Compounds as wrn helicase inhibitors

Abstract

The application discloses a compound shown in formula (I) and a pharmaceutically acceptable salt thereof, and particularly discloses a compound as a WRN helicase inhibitor.

Description

[0001] The present application claims priority to the following:

[0002] Application No. CN2023103018345, Application Date: March 24, 2023;

[0003] Application No. CN202311004828X, Application Date: August 9, 2023;

[0004] Application No. CN2023110500915, Application Date: August 18, 2023;

[0005] Application No. CN2024102950619, Application Date: March 14, 2024. TECHNICAL FIELD

[0006] The present application relates to compounds represented by formula (I) and pharmaceutically acceptable salts thereof, in particular, the present application relates to a kind of as WRN helicase inhibitor compound. BACKGROUND

[0007] There are many short tandem repeat regions in the human genome, these repeat DNA regions are called "microsatellites", which are prone to errors during replication and thus rely heavily on the MMR (mismatch repair) system for repair. When the MMR (mismatch repair) system is abnormal, causing dMMR (deficient mismatch repair), it cannot identify and repair microsatellite replication errors, causing MSI, which may lead to frameshift mutations, thereby causing abnormalities in tumor-related genes, and further inducing the occurrence and development of cancer. In 2017, the Immune Checkpoint Inhibitor (ICI) was approved for the treatment of high microsatellite instability / mismatch repair deficiency (Microsatellite Instability-High / Deficient Mismatch Repair, MSI-H / dMMR) tumors, and MSI-H / dMMR has become the first "pan-tumor" tumor marker. Microsatellite instability-high (MSI-H) cancer cells depend on WRN (Werner syndrome RecQ helicase) helicase activity. Inhibition of WRN can induce DNA double-strand breaks, activate DNA damage response, and induce apoptosis and cell cycle arrest. The usual treatment for dMMR (deficient mismatch repair) / MSI-H cancer patients includes targeted therapy, chemotherapy, and immunotherapy. The clinical effectiveness of targeted therapy and chemotherapy is limited by drug resistance and drug toxicity, and about half of the patients treated with immunotherapy do not respond positively to immune checkpoint inhibitors. 45-60% of MSI-H cancer patients do not respond to immunotherapy, and the primary and secondary drug resistance of targeted therapy, chemotherapy, and immunotherapy needs to be addressed. In 2019, the Broad Institute of Harvard and MIT analyzed the Achilles and drive databases to assess the degree of dependence of each cell line on different targets, and they found that the activity of RecQ DNA helicase WRN is essential for dMMR / MSI-H cell lines in vivo and in vitro, while MSS cells do not depend on WRN for survival. In the MSI-H model, knocking out WRN induces double-strand DNA breaks and selectively promotes apoptosis and cell cycle arrest. This anticancer mechanism is different from targeted drugs (inhibiting cancer cell-specific oncogenic changes) and immunotherapy (inhibiting immune escape and tolerance). In 2021, Dr. Mathew J Garnett's team at the Wellcome Sanger Institute used PDX models to prove that WRN inhibitors can be used as second-line or third-line monotherapy for dMMR patients. In dMMR tumors, tumor mutability is negatively correlated with immune checkpoint blockade response, while WRN sensitivity is not related to mutation load. Due to the different modes of action, combination therapy with checkpoint inhibitors, chemotherapy, or targeted therapy and WRN inhibitors may inhibit cross-resistance and promote tumor eradication.In addition, because the absence of DNA repair modulates the structure of neoantigens, increases the mutational burden, and leads to enhanced immune response, WRN inhibition can also synergize with immunotherapy. Therefore, WRN can serve as a key target for single treatment or combination targeting drugs, chemotherapy or immunotherapy of dMMR / MSI-H tumors.

[0008] MSI-H tumors can occur in multiple sites, among which the incidence in endometrial cancer (31%), colon adenocarcinoma (20%) and gastric cancer (19%) is the highest, and there are about 325,000 new MSI-H tumor patients in the United States and about 300,000 in China every year, and the development of drugs has great market value. In tumors with normal mismatch repair (MSS), the absence (or reduction) of WRN expression does not affect the growth of tumor cells. In tumors with mismatch repair deficiency (MSI-H), if accompanied by the absence (or reduction) of WRN expression, it can cause an increase in the accumulation of cellular DNA double-strand breaks, cell cycle arrest at G1 or G2 / M phase, thereby leading to tumor cell death. This is called synthetic lethal effect. The development of WRN helicase inhibitors is expected to become one of the effective treatment methods for MSI-H cancer. SUMMARY

[0009] In one aspect of the present application, the present application provides a compound represented by formula (I), an optical isomer thereof, or a pharmaceutically acceptable salt thereof,

[0010]

[0011] wherein,

[0012] Ring A is selected from phenyl or 5-10 membered heteroaryl, the phenyl and 5-10 membered heteroaryl being optionally substituted by 1, 2 or 3 R a substituents;

[0013] Ring B is selected from C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 4-20 cycloalkenyl, 4-20 membered heterocycloalkenyl, C 6-20 aryl or 5-20 membered heteroaryl;

[0014] Ring C is selected from C 6-20 aryl or 5-20 membered heteroaryl;

[0015] Ring D is selected from C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, 4-20 membered heterocycloalkenyl, C 6-20 aryl or 5-20 membered heteroaryl;

[0016] Ring E is selected from C 3-20 cycloalkyl, 5-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl;

[0017] L1is selected from a single bond, -N(R b1 )-, -N(R b1 )C(=O)-, -O-, -S-, -(CR b2 R b3 ) t -, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2-, or

[0018] L2is selected from a single bond, -N(R b1 )-, -N(R b1 )C(=O)-, -O-, -S-, -(CR b2 R b3 ) t -, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2-, or

[0019] R1is independently at each occurrence selected from H, F, Cl, Br, OH, N(R b4 )2, CN, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl, or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl, and 5-20 membered heteroaryl being optionally substituted with 1, 2, or 3 R;

[0020] or, two R1are joined together to form a C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl, or 5-20 membered heteroaryl;

[0021] is selected from is selected from

[0022] or, is selected from is selected from

[0023] when is selected from T is selected from C;

[0024] when is selected from T is selected from N or CH;

[0025] when Selected from When R2 is selected from H, F, Cl, Br, OH, N(R b4 )2, CN, SF5, CHO, COOH, C 1-20 Alkyl, C 1-20 Heteroalkyl, C 3-20 Cycloalkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, 3-20 membered heterocycloalkyl, C 4-20 Cycloalkenyl, 4-20 membered heterocycloalkenyl, C 6-20 Aryl or 5-20 membered heteroaryl, the C 1-20 Alkyl, C 1-20 Heteroalkyl, C 3-20 Cycloalkyl, C 2-20 Alkenyl, C 2-20 Alkynyl, 3-20 membered heterocycloalkyl, C 4-20 Cycloalkenyl, 4-20 membered heterocycloalkenyl, C 6-20 Aryl and 5-20 membered heteroaryl are optionally substituted with 1, 2 or 3 R;

[0026] when Selected from When, R2 is selected from O or S;

[0027] R3 and R4 are independently selected from H, F, Cl, Br, OH, N (R b4 )2.CN.C 1-20 Alkyl, C 1-20 Heteroalkyl, C 3-20 Cycloalkyl or 3-20 membered heterocycloalkyl, the C 1-20 Alkyl, C 1-20 Heteroalkyl, C 3-20 Cycloalkyl and 3-20 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0028] Alternatively, R3 and R4 are connected together to form a C 3-20 Cycloalkyl or 3-20 membered heterocycloalkyl, the C 3-20 Cycloalkyl or 3-20 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R;

[0029] R5 are independently selected from H, F, Cl, Br, OH, N (R b4 )2、SF5、CN、CHO、C 1-20 Alkyl, C 1-20 Heteroalkyl, C 3-20 Cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 Aryl or 5-20 membered heteroaryl, the C 1-20alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl, optionally substituted with 1, 2, or 3 R;

[0030] or, two R5are joined together to form a C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 4-20 cycloalkenyl, 4-20 membered heterocycloalkenyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 4-20 cycloalkenyl, 4-20 membered heterocycloalkenyl, C 6-20 aryl and 5-20 membered heteroaryl, optionally substituted with 1, 2, or 3 R;

[0031] R6is independently selected from H, F, Cl, Br, OH, N(R b4 )2, CN, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl, optionally substituted with 1, 2, or 3 R;

[0032] or, two R6are joined together to form a C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl, optionally substituted with 1, 2, or 3 R;

[0033] R7is independently selected from H, F, Cl, Br, OH, N(R b4 )2, CN, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl, optionally substituted with 1, 2, or 3 R;

[0034] R8is selected from H, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl or 3-20 membered heterocycloalkyl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl and 3-20 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0035] R a are each independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl or 3-20 membered heterocycloalkyl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl and 3-20 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0036] R b1 is selected from H, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl are optionally substituted with 1, 2, or 3 R;

[0037] R b2 , R b3 are each independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl or 3-20 membered heterocycloalkyl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl and 3-20 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0038] Alternatively, R b2 and R b3 are joined together to form a C 3-20 cycloalkyl or 3-20 membered heterocycloalkyl;

[0039] R b4 are each independently selected from H, C 1-20 alkyl, C1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl are optionally substituted with 1, 2, or 3 R;

[0040] m, n, p, q, t are each independently selected from 0, 1, 2, or 3;

[0041] R is each independently selected from H, F, Cl, Br, I, OH, NH2, CN, SF5, CHO, COOH, C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl or 5-20 membered heteroaryl, said C 1-20 alkyl, C 1-20 heteroalkyl, C 3-20 cycloalkyl, 3-20 membered heterocycloalkyl, C 6-20 aryl and 5-20 membered heteroaryl are optionally substituted with 1, 2, or 3 R’;

[0042] R’ is selected from H, F, Cl, Br, I, OH, NH2, CH3, CF3, C2H5, CN, SF5, CHO, COOH, or

[0043] the above C 1-6 heteroalkyl, C 1-20 heteroalkyl, 3-20 membered heterocycloalkyl, 4-20 membered heterocycloalkenyl, 5-20 membered heteroaryl, or 5-10 membered heteroaryl comprises 1, 2, or 3 heteroatoms or groups of heteroatoms independently selected from -O-, -NH-, -S-, -C(=O)-, -C(=O)O-, -S(=O)-, -S(=O)2-, and N.

