was dissolved in DMF (1 mL), and potassium carbonate (563.00 mg, 4.07 mmol), potassium iodide (67.62 mg, 407.35 μmol), and p-methoxybenzyl chloride (510.36 mg, 3.26 mmol) were added. The mixture was stirred at 20°C for 3 hours. Water (15 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (50 ml). The mixture was separated, and the organic phase was washed with water, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was separated by flash silica gel column chromatography (PE:EA = 3:1) to obtain intermediate 25e. MS m / z: 362.8, 364.8 [M+1] + , 1 H NMR (400MHz, DMSO-d6) δppm 8.43 (s, 1H), 7.50-7.64 (m, 2H), 7.24 (d, J = 4.2Hz, 2H), 6.85 (d, J = 4.2Hz, 2H), 5.60 (s, 2H), 3.72 (s, 3H).

[0761] Step 6: Synthesis of intermediate 25f

[0762] Tosylmethyl isocyanide (451.57 mg, 2.31 mmol) was dissolved in THF (10 mL) at 0°C and added to sodium hydride (154.19 mg, 3.85 mmol). The mixture was stirred for 10 min, followed by the addition of intermediate 25e (700.00 mg, 1.93 mmol) and stirring at 25°C for 2 hours. Water (5 mL) was added to the reaction solution, which was extracted with ethyl acetate (30 ml). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by flash silica gel column chromatography (PE:EA = 3:1-1:1, then DCM:MeOH = 10:1) to obtain intermediate 25f. MS m / z: 402.0, 404.0 [M+1] + , 1 H NMR (400MHz, CDCl3) δppm 8.44 (s, 1H), 8.13 (s, 1H), 7.40-7.51 (m, 2H), 7.22 (d, J = 4.2Hz, 2H), 6.85 (d, J = 4.2Hz, 2H), 5.61 (s, 2H), 3.79 (s, 3H).

[0763] Step 7: Synthesis of Intermediate 25g

[0764] 1-(tributyltin)-methanol (191.59 mg, 596.69 μmol) and Intermediate 25f (200 mg, 497.24 μmol) were dissolved in 1,4-dioxane (5 mL), and chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (39.12 mg, 49.72 μmol) was added and stirred at 80°C for 1.5 hours. 1-(tributyltin)-methanol (31.93 mg, 99.44 μmol) was then added, and stirring continued at 80°C for 2 hours. The temperature was then raised to 100°C and stirring continued for another 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified on a flash silica gel column (elution ratio: PE:EA = 1:1 - DCM:MeOH = 10:1) to obtain Intermediate 25g. MS m / z:354.1[M+1] + .

[0765] Step 8: Synthesis of Intermediate 25h

[0766] Intermediate 25g (80mg, 226.41μmol) was added to a reaction flask at 15-25°C. Trifluoroacetic acid (3.07g, 26.93mmol, 2.00mL) and trifluoromethanesulfonic acid (4.52mmol, 400.00μL) were added and stirred for 2 hours. The reaction mixture was added with 5mL of water, 5mL of methanol, and solid potassium carbonate to adjust the pH to 10. Stirring was continued for 1 hour. The reaction mixture was extracted with dichloromethane (20mL). The organic phases were combined and washed with water (15mL x 2). The organic phases were dried, filtered, and the filtrate was concentrated under reduced pressure to obtain Intermediate 25h. MS m / z: 234.1[M+1] + .

[0767] Step 9: Synthesis of intermediate 25i

[0768] Intermediate 25h (80 mg, 343.06 μmol) and 1,2-dibromotetrachloroethane (245.77 mg, 754.72 μmol) were added to DCM (1.5 mL), followed by n-tributylphosphine (138.81 mg, 686.11 μmol). The mixture was stirred at 20°C for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude intermediate 25i.

[0769] Step 10: Synthesis of trifluoroacetate salt of compound 25

[0770] Intermediate 25i (30 mg) and compound 9g (28.97 mg, hydrochloride) were dissolved in DMF (1 mL). Et3N (10.25 mg, 101.32 μmol) and KI (16.82 mg, 101.32 μmol) were added, and the mixture was stirred at 50°C for 2 hours. Water (5 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by preparative chromatography (developing ratio: DCM:MeOH = 10:1) and then separated by preparative HPLC (column: Welch Xtimate C18 100 × 40 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain the trifluoroacetate salt of compound 25. MS m / z: 454.2 [M+1] + , 1 H NMR(400MHz,CD3OD)δppm 8.99(s,1H),8.05(d,J=4.4Hz,1H),8.02(s,1H),7.93(d,J=4.4Hz,1H),7.58-7.63(m,1 H),7.46-7.49(m,1H),4.46(s,2H),3.71-3.75(m,4H),3.40-3.51(m,4H),2.92(s,3H).

[0771] Example 26

[0772] Step 1: Synthesis of intermediate 26a

[0773] Methyl 2-pyrrolidone (262.88 mg, 2.10 mmol) and compound 1a (500 mg, 2.10 mmol) were dissolved in DMF (10 mL). Sodium hydride (126.05 mg, 3.15 mmol, 60% purity) was added portionwise and stirred at 20°C for 3 hours. The reaction solution was slowly added dropwise to 20 mL of saturated ammonium chloride solution and extracted with ethyl acetate (2 × 30 mL). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by flash silica gel column chromatography (eluent: PE:EA = 90:10, V / V) to obtain the intermediate 26a (ratio approximately 2:1). MS m / z: 342.8, 344.8 [M+1] + .

[0774] Step 2: Synthesis of intermediate 26b

[0775] The intermediate compound 26a (370 mg) was dissolved in dioxane (10 mL), and 1-(tributyltin)-methanol (415.51 mg, 1.29 mmol) and chloro(2-dicyclohexylphosphino-2,4,6-triisopropyl-1,1-biphenyl)[2-(2-amino-1,1-biphenyl)]palladium(II) (42.42 mg, 53.92 μmol) were added. The mixture was stirred at 110°C for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude product, which was then purified by flash silica gel column chromatography (gradient elution: PE:EA = 100:0 to 60:40) to obtain the intermediate compound 26b. MS m / z: 294.9 [M+1] + .

[0776] Step 3: Synthesis of intermediate 26c

[0777] The mixture, Intermediate 26b (185 mg), was dissolved in MeOH (3 mL), the argon atmosphere was replaced, and palladium hydroxide (8.83 mg, 62.87 μmol) was added. The reaction flask was purged with hydrogen and stirred at 30°C, 50 psi for 16 hours. The mixture was filtered, the filtrate was concentrated under reduced pressure, and 2 mL of DMF and 5 drops of triethylamine were added and stirred at 90°C for 16 hours. DMF was removed using an oil pump to obtain a crude product. A mixture of DCM and MeOH (DCM:MeOH = 1:1, 3 mL) was added to the crude product, and the filter cake, Intermediate 26c, was filtered. MS m / z: 232.8 [M+1] + . 1H NMR (400MHz, DMSO-d6) δppm 11.22 (br s, 1H), 8.18-8.19 (m, 1H), 7.87 (d, J = 4.40Hz 1H), 7.26 (t, J = 8.20Hz, 1H), 7.07-7.08 (m, 1H), 6.70-6.72 (m, 1H), 5.35 (t, J = 5.60Hz, 1H), 4.60 (d, J = 2.80Hz, 2H).

[0778] Step 4: Synthesis of intermediate 26d

[0779] Intermediate 26c (20 mg, 86.13 μmol) and 1,2-dibromotetrachloroethane (28.05 mg, 86.13 μmol) were dissolved in DCM (0.5 mL). n-tributylphosphine (17.43 mg, 86.13 μmol) was added at 0°C. The mixture was stirred at room temperature (20°C) for 16 hours. 1,2-Dibromotetrachloroethane (28.05 mg, 86.13 μmol) and n-tributylphosphine (17.43 mg, 86.13 μmol) were then added and stirring continued for 1 hour. The reaction mixture was filtered to obtain a filter cake, which was Intermediate 26d. 1 H NMR (400MHz, DMSO-d6) δppm 11.41 (br s, 1H), 8.20-8.21 (m, 1H), 7.85 (d, J = 4.40Hz 1H), 7.36 (t, J = 8.20Hz, 1H), 7.10-7.11 (m, 1H), 6.73-6.75 (m, 1H), 4.79 (s, 2H).

[0780] Step 5: Synthesis of compound 26

[0781] Intermediate 9g (11.83 mg, hydrochloride) was dissolved in DMF (1 mL). Triethylamine (17.43 mg, 172.26 μmol) was added, and the mixture was stirred at 25°C for 0.5 hours. Intermediate 26d (10 mg, 43.06 μmol) and potassium iodide (3.57 mg, 21.53 μmol) were then added, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was added with 2 mL of water and extracted with ethyl acetate (3 x 3 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by preparative thin-layer chromatography (DCM:MeOH = 30:1) to afford compound 26. MS m / z: 453.0 [M+1] + .

[0782] Example 27

[0783] Step 1: Synthesis of intermediate 27a

[0784] Intermediate 15e (61.9 mg, 253.46 μmol) and 1,2-dibromotetrachloroethane (82.54 mg, 253.46 μmol) were dissolved in DCM (2 mL). n-Tributylphosphine (51.28 mg, 253.46 μmol) was added at 0°C. The mixture was stirred at room temperature (20°C) for 19 hours. The reaction mixture was filtered to obtain a filter cake, which was Intermediate 27a. 1 H NMR(400MHz,DMSO-d6)δppm 12.02(s,1H)9.05-9.13(m,1H)8.61-8.68(m,1H)8.41-8.49(m,1H)7.77(br d,J=2.01Hz,1H)7.35-7.50(m,1H)4.94(s,2H).

[0785] Step 2: Synthesis of compound 27

[0786] Intermediate 6d (21.23 mg, 78.15 μmol, HCl) was dissolved in DMF (2 mL), and triethylamine (19.77 mg, 195.36 μmol) was added. After stirring at 25°C for 0.5 hours, intermediate 27a (30 mg, 97.68 μmol) and potassium iodide (1.62 mg, 9.77 μmol) were added, and the mixture was stirred at 50°C for 2 hours. 2 mL of water was added to the reaction solution, and the mixture was extracted three times with 5 mL of a mixed solution (DCM:MeOH = 10:1). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was stirred with EA (0.5 mL) for 10 minutes and filtered to obtain a filter cake, compound 27. MS m / z: 462.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 11.90 (s, 1H), 9.09 (dd, J = 4.82, 1.81Hz, 1H), 8.65 (br dd,J=8.00,1.88Hz,2H),8.43(d,J=8.50Hz,1H),8.05-8.12(m,1H),7.88-7.96(m,1H),7.72(dd,J=7.88,4.88Hz,1H),7.39(br t,J=7.44Hz,1H),6.26(br s,1H),3.80(s,2H),3.21-3.22(m,2H),2.79(d,J=4.75Hz,3H),2.72-2.76(m,4H).

[0787] Example 28

[0788] Step 1: Synthesis of compound 28

[0789] Compound 26d (25 mg, 84.72 μmol) was dissolved in DMF (1 mL). Triethylamine (34.29 mg, 338.86 μmol) was added and stirred at 25°C for 0.5 h. Compound 6d (23 mg, hydrochloride) and potassium iodide (7.03 mg, 42.36 μmol) were then added and stirred at 50°C for 2 h. The reaction mixture was added with 2 mL of water and extracted with ethyl acetate (3 × 3 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was stirred with 0.5 mL of ethyl acetate for 10 min and filtered to obtain the filter cake, which was compound 28. MS m / z: 450.0 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δppm 11.32(s,1H),8.64(br d,J=4.77Hz,1H),8.20(s,1H),8.01-8.14(m,1H),7.83-7.97(m,2H),7.26 (t,J=7.78Hz,1H),7.09(d,J=3.01Hz,1H),6.72(t,J=3.26Hz,1H),6.25(br s,1H),3.72(s,2H),3.32(s,2H),3.13-3.22(m,2H),2.79(d,J=4.77Hz,3H),2.66-2.75(m,2H).

[0790] Example 29

[0791] Step 1: Synthesis of intermediate 29a

[0792] Deuterated methylamine hydrochloride (124.71 mg, 1.77 mmol) was added to H2O (1 mL), followed by sodium tert-butoxide (84.95 mg, 884.01 μmol) and stirring at 25°C for 1 hour. A solution of compound 4a (30 mg, 88.40 μmol) in MeOH (2 mL) was added to the reaction mixture, and the reaction was heated to 60°C for 4 hours. The reaction mixture was then concentrated under reduced pressure to yield the crude product, intermediate 29a. MS m / z: 342.1 [M+1] + .

[0793] Step 2: Synthesis of the hydrochloride salt of intermediate 29b

[0794] Intermediate 29a (20 mg) was dissolved in MeOH (0.6 mL), and HCl / dioxane (4 M, 0.2 mL) was added. The mixture was stirred at 20°C for 2 hours. The reaction mixture was directly concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 29b. MS m / z: 242.2 [M+1] + .