[0044] In some embodiments of the application, wherein ring A is selected from phenyl or 5-6 membered heteroaryl, said phenyl and 5-6 membered heteroaryl are optionally substituted with 1, 2, or 3 R a substituents;

[0045] ring B is selected from C 3-6 cycloalkyl, 5-6 membered heterocycloalkyl, C 5-6 cycloalkenyl, 5-6 membered heterocycloalkenyl, C 6-10 aryl or 5-10 membered heteroaryl;

[0046] ring C is selected from C6-10 aryl or 5-10 membered heteroaryl;

[0047] Ring D is selected from C 3-6 cycloalkyl, 5-6 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl, C 6-10 aryl or 5-10 membered heteroaryl;

[0048] Ring E is selected from C 3-6 cycloalkyl, 5-6 membered heterocycloalkyl, C 6-10 aryl or 5-10 membered heteroaryl;

[0049] L1is selected from a single bond, -N(R b1 )-, -O-, -S-, -C(R b2 )2-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2-, or

[0050] L2is selected from a single bond, -N(R b1 )-, -O-, -S-, -C(R b2 )2-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2-, or

[0051] R a are each independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0052] R b1 is selected from H, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0053] R b2 are each independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl or 3-6 membered heterocycloalkyl, said C 1-6 alkyl, C 1-6heteroalkyl, C 3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0054] R1is independently selected from the group consisting of H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl or 3- to 6-membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0055] or, two R1are linked together to form a C 4-6 cycloalkyl or 4- to 6-membered heterocycloalkyl;

[0056] selected from the group consisting of selected from the group consisting of

[0057] or, selected from the group consisting of selected from the group consisting of

[0058] when selected from the group consisting of T is selected from C;

[0059] when selected from the group consisting of T is selected from N or CH;

[0060] when selected from the group consisting of R2is selected from the group consisting of H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl or 3- to 6-membered heterocycloalkyl, said C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl and 3- to 6-membered heterocycloalkyl are optionally substituted with 1, 2, or 3 R;

[0061] when selected from the group consisting of R2is selected from O or S;

[0062] R3, R4are independently selected from the group consisting of H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6Cycloalkyl or 3-6 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0063] Alternatively, R3 and R4 are connected together to form a C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R;

[0064] R5 is independently selected from H, F, Cl, Br, OH, NH2, SF5, CN, HC(=O)-, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0065] Alternatively, two R5s are connected together to form a C 4-6 Cycloalkyl or 4-6 membered heterocycloalkyl, the C 4-6 Cycloalkyl and 4-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0066] R6 are independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0067] Alternatively, two R6s are connected together to form a C 4-6 Cycloalkyl or 4-6 membered heterocycloalkyl, the C 4-6 Cycloalkyl and 4-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0068] R7 are independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6Cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0069] R8 is selected from H, C 1-6 Alkyl, C 3-6 Cycloalkyl or 3-6 membered heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Heteroalkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

[0070] m, n, p, q are each independently selected from 0, 1, 2 or 3;

[0071] R is independently selected from H, F, Cl, Br, I, OH, NH2, CN, COOH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio and C 1-6 Alkylamino, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio or C 1-6 Alkylamino is optionally substituted with 1, 2 or 3 R';

[0072] R' is selected from F, Cl, Br, I, OH, NH2 and CH3;

[0073] The above C 1-6 The heteroalkyl, 5-6 membered heterocycloalkyl, 5-6 membered heterocycloalkenyl, 5-6 membered heteroaryl or 5-10 membered heteroaryl group contains 1, 2 or 3 heteroatoms or heteroatom groups independently selected from -O-, -NH-, -S-, -C(=O)-, -C(=O)O-, -S(=O)-, -S(=O)2- and N.

[0074] In some embodiments of the present invention, R is independently selected from H, F, Cl, Br, I, OH, NH2, CN, SF5, CHO, COOH, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 1-6 Alkylamino, -C 1-6 Alkyl-C 1-6 Alkoxy, -C 1-6 Alkyl-C 1-6 Alkylthio, -C 1-6 Alkyl-C 1-6 Alkylamino, C 1-6 Alkyl-OH, C 1-6 Alkyl-NH2, -C(=O)-C 1-6 Alkyl, C1-6 alkyl-C(=O)-C 1-6 alkyl, -NH-C(=O)-C 1-6 alkyl, C 1-6 alkyl-NH-C(=O)-C 1-6 alkyl, -NH-S(=O)2-C 1-6 alkyl, C 1-6 alkyl-NH-S(=O)2-C 1-6 alkyl, C 3-6 cycloalkyl, phenyl, naphthyl, pyridyl, pyrimidyl, pyridazyl, pyrazyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, piperidyl, piperazinyl, pyranyl, furanyl, thiazolyl, oxazolyl or thiopyranyl,

[0075] the C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino, -C 1-6 alkyl-C 1-6 alkoxy, -C 1-6 alkyl-C 1-6 alkylthio, -C 1-6 alkyl-C 1-6 alkylamino, C 1-6 alkyl-OH, C 1-6 alkyl-NH2, -C(=O)-C 1-6 alkyl, C 1-6 alkyl-C(=O)-C 1-6 alkyl, -NH-C(=O)-C 1-6 alkyl, C 1-6 alkyl-NH-C(=O)-C 1-6 alkyl, -NH-S(=O)2-C 1-6 alkyl, C 1-6 alkyl-NH-S(=O)2-C 1-6 alkyl, C 3-6 cycloalkyl, phenyl, naphthyl, pyridyl, pyrimidyl, pyridazyl, pyrazyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, piperidyl, piperazinyl, pyranyl, furanyl, thiazolyl, oxazolyl and thiopyranyl are optionally substituted with 1, 2 or 3 R', the other variables being as defined in the application.

[0076] In some aspects of the application, R is independently selected from the group consisting of H, F, Cl, Br, I, OH, NH2, CN, SF5, CHO, COOH, CH3, CF3, CHF2, CH2F, CF2Cl, CF2Br, CF2I, the other variables being as defined in the application.

[0077] In some embodiments of the application, R is selected from H, F, Cl, Br, OH, NH2, COOH, Me, CF3, CHF2, CH2F, the other variables are as defined in the application.

[0078] In some embodiments of the application, ring A is selected from phenyl, pyridyl, pyridazinyl, pyrimidinyl, thienyl, thiazolyl, oxazolyl, isoxazolyl, 1H-imidazolyl, 1H-pyrazolyl or 1H-pyrrolyl, optionally substituted with 1, 2 or 3 R a the other variables are as defined in the application.

[0079] In some embodiments of the application, ring A is selected from the other variables are as defined in the application.

[0080] In some embodiments of the application, R a are each independently selected from H, F, Cl, Br, OH, NH2, CN, Me, the Me, is optionally substituted with 1, 2 or 3 R, the other variables are as defined in the application.

[0081] In some embodiments of the application, R a are each independently selected from H, F, Cl, Br, OH, NH2, CN, Me, CF3, the other variables are as defined in the application. In some embodiments of the application, ring A is selected from the other variables are as defined in the application.

[0082] In some embodiments of the application, when is selected from R2is selected from H, F, Cl, Br, OH, NH2, CN, SF5, CHO, COOH, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 4-6 cycloalkenyl, 4-6 membered heterocycloalkenyl, C 6-10 aryl or 5-10 membered heteroaryl, the C 1-6 alkyl, C1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, C 4-6 cycloalkenyl, 4-6 membered heterocycloalkenyl, C 6-10 aryl and 5-10 membered heteroaryl are optionally substituted with 1, 2, or 3 R, the other variables being as defined in the application.

[0083] In some embodiments of the application, when is selected from R2is selected from H, F, Cl, Br, OH, NH2, CN, Me, the Me, is optionally substituted with 1, 2, or 3 R, the other variables being as defined in the application.

[0084] In some embodiments of the application, R2is selected from H, F, Cl, Br, OH, NH2, CN, Me, CF3, the other variables being as defined in the application.

[0085] In some embodiments of the application, the structural unit is selected from the other variables being as defined in the application.

[0086] In some embodiments of the application, the structural unit is selected from the other variables being as defined in the application.

[0087] In some embodiments of the application, R1is independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino, -C 1-6 alkyl-C 1-6 alkoxy, -C 1-6 alkyl-C 1-6 alkylthio, -C 1-6 alkyl-C 1-6 alkylamino, C 1-6 alkyl-OH, C 1-6 alkyl-NH2, C 1-6 alkyl-C(=O)-, C1-6 alkyl-C(=0)-C 1-6 alkyl, C 1-6 alkyl-O-C(=0)-, C 1-6 alkyl-O-C(=0)-C 1-6 alkyl, -NH-C(=0)-C 1-6 alkyl, C 1-6 alkyl-NH-C(=0)-C 1-6 alkyl, C 1-6 alkyl-S(=0)2-, C 1-6 alkyl-S(=0)2-C 1-6 alkyl, -NH-S(=0)2-C 1-6 alkyl, C 1-6 alkyl-NH-S(=0)2-C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 cycloalkyl-S(=0)2- or 3-6 membered heterocycloalkyl,

[0088] the C 1-6 alkyl, C 1-6 alkoxy, C 1-6 alkylthio, C 1-6 alkylamino, -C 1-6 alkyl-C 1-6 alkoxy, -C 1-6 alkyl-C 1-6 alkylthio, -C 1-6 alkyl-C 1-6 alkylamino, C 1-6 alkyl-OH, C 1-6 alkyl-NH2, C 1-6 alkyl-C(=0)-, C 1-6 alkyl-C(=0)-C 1-6 alkyl, C 1-6 alkyl-O-C(=0)-, C 1-6 alkyl-O-C(=0)-C 1-6 alkyl, -NH-C(=0)-C 1-6 alkyl, C 1-6 alkyl-NH-C(=0)-C 1-6 alkyl, C 1-6 alkyl-S(=0)2-, C 1-6 alkyl-S(=0)2-C 1-6 alkyl, -NH-S(=0)2-C 1-6 alkyl, C 1-6 alkyl-NH-S(=0)2-C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6Cycloalkyl-S(=O)2- and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R, and the other variables are as defined herein.

[0089] In some embodiments of the present invention, R1 is independently selected from H, F, Cl, Br, OH, NH2, CN, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 1-3 Alkylamino, C 1-3 Alkyl-C(=O)-, C 1-3 Alkyl-OC(=O)-, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl or C 3-6 Cycloalkyl-S(=O)2-, the C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 1-3 Alkylamino, C 1-3 Alkyl-C(=O)-, C 1-3 Alkyl-OC(=O)-, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl and C 3-6 Cycloalkyl-S(=O)2- is optionally substituted with 1, 2 or 3 R, and the other variables are as defined herein.

[0090] In some embodiments of the present invention, R1 is independently selected from H, F, Cl, Br, OH, NH2, CN, Me, Other variables are as defined in the present invention.

[0091] In some embodiments of the present invention, R1 is independently selected from H, F, Cl, Br, OH, NH2, CN, Me, Other variables are as defined in the present invention.

[0092] In some embodiments of the present invention, ring B is selected from cyclohexyl, 3,6-dihydro-2H-pyranyl, 3,4-dihydro-2H-pyranyl, morpholinyl, cyclohexenyl, piperidinyl, 2,3-dihydro-1,4-dioxinyl, 6-oxa-3-azabicyclo[3.1.1]heptanyl, 1,2,3,4-tetrahydropyridinyl, 1,2,3,6-tetrahydropyridinyl, tetrahydro-2H-pyranyl, 5, 6-dihydro-2H-pyran-2-onyl, phenyl, pyridinyl, pyrrolidinyl, 2-oxa-6-aza-spiro[3,3]heptanyl, 1,1-dioxo-3,6-dihydro-2H-thiopyranyl, oxepinyl, azetidinyl, 2-oxa-7-azaspiro[4.4]nonanyl or hexahydro-1H-furo[3,4-c]pyrrolyl, and the other variables are as defined herein.

[0093] In some embodiments of the present invention, Ring B is selected from cyclohexyl, 3,6-dihydro-2H-pyranyl, 3,4-dihydro-2H-pyranyl, morpholinyl, cyclohexenyl, piperidinyl, 2,3-dihydro-1,4-dioxinyl, 6-oxa-3-azabicyclo[3.1.1]heptanyl, 1,2,3,4-tetrahydropyridinyl, 1,2,3,6-tetrahydropyridinyl, tetrahydro-2H-pyranyl, 5,6-dihydro-2H-pyran-2-onyl, phenyl, pyridinyl or pyrrolidinyl, and the other variables are as defined herein.

[0094] In some embodiments of the present invention, the structural unit Selected from: Other variables are as defined in the present invention.

[0095] In some embodiments of the present invention, the structural unit Selected from: Other variables are as defined in the present invention.

[0096] In some embodiments of the present invention, R5 is independently selected from H, F, Cl, Br, OH, NH2, CN, SF5, CHO, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 1-6 Alkylamino, C 3-6 Cycloalkyl or C 3-6 Heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 1-6 Alkylamino, C 3-6 Cycloalkyl and C 3-6 Heterocycloalkyl is optionally substituted with 1, 2 or 3 R;

[0097] Alternatively, two R5s are connected together to form a C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, C 4-10 Cycloalkenyl, 4-10 membered heterocycloalkenyl, C 6-10 Aryl or 5-10 membered heteroaryl, the C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, C 4-10 Cycloalkenyl, 4-10 membered heterocycloalkenyl, C 6-10 Aryl and 5-10 membered heteroaryl are optionally substituted with 1, 2 or 3 R groups, and the other variables are as defined herein.