[0795] Step 3: Synthesis of compound 29

[0796] Intermediate 29b (19.22 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (162.80 μmol, 22.66 μL) was added. The mixture was stirred at 25°C for 30 minutes. Compound 27a (25 mg, 81.40 μmol) and potassium iodide (1.35 mg, 8.14 μmol) were then added, and the temperature was raised to 50°C and stirred for 3 hours. 2 mL of water was added to the reaction mixture, and the mixture was filtered to obtain a filter cake, compound 29. MS m / z: 468.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=4.63,1.75Hz,1H),8.65(dd,J=8.00,1.75Hz,1H),8.43(d,J=8.13Hz,1H),8.38(s,1H),7.85(d,J=7.88Hz,1H), 7.72(dd,J=8.00,4.50Hz,1H),7.57(dd,J=10.57,8.19Hz,1H),7.37(t,J=7.32Hz,1H),3.75(s,2H),3.15-3.22(m,4H),2.62-2.65(m,4H).

[0797] Example 30

[0798] Step 1: Synthesis of compound 30

[0799] Intermediate 16i (29.46 mg, hydrochloride) was dissolved in DMF (2 mL). Triethylamine (52.72 mg, 520.97 μmol) was added and stirred at 25°C for 0.5 h. Intermediate 27a (40 mg, 130.24 μmol) and KI (21.62 mg, 130.24 μmol) were then added and stirred at 50°C for 2 h. The reaction mixture was added with 2 mL of water and extracted with ethyl acetate (3 × 3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was added with 0.5 mL of ethyl acetate and stirred for 10 min. The filter cake was filtered and purified by preparative thin-layer chromatography (DCM:MeOH = 20:1) to afford compound 30. MS m / z: 473.0 [M+1] + . 1H NMR (400MHz, CDCl3) δppm 8.95-9.02(m,1H),8.73-8.79(m,1H),8.68(dd,J=8.16,1.88Hz,1H),8.46(br d,J=8.03Hz,1H),7.77-7.88(m,1H),7.65-7.75(m,1H),7.48-7.54(m,1H ),7.34-7.43(m,1H),6.98(d,J=8.53Hz,1H),3.63-3.86(m,3H),2.97-3.3 3(m,2H),2.92(d,J=5.27Hz,3H),2.82-2.89(m,2H),2.03-2.43(m,1H),1. 90-1.94(m,1H),1.68-1.83(m,1H),1.33-1.39(m,1H),1.23-1.31(m,2H).

[0800] Example 31

[0801] Step 1: Synthesis of intermediate 31a

[0802] Deuterated methylamine hydrochloride (167.77 mg, 2.38 mmol) was added to H2O (1 mL), and potassium tert-butoxide (133.44 mg, 1.19 mmol) was added to the reaction mixture. The mixture was stirred at 25°C for 1 hour. A solution of compound 6b (40 mg, 118.92 μmol) in MeOH (2 mL) was added to the reaction mixture, and the reaction was heated to 60°C for 16 hours. The reaction mixture was directly concentrated under reduced pressure to obtain the crude product, intermediate 31a. MS m / z: 339.1 [M+1] + .

[0803] Step 2: Synthesis of the hydrochloride salt of intermediate 31b

[0804] Intermediate 31a was dissolved in MeOH (1 mL), and HCl / dioxane (4 M, 99.74 μL) was added. The mixture was stirred at room temperature (25°C) for 2 hours. The reaction mixture was directly concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 31b. MS m / z: 238.9 [M+1] + .

[0805] Step 3: Synthesis of compound 31

[0806] Intermediate 31b (15 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (27.62 mg, 272.99 μmol, 38.00 μL) was added. After stirring at 25°C for 0.5 hour, intermediate 27a (20.96 mg, 68.25 μmol) and potassium iodide (5.66 mg, 34.12 μmol) were added, and the mixture was stirred at 50°C for 2 hours. After cooling, 2 mL of water was added to the reaction mixture, and a solid precipitated. The filter cake was filtered to obtain compound 31. MS m / z: 465.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δppm 11.89(br d,J=1.76Hz,1H),9.02-9.14(m,1H),8.56-8.70(m,2H),8.43(d,J=8.03Hz,1H),8.04-8.14 (m,1H),7.89-7.96(m,1H),7.72(dd,J=8.03,4.52Hz,1H),7.39(t,J=7.53Hz,1H),6.26(br s,1 H),3.80(s,2H),3.31(s,2H),3.18-3.24(m,2H),2.74(br t,J=5.27Hz,2H).

[0807] Example 32

[0808] Step 1: Synthesis of intermediate 32a

[0809] Intermediate 4a (0.2 g, 589.34 μmol) was added to a mixture of THF (5 mL) and H₂O (1 mL). Lithium hydroxide monohydrate (123.65 mg, 2.95 mmol) was added to the reaction mixture with stirring, and the mixture was stirred at 20°C for 4 hours. The pH of the reaction mixture was adjusted to 3-4 with dilute hydrochloric acid (1 mol / L), and the mixture was extracted with ethyl acetate (5 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure at 45°C to yield intermediate 32a. MS m / z: 347.9 [M+23] + .

[0810] Step 2: Synthesis of intermediate 32b

[0811] Under nitrogen, intermediate 32a (0.1 g, 307.38 μmol), 2,2,2-trifluoroethylamine (45.67 mg, 461.06 μmol), benzotriazole tetramethyl tetrafluoroboric acid (197.39 mg, 614.75 μmol), and diisopropylethylamine (119.18 mg, 922.13 μmol) were added to DMF (2 mL) and stirred at 40°C for 4 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product, which was then purified by preparative chromatography (PE / EA = 2 / 1) to obtain intermediate 32b. MS m / z: 351.1 [M-56+1] + .

[0812] Step 3: Synthesis of the hydrochloride salt of intermediate 32c

[0813] Intermediate 32b (57.66 mg, 141.88 μmol) was added to MeOH (2 mL). HCl / dioxane (4 M, 191.54 μL) was added to the reaction mixture with stirring. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure at 45°C to obtain the crude product, the hydrochloride salt of intermediate 32c. MS m / z: 306.9 [M+1] +

[0814] Step 4: Synthesis of compound 32

[0815] Triethylamine (156.29 μmol, 21.75 μL) was added to a solution of intermediate 32c (26.78 mg, hydrochloride) in DMF (1 mL). The reaction was stirred at 20°C for 0.5 hours. Intermediate 27a (30 mg, 78.15 μmol) and potassium iodide (1.30 mg, 7.81 μmol) were added to the reaction mixture. The reaction was stirred at 50°C for 2 hours, filtered, and the filtrate was concentrated under reduced pressure at 60°C to obtain the crude product. The crude product was purified by preparative chromatography (EA: 100%) to obtain compound 32. MS m / z: 555.1 [M+23] + . 1 H NMR (400MHz, CDCl3) δppm 8.92-9.04(m,1H),8.62-8.76(m,2H),8.44(br d,J=8.25Hz,1H),7.93(d,J=8.63Hz,1H),7.71(s,1H),7.45-7.55(m,1H),7.24 -7.35(m,1H),3.92-4.07(m,2H),3.68-3.78(m,2H),3.21(m,4H),2.66(m,4H).

[0816] Example 33

[0817] Step 1: Synthesis of intermediate 33d

[0818] 10a (3 g, 13.89 mmol) was dissolved in toluene (30 mL), followed by the addition of 1-(tert-butyloxycarbonyl)piperazine (2.59 g, 13.89 mmol), RuPhos (648.01 mg, 1.39 mmol), Pd2(dba)3 (381.49 mg, 416.61 μmol), and Cs2CO3 (13.57 g, 41.66 mmol). The mixture was stirred at 100°C under a nitrogen atmosphere for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product, which was then separated by column chromatography (eluent: PE:EA = 100:0 to 90:10 to 75:25, V / V) to afford intermediate 33d. MS m / z: 321.9 [M+1] + .

[0819] Step 2: Synthesis of intermediate 33a

[0820] At 25°C, intermediate 33d (1 g, 3.11 mmol) was added to a solution of methylamineethanol (322.13 mg, 3.11 mmol) and stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was dissolved in 10 ml of dichloromethane and washed with 10 ml of water. The organic phase was separated by extraction, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain intermediate 33a. MS m / z: 320.9 [M+1] + . 1 H NMR (400MHz, CDCl3) δppm 8.16 (d, J = 2.25Hz, 1H), 8.08 (d, J = 8.76Hz, 1H), 7.85 (br s, 1H), 7.24 (dd, J = 2.50, 8.75Hz, 1H), 3.56-3.69 (m, 4H), 3.30 (br d, J=4.63Hz, 4H), 3.01 (d, J=5.00Hz, 3H), 1.49 (s, 9H).

[0821] Step 3: Synthesis of intermediate 33b

[0822] Compound 33a (0.3 g, 936.37 μmol) was dissolved in DMF (1.5 mL). Chlorosuccinimide (137.54 mg, 1.03 mmol) was added at 20°C and stirred for 14 hours. Water (5 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with water (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain intermediate 33b. MS m / z: 355.1, 357.1 [M+1] + . 1H NMR (400MHz, CDCl3) δppm 8.10(d,J=4.2Hz,1H),7.69(br,s,1H),7.40(d,J=4.2Hz,1H),3.63–3.66(m,4H),3.09-3.15(m,4H),3.03(d,J=2.4Hz,3H),1.51(s,9H).

[0823] Step 4: Synthesis of the hydrochloride salt of intermediate 33c

[0824] Intermediate 33b (350 mg, 986.39 μmol) was dissolved in MeOH (6 mL), and HCl / dioxane (4 M, 6 mL) was added. The mixture was stirred at 20°C for 4 hours. The reaction mixture was concentrated under reduced pressure at 60°C to afford the hydrochloride salt of intermediate 33c. MS m / z: 255.1, 257.1 [M+1] + .

[0825] Step 5: Synthesis of compound 33

[0826] Intermediate 27a (20 mg, 65.12 μmol), intermediate 33c (18.96 mg, hydrochloride), potassium iodide (5.41 mg, 32.56 μmol), and triethylamine (26.36 mg, 260.49 μmol) were added sequentially to DMF (1 mL), heated to 50°C, and stirred for 12 hours. Water (3 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. 1.2 mL of methanol was added to the crude product, stirred for 10 minutes, and filtered. The filter cake was compound 33. MS m / z: 481.1 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.90(br s,1H),9.08-9.09(m,1H),8.65(d,J=2.40Hz,2H),8.43(d,J=4.20Hz,1H),7.93(d,J=3.92Hz,1H),7.66-7 .74(m,2H),7.33-7.39(m,1H),3.77(s,2H),3.05-3.15(m,4H),2.79(d,J=2.40Hz,3H),2.65-2.68(m,4H).

[0827] Example 34

[0828] Step 1: Synthesis of intermediate 34a

[0829] Intermediate 2a (200 mg, 854.62 μmol) and (2R,5S)-1-tert-butyloxycarbonyl-2,5-dimethylpiperazine (183.15 mg, 854.62 μmol) were dissolved in toluene (5 mL). RuPhos (79.76 mg, 170.92 μmol), Cs2CO3 (556.90 mg, 1.71 mmol), and Pd2(dba)3 (78.26 mg, 85.46 μmol) were added sequentially. The mixture was stirred at 100°C under nitrogen for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by column chromatography (eluent: PE:EA = 100:0 to 90:10 to 75:25, V / V) to obtain intermediate 34a. MS m / z: 368.0 [M+1] + , 1 H NMR (400MHz, CDCl3) δppm 7.97 (dd, J=8.03, 1.25Hz, 1H), 7.20 (dd, J=10.04, 8.03Hz, 1H), 4.38-4.45 (m, 1H), 4.05 (br d,J=2.26Hz,1H),3.97(s,3H),3.74-3.83(m,1H),3.47-3.52(m,2H),3.10( d,J=13.05Hz,1H),1.51(s,9H),1.29-1.32(m,3H),1.12(d,J=5.27Hz,3H).

[0830] Step 2: Synthesis of intermediate 34b

[0831] Intermediate 34a (187.4 mg, 510.05 μmol) was dissolved in EtOH (2 mL), and ethanolic methylamine (792.03 mg, 7.63 mmol, 30%) was added. The mixture was stirred at 25°C for 16 hours. The reaction mixture was directly concentrated under reduced pressure to obtain intermediate 34b. MS m / z: 367.0 [M+1] + , 1 H NMR (400MHz, CDCl3) δppm 7.99(dd,J=8.03,1.26Hz,1H),7.47-7.54(m,1H),7.24(dd,J=10.16,8.16Hz,1H),4.39-4.52(m,1H),3.93-4.02(m,1H),3.73 -3.78(m,1H),3.50(dd,J=12.17,3.89Hz,2H),2.95-3.02(m,4H),1.51(s,9H),1.31(d,J=6.78Hz,3H),1.09(d,J=6.53Hz,3H).

[0832] Step 3: Synthesis of the hydrochloride salt of intermediate 34c

[0833] Intermediate 34b (170 mg, 463.94 μmol) was dissolved in MeOH (2 mL), and HCl / dioxane (4 M, 0.62 mL) was added. The mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated under reduced pressure to afford the hydrochloride salt of intermediate 34c. MS m / z: 266.9 [M+1] + .