[0098] In some embodiments of the present invention, R5 is independently selected from H, F, Cl, Br, OH, NH2, CN, SF5, CHO, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 1-6 Alkylamino, C 3-6 Cycloalkyl or C 3-6 Heterocycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 1-6 Alkylamino, C 3-6 Cycloalkyl and C 3-6 Heterocycloalkyl is optionally substituted with 1, 2 or 3 R;

[0099] Alternatively, two R5s are connected together to form a C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, C 4-10 Cycloalkenyl, 4-10 membered heterocycloalkenyl, C 6-10 Aryl or 5-10 membered heteroaryl, the C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, C 4-10 Cycloalkenyl, 4-10 membered heterocycloalkenyl and C 6-10 The aryl group is optionally substituted with 1, 2 or 3 R groups, and the other variables are as defined herein.

[0100] In some embodiments of the present invention, R5 is independently selected from H, F, Cl, Br, OH, NH2, CN, SF5, Me, CHO, The Me, Optionally substituted with 1, 2 or 3 R;

[0101] Alternatively, two R5s are connected together to form described It is optionally substituted with 1, 2 or 3 R, and the other variables are as defined herein.

[0102] In some embodiments of the present invention, R5 is independently selected from H, F, Cl, Br, OH, NH2, CN, SF5, Me, HC(=O)-, The Me, It is optionally substituted with 1, 2 or 3 R, and the other variables are as defined herein.

[0103] In some embodiments of the application, R5is independently selected from H, F, CI, Br, OH, NH2, SF5, CN, Me, CF3, HC(=0)-,

[0104] Alternatively, two R5are joined together to form The other variables are as defined in the application.

[0105] In some embodiments of the application, R5is independently selected from H, F, CI, Br, OH, NH2, SF5, CN, Me, CF3, HC(=0)-, The other variables are as defined in the application.

[0106] In some embodiments of the application, ring C is selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl or thienyl, and the other variables are as defined in the application.

[0107] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0108] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0109] In some embodiments of the application, R6is independently selected from H, F, CI, Br, OH, NH2, CN, Me, said Me, optionally substituted with 1, 2 or 3 R, and the other variables are as defined in the application.

[0110] In some embodiments of the application, R6is independently selected from H, F, CI, Br, OH, NH2, CN, Me, CF3, The other variables are as defined in the application.

[0111] In some embodiments of the application, ring D is selected from

[0112] T1, T2, T3, T4are independently selected from N or CH;

[0113] X1, X2, X3, X4are independently selected from a single bond or CH2;

[0114] X5, X6are independently selected from a single bond, CH2, or CH2CH2, and X5, X6are not simultaneously selected from a single bond;

[0115] X7, X8, X9, X 10 are independently selected from a single bond, NH, O, S, CH2, or X7, X8, X9, X 10 at most 3 are simultaneously selected from a single bond;

[0116] L a is selected from C 1-6 alkyl, C 2-6 alkenyl, or C 1-6 heteroalkyl, said C 1-6 alkyl and C 1-6 heteroalkyl are optionally substituted with 1, 2, or 3 R, the other variables being as defined in the application.

[0117] In some embodiments of the application, ring D is selected from the other variables being as defined in the application.

[0118] In some embodiments of the application, ring D is selected from the other variables being as defined in the application.

[0119] In some embodiments of the application, ring D is selected from piperidinyl, piperazinyl, or 1,2,3,6-tetrahydropyridinyl, the other variables being as defined in the application.

[0120] In some embodiments of the application, structural element is selected from the other variables being as defined in the application.

[0121] In some embodiments of the application, structural element is selected from the other variables being as defined in the application.

[0122] In some embodiments of the application, R7is independently selected from H, F, Cl, Br, OH, NH2, CN, Me, said Me, is optionally substituted with 1, 2, or 3 R, the other variables being as defined in the application.

[0123] In some embodiments of the application, R7is independently selected from H, F, Cl, Br, OH, NH2, CN, Me, CF3, the other variables are as defined in the application.

[0124] In some embodiments of the application, ring E is selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl or thienyl, the other variables being as defined in the application.

[0125] In some embodiments of the application, the structural unit is selected from the other variables are as defined in the application.

[0126] In some embodiments of the application, the structural unit is selected from the other variables are as defined in the application.

[0127] In some embodiments of the application, R3, R4are each independently selected from H, F, Cl, Br, OH, NH2, CN, Me, the Me, optionally substituted with 1, 2 or 3 R, the other variables being as defined in the application.

[0128] In some embodiments of the application, R3, R4are each independently selected from H, F, Cl, Br, OH, NH2, CN, Me, CF3, the other variables are as defined in the application.

[0129] In some embodiments of the application, R8is selected from H, Me, the Me, optionally substituted with 1, 2 or 3 R, the other variables being as defined in the application.

[0130] In some embodiments of the application, R8is selected from H, Me, CF3, the other variables are as defined in the application.

[0131] The application also provides a compound of the formula

[0132]

[0133]

[0134]

[0135]

[0136]

[0137]

[0138]

[0139]

[0140]

[0141]

[0142] In still another aspect of the present application, the present application also provides a pharmaceutical composition. In some embodiments of the present application, the above-mentioned pharmaceutical composition protects the above-mentioned compound, optical isomer thereof or pharmaceutically acceptable salt thereof.

[0143] In some embodiments of the present application, the above-mentioned pharmaceutical composition further comprises a pharmaceutical excipient.

[0144] In still another aspect of the present application, the present application also provides the use of the above-mentioned compound, optical isomer thereof or pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating a tumor-related disease.

[0145] The object of the present application is to provide a compound as a WRN inhibitor or a stereoisomer, deuterated compound, solvate, prodrug, metabolite, pharmaceutically acceptable salt or co-crystal thereof, and intermediates and preparation methods thereof, and the use thereof in the preparation of a medicament for treating a microsatellite instability tumor-related disease.

[0146] In some embodiments of the present application, the tumor-related disease is one or more of the diseases related to solid tumors.

[0147] The compound of the present application can be used alone or in combination with other chemotherapeutic drugs, targeted therapy drugs or immunotherapy drugs for the treatment of various tumors, especially microsatellite instability (MSI) malignant tumors, or mismatch repair deficient (dMMR) malignant tumors, or malignant tumors detected with a large number of (TA) n repeat sequences, including but not limited to colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, etc.

[0148] Definitions and explanations

[0149] 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 being indefinite or unclear if it is not specifically defined, but should be understood according to its ordinary meaning. When a trade name appears in the present text, it is intended to refer to its corresponding product or active ingredient thereof.

[0150] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0151] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds of the present invention with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in neat solution or in 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 neutral form of such compounds with a sufficient amount of acid in solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, trifluoroacetic acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; and salts of amino acids (such as arginine) and organic acids such as glucuronic acid. 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.

[0152] The 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.

[0153] 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.

[0154] Unless otherwise indicated, the term "tautomer" or "tautomer forms" refers to different functional group isomers that are in dynamic equilibrium at room temperature and can rapidly interconvert. If tautomers are possible (e.g., in solution), a chemical equilibrium of tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions by 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, two tautomers.

[0155] The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be radiolabeled with radioactive isotopes, such as for example tritium ( 3 H), iodine-125 ( 125 I) or carbon-14 ( 14 C). For example, deuterium can be substituted for hydrogen to form deuterated drugs. Deuterium is a heavier isotope of hydrogen, and the bond between a carbon and a deuterium is stronger than the bond between carbon and protium (ordinary hydrogen). Deuterated drugs have advantages over non-deuterated drugs, such as reduced toxicity, increased stability, increased efficacy, increased biological half-life, and the like. All isotopic variations of the compounds of the present application, whether radioactive or not, are encompassed within the scope of the present application.

[0156] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0157] The term "substituted" means that any one or more hydrogen atoms on the particular atom is replaced with a substituent group, which can include variants of deuterium and hydrogen, as long as the valency of the particular atom is normal and the substituted compound is stable. The term "optionally substituted" means that it can or can not be substituted and, unless otherwise specified, the types and number of substituents can be any that are chemically possible.

[0158] When any variable (e.g., R) occurs more than one time in a compound; each definition is independent. Thus, if a group is substituted with 1, 2, or 3 R's, then said group can optionally be substituted up to three times with R, and each occurrence of R is selected independently. may be selected from etc.

[0159] When one of the variables is selected from a single bond, it indicates that the two groups to which it is attached are directly connected, such as where L2 represents a single bond, it indicates that the structure is actually A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CH2- indicates the 1-6 alkylcarbonyl- refers to a C 1-6 alkyl group attached to the remainder of the molecule through a carbonyl group. However, when the point of attachment of a substituent is apparent to one of ordinary skill in the art, such as halo substituents, the "-" can be omitted.

[0160] Unless otherwise indicated, a dashed line indicates a point of attachment for a group, such as in .

[0161] When a recited substituent does not indicate through which atom of the recited group it is attached to the rest of the molecule, the substituent can be bonded through any atom of the group, for example, a pyridyl group as a substituent can be attached to the rest of the molecule through any one of the carbon atoms of the pyridyl ring.

[0162] When a recited linking group does not indicate its direction of attachment, its direction of attachment is arbitrary, for example, where the linking group L is in this case the phenyl and cyclopentyl groups can be attached in the same direction as the reading order from left to right to form or in the opposite direction to the reading order from left to right to form The combination of substituents and / or variations thereof are only permitted if such combination would result in a stable compound.

[0163] Unless otherwise specified, the number of atoms in a ring refers to the number of atoms that are bonded together to form the ring itself (e.g., monocyclic compounds, fused ring compounds, spirocyclic compounds, bridged ring compounds, crosslinked compounds, carbocyclic compounds, heterocyclic compounds). The number of atoms in a ring is often defined as the ring size, e.g., a "4-6 membered ring" refers to a "ring" that is arranged around 4-6 atoms. When a ring is substituted, the atoms included in the substituent are not included in the ring atoms. Unless otherwise specified, benzene is a 6-membered ring, naphthalene is a 10-membered ring, and thiophene is a 5-membered ring.

[0164] Unless otherwise specified, the term "alkyl" refers to a saturated hydrocarbon group including primary (normal), or secondary carbon atoms, or tertiary carbon atoms, or quaternary carbon atoms, or a combination thereof, which can be straight-chained and / or branched, which can be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Unless otherwise specified, an alkyl group can be optionally substituted.

[0165] Unless otherwise specified, the term "C 1-20 "alkyl" is used to denote a straight-chained or branched saturated carbon hydride group consisting of 1 to 20 carbon atoms. The C 1-20 alkyl group includes C 1-19 , C 1-15 , C 1-10 , C 1-5 , C 1-4 , C 2-20 , C 2-12 , C 2-6 alkyl group; which can be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Unless otherwise specified, an alkyl group can be optionally substituted. 1-20 Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, sec-butyl, n-pentyl, n-hexyl, 1-methylhexyl, n-nonyl, n-decyl, adamantyl, n-undecyl, n-dodecyl, 2- ethyldodecyl, 2-butyldodecyl, n-eicosyl, methylene, 1,2-ethanediyl, 1,3-propanediyl, 1,4- butanediyl, 1,5-pentanediyl, 1,6-hexanediyl, 1,7-heptanediyl, 1,8-octanediyl, 1,9-nonanediyl, 1,10-decanediyl, 1,12-dodecanediyl, 1,14-tetradecanediyl, 1,16-hexadecanediyl, 1,18- octadecanediyl, 1,20-icosanediyl, and the like.

[0166] Unless otherwise specified, the term "C 1-6 "alkyl" is used to denote a straight-chained or branched saturated carbon hydride group consisting of 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 2-6Alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl ("Me"), ethyl ("Et"), propyl such as n-propyl ("n-Pr") or isopropyl ("i-Pr"), butyl such as n-butyl ("n-Bu"), isobutyl ("i-Bu"), sec-butyl ("s-Bu"), or tert-butyl ("t-Bu"), pentyl, hexyl, methylene, 1,2-ethylene, 1,3-propylene, 1,4-butylene, 1,5-pentylene, 1,6-hexylene, and the like.