[0834] Step 4: Synthesis of trifluoroacetate salt of compound 34

[0835] Intermediate 34c (26.51 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (39.54 mg, 390.73 μmol) was added. After stirring at 25°C for 0.5 hour, Intermediate 27a (30 mg, 97.68 μmol) and potassium iodide (8.11 mg, 48.84 μmol) were added, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was filtered, and the filtrate was separated by preparative HPLC (column: Xtimate C18 100×30mm×3μm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 14%-44% acetonitrile) to obtain the trifluoroacetate salt of compound 34. MS m / z: 493.1 [M+1] + . 1 H NMR (400MHz, CD3OD) δppm9.11 (dd, J=4.52, 1.76Hz, 1H), 8.78 (dd, J=8.03, 1.76Hz, 1H), 8.69 (d, J=8.53Hz, 1H),8.01(d,J=7.03Hz,1H),7.85-7.92(m,1H),7.76(dd,J=8.03,4.52Hz,1H),7.53-7.56(m,1H),4.98(br d,J=16.56Hz,2H),4.47(br d,J=14.31Hz,1H),3.74-3.90(m,1H),3.49-3.57(m,2H),3.07-3.20(m,2H),2.95(s,3H),1.65(d,J=6.27Hz,3H),1.02(d,J=6.02Hz,3H).

[0836] Example 35

[0837] Step 1: Synthesis of intermediate 35a

[0838] (2S,5R)-1-tert-Butyloxycarbonyl-2,5-dimethylpiperazine (180.00 mg, 839.93 μmol) and intermediate 2a (196.56 mg, 839.93 μmol) were dissolved in toluene (10 mL). Cs2CO3 (821.00 mg, 2.52 mmol), RuPhos (39.19 mg, 83.99 μmol), and Pd2(dba)3 (38.46 mg, 42.00 μmol) were added sequentially. The mixture was stirred at 100°C for 16 hours under nitrogen. The reaction mixture was filtered while hot, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was added to 2 mL of a mixture of petroleum ether and ethyl acetate (PE / EA = 5 / 1), stirred at 20°C for 0.5 hour, filtered, and the filter cake was concentrated under reduced pressure to obtain intermediate 35a. MS m / z: 368.1 [M+1] + .

[0839] Step 2: Synthesis of intermediate 35b

[0840] Intermediate 35a (100 mg, 272.17 μmol) was dissolved in EtOH (2 mL), and ethanolic methylamine (1.13 g, 10.89 mmol, 30%) was added. The mixture was stirred at room temperature (25°C) for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain Intermediate 35b. MS m / z: 367.1 [M+1] + .

[0841] Step 3: Synthesis of the hydrochloride salt of intermediate 35c

[0842] Intermediate 35b (100 mg, 272.90 μmol) was dissolved in MeOH (2 mL), and HCl / dioxane (4 M, 0.38 mL) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to afford the hydrochloride salt of Intermediate 35c. MS m / z: 267.0 [M+1] + .

[0843] Step 4: Synthesis of trifluoroacetate salt of compound 35

[0844] Triethylamine (15.8.2 mg, 156.29 μmol) was added to a solution of intermediate 35c (23.66 mg, hydrochloride) in DMF (1 mL). The reaction was stirred at 20°C for 0.5 hours. Intermediate 27a (30 mg, 78.15 μmol) and KI (1.30 mg, 7.815 μmol) were added to the reaction mixture, stirred at 50°C for 0.5 hours, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative HPLC (column: Xtimate C18 100 × 30 mm × 3 μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 14% to 44% acetonitrile) to obtain the trifluoroacetate salt of compound 35. MS m / z: 493.2 [M+1]+1 H NMR(400MHz,CD3OD)δppm 9.11(dd,J=4.52,1.76Hz,1H),8.77(dd,J=8.03,1.76Hz,1H),8.69(d,J=7.78Hz,1H),8.01(d,J=7.78Hz,1H) ,7.86-7.93(m,1H),7.76(dd,J=8.03,4.77Hz,1H),7.55(dd,J=8.28,6.78Hz,1H),4.91-5.02(m,2H),4.50(br d,J=13.55Hz,1H),3.75-3.83(m,1H),3.48-3.58(m,2H),3.10-3.23(m,2H),2.94(s,3H),1.66(d,J=6.53Hz,3H),1.01(d,J=6.02Hz,3H).

[0845] Example 36

[0846] Step 1: Synthesis of intermediate 36a

[0847] (S)-1-N-tert-Butyloxycarbonyl-2-methylpiperazine (128.37 mg, 640.96 μmol) and intermediate 2a (150 mg, 640.96 μmol) were dissolved in toluene (10 mL). RuPhos (59.82 mg, 128.19 μmol), Cs2CO3 (417.68 mg, 1.28 mmol), and Pd2(dba)3 (58.69 mg, 64.10 μmol) were added sequentially. The mixture was stirred at 100°C under nitrogen for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by column chromatography (eluent: PE:EA = 90:10 to 75:25, V / V) to obtain intermediate 36a. MS m / z: 354.1 [M+1] + . 1 H NMR (400MHz, CDCl3) δppm 7.99 (dd, J=8.03, 1.00Hz, 1H), 7.26 (dd, J=10.04, 8.28Hz, 1H), 4.38 (br d, J=5.02Hz, 1H), 4.02 (br s,1H),3.98(s,3H),3.44-3.57(m,2H),3.31(td,J=12.80,3.26Hz,1H),3.02(dd ,J=11.92,3.64Hz,1H),2.84-2.94(m,1H),1.51(s,9H),1.35(d,J=6.78Hz,3H).

[0848] Step 2: Synthesis of intermediate 36b

[0849] Intermediate 36a (100 mg, 272.17 μmol) was dissolved in EtOH (2 mL), and ethanolic methylamine (0.65 g, 6.29 mmol, 30%) was added. The mixture was stirred at room temperature (25°C) for 16 hours. The reaction mixture was directly concentrated under reduced pressure to obtain intermediate 36b. MS m / z: 353.1 [M+1] + . 1 H NMR (400MHz, CDCl3) δppm 7.94-8.06 (m, 1H), 7.52 (br d, J = 4.27Hz, 1H), 7.29-7.34 (m, 1H), 4.39 (br s, 1H), 4.01 (br d,J=13.80Hz,1H),3.38-3.49(m,2H),3.30(td,J=12.74,3.39Hz,1H),3.02(d,J=5.02Hz,3 H), 2.97 (dd, J = 11.80, 3.51Hz, 1H), 2.81-2.89 (m, 1H), 1.51 (s, 9H), 1.36 (d, J = 6.78Hz, 3H).

[0850] Step 3: Synthesis of the hydrochloride salt of intermediate 36c

[0851] Intermediate 36b (51 mg, 428.49 μmol) was dissolved in MeOH (2 mL), and HCl / dioxane (4 M, 0.54 mL) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to afford the hydrochloride salt of intermediate 36c. MS m / z: 253.0 [M+1] + .

[0852] Step 4: Synthesis of compound 36

[0853] Triethylamine (32.95 mg, 325.61 μmol) was added to a solution of intermediate 36c (21.18 mg, hydrochloride) in DMF (1 mL). The reaction mixture was stirred at 20°C for 0.5 hours. Intermediate 27a (25 mg, 81.40 μmol) and KI (6.76 mg, 40.70 μmol) were added to the reaction mixture, and the reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was added with 2 mL of water and extracted with ethyl acetate (3 × 3 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was stirred with 1 mL of methanol for 10 minutes, and filtered to obtain the filter cake, which was compound 36. MS m / z: 479.0 [M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 11.87(s,1H),9.08(dd,J=4.64,1.88Hz,1H),8.64(dd,J=8.03,1.76Hz,1H),8.34-8.49(m, 2H),7.80-7.89(m,1H),7.71(dd,J=8.03,4.77Hz,1H),7.56(dd,J=10.54,8.28Hz,1H),7.3 3-7.44(m,1H),4.12(d,J=13.55Hz,1H),3.50(d,J=13.80Hz,1H),3.36-3.43(m,1H),2.87- 2.98(m,1H),2.74-2.86(m,5H),2.65-2.73(m,1H)2.28-2.47(m,2H)1.22(d,J=6.02Hz,3H).

[0854] Example 37

[0855] Step 1: Synthesis of intermediate 37a

[0856] (R)-1-N-tert-Butyloxycarbonyl-2-methylpiperazine (0.13 g, 649.10 μmol) and intermediate 2a (151.90 mg, 649.10 μmol) were dissolved in toluene (10 mL). Cs2CO3 (634.47 mg, 1.95 mmol), RuPhos (30.29 mg, 64.91 μmol), and Pd2(dba)3 (29.72 mg, 32.46 μmol) were added sequentially. The mixture was stirred at 100°C for 16 hours under nitrogen. The reaction mixture was filtered while hot, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was added with 2 mL of a mixture of PE and EA (PE / EA = 5 / 1), stirred at 20°C for 0.5 hour, filtered under reduced pressure, and the filter cake was dried under reduced pressure to obtain intermediate 37a. MS m / z: 376.1 [M+23] + .

[0857] Step 2: Synthesis of intermediate 37b

[0858] Intermediate 37a (96.18 mg, 272.17 μmol) was dissolved in EtOH (2 mL), and ethanolic methylamine (1.13 g, 10.89 mmol, 30%) was added. The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to afford Intermediate 37b. MS m / z: 375.1 [M+23] + .

[0859] Step 3: Synthesis of the hydrochloride salt of intermediate 37c

[0860] Intermediate 37b (100 mg, 283.77 μmol) was dissolved in MeOH (2 mL), and HCl / dioxane (4 M, 0.38 mL) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to afford the hydrochloride salt of Intermediate 37c. MS m / z: 253.0 [M+1] + .

[0861] Step 4: Synthesis of compound 37

[0862] Triethylamine (10.54 mg, 104.19 μmol) was added to a solution of intermediate 37c (15.04 mg, hydrochloride) in DMF (1 mL). The mixture was stirred at 20°C for 0.5 hours. Intermediate 27a (20 mg, 52.10 μmol) and KI (864.82 μg, 5.21 μmol) were added to the reaction mixture, and the mixture was stirred at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative chromatography (EA, 100%) to afford compound 37. MS m / z: 479.1 [M+1] + .

[0863] Example 38

[0864] Step 1: Synthesis of intermediate 38b

[0865] Compound 38a (1 g, 5.70 mmol), methyl 3-amino-2-fluorobenzoate (867.23 mg, 5.13 mmol), and pyridine (2.70 g, 34.18 mmol, 2.76 mL) were added to DCM (10 mL). Phosphorus oxychloride (960.81 mg, 6.27 mmol) was added at 0-5°C and stirred at this temperature for 2 hours. 10 mL of water was added to the reaction mixture at 0-5°C, and the mixture was extracted with DCM (10 mL x 2). The organic phases were combined and concentrated under reduced pressure to obtain the crude product, which was then purified by column chromatography (eluent: PE:EA = 100:0-90:10, V / V) to afford Intermediate 38b. MS m / z: 327.0, 329.0 [M+1] + , 1 H NMR (400MHz, CDCl3) δppm 8.91 (br s,1H),8.64-8.72(m,1H),8.45(d,J=3.00Hz,1H),8.11(dd,J=7.75,3.00Hz,1H),7.75-7.80(m,1H),7.29-7.34(m,1H),3.98(s,3H).

[0866] Step 2: Synthesis of intermediate 38c

[0867] Intermediate 38b (1.3 g, 3.98 mmol) and potassium carbonate (1.65 g, 11.94 mmol) were added to DMF (15 mL), followed by p-methoxybenzyl chloride (747.85 mg, 4.78 mmol). The mixture was heated to 90°C and stirred for 2 hours. The mixture was filtered, and 15 mL of water was added to the filtrate. The mixture was extracted with EA (30 mL x 2). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was isolated and purified by column chromatography (eluent: PE:EA = 100:0 to 75:25, V / V) to obtain Intermediate 38c. MS m / z: 447.1, 449.1 [M+1] + .

[0868] Step 3: Synthesis of intermediate 38d

[0869] Under nitrogen, intermediate 38c (300 mg, 671.40 μmol), tributylphosphine (135.83 mg, 671.40 μmol), 1,3-bis(diphenylphosphino)propane (110.76 mg, 268.56 μmol), Pd(OAc)2 (60.29 mg, 268.56 μmol), and potassium carbonate (185.58 mg, 1.34 mmol) were added to DMF (8 mL) and microwaved at 120°C for 10 min. The reaction mixture was filtered, the filter cake was washed with EA (30 mL), and the filtrate was concentrated to obtain the crude product. The crude product was separated and purified by column chromatography (eluent: PE:EA = 100:0-90:10, V / V) to obtain intermediate 38d, MS m / z: 411.0 [M+1]. + . 1 H NMR(400MHz,DMSO-d6)δppm 9.22(d,J=2.88Hz,1H),8.69(d,J=8.63Hz,1H),8.51(dd,J=8.38,2.88Hz,1H),7.80(dd,J=8.38,6 .13Hz, 1H), 7.15 (d, J = 8.50Hz, 2H), 6.87 (d, J = 8.76Hz, 2H), 5.60 (s, 2H), 3.86 (s, 3H), 3.71 (s, 3H).