[0167] 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). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), methylene, 1,2-ethylene, 1,3-propylene, and the like.

[0168] Unless otherwise specified, the term "alkenyl" refers to a group containing a carbon-carbon sp 2 A double-bonded hydrocarbon group may represent a straight and / or branched alkenyl group, wherein a branched group refers to one or more alkyl groups such as methyl, ethyl or propyl groups attached to a straight alkenyl chain. It may be monovalent, divalent or polyvalent. Unless otherwise specifically stated in the specification, an alkenyl group may be optionally substituted.

[0169] Unless otherwise specified, “C 2-20 "Alkenyl" is used to represent a linear or branched hydrocarbon group consisting of 2 to 20 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. 2-20 Alkenyl groups include C 2-19 、C 2-15 、C 2-10 、C 2-5 、C 2-4 、C 3-20 、C 4-12 、C 5-6 It can be monovalent, divalent or polyvalent. 2-20 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, pentenyl, hexenyl, butadienyl, pentyladienyl, hexadienyl, octenyl, decenyl, n-undecenyl, ethenylene, propenylene, sec-butenylene, 2-methylbutenylene, and the like.

[0170] Unless otherwise specified, "C 2-6 "Alkylene" is used to denote a straight or branched divalent hydrocarbon group of 1 to 20 carbon atoms, which can be saturated or unsaturated. The C 2-6 Alkylene includes C 2-4 , C4, C3, and C2 alkylene, etc.; which can be monovalent, divalent, or multivalent. C 2-3 Examples of alkylene groups include, but are not limited to, methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, ethylethylene, propylethylene, butylethylene, butadiene, pentadiene, hexadiene, etc. 2-6 "Alkenyl" is used to denote a straight or branched hydrocarbon group of 2 to 6 carbon atoms which contains at least one carbon-carbon double bond, which can be located in any position on the group. The C 2-3 Alkenyl includes C3and C2alkenyl; the C 2-3 Alkenyl can be monovalent, divalent, or multivalent. C 2-3 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, ethylethenyl, propylethenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl, hexadienyl, etc.

[0171] Unless otherwise specified, "C 2-3 "Alkenyl" is used to denote a straight or branched hydrocarbon group of 2 to 3 carbon atoms which contains at least one carbon-carbon double bond, which can be located in any position on the group. The C 2-20 Alkenyl includes C3and C2alkenyl; the C 2-20 Alkenyl can be monovalent, divalent, or multivalent. C 2-19 Examples of alkenyl groups include, but are not limited to, ethenyl, propenyl, ethylethenyl, propylethenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl, hexadienyl, etc.

[0172] Unless otherwise specified, the term "alkynyl" refers to a hydrocarbon group containing at least one unsaturated site, i.e., a carbon-carbon sp3 bond, which can represent straight chain and / or branched chain alkynyl groups, branched meaning one or more alkyl groups such as methyl, ethyl, or propyl are attached to a straight chain alkynyl chain. It can be monovalent, divalent, or multivalent. Unless otherwise specified in the specification, the alkynyl group can be optionally substituted.

[0173] Unless otherwise specified, the term "C 2-20 "Alkynyl" is used to denote a straight or branched hydrocarbon group of 2 to 20 carbon atoms which contains at least one carbon-carbon triple bond, which can be located in any position on the group. The C 2-20 Alkynyl includes C 2-19 , C 2-15 , C 2-10 , C 2-5 , C 2-4 , C 3-20 , C 4-12 , C 5-6 Alkynyl; which can be monovalent, divalent, or multivalent. C 2-20 Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, ethylethynyl, propylethynyl, butynyl, butadiynyl, pentynyl, pentadiynyl, hexynyl, hexadiynyl, etc.

[0174] The term "C 2-6 Alkynyl" is used to denote a straight or branched hydrocarbon group containing at least one carbon-carbon triple bond consisting of from 2 to 6 carbon atoms, the carbon-carbon triple bond can be located in any position of the group. It can be monovalent, divalent or multivalent. Said C 2-6 Alkynyl includes C 2-5 , C 2-4 , C 2-3 , C2, C 2-6 , C6and C5alkynyl, etc. Examples of C 2-6 Alkynyl include, but are not limited to, ethynyl, propynyl, ethynylene, propynylene, pentynyl, pentynylene, etc.

[0175] Unless otherwise specified, "C 2-3 Alkynyl" is used to denote a straight or branched hydrocarbon group containing at least one carbon-carbon triple bond consisting of from 2 to 3 carbon atoms, the carbon-carbon triple bond can be located in any position of the group. It can be monovalent, divalent or multivalent. Said C 2-3 Alkynyl includes C3and C2alkynyl. C 2-3 Examples of C

[0176] Unless otherwise specified, the term "heteroalkyl", by itself or in combination with another term, represents a stable straight or branched chain, or combination thereof, of the indicated number of carbon atoms, and at least one heteroatom or heteroatom group, wherein the "alkyl" of the "heteroalkyl" is the same as defined herein. In some embodiments, the heteroatom is selected from B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen heteroatom is optionally quaternized. In other embodiments, the heteroatom group is selected from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)-, and -S(=O)N(H)-. In some embodiments, the heteroalkyl is C 1-20 heteroalkyl; in some embodiments, the heteroalkyl is C 1-6 heteroalkyl; in some embodiments, the heteroalkyl is C 1-3Heteroalkyl. The heteroatom or heteroatom group can be located at any internal position of the heteroalkyl group, including the position at which the alkyl group is attached to the rest of the molecule. Examples of heteroalkyl include, but are not limited to, -OCH3, -OCH2CH3, -OCH2CH2CH3, -OCH2(CH3)2, -CH2-CH2-O-CH3, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)(CH2CH3), -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, -CH2-S-CH2-CH3, -CH2-CH2, -S(=O)-CH3, -CH2-CH2-S(=O)2-CH3, and the like; up to two of the heteroatoms thereof can be consecutive, e.g., -CH2-NH-OCH3. Unless otherwise expressly indicated, a heteroalkyl group is optionally substituted. Unless otherwise specified, the term "alkoxy" denotes an alkyl group comprising 1 to 20 carbon atoms attached to the rest of the molecule through an oxygen atom, wherein "alkyl" in the "alkyl group" is defined herein as above. Unless otherwise expressly indicated, an alkoxy group is optionally substituted.

[0177] Unless otherwise specified, the term "C 1-20 alkyl group" denotes those alkyl groups comprising 1 to 20 carbon atoms. The C 1-20 alkyl groups include C 1-19 , C 1-10 , C 1-5 , C 2-20 , C 2-8 , C6, C5, and C4alkyl groups, etc. Examples of C 1-20 alkyl groups include, but are not limited to, methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl, and t-butyl), pentyl (including n-pentyl, isopentyl, and neopentyl), hexyl, n-hexyl, 1-methylhexyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, 2- ethyldodecyl, n-icosyl, methyleneoxy, ethyleneoxy, propyleneoxy, butyleneoxy, pentyleneoxy, heptyleneoxy, dodecyleneoxy, and the like.

[0178] Unless otherwise specified, the term "C 1-6 alkyl group" denotes those alkyl groups comprising 1 to 6 carbon atoms. The C 1-6 alkyl groups include C 1-4 , C 1-3 , C 1-2 , C 2-6、C 2-4 , C6, C5, C4 and C3 alkoxy, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentoxy and neopentoxy), hexyloxy, methyleneoxy, ethyleneoxy, propyleneoxy, butyleneoxy, pentyleneoxy, and the like.

[0179] Unless otherwise specified, the term “C 1-4 "Alkoxy" refers to those alkyl groups containing 1 to 4 carbon atoms which are attached to the rest of the molecule via an oxygen atom. 1-4 Alkoxy groups include C 1-3 、C 1-2 、C 2-4 , C4 and C3 alkoxy, etc. 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), methyleneoxy, ethyleneoxy, propyleneoxy, butyleneoxy, and the like.

[0180] 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), methyleneoxy, ethyleneoxy, propyleneoxy, and the like.

[0181] Unless otherwise specified, the term "amino" may be monovalent - NH2, divalent or multi-price

[0182] Unless otherwise specified, the term "alkylamino" refers to an alkyl group attached to the rest of the molecule via an amino group as defined above, wherein "alkyl" in "alkyl group" has the same meaning as described above in the present invention. Unless otherwise specified in the specification, an alkylamino group may be optionally substituted.

[0183] Unless otherwise specified, the term “C 1-20 "Alkylamino" means an alkyl group containing 1 to 20 carbon atoms which is attached to the rest of the molecule via an amino group. 1-20 Alkylamino groups include C 1-19 、C 1-14 、C1-12 、C 2-6 、C 2-4 、C 15 、C 10 , C8, C5 and C 20 Alkylamino, etc. C 1-20 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)(CH2CH3), -NHCH2CH2CH3, -NHCH2(CH3)2, -NHCH2CH2CH2CH3, -NHCH2CH2CH2CH2CH3, -NHCH2CH2CH2CH2CH3, -NHCH2CH2CH2CH2CH3, -N(CH2CH2CH3)(CH2CH2CH2CH3), and the like.

[0184] Unless otherwise specified, the term “C 1-6 "Alkylamino" means an alkyl group containing 1 to 6 carbon atoms which is attached to the rest of the molecule via an amino group. 1-6 Alkylamino groups include C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4, C3 and C2 alkylamino, etc. 1-6 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)(CH2CH3), -NHCH2CH2CH3, -NHCH2(CH3)2, -NHCH2CH2CH2CH3, and the like.

[0185] Unless otherwise specified, the term “C 1-4 "Alkylamino" means an alkyl group containing 1 to 4 carbon atoms which is attached to the rest of the molecule via an amino group. 1-4 Alkylamino groups include C 1-3 、C 1-2 、C 2-4 , C4, C3 and C2 alkylamino, etc. 1-4 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -N(CH2CH3)(CH2CH3), -NHCH2CH2CH3, -NHCH2(CH3)2, -NHCH2CH2CH2CH3, and the like.

[0186] Unless otherwise specified, the term “C 1-3"Alkylamino" means an alkyl group containing 1 to 3 carbon atoms which is attached to the rest of the molecule via an amino group. 1-3 Alkylamino groups include C 1-2 , C3 and C2 alkylamino, etc. 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, and the like.

[0187] Unless otherwise specified, the term "alkylthio" refers to an alkyl group attached to the rest of the molecule via a sulfur atom, wherein "alkyl" in "alkyl group" has the same meaning as described above in the present invention. Unless otherwise specified in the specification, an alkylthio group may be optionally substituted.

[0188] Unless otherwise specified, the term “C 1-20 "Alkylthio" refers to those alkyl groups containing 1 to 20 carbon atoms which are attached to the rest of the molecule via a sulfur atom. 1-20 Alkylthio includes C 1-19 、C 1-14 、C 1-12 、C 2-6 、C 2-4 、C 15 、C 10 , C8, C5 and C 20 Alkylthio, etc. 1-20 Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, -SCH2CH2CH2CH3, -SCH2CH2(CH3)2, -SCH2CH2CH2CH2CH3, -SCH2CH2CH2CH2CH3, -SCH2(CH2CH2CH3)(CH2CH2CH2CH3), and the like.

[0189] Unless otherwise specified, the term “C 1-6 "Alkylthio" refers to those alkyl groups containing 1 to 6 carbon atoms which are linked to the rest of the molecule via a sulfur atom. 1-6 Alkylthio includes C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-4 , C6, C5, C4, C3 and C2 alkylthio, etc. 1-6 Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, and the like.