[0870] Step 4: Synthesis of intermediate 38e

[0871] Intermediate 38d (140 mg, 341.16 μmol) was added to a mixture of trifluoroacetic acid (1 mL) and trifluoromethanesulfonic acid (0.2 mL) and stirred at 25°C for 1 hour. 4 mL of water was added to the reaction mixture, and a solid precipitated. The solid was filtered to obtain a filter cake, Intermediate 38e. 1H NMR (400MHz, DMSO-d6) δppm12.23 (br s, 1H), 9.18 (d, J = 2.26Hz, 1H), 8.44-8.46 (m, 2H), 7.70-7.84 (m, 1H), 3.93 (s, 3H).

[0872] Step 5: Synthesis of Intermediate 38f

[0873] Under nitrogen, LiAlH4 (23.54 mg, 620.22 μmol) was added to a solution of intermediate 38e (120 mg, 413.48 μmol) in THF (4 mL) at 0°C and stirred at this temperature for 1 hour. 24 μL of water, 24 μL of 15% sodium hydroxide solution, and 72 μL of ice water were slowly added to the reaction solution to quench the reaction. 2 mL of water was added to the reaction solution and extracted with EA (3 mL x 2). The organic phases were combined, dried, and concentrated to obtain the crude product. The mixture was stirred with 3 mL of a mixed solution (PE:EA = 1:1) for 10 minutes and filtered to obtain the filter cake, intermediate 38f. MS m / z: 263.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6)δppm 12.01(br s,1H),9.12(d,J=2.76Hz,1H),8.41(dd,J=8.66,2.89Hz,1H),8.37(d,J=8.0 3Hz, 1H), 7.37-7.46 (m, 1H), 5.46 (t, J = 5.65Hz, 1H), 4.68 (d, J = 5.27Hz, 2H).

[0874] Step 6: Synthesis of Intermediate 38g

[0875] Intermediate 38f (50 mg, 190.69 μmol) and 1,2-dibromotetrachloroethane (136.61 mg, 419.51 μmol) were dissolved in DCM (0.5 mL). Tributylphosphine (77.16 mg, 381.37 μmol, 94.10 μL) was added at 0°C. The mixture was warmed to 25°C and stirred for 3 hours. The reaction mixture was filtered to obtain the filter cake, which was Intermediate 38g. 1 H NMR(400MHz,DMSO-d6)δppm 12.15(s,1H),9.14(d,J=3.01Hz,1H),8.42(dd,J=8.53,3.01Hz,1H),8.36(d,J=8.28Hz,1H),7.38-7.51(m,1H),4.84(s,2H).

[0876] Step 7: Synthesis of compound 38

[0877] Triethylamine (15.56 mg, 153.80 μmol) was added to a solution of intermediate 9g (16.90 mg, hydrochloride) in DMF (1 mL) at 25°C. The mixture was stirred for 30 minutes, followed by the addition of intermediate 38g (25 mg, 76.90 μmol) and KI (1.28 mg, 7.69 μmol). The mixture was stirred at 50°C for 3 hours. 2 mL of water was added to the reaction mixture with stirring, and the mixture was stirred for approximately 10 minutes. The mixture was filtered to obtain a filter cake, compound 38. MS m / z: 483.1 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.05(s,1H),9.14(d,J=2.76Hz,1H),8.35-8.43(m,3H),7.81-7.88(m,1H),7.57(dd,J=10.54,8.03 Hz, 1H), 7.35-7.41 (m, 1H), 3.74 (s, 2H) 3.15-3.23 (m, 4H), 2.76 (d, J = 4.77Hz, 3H), 2.60-2.65 (m, 4H).

[0878] Example 39

[0879] Step 1: Synthesis of intermediate 39a

[0880] Compound 2a (200 mg, 854.62 μmol) and (R)-4-tert-butyloxycarbonyl-2-methylpiperazine (171.16 mg, 854.62 μmol) were dissolved in toluene (5 mL). RuPhos (79.76 mg, 170.92 μmol), Cs2CO3 (556.90 mg, 1.71 mmol), and Pd2(dba)3 (78.26 mg, 85.46 μmol) were added sequentially. The mixture was stirred at 100°C under nitrogen for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (eluent: PE:EA = 100:0 to 90:10 to 75:25, V / V) to obtain intermediate 39a. MS m / z: 353.9 [M+1] + .

[0881] Step 2: Synthesis of intermediate 39b

[0882] Intermediate 39a (150 mg, 424.46 μmol) was dissolved in EtOH (2 mL), and ethanolic methylamine (659.13 mg, 21.22 mmol) was added. The mixture was stirred at 25°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain Intermediate 39b. MS m / z: 353.0 [M+1] + .

[0883] Step 3: Synthesis of the hydrochloride salt of intermediate 39c

[0884] Intermediate 39b (150 mg, 425.65 μmol) was dissolved in MeOH (3 mL), and hydrogen chloride / dioxane (4 M, 3 mL) was added. The mixture was stirred at room temperature (25°C) for 3 hours. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 39c. MS m / z: 253.1 [M+1] +

[0885] Step 4: Synthesis of trifluoroacetate salt of compound 39

[0886] Intermediate 27a (25 mg, 81.40 μmol), intermediate 39c (20.54 mg, hydrochloride salt), KI (6.76 mg, 40.70 μmol), and triethylamine (32.95 mg, 325.61 μmol) were dissolved in DMF (1 mL) and heated to 50°C with stirring for 2 hours. The mixture was cooled, 3 mL of water was added, and extraction was performed with ethyl acetate (4 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. This was separated by preparative HPLC (column: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 3% to 33% acetonitrile) to afford the trifluoroacetate salt of compound 39. MS m / z: 479.2 [M+1] + . 1 H NMR(400MHz,CD3OD)δppm 9.08(dd,J=1.76,4.77Hz,1H),8.74(d,J=4.82Hz,1H),8.64(d,J=8.53Hz, 1H),7.96(d,J=8.03Hz,1H),7.73(dd,J=4.52,8.03Hz,1H),7.52(dd,J=6. 78,8.28Hz,1H),7.35(d,J=8.03Hz,1H),4.54(s,2H),3.85-4.12(m,2H),3 .32-3.51(m,4H),3.23-3.28(m,1H),2.93(s,3H),1.13(d,J=6.27Hz,3H).

[0887] Example 40

[0888] Step 1: Synthesis of trifluoroacetate salt of compound 40

[0889] Triethylamine (15.56 mg, 153.80 μmol) was added to a solution of intermediate 6d (16.72 mg, hydrochloride) in DMF (1 mL) at 25°C. The mixture was stirred for 30 minutes, followed by the addition of intermediate 38g (25 mg, 76.90 μmol) and KI (1.28 mg, 7.69 μmol). The mixture was stirred at 50°C for 3 hours. The reaction mixture was filtered and separated by preparative HPLC (column: C18 100×40 mm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 6%-36% acetonitrile) to afford the trifluoroacetate salt of compound 40. MS m / z: 480.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.29(br s,1H),10.26(s,1H),9.18(d,J=2.63Hz,1H),8.66(d,J=5.13Hz,1H),8.42-8.56(m,2H),8.13(t,J=8.63Hz,1H),7.98(d,J=7.63Hz,1H),7.45 -7.61(m,1H),6.27(s,1H),4.66(s,2H),3.90-4.12(m,2H),3.66-3.83 (m,1H),2.80(d,J=4.75Hz,3H),2.74-2.79(m,1H),2.22-2.32(m,2H).

[0890] Example 41

[0891] Step 1: Synthesis of intermediate 41b

[0892] Compound 41a (2 g, 9.90 mmol) and methyl 3-amino-2-fluorobenzoate (867.23 mg, 5.13 mmol) were added to DMF (20 mL), followed by triethylamine (3.01 g, 29.70 mmol) and HATU (4.52 g, 11.88 mmol), and stirred at 25°C for 16 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted with EA (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. Intermediate 41b was obtained by silica gel column chromatography (eluent: DCM:MeOH = 100:0-99:1, V / V) to obtain intermediate 41b. MS m / z: 352.9, 354.9 [M+1] + , 1H NMR(400MHz,DMSO-d6)δppm 10.67-10.84(m,1H),8.89(s,1H),8.70(d,J=5.02Hz,1H),8.06-8.12(m,1H),7 .74-7.78(m,1H),7.65(d,J=4.77Hz,1H),7.38(t,J=7.91Hz,1H),3.88(s,3H).

[0893] Step 2: Synthesis of Intermediate 41c

[0894] Intermediate 41b (1.8 g, 5.10 mmol) and potassium carbonate (2.11 g, 15.29 mmol) were added to DMF (20 mL), followed by p-methoxybenzyl chloride (957.90 mg, 6.12 mmol). The mixture was heated to 90°C and stirred for 3 hours. The mixture was filtered, and 20 mL of water was added to the filtrate. The mixture was extracted with EA (30 mL x 2). The organic phases were combined and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: PE:EA = 2:1, V / V) to obtain Intermediate 41c. MS m / z: 473.0, 475.0 [M+1] + .

[0895] Step 3: Synthesis of Intermediate 41d

[0896] Under nitrogen, intermediate 41c (300 mg, 633.86 μmol), tributylphosphine (135.83 mg, 671.40 μmol), 1,3-bis(diphenylphosphino)propane (104.57 mg, 253.54 μmol), Pd(OAc)2 (56.92 mg, 253.54 μmol), and potassium carbonate (175.21 mg, 1.27 mmol) were added to DMF (10 mL) and microwave-treated at 140°C for 10 min. The reaction mixture was filtered, and the filtrate was added with 20 mL of water and extracted three times with EA (30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain the crude product. The crude product was purified by column chromatography (eluent: PE:EA = 100:0 to 60:40, v / v) to obtain intermediate 41d. MS m / z: 393.1 [M+1] + , 1H NMR (400MHz, DMSO-d6) δ9.99(s,1H),8.95(d,J=5.13Hz,1H),8.66(d,J=8.50Hz,1H),8.22(d,J=5.13Hz,1H),7.76 (dd,J=8.38,6.25Hz,1H),7.13(d,J=8.50Hz,2H),6.86(d,J=8.63Hz,2H),5.58(s,2H),3.86(s,3H),3.71(s,3H).

[0897] Step 4: Synthesis of Intermediate 41e

[0898] Intermediate 41d (250 mg, 637.14 μmol) was added to a mixture of trifluoroacetic acid (1 mL) and trifluoromethanesulfonic acid (0.2 mL) and stirred at 25°C for 1 hour. 2 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (8 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to afford Intermediate 41e. MS m / z: 273.1 [M+1] + , 1 H NMR(400MHz,DMSO-d6)δppm12.17(br s,1H),9.95(s,1H),8.93(d,J=5.13Hz,1H),8.52(d,J=8.50Hz,1H),8.19(d,J=5.25Hz,1H),7.72(dd,J=8.44,6.69Hz,1H),3.93(s,3H).

[0899] Step 5: Synthesis of Intermediate 41f

[0900] Under nitrogen, LiAlH4 (2.5 M in tetrahydrofuran, 205.71 μL) was added to a solution of intermediate 41e (140 mg, 514.27 μmol) in THF (5 mL) at 0°C and stirred at this temperature for 1 hour. 30 μL of water, 30 μL of 15% sodium hydroxide solution, and 90 μL of ice water were slowly added to the reaction solution to quench the reaction. 10 mL of water was added to the reaction solution, and the mixture was extracted with EA (20 mL x 3). The organic phases were combined, dried, and concentrated to yield the crude product, intermediate 41f. MS m / z: 245.1 [M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 11.96(br s,1H),9.89(s,1H),8.85(d,J=5.02Hz,1H),8.41(d,J=8.53Hz,1H),8.14(d,J=5. 02Hz, 1H), 7.39 (t, J = 7.65Hz, 1H), 5.46 (t, J = 5.77Hz, 1H), 4.68 (d, J = 4.77Hz, 2H).

[0901] Step 6: Synthesis of Intermediate 41g

[0902] Intermediate 41f (50 mg, 204.73 μmol) and 1,2-dibromotetrachloroethane (146.67 mg, 450.41 μmol) were dissolved in DCM (0.5 mL). Tributylphosphine (82.84 mg, 409.47 μmol) was added at 0°C. The mixture was warmed to 25°C and stirred for 3 hours. The reaction mixture was filtered to obtain the filter cake, which was Intermediate 41g. 1 H NMR(400MHz,DMSO-d6)δppm 12.16(s,1H),9.90-9.98(m,1H),8.82-9.06(m,1H),8.38-8.48(m,1H),8.21(d,J=5.02Hz,1H),7.46(t,J=7.65Hz,1H),4.94(s,2H).

[0903] Step 7: Synthesis of trifluoroacetate salt of compound 41

[0904] Triethylamine (153.80 μmol, 21.41 μL) was added to a solution of intermediate 9g (16.90 mg, hydrochloride) in DMF (1 mL) at 25°C. The mixture was stirred for 30 minutes, followed by the addition of intermediate 41g (25 mg, 81.40 μmol) and KI (1.35 mg, 8.14 μmol). The mixture was stirred at 50°C for 2 hours. The reaction mixture was filtered and purified by preparative liquid chromatography (HPLC column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain the trifluoroacetate salt of compound 41. MS m / z: 465.2 [M+1] + . 1H NMR(400MHz,CD3OD)δppm 9.86(s,1H),8.91(d,J=5.27Hz,1H),8.49(d,J=8.53Hz,1H),8.33(d,J=5.27Hz,1H),7.96(d,J=8.28Hz,1H), 7.64(dd,J=10.29,8.28Hz,1H),7.56(dd,J=8.28,7.03Hz,1H),4.66(s,2H),3.47-3.72(m,8H),2.93(s,3H).

[0905] Example 42

[0906] Step 1: Synthesis of trifluoroacetate salt of compound 42

[0907] Triethylamine (16.47 mg, 162.80 μmol) was added to a solution of intermediate 6d (17.69 mg, hydrochloride) in DMF (1 mL). The mixture was stirred at 25°C for 30 min. Intermediate 41g (25 mg, 81.40 μmol) and KI (1.35 mg, 8.14 μmol) were then added. The mixture was heated to 50°C and stirred for 2 hours. The reaction mixture was filtered and purified by preparative HPLC (column: C18 100×40 mm; mobile phase: [water(trifluoroacetic acid)-acetonitrile]; gradient: 0%-30% acetonitrile) to obtain the trifluoroacetate salt of compound 42. MS m / z: 462.1 [M+1] + . 1 H NMR (400MHz, CD3OD) δ9.86 (s, 1H), 8.91 (d, J = 5.27Hz, 1H), 8.50 (d, J = 7.78Hz, 1H), 8.34 (d, J = 5.77Hz, 1H), 8.08-8.15 (m, 1H), 8.02-8 .07(m,1H),7.58(dd,J=8.28,7.03Hz,1H),6.31(s,1H),4.74(s,2H),4.11(s,2H),3.58-3.85(m,2H),2.96-3.01(m,2H),2.96(s,3H).

[0908] Example 43

[0909] Step 1: Synthesis of intermediate 43b

[0910] Under nitrogen, 43e (800 mg, 3.19 mmol), compound 43a (1.09 g, 3.51 mmol), Pd(dppf)Cl2 (233.70 mg, 319.39 μmol), and potassium acetate (626.91 mg, 6.39 mmol) were added to a mixture of H2O (1.6 mL) and THF (8 mL). The mixture was heated to 70°C and stirred for 16 hours. The reaction mixture was filtered, the filter cake was rinsed with EA (15 mL), and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by flash column chromatography (eluent: PE:EA = 100:0-90:10, V / V) to obtain intermediate 43b. MS m / z: 353.1 [M+1] + . 1 H NMR (400MHz, CDCl3) δppm 8.07 (dd, J=7.78, 1.00Hz, 1H), 7.64-7.76 (m, 1H), 5.83 (s, 1H), 4.11 (s, 2H), 3.67 (t, J=5.27Hz, 2H), 2.50 (br s,2H),1.52(s,9H),1.26(s,3H).

[0911] Step 2: Synthesis of intermediate 43c

[0912] Methylamine ethanol solution (9.05 g, 87.42 mmol, 30%) was added to a solution of intermediate 43b (300 mg, 850.31 μmol) in MeOH (1 mL) and stirred at 40°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain intermediate 43c. MS m / z: 352.1 [M+1] + . 1 H NMR (400MHz, CD3OD) δppm7.92(d,J=7.78Hz,1H),7.74(d,J=7.53Hz,1H),5.75(s, 1H),3.98(s,2H),3.53-3.61(m,2H),2.85(s,3H),2.33-2.41(m,2H),1.40(s,9H).

[0913] Step 3: Synthesis of the hydrochloride salt of intermediate 43d

[0914] Hydrogen chloride / dioxane (4M, 1.06 mL) was added to a solution of intermediate 43c (298 mg, 847.01 μmol) in MeOH (0.5 mL). The mixture was stirred at 25°C for 1 hour. Hydrogen chloride / dioxane (4M, 635.26 μL) was added and stirring continued for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 43d. MS m / z: 252.1 [M+1] + .

[0915] Step 4: Synthesis of compound 43

[0916] Triethylamine (32.95 mg, 325.61 μmol) was added to a solution of intermediate 43d (37.53 mg, hydrochloride) in DMF (1 mL) and stirred at 25°C for 30 min. Intermediate 27a (50 mg, 162.80 μmol) and KI (2.70 mg, 16.28 μmol) were then added, and the mixture was heated to 50°C and stirred for 2.5 hours. DMF (2 mL) and 1 mL of water were added to the reaction solution, and a solid precipitated. Filtering afforded a filter cake, compound 43. MS m / z: 478.1 [M+1] + . 1 H NMR (400 MHz, DMSO-d6) δ ppm 11.90(s,1H),9.09(dd,J=4.57,1.81Hz,1H),8.65(dd,J=8.07,1.81Hz,1H),8 .61(d,J=4.88Hz,1H),8.43(d,J=8.51Hz,1H),7.88-7.93(m,2H),7.72(dd,J= 8.07,4.57Hz,1H),7.36-7.43(m,1H),5.86(s,1H),3.82(s,2H),3.67-3.74(m , 2H), 3.18 (d, J = 2.50Hz, 2H), 2.81 (d, J = 4.75Hz, 3H), 2.45 (d, J = 1.63Hz, 2H).

[0917] Example 44

[0918] Step 1: Synthesis of intermediate 44a

[0919] Intermediate 15d (100 mg, 367.34 μmol) was dissolved in THF (1 mL). Deuterated lithium aluminum hydride powder (20.91 mg, 551.00 μmol) was added at 0°C and stirred at 0°C for 1 hour. The reaction mixture was added to a saturated sodium potassium tartrate solution (5 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to yield Intermediate 44a. MS m / z: 246.9 [M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 11.87(s,1H),9.08(dd,J=1.8,4.5Hz,1H),8.64(dd,J=1.9,7.9Hz,1H),8.43(d,J= 7.5Hz, 1H), 7.71 (dd, J=4.8, 8.0Hz, 1H), 7.40 (dd, J=6.7, 8.2Hz, 1H), 5.41 (s, 1H).

[0920] Step 2: Synthesis of intermediate 44b

[0921] Intermediate 44a (56 mg, 227.43 μmol) was dissolved in DCM (1 mL), and 1,2-dibromotetrachloroethane (162.93 mg, 500.34 μmol) was added. Tributylphosphine (92.02 mg, 454.85 μmol) was added at 0°C, and the mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated under reduced pressure, methanol (2 mL) was added, and the mixture was stirred for 5 minutes. The filter cake was filtered to obtain Intermediate 44b. 1 H NMR (400MHz, DMSO-d6) δppm 12.00 (s, 1H), 9.09 (d, J = 4.5Hz, 1H), 8.71-8.61 (m, 1H), 8.49-8.38 (m, 1H), 7.80-7.69 (m, 1H), 7.51-7.36 (m, 1H).

[0922] Step 3: Synthesis of compound 44

[0923] Intermediate 9g (31.99 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (58.92 mg, 582.28 μmol) was added. After stirring at 25°C for 0.5 hour, Intermediate 44b (45 mg, 145.57 μmol) and KI (12.08 mg, 72.78 μmol) were added, and the mixture was stirred at 50°C for 2 hours. Water (1.5 mL) was added to the reaction solution and stirred for 10 minutes. A solid precipitated, which was filtered to obtain the filter cake, which was Compound 44. MS m / z: 489.0 [M+23] + . 1H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=1.8,4.5Hz,1H),8.64(dd,J=1.8,8.0Hz,1H),8.47-8.37(m,2H),7.84(d,J=7.8Hz,1H),7.72(dd,J=4.5, 8.0Hz, 1H), 7.57 (dd, J = 8.3, 10.5Hz, 1H), 7.37 (dd, J = 6.7, 8.2Hz, 1H), 3.17-3.20 (m, 4H), 2.76 (d, J = 4.8Hz, 3H), 2.57-2.65 (m, 4H).

[0924] Example 45

[0925] Step 1: Synthesis of compound 45

[0926] Intermediate 33c (37.68 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (65.47 mg, 646.98 μmol) was added. After stirring at 25°C for 0.5 hr, intermediate 44b (50 mg, 161.74 μmol) and KI (13.42 mg, 80.87 μmol) were added, and the mixture was stirred at 50°C for 1 hour. After cooling, the reaction mixture was added with water (1 mL) and stirred for 10 min. A solid precipitated, which was filtered to obtain the filter cake, compound 45. MS m / z: 483.1 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=1.8,4.5Hz,1H),8.65(dd,J=1.8,8.0Hz,1H),8.48-8.38(m,2H),7.94(d,J=8.3Hz,1H),7.72(dd,J=4 .5,8.0Hz,1H),7.66(d,J=8.3Hz,1H),7.37(dd,J=6.7,7.9Hz,1H),3.10-3.15(m,4H),2.79(d,J=4.8Hz,3H),2.62-2.67(m,4H).

[0927] Example 46

[0928] Step 1: Synthesis of intermediate 46b

[0929] Intermediate 46a (2 g, 6.22 mmol) was dissolved in THF (20 mL) and H₂O (4 mL). Lithium hydroxide monohydrate (1.31 g, 31.12 mmol) was added and stirred at room temperature (25°C) for 2 hours. The reaction solution was adjusted to pH 3 with dilute hydrochloric acid (1 mol / L) and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain Intermediate 46b. MS m / z: 307.9 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 8.37(d,J=2.8Hz,1H),7.88(d,J=8.8Hz,1H),7.36(dd,J=3.0,8.8Hz,1H),3.50-3.46(m,4H),3.39-3.36(m,4H),1.43(s,9H).

[0930] Step 2: Synthesis of intermediate 46c

[0931] Intermediate 46b (400 mg, 1.30 mmol) and deuterated methylamine hydrochloride (100.98 mg, 1.43 mmol) were dissolved in EA (4 mL). DIEA (336.41 mg, 2.60 mmol) and n-butylphosphonic anhydride (50% ethyl acetate solution) (937.74 mg, 2.60 mmol) were added and stirred at 25°C for 3 hours. An additional 937.74 mg of n-butylphosphonic anhydride was added and stirring continued for 16 hours. The reaction mixture was adjusted to pH 10, 5 mL of water was added, and extraction was performed with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield Intermediate 46c. MS m / z: 324.0 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 8.43-8.37(m,1H),8.28(d,J=2.8Hz,1H),7.84(d,J=8.8Hz,1H),7.41(dd,J=2.8,8.8Hz,1H),3.52-3.45(m,4H),3.32-3.30(m,4H),1.43(s,9H).

[0932] Step 3: Synthesis of intermediate 46d

[0933] Intermediate 46c (314 mg, 970.92 μmol) was dissolved in DMF (3.5 mL), and chlorosuccinimide (142.61 mg, 1.07 mmol) was added. The mixture was stirred at 50°C for 1 hour. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield Intermediate 46d. MS m / z: 358.0 [M+1] + ,380.0[M+23] + . 1 H NMR (400MHz, DMSO-d6) δppm 8.44 (s, 1H), 7.95 (d, J = 8.0 Hz, 1H), 7.68 (d, J = 8.0 Hz, 1H), 3.52-3.47 (m, 4H), 3.09-3.01 (m, 4H), 1.43 (s, 9H).

[0934] Step 4: Synthesis of the hydrochloride salt of intermediate 46e

[0935] Intermediate 46d (355 mg, 992.04 μmol) was dissolved in MeOH (4 mL), and hydrogen chloride / dioxane (4 M, 1.34 mL) was added. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to give the hydrochloride salt of intermediate 46e. MS m / z: 257.9 [M+1] + .

[0936] Step 5: Synthesis of compound 46

[0937] Intermediate 46e (38.32 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (65.90 mg, 651.22 μmol) was added. After stirring at 25°C for 0.5 h, KI (13.51 mg, 81.40 μmol) and intermediate 27a (50 mg, 162.80 μmol) were added, and the mixture was stirred at 50°C for 1 h. After cooling, the reaction mixture was added with 2 mL of water and stirred for 10 min. A solid precipitated, which was filtered to obtain the filter cake, compound 46. MS m / z: 484.0 [M+1] + , 506.0[M+23] + . 1H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=1.8,4.5Hz,1H),8.64(dd,J=1.8,8.0Hz,1H),8.47-8.38(m,2H),7.94(d,J=8.3Hz,1H),7.72(dd ,J=4.5,8.0Hz,1H),7.67(d,J=8.0Hz,1H),7.37(dd,J=6.8,8.0Hz,1H),3.76(s,2H),3.16-3.12(m,4H),2.69-2.63(m,4H).

[0938] Example 47

[0939] Step 1: Synthesis of intermediate 47a

[0940] Deuterated methylamine hydrochloride (399.85 mg, 5.67 mmol) was added to H2O (2 mL), followed by sodium tert-butoxide (272.39 mg, 2.83 mmol), and the mixture was stirred at 25°C for 1 hour. A solution of intermediate 43b (100 mg, 283.44 μmol) in MeOH (4 mL) was added, the temperature was raised to 60°C, and the mixture was stirred for 4 hours. The mixture was cooled to room temperature and extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product. Intermediate 47a was isolated and purified by preparative chromatography (developing solvent: PE:EA = 2:1) to obtain intermediate 47a. MS m / z: 355.1, 357.1 [M+1] + .