[0190] The term "C 1-4 Alkylthio" means those alkyl groups connected to the rest of the molecule by a sulfur atom. The C 1-4 alkylthio groups include C 1-3 , C 1-2 , C 2-4 , C4, C3, and C2 alkylthio groups, and the like. Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, and the like. 1-4

[0191] The term "C 1-3 alkylthio" means those alkyl groups connected to the rest of the molecule by a sulfur atom. The C 1-3 alkylthio groups include C 1-3 , C 1-2 , and C3 alkylthio groups, and the like. Examples of alkylthio groups include, but are not limited to, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH2(CH3)2, and the like. 1-3

[0192] The term "cycloalkyl" means, unless otherwise specified, a stable non-aromatic, mono- or polycyclic, saturated hydrocarbon group composed of carbon and hydrogen atoms, which can include fused, spiro, and / or bridged ring ring systems. Monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, but are not limited to, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, and the like. The term "C 4-6 cycloalkyl" means a cycloalkyl group having 4-6 ring carbon atoms. Similarly, the term "C 3-4 cycloalkyl" means a cycloalkyl group having 3-4 ring carbon atoms. Unless otherwise specified, a cycloalkyl group can be optionally substituted.

[0193] The term "C 3-20 cycloalkyl" means a saturated monocyclic or polycyclic hydrocarbon group having 3-20 ring carbon atoms, for example, having 3-15 ring carbon atoms, for example, 3-6 ring carbon atoms; which can be monovalent, divalent, or multivalent. Examples of C 3-20 cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl, and the like.

[0194] The term "C 3-6 cycloalkyl" means a saturated monocyclic or bicyclic hydrocarbon group having 3-6 ring carbon atoms, for example, having 3-5 ring carbon atoms, for example, 3-4 ring carbon atoms; which can be monovalent, divalent, or multivalent. Examples of C 3-6 ​​Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0195] Unless otherwise specified, “C 4-6 "Cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 4 to 6 carbon atoms, which is a monocyclic or bicyclic system. 4-6 Cycloalkyl groups include C 4-5 、C 5-6 , C4, C5 and C6 cycloalkyl, etc.; which may be monovalent, divalent or polyvalent. 4-6 Examples of cycloalkyl groups include, but are not limited to, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0196] Unless otherwise specified, the term "heterocycloalkyl" refers to a non-aromatic saturated cyclic group that exists as a monocyclic, fused, spirocyclic and / or bridged ring, wherein at least one of the ring atoms is a heteroatom or heteroatom group, and the rest are carbon atoms; in some embodiments, each occurrence of the heteroatom is independently selected from B, O, N and S, wherein the nitrogen and sulfur atoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2), the nitrogen heteroatom is optionally quaternized, and in other embodiments, each occurrence of the heteroatom group is independently selected from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)-, and -S(=O)N(H)-. The heteroatom or heteroatom group may be positioned at any interior position of the heterocycloalkyl group, including the position at which the heterocycloalkyl group is attached to the rest of the molecule. In some embodiments, the heterocycloalkyl group is a 3-20 membered heterocycloalkyl group; in some embodiments, the heterocycloalkyl group is a 3-10 membered heterocycloalkyl group; in other embodiments, the heterocycloalkyl group is a 3-6 membered heterocycloalkyl group. Unless stated otherwise specifically in the specification, the heterocycloalkyl group may be optionally substituted. Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from B, O, S and N or heteroatoms as described above, and the rest are carbon atoms, wherein 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). It includes monocyclic and bicyclic ring systems, where the bicyclic ring systems include spiro, fused, and bridged rings. Further, with respect to this "3-6 membered heterocycloalkyl," the heteroatom or heteroatom group can be positioned at any interior position of the heterocycloalkyl group, including can occupy the position of attachment of the heterocycloalkyl group to the remainder of the molecule. The 3-6 membered heterocycloalkyl group includes 5-6 membered, 4 membered, 5 membered, and 6 membered heterocycloalkyl groups, and the like. Examples of 3-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, or homopiperidinyl, and the like.

[0197] Unless otherwise specified, the term "cycloalkenyl" as used herein refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon group consisting of carbon and hydrogen atoms, having one or more carbon-carbon sp 2 double bonds, which can include fused, spiro, and / or bridged ring systems. Monocyclic cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, and the like. Polycyclic cycloalkenyl groups include, but are not limited to, bicyclo[2.2.1]hept-2-enyl, and the like. Unless otherwise specified, a cycloalkenyl group can be optionally substituted. "C 3-7 Cycloalkenyl" includes C3, C4, C5, C6, and C7 cycloalkenyl groups. Examples of cycloalkenyl groups include, but are not limited to, cyclobutenyl, cyclopentenyl, and cyclohexenyl.

[0198] Unless otherwise specified, the term "heterocycloalkenyl" as used herein refers to a cyclic alkenyl group comprising several heteroatoms or heteroatom groups, which in some embodiments are independently selected at each occurrence from B, O, N, and S, wherein the nitrogen and sulfur atoms are optionally oxidized (i.e., NO and S(O) p, p is 1 or 2), the nitrogen atom is optionally quaternized, in other embodiments, the heteroatom group is independently selected for each occurrence from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O)2-, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O)2N(H)-, and -S(=O)N(H)-. "5-6 membered heterocycloalkenyl" by itself or in combination with other terminology, means an unsaturated cyclic group consisting of 5 to 6 ring atoms, 1, 2, 3, or 4 of which are heteroatoms independently selected from B, O, S, and N, or a heteroatom group as described above, and the remainder of which are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms can be optionally oxidized (i.e., NO and S(O) p Examples of heterocycloalkenyl groups include, but are not limited to Unless otherwise specified, a heterocycloalkenyl group can be optionally substituted.

[0199] Unless otherwise specified, when a substituent attached to ring A can be connected to ring A to form a ring, it means that the substituent can be connected to any site of ring A to form a new ring, including fused, spiro, or bridged rings; wherein ring A can be selected from cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl, etc. as described above. For example, R in may be connected to to form a 6-membered ring, examples of which include, but are not limited to

[0200] Unless otherwise specified, C n-n+m or C n -C n+m includes any specific instance of n to n+m carbons, for example C 1-12 includes C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 also includes any range of n to n+m, for example C 1-12 includes 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 ​and the like. Similarly, n- to n+m-membered means the number of atoms in the ring is n to n+m, for example, 3- to 12-membered rings include 3-membered rings, 4-membered rings, 5-membered rings, 6-membered rings, 7-membered rings, 8-membered rings, 9-membered rings, 10-membered rings, 11-membered rings, and 12-membered rings, and also include any range within n to n+m, for example, 3- to 6-membered rings, 3- to 9-membered rings, 5- to 6-membered rings, 5- to 7-membered rings, 6- to 7-membered rings, 6- to 8-membered rings, and 6- to 10-membered rings, and the like.

[0201] Unless otherwise specified, the term "aryl" means a hydrocarbon ring system group comprising at least one aromatic ring. In the present application, aryl groups can be monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which can include fused, spiro, and / or bridged ring systems. Aryl groups include, but are not limited to, benzene, naphthalene, anthracene, fluoranthene, phenanthrene, benzophenanthrene, pyrene, tetracene, chrysene, benzochrysene, azulene, fluorene, and derivatives thereof. Unless otherwise specified in the specification, aryl groups can be optionally substituted.

[0202] Unless otherwise specified, the term "heteroaryl" means a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 20 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl groups are preferably 5- to 10-membered, containing 1 to 3 heteroatoms; more preferably 5- or 6-membered, containing 1 to 3 heteroatoms; non-limiting examples include pyrazolyl, imidazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, and the like. Heteroaryl groups can be attached to the rest of the molecule through a heteroatom or a carbon atom. The heteroaryl ring can be fused to an aryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring that is attached to the parent structure is the heteroaryl ring, non-limiting examples of which include: Non-limiting examples of heteroaryl groups also include triazine, pyridine, pyrimidine, imidazole, furan, thiophene, benzofuran, benzothiophene, indole, carbazole, pyrroloimidazole, pyrrolopyrrole, thienopyrrole, thienothiophene, furanopyrrole, furanofuran, thienofuran, benzisoxazole, benzisothiazole, benzimidazole, quinoline, isoquinoline, perylene, quinoxaline, phenanthridine, berberine, quinazoline, quinazolinone, dibenzothiophene, dibenzofuran, carbazole, and derivatives thereof. Unless otherwise specified in the specification, heteroaryl groups can be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, or heterocycloalkylthio.

[0203] The term "substituted" as used herein means that at least one hydrogen atom in any of the above groups (i.e., alkyl, alkenyl, alkynyl, heteroalkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl) is replaced with a bond to a non-hydrogen atom including, but not limited to, halogen atoms (e.g., F, Cl, Br, I), oxygen-containing groups (e.g., hydroxyl, alkoxy, ester), sulfur-containing groups (e.g., thiol, thioalkyl, sulfone, sulfonyl, sulfoxide), nitrogen-containing groups (e.g., amine, amide, dialkylamine, arylamine, aryl-alkyl-amine, diarylamine, N-oxide, imide, enamine), silicon-containing groups (e.g., trialkylsilyl, dialkylarylsilyl, alkylbaryl silyl, triarylsilyl), and other heteroatoms in various other groups.

[0204] The term "substituted" as used herein also means that one or more hydrogen atoms in any of the above groups (i.e., alkyl, alkenyl, alkynyl, heteroalkyl, alkoxy, alkylamino, alkylthio, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, aryl, heteroaryl) is replaced with a higher order bond (e.g., a double or triple bond) to a heteroatom, such as the oxygen in carbonyl, carboxyl, and ester groups, and the nitrogen in imine, oxime, hydrazone, and nitrile groups. For example, "substituted" means that one or more hydrogen atoms in any of the above groups is replaced with -NR g R h , -NR g C(=O)R h , -NR g C(=O)NR g R h , -NR g C(=O)OR h , -NR g SO2R h , -OC(=O)NR g R h , -OR g , -SR g , -SOR g , SO2R g , -OSO2R g , -SO2OR g ,

[0205] =NSO2R g , and -SO2NR g R h . "Substituted" can also mean that one or more hydrogen atoms in any of the above groups is replaced with -C(=O)R g , -C(=O)OR g , -C(=O)NR g R h , -CH2SO2Rg -CH2SO2NR g R h substituted. The R g and R h are the same or different, and are independently selected from hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkyl-alkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocycloalkyl-alkyl, heteroaryl, N-heteroaryl, heteroaryl-alkyl. "Substituted" can also mean that one or more hydrogen atoms of any of the above groups are replaced by an amino, cyano, hydroxy, imino, nitro, oxo, thioxo, halogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkyl-alkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocycloalkyl-alkyl, heteroaryl, N-heteroaryl, heteroaryl-alkyl. Additionally, each of the above substituents can be optionally substituted with one or more of the above substituents.

[0206] It will be understood by those skilled in the art that some of the compounds of Formula (I) can comprise one or more chiral centers and therefore exist as two or more stereoisomers. Accordingly, the compounds of the present application can exist as individual stereoisomers (e.g., enantiomers, diastereomers) and mixtures thereof in all proportions, e.g., racemates, and where appropriate, as tautomers and geometric isomers.

[0207] The term "stereoisomers" as used herein refers to compounds which have the same chemical constitution, but differ in the arrangement of atoms or groups in space.

[0208] The term "enantiomers" as used herein refers to two stereoisomers of a compound which are non-superimposable mirror images of one another.

[0209] The term "diastereomers" as used herein refers to stereoisomers which have two or more chiral centers and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g., melting points, boiling points, spectral properties, or biological activity. Mixtures of diastereomers can be separated by high resolution analytical methods, e.g., electrophoresis and chromatography, e.g., HPLC.