[0941] Step 2: Synthesis of intermediate 47b hydrochloride

[0942] At 25°C, add hydrogen chloride / dioxane (4M, 211.36 μL) to a solution of intermediate 47a (60 mg, 169.09 μmol) in MeOH (0.5 mL) and stir for 2 hours. The reaction mixture is concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 47b. MS m / z: 255.1, 257.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.36 (br s, 2H), 8.63 (s, 1H), 8.03 (d, J = 7.78Hz, 1H), 7.89 (d, J = 7.78Hz, 1H), 5.93 (br s, 1H), 3.76 (br d,J=1.51Hz,2H), 3.30(br d,J=4.52Hz,2H), 2.63(br d,J=1.51Hz,2H).

[0943] Step 3: Synthesis of compound 47

[0944] Triethylamine (32.95 mg, 325.61 μmol) was added to a solution of intermediate 47b (37.93 mg, hydrochloride) in DMF (1.5 mL) and stirred at 25°C for 0.5 h. Intermediate 27a (50 mg, 162.80 μmol) and KI (2.70 mg, 16.28 μmol) were then added and stirred at 50°C for 2.5 h. After cooling, 1.5 mL of water was added to the reaction mixture. Solid precipitated and was filtered to obtain a filter cake, compound 47. MS m / z: 481.0, 483.0 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.89(s,1H),9.09(dd,J=4.50,1.75Hz,1H),8.65(dd,J=8.00,1.75Hz, 1H),8.57(s,1H),8.44(d,J=8.25Hz,1H),7.95-7.99(m,1H),7.88-7.93( m,1H),7.72(dd,J=8.00,4.63Hz,1H),7.36-7.43(m,1H),5.87(s,1H),3. 82(s,2H),3.19(d,J=2.13Hz,2H),2.72-2.78(m,2H),2.44-2.47(m,2H).

[0945] Example 48

[0946] Step 1: Synthesis of intermediate 48a

[0947] (R)-1-tert-Butyloxycarbonyl-2-methylpiperazine (0.2 g, 998.62 μmol), 43e (250.13 mg, 998.62 μmol), Cs2CO3 (650.74 mg, 2.00 mmol), RuPhos (93.20 mg, 199.72 μmol), and Pd2(dba)3 (91.44 mg, 99.86 μmol) were added to toluene (4 mL). The atmosphere was purged with nitrogen three times and stirred at 100°C for 6 hours. The reaction mixture was filtered under reduced pressure while hot, and the filtrate was concentrated under reduced pressure at 50°C to obtain the crude product. The crude product was purified by column chromatography (eluent: PE / EA = 4 / 1, V / V) to obtain intermediate 48a. MS m / z: 370.0, 372.0 [M+1] + .

[0948] Step 2: Synthesis of intermediate 48b

[0949] Intermediate 48a (183.15 mg, 495.21 μmol) was added to EtOH (2 mL). Methylamine ethanol solution (2.27 g, 21.93 mmol) was added to the reaction mixture with stirring. The reaction mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure at 45°C to obtain the crude product, Intermediate 48b. MS m / z: 312.9, 314.9 [M-56] + .

[0950] Step 3: Synthesis of the hydrochloride salt of intermediate 48c

[0951] Intermediate 48b (100 mg, 271.11 μmol) was added to MeOH (2 mL). Hydrogen chloride / dioxane (4 M, 383.08 μL) was added to the reaction mixture with stirring. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure at 45°C to obtain the hydrochloride salt of Intermediate 48c. MS m / z: 268.9, 270.9 [M+1] + .

[0952] Step 4: Synthesis of compound 48

[0953] Triethylamine (26.36 mg, 260.49 μmol) was added to a solution of intermediate 48c (39.75 mg, hydrochloride) in DMF (2 mL). The reaction was stirred at 20°C for 0.5 hours. Intermediate 27a (50 mg, 130.24 μmol) and KI (2.16 mg, 13.02 μmol) were added to the reaction mixture, and the reaction was stirred at 50°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative chromatography (100% EA) to obtain compound 48. MS m / z: 495.2, 497.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.86(s,1H),9.08(dd,J=4.57,1.69Hz,1H),8.64(dd,J=8.00,1.75Hz,1H),8.36-8.48 (m,2H),7.94(d,J=8.13Hz,1H),7.60-7.76(m,2H),7.33-7.47(m,1H),4.12(d,J=13.38H z,1H),3.53(d,J=13.88Hz,1H),3.20-3.26(m,1H),2.85-2.95(m,1H),2.80-2.85(m,1H ), 2.79 (d, J = 4.75Hz, 3H), 2.67-2.77 (m, 2H), 2.38-2.45 (m, 2H), 1.24 (d, J = 5.50Hz, 3H).

[0954] Example 49

[0955] Step 1: Synthesis of intermediate 49a

[0956] Deuterated methylamine hydrochloride (190.72 mg, 2.70 mmol) was added to H2O (1 mL), and sodium tert-butoxide (129.92 mg, 1.35 mmol) was added to the reaction mixture, which was stirred at 25°C for 1 hour. Intermediate 48a (50 mg, 135.19 μmol) in MeOH (2 mL) was added to the reaction mixture, and the reaction was heated to 60°C for 4 hours. The mixture was extracted with ethyl acetate (5 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain intermediate 49a. MS m / z: 394.1, 396.1 [M+23] + .

[0957] Step 2: Synthesis of the hydrochloride salt of intermediate 49b

[0958] Intermediate 49a (50 mg, 134.45 μmol) was added to MeOH (2 mL). Hydrogen chloride / dioxane (4 M, 383.08 μL) was added to the reaction mixture with stirring. The mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of intermediate 49b. MS m / z: 272.0, 274.0 [M+1] + .

[0959] Step 3: Synthesis of compound 49

[0960] Triethylamine (26.36 mg, 260.49 μmol) was added to Intermediate 49b (40.14 mg, hydrochloride) in DMF (2 mL), and the reaction was stirred at 20°C for 0.5 hours. Intermediate 27a (50 mg, 130.24 μmol) and KI (2.16 mg, 13.02 μmol) were added to the reaction mixture, and the reaction was stirred at 50°C for 2 hours. The crude product was concentrated under reduced pressure and purified by preparative chromatography (100% EA) to afford Compound 49. MS m / z: 498.2, 500.2 [M+1] + . 1H NMR (400 MHz, DMSO-d6) δ ppm 11.86(s,1H),9.08(dd,J=4.57,1.69Hz,1H),8.64(dd,J=7.94,1.81Hz,1H), 8.33-8.50(m,2H),7.94(d,J=8.25Hz,1H),7.58-7.78(m,2H),7.31-7.48(m, 1H),4.12(d,J=13.38Hz,1H),3.53(d,J=13.76Hz,1H),3.24(s,1H),2.80-2. 93(m,2H),2.70-2.79(m,2H),2.43(t,J=9.01Hz,2H),1.24(d,J=5.50Hz,3H).

[0961] Example 50

[0962] Step 1: Synthesis of compound 50

[0963] Triethylamine (31.12 mg, 307.59 μmol) was added to a solution of intermediate 33c (35.83 mg, hydrochloride) in DMF (1 mL) and stirred at 25°C for 0.5 h. Intermediate 38g (50 mg, 153.80 μmol) and KI (2.55 mg, 15.38 μmol) were then added and stirred at 50°C for 2.5 h. After cooling, water (1 mL) was added to the reaction mixture, resulting in the precipitation of a solid which was directly filtered to obtain the filter cake, compound 50. MS m / z: 499.1, 501.1 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.03(br s,1H),9.13(d,J=2.88Hz,1H),8.39-8.46(m,2H),8.36(d,J=8.25Hz,1H),7.93(d,J=8.25Hz,1H),7.66(d,J=8.25 Hz,1H),7.38(dd,J=8.00,6.75Hz,1H),3.76(s,2H),3.08-3.16(m,4H),2.79(d,J=4.88Hz,3H),2.63-2.69(m,4H).

[0964] Example 51

[0965] Step 1: Synthesis of compound 51

[0966] Triethylamine (18.67 mg, 184.55 μmol) was added to a solution of intermediate 29b (23.07 mg, hydrochloride) in DMF (1 mL) and stirred at 25°C for 0.5 h. Intermediate 38g (30 mg, 92.28 μmol) and KI (1.53 mg, 9.23 μmol) were then added and stirred at 50°C for 2.5 h. After cooling to room temperature, 1 mL of water was added to the reaction mixture. Solid precipitated and was filtered to obtain a filter cake, compound 51. MS m / z: 486.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.05(s,1H),9.13(d,J=3.00Hz,1H),8.42(dd,J=8.57,2.94Hz,1H),8.34-8.39(m,2H),7.84(d,J=7.50Hz, 1H),7.56(dd,J=10.57,8.19Hz,1H),7.34-7.41(m,1H),3.74(s,2H),3.15-3.22(m,4H),2.59-2.65(m,4H).

[0967] Example 52

[0968] Step 1: Synthesis of compound 52

[0969] Intermediate 46e (21.72 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (37.35 mg, 369.11 μmol) was added. After stirring at 25°C for 0.5 hours, KI (7.66 mg, 46.14 μmol) and intermediate 38e (30 mg, 92.28 μmol) were added, and the mixture was stirred at 50°C for 2 hours. After cooling to room temperature, 1 mL of water was added to the reaction mixture, and a solid precipitated. The filter cake was filtered to obtain compound 52. MS m / z: 502.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.06(s,1H),9.14(d,J=3.0Hz,1H),8.44-8.40(m,2H),8.37(d,J=8.5Hz,1H),7.94(d,J=8.0Hz ,1H),7.66(d,J=8.3Hz,1H),7.39(t,J=7.5Hz,1H),3.76(s,2H),3.12(m,4H),2.69-2.62(m,4H).

[0970] Example 53

[0971] Step 1: Synthesis of intermediate 53a

[0972] Intermediate 38e (150 mg, 516.85 μmol) was dissolved in THF (2 mL). Deuterated lithium aluminum hydride powder (29.42 mg, 775.27 μmol) was added at 0°C and stirred at 0°C for 1 hour. 30 μL of water, 30 μL of 15% sodium hydroxide solution, and 90 μL of ice water were slowly added to the reaction solution to quench the reaction. Water (5 mL) was added to the reaction solution, followed by extraction with ethyl acetate (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure at 45°C to afford Intermediate 53a. MS m / z: 265.1 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δppm 9.10 (d, J = 2.8 Hz, 1H), 8.34-8.41 (m, 2H), 7.36 (t, J = 8.0 Hz, 1H), 5.38 (s, 1H).

[0973] Step 2: Synthesis of intermediate 53b

[0974] Intermediate 53a (130 mg, 492.01 μmol) was dissolved in DCM (3 mL), and 1,2-dibromotetrachloroethane (352.48 mg, 1.08 mmol) was added. Tributylphosphine (199.09 mg, 984.01 μmol) was added at 0°C, and the mixture was stirred at 25°C for 3 hours. The reaction mixture was concentrated under reduced pressure, methanol (5 mL) was added, and the mixture was stirred for 5 minutes. The filter cake was filtered to obtain Intermediate 53b. 1 H NMR (400MHz, DMSO-d6) δppm 12.16 (s, 1H), 9.13 (d, J = 2.8Hz, 1H), 8.41-8.43 (m, 1H), 8.34-8.37 (m, 1H), 7.44 (t, J = 8.0Hz, 1H).

[0975] Step 3: Synthesis of compound 53

[0976] Intermediate 9g (22.68 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (18.56 mg, 183.42 μmol) was added. After stirring at 25°C for 0.5 hour, Intermediate 53b (30 mg, 91.71 μmol) and KI (1.52 mg, 9.17 μmol) were added, and the mixture was stirred at 50°C for 2 hours. Water (1 mL) was added to the reaction solution and stirred for 10 minutes. A solid precipitated, which was filtered to obtain the filter cake, which was Compound 53. MS m / z: 485.1 [M+1] + . 1H NMR(400MHz,DMSO-d6)δppm 12.05(s,1H),9.14(d,J=3.01Hz,1H),8.39-8.45(m,2H),8.37(d,J=8.03Hz,1H),7.84(d,J=7.28Hz,1H),7.56( dd,J=10.54,8.28Hz,1H),7.38(t,J=7.15Hz,1H),3.14-3.24(m,4H),2.76(d,J=4.77Hz,3H),2.60-2.66(m,4H).

[0977] Example 54

[0978] Step 1: Synthesis of compound 54

[0979] Intermediate 29b (23.87 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (43.49 mg, 429.80 μmol) was added. After stirring at 25°C for 0.5 hour, KI (8.92 mg, 53.73 μmol) and intermediate 44b (33.22 mg, 107.45 μmol) were added, and the mixture was stirred at 50°C for 1 hour. 1 mL of water was added to the reaction mixture, and a solid precipitated. Filtering gave a filter cake, Compound 54. MS m / z: 470.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 11.90(s,1H),9.09(dd,J=1.8,4.5Hz,1H),8.64(dd,J=1.6,7.9Hz,1H),8.43(d,J=8.3Hz,1H),8.38(s,1H),7.84(d,J=8.5H z,1H),7.72(dd,J=4.5,8.0Hz,1H),7.57(dd,J=8.2,10.7Hz,1H),7.39-7.34(m,1H),3.17-3.21(m,4H),2.59-2.65(m,4H).