[0210] Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane-polarized light. In describing optical activity, the prefixes D and L or R and S are used to denote the absolute configuration of the molecule about its chiral center. The prefixes d and 1 or (+) and (-) are employed to designate the sign of the rotation of plane-polarized light by the compound, with (-) or 1 indicating that the compound is levorotatory. A compound with the (+) or d prefix is dextrorotary. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer can also be referred to as an enantiomer when there are two stereoisomers and a mixture of such isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or racmate, which can occur where there has been no stereoselection or stereospecificity in a chemical reaction or process. The terms "racemic mixture" and "racmate" refer to an equimolar mixture of two enantiomeric forms lacking optical activity.

[0211] Racemic mixtures can be used as such or resolved into their individual isomers. A stereochemically pure compound or a mixture enriched in one or more isomers can be obtained by resolution. Methods for separating isomers are well known, including physical methods such as chromatography employing a chiral adsorbant. Individual isomers can be prepared in chiral form from chiral precursors. Alternatively, individual isomers can be obtained chemically from mixtures by forming diastereomeric salts with a chiral acid (such as the individual enantiomers of 10-camphorsulfonic acid, camphoric acid, a-bromocamphoric acid, tartaric acid, diacetyltartaric acid, malic acid, pyrrolidine-5-carboxylic acid, and the like), fractional crystallization of the salts, and liberation of the resolved base. This process can be repeated to obtain one or both isomers substantially free of the other, i.e., optical purity of, for example, at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% by weight of the desired stereoisomer. Alternatively, as is well known to those of ordinary skill in the art, the racmate can be covalently bonded to a chiral compound (chiral auxiliary) to give a diastereomeric mixture.

[0212] The term "tautomer" or "tautomeric form" as used herein refers to different energy structures that are interconvertible via a low energy barrier. For example, prototropic tautomers (also known as proton-shift tautomers) include interconversions by proton migration, such as keto-enol and imine-enamine isomerization. Valence tautomers include interconversions by reorganization of some of the bonding electrons.

[0213] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the specific embodiments set forth below with other chemical synthetic methods well known in the art, and equivalents thereof as appreciated by those skilled in the art, preferred embodiments including but not limited to the examples of the present application.

[0214] Technical and scientific terms used herein that are not specifically defined have the meanings that would be given to them by one of ordinary skill in the art to which this application pertains. DETAILED DESCRIPTION

[0215] The present application is described in detail below by way of examples, but it is not meant to present any limitations on the present application. The present application has been described in detail by specific embodiments, and the specific embodiments of the present application are disclosed, it would be apparent to those skilled in the art that various changes and modifications of the specific embodiments of the present application can be made without departing from the spirit and scope of the present application.

[0216] Example 1: Synthesis of Compound 1

[0217]

[0218] Step 1: Preparation of Compound 1-2

[0219] Compound 1-1 (1.00 g, 5.81 mmol) and triphosgene (1.72 g, 5.81 mmol) were dissolved in super dry tetrahydrofuran (50 mL). Under ice water bath and nitrogen protection, triethylamine (2.35 g, 23.2 mmol) was added dropwise into the reaction system. After dropwise, the reaction system was continued to be stirred for 1 hour under ice water bath. Then 4-amino-1-tert-butoxycarbonylpiperidine (2.02 g, 10.1 mmol) was added. It was naturally raised to room temperature and stirred for 16 hours. LCMS showed that the reaction was complete. It was quenched by saturated aqueous ammonium chloride solution (100 mL), extracted with ethyl acetate (70 mL x 3). The organic phase was combined and concentrated under reduced pressure, and the obtained residue was purified by reverse phase column chromatography (water / acetonitrile = 3 / 2) to obtain compound 1-2 (1.94 g, yield: 84.0%). LC-MS (ESI) [M+H] + 399.1.

[0220] Step 2: Preparation of Compound 1-3

[0221] To a solution of compound 1-2 (800 mg, 2.01 mmol) in tetrahydrofuran (10 mL) was added potassium tert-butoxide (1.35 g, 12.0 mmol) under nitrogen protection. The reaction system was stirred at 60 °C for 2 hours. LCMS showed that the reaction was complete. Concentration under reduced pressure, the resulting residue was purified by column chromatography (dichloromethane / methanol = 10 / 1) to give compound 1-3 (338 mg, yield: 47.8%). LC-MS (ESI) [M+H] + 297.0.

[0222] Step 3: Preparation of compound 1-4

[0223] To a solution of compound 1-3 (238 mg, 0.675 mmol) in acetonitrile (10 mL) was added N-bromosuccinimide (132 mg, 0.743 mmol) under nitrogen protection. The reaction system was stirred at room temperature for 3 hours. LCMS showed that the reaction was complete. Concentration under reduced pressure, the resulting residue was purified by reversed-phase column chromatography (water / acetonitrile = 1 / 1) to give compound 1-4 (212 mg, yield: 72.9%). LC-MS (ESI) [M+H] + 374.9 / 376.9.

[0224] Step 4: Preparation of compound 1-5

[0225] To a solution of compound 1-4 (200 mg, 0.464 mmol) in N,N-dimethylformamide (5 mL) was added N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (202 mg, 0.556 mmol) and N,N-diisopropylethylamine (120 mg, 0.927 mmol) under ice water bath and nitrogen protection. The reaction system was stirred at ice water bath for 2 hours. LCMS showed that the reaction was complete. Quenching with water (20 mL), extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with saturated brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the resulting residue was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give compound 1-5 (203 mg, yield: 65.7%). LC-MS (ESI) [M+H-56] + 609.9 / 611.9.

[0226] Step 5: Preparation of compound 1-6

[0227] Compound 1-5 (467 mg, 0.700 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 3,6-dihydro-2H-pyran (177 mg, 0.840 mmol), [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (14.0 mg, 0.070 mmol) and sodium carbonate (223 mg, 2.10 mmol) were dissolved in 1,4-dioxane / water (10 / 1 mL). The reaction system was stirred at 80 °C for 4 h. LCMS showed the reaction was complete. Diluted with water (30 mL), extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, the residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 1-6 (303 mg, yield: 70.5%). LC-MS (ESI) [M+H] + 670.2.

[0228] Step 6: Preparation of compound 1-7

[0229] To a solution of compound 1-6 (118 mg, 176 μmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL) at room temperature. The reaction system was stirred at room temperature for 2 h. LCMS showed the reaction was complete. Concentrated under reduced pressure to give compound 1-7 crude (120 mg). The crude was used directly in the next step. LC-MS (ESI) [M+H] + 570.4.

[0230] Step 7: Preparation of compound 1-8

[0231] To a solution of compound 1-7 crude (120 mg), 5-benzyloxy-6-methylpyrimidine-4- carboxylic acid (51.5 mg, 211 μmol) and N,N-diisopropylethylamine (68.1 mg, 527 μmol) in dichloromethane (10 mL) was added 2-(7-azabenzotriazol-1-yl)-1,1,3,3- tetramethyluronium hexafluorophosphate (79.5 mg, 211 μmol) at room temperature. The reaction system was stirred at room temperature for 4 h. LCMS showed the reaction was complete. Concentrated under reduced pressure, the residue was purified by column chromatography (dichloromethane / methanol = 20 / 1) to give compound 1-8 (120 mg, total yield of two steps: 85.6%). LC-MS (ESI) [M+H] + 796.2.

[0232] Step 8: Preparation of compound 1

[0233] To a solution of compound 1-8 (100 mg, 126 μmol) in dichloromethane (5 mL) was added boron trichloride solution in dichloromethane (1.00 mol / L, 251 μL, 251 μmol) dropwise under ice water bath and nitrogen protection. The reaction system was stirred at room temperature for 3 hours. LCMS showed that the reaction was complete. Quenching with water (10 mL), extraction with dichloromethane (30 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by prep-HPLC to give compound 1 (36.0 mg, yield: 40.6%). LC-MS (ESI) [M+H] + 706.2.

[0234] 1 H NMR (400 MHz, DMSO-d6) δ 10.36 (s, 1H), 10.18 (s, 1H), 8.56 (s, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.98 (d, J = 2.1 Hz, 1H), 7.73 (dd, J = 9.0, 2.1 Hz, 1H), 6.64 (dd, J = 3.1, 1.6 Hz, 1H), 5.09 (s, 1H), 4.96 (d, J = 5.1 Hz, 2H), 4.63 (d, J = 12.8 Hz, 1H), 4.25 (d, J = 2.9 Hz, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.56 (d, J = 13.2 Hz, 1H), 3.16 (t, J = 12.9 Hz, 1H), 2.89 (s, 1H), 2.55 (s, 4H), 2.43 (s, 3H), 1.73 (d, J = 12.0 Hz, 1H), 1.56 (d, J = 11.9 Hz, 1H).

[0235] Example 2: Synthesis of compound 2

[0236]

[0237] Step 1: Preparation of compound 2-2

[0238] To a solution of compound 2-1 (800 mg, 1.86 mmol), 4-(4,4,5,5-tetramethyl-1,3,2- dioxaborolan-2-yl)-3,6-dihydro-2H-pyran (469 mg, 2.23 mmol), [1,1'-bis(diphenylphosphino) ferrocene]dichloropalladium (136 mg, 186 µmol) and cesium carbonate (1.82 g, 5.58 mmol) in 1,4-dioxane / water (30 / 5 mL) was stirred at 100 ℃ for 12 h. LCMS showed the reaction was complete. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane / methanol = 20 / 1) to give compound 2-2 (750 mg, yield: 93.1%). LC-MS (ESI) [M+H] + 378.0.

[0239] Step 2: Preparation of compound 2-3

[0240] To a solution of compound 2-2 (750 mg, 1.73 mmol) in N,N-dimethylformamide (5 mL) was added N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (629 mg, 1.73 mmol) and N,N-diisopropylethylamine (448 mg, 3.46 mmol) under nitrogen protection successively. The reaction mixture was stirred at 40 ℃ for 4 h. LCMS showed the reaction was complete. Water (20 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with saturated brine (40 mL x 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 2-3 (770 mg, yield: 66.5%). LC-MS (ESI) [M+H-56] + 612.8.

[0241] Step 3: Preparation of compound 2-4

[0242] To a solution of compound 2-3 (400 mg, 598 µmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 h. LCMS showed the reaction was complete. Concentration under reduced pressure gave compound 2-4 (340 mg). The crude product was used directly in the next step. LC-MS (ESI) [M+H] + 569.0.

[0243] Step 4: Preparation of compound 2-5

[0244] To a solution of compound 2-4 crude (340 mg), 5-benzyloxy-6-methylpyrimidine-4- carboxylic acid (175 mg, 717 μmol) and N,N-diisopropylethylamine (312 μL, 1.79 μmol) in dichloromethane (10 mL) was added 2-(7-azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (232 mg, 717 μmol) at room temperature. The reaction was stirred at room temperature for 2 h. LCMS showed the reaction was complete. Concentration under reduced pressure, the residue was purified by column chromatography (dichloromethane / methanol = 20 / 1) to give compound 2-5 (450 mg, total yield: 94.7% over two steps). LC-MS (ESI) [M+H] + 795.2.

[0245] Step 5: Preparation of compound 2

[0246] To a solution of compound 2-5 (350 mg, 440 μmol) in dichloromethane (10 mL) was added boron trichloride solution in dichloromethane (1.00 mol / L, 880 μL, 880 μmol) dropwise under ice water bath and nitrogen protection. The reaction was stirred at room temperature for 3 h. LCMS showed the reaction was complete. Quenching with water (10 mL), extraction with dichloromethane (30 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, the residue was purified by preparative-HPLC to give compound 2 (85.0 mg, yield: 27.4%). LC-MS (ESI) [M+H] + 705.0.