[0980] Example 55

[0981] Step 1: Synthesis of compound 55

[0982] Intermediate 29b (25 mg, hydrochloride) was dissolved in DMF (1 mL), and triethylamine (40.48 mg, 400.05 μmol) was added. After stirring at 25°C for 0.5 hour, KI (8.30 mg, 50.01 μmol) and intermediate 53b (32.72 mg, 100.01 μmol) were added, and the mixture was stirred at 50°C for 1 hour. After cooling, 1 mL of water was added to the reaction mixture, and a solid precipitated. The filter cake was filtered to obtain compound 55. MS m / z: 488.2 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δppm 12.06(s,1H),9.14(d,J=2.8Hz,1H),8.42(dd,J=3.0,8.5Hz,1H),8.39-8.34(m,2H),7.84(d,J=8.0Hz ,1H),7.56(dd,J=8.0,10.5Hz,1H),7.38(dd,J=6.5,8.3Hz,1H),3.15-3.19(m,4H),2.56-2.64(m,4H).

[0983] Biological test data:

[0984] Experimental Example 1: Fluorescence polarization assay to determine the binding activity of compounds to PARP1

[0985] Experimental methods:

[0986] Fluorescence polarization experiments were performed in 96-well black-walled plates (Greiner) using a reaction buffer of 50 mM Tris pH 8, 0.001% Triton X100, 10 mM MgCl2, and 150 mM NaCl. The fluorescent probe and PARP1 protein carrying the His-Avi tag were both produced by Shanghai WuXi AppTec Pharmaceutical Development Co., Ltd. In a 100 μL reaction system, 8 nM PARP1, 5 nM fluorescent probe, and compound were added, respectively. The compound concentrations were based on the starting concentration and dilution gradient listed in Table 1. For example, compound 1 started at 100 nM and was diluted 3-fold. The cells were incubated at room temperature in the dark for 4 h and read using Envision. The fluorescence polarization values ​​and compound concentrations were nonlinearly fitted using Graphpad 8.0 software to obtain the IC 50 value.

[0987] Experimental results:

[0988] Table 1. PARP1 binding activity test of the compounds of the present invention

[0989] Conclusion: The compounds of the present invention have excellent binding activity to PARP1.

[0990] Experimental Example 2: Fluorescence polarization assay to determine compound binding activity to PARP2

[0991] Experimental methods:

[0992] Fluorescence polarization experiments were performed in 96-well black-walled plates (Greiner) using a reaction buffer of 50 mM Tris pH 8, 0.001% Triton X100, 10 mM MgCl2, and 150 mM NaCl. The fluorescent probe and PARP2 protein carrying the His-Avi tag were both produced by Shanghai WuXi AppTec Pharmaceutical Development Co., Ltd. 3 nM PARP2, 5 nM fluorescent probe, and compound were added to a 100 μL reaction system, respectively. The compound concentrations were used as the starting concentration and dilution multiple according to Table 2. The cells were incubated at room temperature in the dark for 4 h and read using Envision. The fluorescence polarization values ​​and compound concentrations were nonlinearly fitted using Graphpad 8.0 software to obtain the IC 50 value.

[0993] Experimental results:

[0994] Table 2. PARP2 binding activity test of the compounds of the present invention

[0995] Conclusion: The compounds of the present invention have weak binding activity to PARP2.

[0996] Experimental Example 3: Anti-proliferation experiment of compounds in MDA-MB-436 cells (BRCA1 mutation)

[0997] Experimental methods:

[0998] MDA-MB-436 cells (BRCA1 mutation) were seeded in a black (transparent bottom) 96-well plate with 135 μL of cell suspension per well, containing 3500 MDA-MB-436 cells (BRCA1 mutation). The cell plate was cultured in a carbon dioxide incubator overnight. A 400X stock solution of the test compound was prepared and the test compound was diluted 5-fold to the 9th concentration, from 4 mM to 104 nM, using a dispenser. A double-well experiment was set up. 78 μL of culture medium was added to the middle plate, and then 2 μL of the gradient dilution compound per well was transferred to the middle plate according to the corresponding position. 2 μL of DMSO was added to the vehicle control and blank control, mixed, and then 15 μL was transferred to each well of the cell plate. The concentration range of the compound transferred to the cell plate was 10 μM to 0.26 nM, and the final DMSO concentration was 0.25%. The cell plate was cultured in a carbon dioxide incubator for 7 days. Remove the cell plate and allow it to equilibrate to room temperature for 30 minutes. Add 75 μL of cell viability chemiluminescent detection reagent to each well and shake the plate on an orbital shaker for 3 minutes to induce cell lysis. Incubate at room temperature for 10 minutes to allow the luminescent signal to stabilize. Luminescent signals were detected on a 2104 EnVision plate reader. The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1 – (RLU compound – RLU blank control) / (RLU vehicle control – RLU blank control)) × 100%. Inhibition rates at different compound concentrations were calculated in Excel, and inhibition curves and related parameters were plotted using GraphPad Prism software.

[0999] Experimental results: as shown in Table 3:

[1000] Table 3. MDA-MB-436 cell (BRCA1 mutation) proliferation inhibition experiment

[1001] Experimental conclusion: The compound of the present invention has excellent proliferation inhibitory activity on MDA-MB-436 cells (BRCA1 mutation).

[1002] Experimental Example 4: DLD1 (BRCA2KO) cell anti-proliferation experiment of compounds

[1003] Experimental methods:

[1004] DLD1 (BRCA2 KO) cells were seeded in a white 96-well plate, with 80 μL of cell suspension per well containing 1000 DLD1 (BRCA2 KO) cells. The plate was incubated overnight in a CO2 incubator. The test compound was diluted 5-fold using a pipette to the eighth concentration, from 2 mM to 0.0256 μM, in duplicate. 78 μL of culture medium was added to the middle plate. 2 μL of the serially diluted compound was transferred to each well of the plate, mixed, and then 20 μL was transferred to each well of the plate. The compound concentrations transferred to the plate ranged from 10 μM to 0.128 nM. The plate was incubated in a CO2 incubator for 7 days. A separate plate was prepared, and the signal value was read on the day of drug addition, which was used as the maximum value (Max in the equation below) for data analysis. 25 μL of cell viability chemiluminescent detection reagent was added to each well of this plate and incubated at room temperature for 10 minutes to allow the luminescent signal to stabilize. The data were read using a multilabel analyzer. After the incubation of the cell plates with the added compounds, 25 μL of cell viability chemiluminescent detection reagent was added to each well of the cell plates and incubated at room temperature for 10 minutes to allow the luminescent signal to stabilize. The readings were read using a multi-label analyzer. The raw data were converted to inhibition rate, IC, using the equation (Sample-Min) / (Max-Min)×100%. 50 The value can be obtained by four-parameter curve fitting (obtained in "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism).

[1005] Experimental results: as shown in Table 4.

[1006] Table 4. DLD1 (BRCA2KO) cell anti-proliferation experiment

[1007] Experimental conclusion: The compound of the present invention has a good anti-proliferation inhibitory effect on DLD1 (BRCA2KO) cells.

[1008] Experimental Example 5: Pharmacokinetic Evaluation and Brain Penetration Evaluation of the Compounds of the Invention in Mice

[1009] Experimental methods:

[1010] The trifluoroacetic acid salt of compound 16 was tested in female BALB / C mice, while the trifluoroacetic acid salt of compound 15 was tested in male CD-1 mice. Mice (7-9 weeks of age, fasted overnight) were injected with a 0.4 mg / mL clear solution of the test compound in 90% water (10% 2-hydroxypropyl-β-cyclodextrin) via the tail vein at a dose of 2 mg / kg. Mice (7-9 weeks of age, fasted overnight) were administered a 1 mg / mL clear solution of the test compound in 0.5% methylcellulose water via oral gavage at a dose of 10 mg / kg. The two groups of animals were respectively given blood from the jugular vein at 0.0833, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, and 24 hours after administration, and from the tail vein at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, and 24 hours. About 30 μL of blood was placed in an anticoagulant tube with EDTA-K2, and the plasma was separated by centrifugation. Among them, the trifluoroacetic acid salt of compound 15 was intravenously administered (IV, 2 mg / kg) in one group (blood was collected at the above time points), and orally administered (PO, 10 mg / kg) in one group (blood was collected at the above time points). In addition, a group of oral administration (PO, 10 mg / kg) was set up to collect plasma and brain tissue homogenate at 2 hours. The blood drug concentration was determined by LC-MS / MS, and WinNonlin was used to determine the blood drug concentration. TM Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software was used to calculate relevant pharmacokinetic parameters using the non-compartmental linear logarithmic trapezoidal method.

[1011] Experimental results: The pharmacokinetic data of mice are shown in Table 5, and the brain penetration data of mice are shown in Table 6:

[1012] Table 5. Pharmacokinetic data in mice Note: “-” indicates that the parameter cannot be obtained by calculation; C0 represents the initial concentration; C max represents the peak concentration; T max T stands for time to peak 1 / 2 Vd stands for elimination half-life; ss represents the apparent volume of distribution at steady state; Cl represents the total clearance; T last represents the time point at which the last quantifiable drug concentration can be measured; AUC 0-last It represents the area under the plasma concentration-time curve from time 0 to the last quantifiable time point.

[1013] Table 6. Brain penetration data of the compounds of the present invention in mice

[1014] Experimental conclusion: The compound of the present invention has excellent metabolic stability in vivo and excellent oral absorption drug exposure; oral administration has excellent brain tissue drug concentration and high brain-blood ratio.

[1015] Experimental Example 6: Permeability Evaluation of the Compounds of the Invention

[1016] Cell lines:

[1017] In this study, the MDR1-MDCK II cell line, which was authorized by the laboratory of Piet Borst, Netherlands Cancer Institute, was used as an in vitro model for permeability evaluation. 5 cells / cm 2 The cells were seeded into Transwell-96 well cell plates and cultured in a carbon dioxide incubator for 4-7 days before use in transport experiments.

[1018] Experimental conditions:

[1019] Test sample concentration: 2.00 μM

[1020] Test direction and number of parallel samples: bidirectional AB and BA, 2 parallel

[1021] Transport buffer (TB): HBSS solution containing 10 mM HEPES (pH 7.40 ± 0.05)

[1022] Incubation conditions: 37 ± 1 ° C, 5% CO2, incubate for 150 minutes

[1023] Control compounds: Nadolol and metoprolol were used as low-osmotic and high-osmotic control compounds, respectively, and digoxin was used as a substrate for P-glycoprotein. Nadolol and metoprolol were administered at a concentration of 2.00 μM, and digoxin was administered at a concentration of 10.0 μM.

[1024] Monolayer cell membrane integrity test:

[1025] After the transport experiment, the integrity of the MDR1-MDCK II cell layer was assessed using the Lucifer Yellow Rejection Assay. The remaining solution in the apical and basolateral wells was removed, and 75 μL of TB containing 100 μM Lucifer Yellow was added to the apical well and 250 μL of TB was added to the basolateral well. The cell plate was incubated in a cell culture incubator at 37 ± 1°C, 5% CO2, and saturated humidity for 30 minutes. A 20 μL sample was then taken from the apical well and mixed with 60 μL of TB. An 80 μL sample was taken from the basolateral well and the relative fluorescence intensity (RFU) was measured using a microplate reader at 425 / 528 nm (excitation / emission).

[1026] Sample analysis:

[1027] In this study, sample analysis of the test and reference compounds, nadolol, metoprolol, and digoxin, was performed using liquid chromatography-tandem mass spectrometry (LC-MS / MS). Retention times of the analytes and internal standards, chromatogram acquisition, and chromatogram integration were performed using Analyst software (Sciex, Framingham, MA, USA). Semiquantitative analysis was performed using the comparison of analyte peak areas with the internal standard peak area.

[1028] Data Analysis:

[1029] V R is the volume of the receiving solution (0.075 mL for surface A and 0.25 mL for surface B); Area is the relative surface area of ​​the cell monolayer (0.0804 cm2); Time is the incubation time (9000 s); C0 is the peak area ratio of the compound at the dosing end; V D is the volume of the dosing end (0.075 mL for side A and 0.25 mL for side B); C D and C R are the peak area ratios of the compounds at the dosing end and the receiving end, respectively.

[1030] The %Lucifer Yellow transmittance is calculated using the following formula:

[1031] RFU Apical and RFU Basolateral are the relative fluorescence intensities of fluorescein at the top and bottom, respectively. Apical With V Basolateral The loading volumes are apical and basolateral (0.0750 and 0.250 mL, respectively).

[1032] Experimental results:

[1033] The results of the permeability test of the compounds of the present invention on the MDR1-MDCK II cell line are shown in Table 7.

[1034] Table 7 Summary of permeability evaluation of inventive compounds Note: Low permeability: P app ≤1.0(×10 -6 cm / s); medium permeability 1.0 <P app <5.5(×10 -6 cm / s); high permeability: P app ≥5.5 (×10 -6 cm / s).

[1035] Experimental conclusion: The compound of the present invention exhibits excellent membrane permeability in the cell membrane permeability study.

[1036] Experimental Example 7: Pharmacokinetic Evaluation and Brain Penetration Evaluation of the Compounds of the Invention in Rats

[1037] Experimental methods:

[1038] A 0.2 mg / mL clear solution of the test compound in 90% 2-hydroxypropyl-β-cyclodextrin was injected into the tail vein of male SD rats (200-230 g) overnight fasted at a dose of 1 mg / kg. A 0.5 mg / mL solution of the test compound in 90% 2-hydroxypropyl-β-cyclodextrin was administered orally to male SD rats (200-230 g) overnight fasted at a dose of 5 mg / kg. Blood was collected from the jugular vein and the tail vein at 0.0833, 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, and 24 hours after administration, respectively, and approximately 30 μL of blood was collected from the animals in EDTA-K2-containing anticoagulant tubes. Plasma was separated by centrifugation. Compound 15 trifluoroacetate was administered intravenously (IV, 1 mg / kg) in one group (blood was collected at the same time points as above) and orally (PO, 5 mg / kg) in one group (blood was collected at the same time points as above). In addition, plasma and brain homogenate were collected at 2 hours after oral administration (PO, 5 mg / kg). The blood drug concentration was determined by LC-MS / MS. WinNonlin TM Version 6.3 (Pharsight, Mountain View, CA) pharmacokinetic software was used to calculate relevant pharmacokinetic parameters using the non-compartmental linear logarithmic trapezoidal method.

[1039] Experimental results: The pharmacokinetic data of rats are shown in Table 8, and the brain penetration data of rats are shown in Table 9:

[1040] Table 8. Pharmacokinetic data in rats Note: “-” indicates that the parameter cannot be obtained by calculation; C0 represents the initial concentration; C max represents the peak concentration; T max T stands for time to peak 1 / 2 Vd stands for elimination half-life; ss represents the apparent volume of distribution at steady state; Cl represents the total clearance; T last represents the time point at which the last quantifiable drug concentration can be measured; AUC 0-last represents the area under the plasma concentration-time curve from time 0 to the last quantifiable time point; F (%) represents bioavailability, using AUC 0-last calculate.

[1041] Table 9. Brain penetration data of the compounds of the present invention in rats

[1042] Experimental conclusion: The compound of the present invention has excellent metabolic stability in vivo and excellent oral absorption drug exposure; oral administration has excellent brain tissue drug concentration and high brain-blood ratio.

[1043] Experimental Example 8: In vitro liver microsomal stability of the compound of the present invention

[1044] Experimental purpose: To investigate the metabolic stability of the invented compound in vitro through the metabolic stability of human liver microsomes.

[1045] Experimental conditions: 1 μM of the compound was incubated with human liver microsomes supplemented with NADPH regeneration system at 37°C for a certain period of time up to 60 minutes. The concentration of the compound in the resulting sample was determined by LC-MS / MS.

[1046] Experimental Procedure: 96-well incubation plates were designated T0, T5, T15, T30, T45, T60, Blank60, and NCF60. The corresponding reaction time points for each plate were 0, 5, 15, 30, 45, and 60 minutes. Blank60 plates were not incubated with test or control compounds, and samples were taken after 60 minutes of incubation. NCF60 plates were incubated for 60 minutes with potassium phosphate buffer instead of NADPH regeneration system solution. Samples were collected from a single well at all time points.

[1047] Add 5 μL of test or control working solution and 100 μL of microsomal working solution (liver microsomal protein concentration is 0.5 mg / mL) to the T0, T5, T15, T30, T45, T60 and NCF60 plates, respectively. Add only microsomal working solution to the Blank60 plate. Then, place the incubation plates Blank60, T5, T15, T30, T45 and T60 except T0 and NCF60 in a 37°C water bath for preincubation for approximately 10 minutes.

[1048] T0 plate samples were first added with 180 μL of stop solution (the stop solution was an acetonitrile:methanol (95:5, V / V) solution containing 100 ng / mL tolbutamide) and then the NADPH regeneration system working solution was added.

[1049] Add 50 μL potassium phosphate buffer to each well of the NCF60 plate and incubate for 60 minutes.

[1050] After preincubation of Blank60, T5, T15, T30, T45, and T60 plates, 44 μL of NADPH regeneration system working solution was added to each well to initiate the reaction. Therefore, for wells containing the test or control working solution, the final reaction concentration was 1 μM, the concentration of liver microsomes was 0.5 mg / mL, and the final concentrations of DMSO and acetonitrile in the reaction system were 0.01% (v / v) and 0.99% (v / v), respectively.

[1051] After incubation for an appropriate time (e.g., 5, 15, 30, 45, and 60 minutes), add 180 μL of stop solution to each test sample well and control sample well on Blank60, T5, T15, T30, T45, T60, and NCF60 plates, respectively, to terminate the reaction.

[1052] All sample plates were shaken and centrifuged for 10 minutes, and 80 μL of the test sample supernatant was diluted into 240 μL of acetonitrile: water (1:9, V / V) solution containing 0.1% formic acid for LC-MS / MS analysis.

[1053] The half-life of the compound in human liver microsome metabolism was obtained by calculating the corresponding time points and the remaining percentage of the compound. 1 / 2 and clearance CL int(liver) .

[1054] Experimental results: as shown in Table 10.

[1055] Table 10 Results of in vitro metabolic stability studies of compounds of the present invention in human or mouse liver microsomes

[1056] NA: Not tested

[1057] Conclusion: The compounds of the present invention exhibit excellent stability in human liver microsomes and mouse liver microsomes in vitro.

[1058] Experimental Example 9: In vivo pharmacodynamic study of the compounds of the present invention on a BALB / c Nude mouse model of subcutaneous xenograft tumors of human breast cancer MDA-MB-436 cells

[1059] Purpose of the experiment:

[1060] The inhibitory effect of the compound of the present invention on the growth of human breast cancer MDA-MB-436 nude mouse transplanted tumor model in vivo was studied.

[1061] Experimental methods:

[1062] 1) Cell culture:

[1063] Conventional cell culture was performed in 5% CO2, 37°C and MEM medium containing 10% fetal bovine serum; the cells were digested and passaged with 0.25% trypsin; depending on cell growth, the cells were passaged 2 to 3 times per week with a passage ratio of 1:3 to 1:6.

[1064] 2) Animal model preparation

[1065] Animals: Female Balb / c nude mice, 6-8 weeks old.

[1066] MDA-MB-436 cells in the logarithmic growth phase were collected, counted, and resuspended in 50% serum-free MEM medium and 50% matrigel to adjust the cell concentration to 5.0×10 7 cells / mL; pipette the cells to disperse them evenly and then put them into a 50mL centrifuge tube, which was placed in an ice box; use a 1mL syringe to draw up the cell suspension and inject it into the subcutaneous part of the right axilla of the nude mice. Each animal was inoculated with 200μL (1.1×10 7 Cells / mouse) were used to establish an MDA-MB-436 nude mouse xenograft tumor model. After inoculation, the animal status and tumor growth were regularly observed. The tumor diameter was measured using an electronic vernier caliper, and the data were directly entered into an Excel spreadsheet to calculate the tumor volume. When the tumor volume reached 100-200 mm 3 Animals in good health and with similar tumor volumes were selected and randomly divided into groups (n=6) based on tumor volume, while ensuring that the average body weight of each group remained consistent. The day of grouping was designated as the first day of the experiment (D1). After the start of the experiment, tumor diameters were measured twice weekly, and tumor volumes were calculated. Animal body weights were also recorded.

[1067] The formula for calculating tumor volume (TV) is as follows: TV (mm 3)=a×w 2 / 2; where a represents the long diameter of the tumor (mm); w represents the short diameter of the tumor (mm).

[1068] 3) Dosage: 1.0 mg / kg; Administration: Oral administration (po), once daily (QD)

[1069] 4) Data Recording

[1070] The calculation formula of relative tumor volume (RTV) is: RTV=TVt / TVinitial; where TVinitial is the tumor volume measured at the time of group administration; TVt is the tumor volume measured at each time during the administration period.

[1071] The relative tumor proliferation rate (%T / C) was calculated as follows: %T / C = 100% × (RTVT / RTVC); where RTVT represents the RTV of the treatment group; and RTVC represents the RTV of the solvent control group.

[1072] The calculation formula for tumor growth inhibition rate (TGI) is: TGI = 100% × [1-(TVt(T)-TVinitial(T)) / (TVt(C)-TVinitial(C))]; where TVt(T) represents the tumor volume of the treatment group measured at each time; TVinitial(T) represents the tumor volume of the treatment group when the drugs were administered in groups; TVt(C) represents the tumor volume of the vehicle group measured at each time; TVinitial(C) represents the tumor volume of the vehicle group when the drugs were administered in groups.

[1073] Experimental results: The tumor growth volume is shown in FIG1 , the changes in mouse body weight during administration are shown in FIG2 , and the tumor growth inhibition rate and relative tumor proliferation rate are shown in Table 11 .

[1074] Table 11 Summary of MDA-MB-436 tumor growth inhibition rate and relative tumor proliferation rate 1 The comparison between the two groups was analyzed using T-test, and p < 0.05 was considered to be a significant difference, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

[1075] Experimental conclusion: The compound of the present invention exhibits significant anti-tumor activity.

Claims

1. A compound represented by formula (XII) or a pharmaceutically acceptable salt thereof, in, X is selected from O and S; Structural unit Selected from Ring A is selected from phenyl and 6-membered heteroaryl; Ring B is selected from 6-membered heteroaryl; L is selected from a single bond, and ring C is selected from is selected from a single bond or a double bond; T2 is selected from C, N and CH; T3 and T4 are independently selected from N and CR 10 ; R1 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens; R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens; R3 is selected from H and halogen; R9 is absent or selected from H and halogen; R4 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy; R5 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy; R6 and R7 are independently selected from H, halogen, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens; R8 is selected from C 1-3 Alkyl and C 3-5 Cycloalkyl, the C 1-3 Alkyl and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 R groups, each independently; R 10 is selected from H and halogen; R 13 and R 14 are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 The cycloalkyl groups are each independently optionally substituted with 1, 2 or 3 halogens; R 15 and R 16 are independently selected from H, D, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 halogens; Alternatively, R2 and R4 form a ring, so that the structural unit Selected from Alternatively, R3 and R5 form a ring, so that the structural unit Selected from Each R a are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens; Alternatively, two R on adjacent atoms a Together with the atoms to which they are attached, they form a double bond or a cyclopropyl group; Each R b are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens; Each R c are independently selected from H, halogen, C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl, the C 1-3 Alkyl, C 1-3 Alkoxy and C 3-5 Cycloalkyl is optionally substituted with 1, 2 or 3 halogens; Each R is independently selected from halogen and D; n is selected from 0, 1, 2, 3 and 4; The condition is that when the structural unit Selected from L is selected from a single bond, and ring C is selected from When R2 and R4 form a ring or R3 and R5 form a ring; The halogen represents F, Cl, Br and I atoms; The "hetero" in the 5-6 membered heteroaryl group represents 1, 2, 3 or 4 heteroatoms or heteroatom groups independently selected from -O-, -S- and -N-.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure shown in formula (XII-1), in, Ring A is selected from 6-membered heteroaryl groups; R2, R3, R4, R5, R6, R7, R8, R9, R 13 , R 14 , R 15 , R 16 , R c , X, T2, T3, T4, n and As defined in claim 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein X is selected from O.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Each R a , each R b and each R c are independently selected from H, F and CH3.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein T3 is selected from N, CH and CF; alternatively, T3 is selected from N.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein T4 is selected from N and CH.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R1 is selected from CH3 and CH2CH3; alternatively, R1 is selected from CH3.

8. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R2 is selected from H and CH3.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R6 and R7 are independently selected from H, F and CH3; or, R6 and R7 are independently selected from H and F.

10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R4 and R5 are each independently selected from H, F, Cl, CN and CH3.

11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R8 is selected from CH3, CH2CH3, CH2CF3, cyclopropyl and CD3.

12. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R3 and R9 are independently selected from H and F.

13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 10 Selected from H and F.

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 13 and R 14 are independently selected from H and CH3.

15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: R 15 and R 16 are independently selected from H, D and CH3.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Ring A is selected from phenyl, pyridyl, pyrazinyl and pyrimidinyl.

17. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Ring B is selected from pyridyl, pyrazinyl and pyrimidinyl.

18. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: Structural unit 19. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 20. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 21. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: L is selected from a single bond, a structural unit Selected from 22. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (P-1): in, Ring A is selected from pyridyl; R2, R3, R4, R5, R7, R8, R9, R 13 、R 14 、R 15 、R 16 、R c , T2, n and As defined in claim 1.

23. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (P-2): in, is selected from a single bond or a double bond; T2 is selected from C and N; R2 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 halogens; R3 is selected from H and halogen; R4 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy; R5 is selected from H, halogen, CN, C 1-3 Alkyl and C 1-3 alkoxy; R9 is absent or selected from H and halogen; R7, R8, R 13 、R 14 、R 15 、R 16 、R c and n as defined in claim 1.

24. The following compound or a pharmaceutically acceptable salt thereof:

25. The compound according to claim 24 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the group consisting of:

26. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to any one of claims 1 to 25 or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier, diluent or excipient.

27. Use of the compound according to any one of claims 1 to 25 or a pharmaceutically acceptable salt thereof, or the composition according to claim 26, in the preparation of a drug for treating solid tumors.

28. The use according to claim 27, wherein the solid tumor refers to solid tumors such as ovarian cancer, breast cancer, prostate cancer and brain glioma.

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