[0247] 1 H NMR (400 MHz, DMSO-d6) δ 10.20 (d, J = 17.6 Hz, 2H), 8.56 (s, 1H), 8.10 (d, J = 8.6 Hz, 1H), 7.97 (d, J = 2.1 Hz, 1H), 7.72 (dd, J = 8.7, 2.2 Hz, 1H), 7.46 (s, 1H), 6.52 (s, 1H), 5.04 (d, J = 17.4 Hz, 3H), 4.62 (d, J = 12.8 Hz, 1H), 4.24 (d, J = 3.2 Hz, 2H), 3.81 (t, J = 5.4 Hz, 2H), 3.55 (d, J = 13.2 Hz, 1H), 3.13 (t, J = 12.9 Hz, 1H), 2.86 (t, J = 12.9 Hz, 1H), 2.62 - 2.53 (m, 2H), 2.48 - 2.44 (m, 2H), 2.43 (s, 3H), 1.72 (d, J = 11.9 Hz, 1H), 1.55 (d, J = 12.1 Hz, 1H).

[0248] Example 3: Synthesis of compound 3

[0249]

[0250] Step 1: Preparation of compound 3-2

[0251] Compound 3-1 (2.50 g, 13.5 mmol) and triphosgene (1.40 g, 4.72 mmol) were dissolved in dichloromethane (25 mL). Under ice water bath and nitrogen protection, triethylamine (4.10 g, 40.5 mmol) was added dropwise into the reaction system. After dropwise, the reaction system was continuously stirred for 1 hour under ice water bath. Then 4-amino-1-tert-butoxycarbonylpiperidine (2.52 g, 12.58 mmol) was added. The reaction was allowed to naturally rise to room temperature and reacted for 4 hours. LCMS showed that the reaction was complete. Water (100 mL) was added for quenching, and dichloromethane (70 mL x 3) was used for extraction. The organic phases were combined and concentrated under reduced pressure. The obtained residue was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain compound 3-2 (2.60 g, yield: 51.7%). LC-MS (ESI) [M+H-56] + 356.2.

[0252] Step 2: Preparation of compound 3-3

[0253] Under nitrogen protection, potassium tert-butoxide (2.13 g, 18.9 mmol) was added into a solution of compound 3-2 (2.60 g, 6.32 mmol) in super dry tetrahydrofuran (30 mL). The reaction system was stirred at 60°C for 6 hours. LCMS showed that the reaction was complete. Water (50 mL) was added for quenching, and ethyl acetate (70 mL x 3) was used for extraction. The combined organic phases were washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the obtained residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound 3-3 (1.10 g, yield: 47.6%). LC-MS (ESI) [M+H-56] + 310.4.

[0254] Step 3: Preparation of compound 3-4

[0255] To a solution of compound 3-3 (1.00 g, 2.74 mmol) in dichloromethane (20 mL) was added N-bromosuccinimide (487 mg, 2.74 mmol) under ice water bath and nitrogen protection. The reaction system was stirred for 1 hour under ice water bath. LCMS showed the reaction was complete. Quench with water (50 mL), extracted with ethyl acetate (70 mL x 3). The combined organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 3-4 (950 mg, yield: 78.1%). LC-MS (ESI) [M+H-56] + 388.0 / 390.0.

[0256] Step 4: Preparation of compound 3-5

[0257] Compound 3-4 (900 mg, 2.03 mmol), 4-(4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl)-3, 6-dihydro-2H-pyran (511 mg, 2.43 mmol), [1, 1'-bis(diphenylphosphino)ferrocene]dichloropalladium (148 mg, 202 μmol) and cesium carbonate (1.98 g, 6.08 mmol) were dissolved in 1, 4-dioxane / water (10 / 3 mL). The reaction system was stirred at 100 °C for 12 hours. LCMS showed the reaction was complete. The reaction system was cooled to room temperature, quenched with water (50 mL), extracted with ethyl acetate (70 mL x 3). The combined organic phase was washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, the obtained residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 3-5 (300 mg, yield: 33.1%). LC-MS (ESI) [M+H-56] + 392.2.

[0258] Step 5: Preparation of compound 3-6

[0259] To a solution of compound 3-5 (260 mg, 581 μmol) in N,N-dimethylformamide (5 mL) was added N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (211 mg, 581 μmol) and N,N-diisopropylethylamine (150 mg, 1.16 mmol) successively under ice water bath and nitrogen protection. The reaction system was stirred at 40 °C for 3 h. LCMS showed that the reaction was complete. Quenched by water (20 mL), extracted by ethyl acetate (50 mL x 3). The combined organic phase was washed by saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 3-6 (250 mg, yield: 63.0%). LC-MS (ESI) [M+H-56] + 627.2.

[0260] Step 6: Preparation of compound 3-7

[0261] To a solution of compound 3-6 (250 mg, 366 μmol) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL) at room temperature. The reaction system was stirred at room temperature for 2 h. LCMS showed that the reaction was complete. Concentrated under reduced pressure to give compound 3-7 crude (250 mg). The crude was used directly in the next step reaction. LC-MS (ESI) [M+H] + 583.2.

[0262] Step 7: Preparation of compound 3-8

[0263] To a solution of compound 3-7 crude (250 mg), 5-benzyloxy-6-methylpyrimidine-4- carboxylic acid (105 mg, 429 μmol) and N,N-diisopropylethylamine (55.4 mg, 429 μmol) in dichloromethane (10 mL) was added 2-(7-azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (162 mg, 429 μmol) at room temperature. The reaction system was stirred at room temperature for 12 h. LCMS showed that the reaction was complete. Concentrated under reduced pressure, the obtained residue was purified by column chromatography (dichloromethane / methanol = 20 / 1) to give compound 3-8 (200 mg, total yield of two steps: 67.6%). LC-MS (ESI) [M+H] + 809.4.

[0264] Step 8: Preparation of compound 3

[0265] To a solution of compound 3-8 (100 mg, 123 μmol) in dichloromethane (10 mL) was added boron trichloride solution in dichloromethane (1.00 mol / L, 247 μL, 247 μmol) dropwise under ice water bath and nitrogen protection. The reaction system was stirred at room temperature for 2 hours. LCMS showed that the reaction was complete. Quench with water (20 mL), extracted with dichloromethane (30 mL x 3). The combined organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by prep-HPLC to give compound 3 (13.6 mg, yield: 15.3%). LC-MS (ESI) [M+H] + 719.2.

[0266] 1 H NMR (400 MHz, DMSO-d6) δ 10.33 (s, 1H), 10.18 (s, 1H), 8.56 (s, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.97 (d, J = 2.1 Hz, 1H), 7.73 (dd, J = 8.8, 2.1 Hz, 1H), 6.04 - 5.92 (m, 1H), 5.06 (s, 1H), 4.88 (d, J = 3.6 Hz, 2H), 4.63 (d, J = 12.9 Hz, 1H), 4.21 (m, 2H), 3.78 (t, J = 5.4 Hz, 2H), 3.56 (d, J = 13.3 Hz, 1H), 3.13 (t, J = 12.9 Hz, 1H), 2.86 (t, J = 12.7 Hz, 1H), 2.59 (d, J = 13.6 Hz, 1H), 2.45 (s, 3H), 2.43 (s, 3H), 2.36 - 2.30 (m, 3H), 1.70 (d, J = 12.0 Hz, 1H), 1.53 (d, J = 12.0 Hz, 1H).

[0267] Example 4: Synthesis of compound 4

[0268]

[0269] Step 1: Preparation of compound 4-2

[0270] To a solution of compound 4-1 (3.80 g, 33.31 mmol) and PMBC1 (7.85 g, 49.97 mmol) in DMF (40.0 mL) was added potassium carbonate (6.90 g, 49.97 mmol) at room temperature. The reaction mixture was stirred at 60 °C for 5 h, then cooled, quenched with water (50 mL), stirred for 5 min with petroleum ether: ethyl acetate (5:1) 40 mL and filtered. The filter cake was washed with petroleum ether: ethyl acetate (5:1) to give compound 4-2 (3.90 g) in 50% yield.

[0271] Step 2: Preparation of compound 4-3

[0272] To a solution of compound 4-2 (3.40 g, 14.52 mmol) in a mixture of methanol and water (40 mL, 3:1) was added iron powder (2.44, 43.55 mmol) and ammonium chloride (3.92 g, 72.58 mmol). The reaction mixture was stirred at 70 °C for 2 h, then cooled, filtered through celite, and the filtrate was concentrated to dryness in vacuo. The residue was dissolved in ethyl acetate, washed with saturated brine, separated, dried, and the solvent was removed in vacuo to give compound 4-3 (2.55 g) in 86.0% yield. LC-MS (ESI) [M+H] + : 205.2.

[0273] Step 3: Preparation of compound 4-4

[0274] To a solution of compound 4-3 (2 g, 9.79 mmol), 4-(1-methoxy-1,3-dioxolan-2-yl)piperazine-1- carboxylic acid tert-butyl ester (3.5 g, 11.13 mmol) and TsOH (168.44 mg, 979.30 μmol) in EtOH (35 mL) was added. The reaction mixture was heated to 100 °C and stirred for 12 h. TLC showed that the starting material was consumed. The reaction mixture was quenched with water and the product was extracted with ethyl acetate. The organic phase was washed with aqueous sodium carbonate, separated, dried, and concentrated to dryness in vacuo to give compound 4-4 (4.91 g). The residue was used directly in the next step. The crude product was obtained in 100% yield. LC-MS (ESI) [M+H] + : 501.2.

[0275] Step 4: Preparation of compound 4-5

[0276] Compound 4-4 (4 g, 7.99 mmol) was dissolved in diphenyl ether: dichloromethane (5 mL, 4: 1), diphenyl ether (43 mL) was added slowly dropwise to the solution which was previously heated to an internal temperature of 220 °C, and the system was stirred at 220 °C for 4.5 minutes. After the system was cooled to room temperature, the reaction was directly quenched with cyclohexane, and the solid was filtered and washed with cyclohexane to obtain compound 4-5 (1.1 g) with a yield of 29.4%. LC-MS (ESI) [M+H] + : 469.2.

[0277] Step 5: Preparation of compound 4-6

[0278] Compound 4-5 (1 g, 2.13 mmol) was dissolved in DMF (7 mL), ethyl bromoacetate (712.84 mg, 4.27 mmol) and cesium carbonate (2.10 g, 6.40 mmol) were added to the solution, and the system was stirred at room temperature for 2 hours. The system was directly quenched with water, and the product was extracted with ethyl acetate, the organic phase was separated and dried, and then concentrated to dryness under vacuum. The residue was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain compound 4-6 (0.27 g) with a yield of 22.9%. LC-MS (ESI) [M+H] + : 555.2.

[0279] Step 6: Preparation of compound 4-7

[0280] Compound 4-6 (170 mg, 306.51 μmol) was dissolved in DCM (2 mL), and TFA (1 mL) was added to the solution at room temperature, and the system was stirred at room temperature for 0.5 hours. TLC detection showed that the raw material disappeared. The system was directly concentrated to dryness under vacuum. The residue was directly used in the next step, and the crude product yield was 100%. LC-MS (ESI) [M+H] + : 455.2.

[0281] Step 7: Preparation of compound 4-8

[0282] Compound 4-7 (150 mg, 330.02 μmol) was dissolved in TFA (2 mL), and TfOH (0.05 mL) was added to the solution, and the system was stirred at room temperature for 10 minutes. TLC detection showed that the raw material disappeared. The system was directly concentrated to dryness under vacuum. The residue was directly used in the next step, and the crude product yield was 100%. LC-MS (ESI) [M+H] + : 335.2.

[0283] Step 8: Preparation of compound 4-9

[0284] Compound 4-8 (200 mg, 598.13 μmol), 5-(benzyloxy)-6-methylpyrimidine-4- carboxylic acid (146.09 mg, 598.13 μmol) were dissolved in DMF (3.96 mL), to which DIEA (773.05 mg, 5.98 mmol, 1.04 mL) and HATU (293.36 mg, 777.58 μmol) were added, after which the system was stirred at room temperature for 2 hours. The system was directly quenched with water, and the byproduct was extracted with ethyl acetate, and the aqueous phase was washed with citric acid, and after the organic phase was separated, dried, and concentrated to dryness under vacuum. The residue was purified by HPLC to obtain compound 4-9 (110 mg) at a yield of 32.83%. LC-MS (ESI) [M+H] + : 561.2.

[0285] Step 9: Preparation of compound 4-10

[0286] Compound 4-9 (16 mg, 28.54 μmol), (3,6-dihydro-2H-pyran-4-yl)boronic acid (12.78 mg, 99.89 μmol), PhenCuPPh3Br (6.70 mg, 11.42 μmol) were dissolved in DMSO (1 mL), after which the system was stirred at 90°C under air ball protection for 3 hours. The system was directly quenched with water, and the product was extracted with ethyl acetate, and the organic phase was separated, dried, and concentrated to dryness under vacuum. The residue was purified by a medium-pressure preparative column (DCM:MeOH=10:1) to obtain compound 4-10 (11 mg) at a yield of 59.88%. LC-MS (ESI) [M+H] + : 643.2. Step 10: Preparation of compound 4-11

[0287] Compound 4-10 (17 mg, 26.45 μmol) was dissolved in a mixed solvent of water (1.5 mL), MeOH (1.5 mL), and THF (1.5 mL), to which NaOH (10.58 mg, 264.51 μmol) was added, after which the system was stirred at room temperature for 0.25 hours. The pH was adjusted to 4.0 with 3N hydrochloric acid, and the product was extracted with ethyl acetate, and the organic phase was separated, dried, and rotary evaporated to obtain crude product 4-11 (16 mg) at a crude product yield of 100%. LC-MS (ESI) [M+H] + : 615.2.

[0288] Step 11: Preparation of compound 4-12

[0289] Compound 4-11 (16 mg, 26.05 μmol), 2-chloro-4-trifluoromethylaniline (15.20 mg, 78.17 μmol), pyridine (20.37 mg, 261 μmol) were dissolved in DCM (0.5 mL), to which phosphorous oxychloride (20.29 mg, 130 μmol) was added, after addition, the system was stirred at room temperature for 0.5 hours. TLC showed that the product point, the system was directly quenched with water, the product was extracted with ethyl acetate, the organic phase was separated, dried and concentrated to dryness under vacuum. The residue was purified by medium pressure preparative column (dichloromethane:methanol = 10:1) to obtain compound 4-12 (16 mg) with a yield of 77.88%. LC-MS (ESI) [M+H] + : 792.2.

[0290] Step 12: Preparation of compound 4

[0291] Compound 4-12 (18 mg, 22.75 μmol) was dissolved in DCM (2 mL), to which boron trichloride (180.00 μL, IN) was added at room temperature, after addition, the system was stirred at room temperature for 0.25 hours. The system was directly quenched with methanol, the residue was purified by preparative HPLC to obtain compound 4 crude product (10.63 mg, containing isomers), the crude product 4 was further purified by SFC (column ChiralPak AD, 250 x 30 mm I.D., 10 μm; mobile phase [A: carbon dioxide, B: isopropanol (containing 0.1% ammonia water)]; component B%: 40%, flow rate: 150 mL / min, column temperature: 38 °C, wavelength: 220 nm, cycle time: ~ 5 min) to obtain compound 4 (4.11 mg) with a yield of 38.66% (the retention time of the target product was 0.788 minutes, the analysis method: column: ChiralPak AD, 50 x 4.6 mm I.D., 3 μm, mobile phase [A: carbon dioxide, B: isopropanol (containing 0.05% DEA)], 40% B, flow rate: 3 mL / min, column temperature: 35 °C). LC-MS (ESI) [M+H]+: 702.2.

[0292] 1H NMR (400 MHz, Methanol-d4) δ 8.31 (s, 1H), 8.04 (d, J = 8.8 Hz, 1H), 7.74-7.68 (m, 1H), 7.55-7.47 (m, 1H), 6.79-6.73 (m, 1H), 5.29 (s, 2H), 4.30-4.22 (m, 2H), 3.94-3.85 (m, 2H), 3.84-3.72 (m, 2H), 3.42-3.28 (m, 1H), 3.10-2.95 (m, 3H), 2.81 (s, 3H), 2.71-2.60 (m, 1H), 2.38 (s, 3H), 2.09 (t, J = 7.6 Hz, 1H), 1.99-1.87 (m, 1H), 0.80 (t, J = 6.4 Hz, 3H).

[0293] Example 5: Synthesis of compound 5

[0294]

[0295] Step 1: Synthesis of compound 5-2

[0296] Compound 5-1 (1.0 g, 8.84 mmol) in THF (30 mL) was added dropwise to a solution of NaH (1.06 g, 26.54 mmol, 60% purity) in THF (30 mL) at 0 °C, stirred for 0.5 h at 0 °C, dropwise added 2- (trimethylsilyl)ethoxymethyl chloride (1.77 g, 10.62 mmol, 1.88 mL) mixture continued to react for 1.5 h at 0 °C. The mixture was slowly added to saturated ammonium chloride solution (50 mL), extracted with ethyl acetate (50 mL x 2), the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated to give compound 5-2 (2.1 g, 8.63 mmol) with a yield of 97.58%.

[0297] Step 2: Synthesis of compound 5-3

[0298] Compound 5-2 (1.0 g, 4.11 mmol) was dissolved in MeOH (10 mL), Pd / C (200 mg, 10% purity) mixture was reacted under hydrogen for 4 h at 25 °C until the raw material was completely reacted. The mixture was filtered and concentrated under reduced pressure to give compound 5-3 (870 mg, 4.08 mmol) with a yield of 99.2%. LC-MS (ESI) [M+H] + 214.2.

[0299] Step 3: Synthesis of compound 5-4

[0300] Compound 5-3 (1.0 g, 4.69 mmol), compound 4-(1-methoxy-1,3-dioxolan-2-yl)piperazine-1- carboxylic acid tert-butyl ester (1.47 g, 4.69 mmol) were dissolved in toluene (10 mL), p-toluenesulfonic acid (80.71 mg, 468.71 μmol) was added, the mixture was reacted at 120 °C for 16 hours until the raw material was completely reacted, the mixture was poured into saturated sodium bicarbonate solution (30 mL), extracted with ethyl acetate (30 mL x 2), the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give compound 5-4 (1.28 g) with a yield of 53.58%. LC-MS (ESI) [M+H] + : 510.4.

[0301] Step 4: Synthesis of compound 5-5

[0302] Compound 5-4 (680 mg, 1.33 mmol) was added to diphenyl ether (8 mL) heated to 280 °C under reflux state for 5 minutes until the product was generated. The mixture was cooled to room temperature, poured into cyclohexane, stirred, and filtered. The filtrate was concentrated and the filter cake was separated by column chromatography (PE / EA = 10 / 1) to give compound 5-5 (100 mg) with a yield of 15.69%. LC-MS (ESI) [M+H] + : 478.4.

[0303] Step 5: Synthesis of compound 5-6

[0304] Compound 5-5 (160 mg, 334.96 μmol) was dissolved in ACN (3 mL), cesium carbonate (327.41 mg, 1.00 mmol), ethyl bromoacetate (83.91 mg, 502.44 μmol) were added, and the mixture was reacted at 25 °C for 1 hour until the reaction was complete. The reaction mixture was poured into water (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (PE / EA = 1 / 1) to give compound 5-6 (83 mg) with a yield of 43.9%. LC-MS (ESI) [M+H] + : 564.4.

[0305] Step 6: Synthesis of compound 5-7

[0306] Compound 5-6 (30 mg, 53.21 μmol) was dissolved in MeOH (3 mL), HCl (4 M, 4.00 mmol, 1 mL) was added, and the mixture was reacted at 25 °C for 4 hours until the raw material was completely reacted. The mixture was poured into a saturated sodium bicarbonate solution (10 mL), extracted with ethyl acetate (10 mL x 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain compound 5-7 (24 mg) with a yield of 97.3%. LC-MS (ESI) [M+H] + : 464.3.

[0307] Step 7: Synthesis of compound 5-8

[0308] Compound 5-7 (177.00 mg, 724.69 μmol) was dissolved in DMF (5 mL), HATU (455.67 mg, 1.21 mmol), DIEA (234.15 mg, 1.81 mmol, 315.57 μL), and compound 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (280 mg, 603.91 μmol) were added, and the resulting mixture was reacted at 25 °C for 2 hours until the reaction was complete. The reaction mixture was poured into water (10 mL), extracted with ethyl acetate (10 mL x 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain compound 5-8 (360 mg) with a yield of 86.4%. LC-MS (ESI) [M+H] + : 690.2.

[0309] Step 8: Synthesis of compound 5-9

[0310] Compound 5-8 (350 mg, 507.34 μmol) was dissolved in a solution of HCl (4 M, 24.00 mmol, 6 mL) in dioxane (6 mL), and the mixture was reacted at 25 °C for 4 hours until the raw material was completely reacted. The reaction mixture was poured into a saturated sodium bicarbonate solution (15 mL), extracted with ethyl acetate (15 mL x 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 5-9 (200 mg) with a yield of 70.4%. LC-MS (ESI) [M+H] + : 690.2.

[0311] Step 9: Synthesis of compound 5-10

[0312] Compound 5-9 (300 mg, 536.08 μmol) was dissolved in DMSO (1 mL), compound (3,6-dihydro-2H-pyran-4-yl)boronic acid (102.87 mg, 804.12 μmol) and PhenCuPPh (30.45 mg, 53.61 μmol) were added, and the resulting reaction system was stirred under air until the reaction was complete. The reaction solution was quenched with water, extracted with ethyl acetate (5 mL x 2), and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was purified by silica gel column chromatography (MeOH / DCM = 3%) to give compound 5-10 (135 mg, 204.06 μmol) with a yield of 38.1%. LC-MS (ESI) [M+H] + : 642.2.

[0313] Step 10: Synthesis of compound 5-11

[0314] Compound 5-10 (130.00 mg, 202.58 μmol) was dissolved in MeOH (0.5 mL), THF (1.5 mL), and water (0.5 mL), and LiOH (16.98 mg, 709.04 μmol) was added. The resulting mixture was reacted at 25°C for 1 hour until the starting material was completely reacted. The mixture was adjusted to pH < 4 with hydrochloric acid, extracted with ethyl acetate (10 mL x 2), and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give compound 5-11 (120 mg, 195.55 μmol) with a yield of 96.53%. LC-MS (ESI) [M+H] + : 612.2.

[0315] Step 11: Synthesis of compound 5-12

[0316] Compound 5-11 (110.00 mg, 179.25 μmol), 2-chloro-4-trifluoromethylaniline (52.58 mg, 268.88 μmol), and pyridine (70.89 mg, 896.26 μmol, 72.20 μL) were dissolved in DCM (1.93 mL), and phosphorus oxychloride (41.23 mg, 268.88 μmol) was added. The resulting reaction mixture was stirred at 25°C for 1 hour until the starting material was completely reacted. The reaction was quenched with water (5 mL), and the product was extracted with ethyl acetate. The organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under vacuum to give a crude product. The crude product was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 5-12 (130 mg, 164.30 μmol) with a yield of 91.66%. LC-MS (ESI) [M+H] + : 791.2.

[0317] Step 12: Synthesis of compound 5

[0318] Compound 5-12 (60 mg, 75.83 μmol) was dissolved in DCM (2 mL), and boron trichloride (88.85 mg, 758.33 μmol) was added. The mixture was reacted at 25 °C for 0.5 hours until the raw material was completely reacted. The mixture was separated by HPLC preparation to obtain compound 5 (5 mg, 7.13 μmol) with a yield of 9.40%. LC-MS (ESI) [M+H] + : 701.2.

[0319] 1 H NMR (400 MHz, DMSO-d6) δ 10.24 (s, 1H), 10.20 (s, 1H), 8.65 (s, 1H), 8.58 (s, 1H), 8.15 (d, J = 8.5 Hz, 1H), 7.98 (d, J = 2.0 Hz, 1H), 7.72 (d, J = 8.6 Hz, 1H), 6.41 (d, J = 2.4 Hz, 1H), 5.16 (s, 2H), 4.49 (d, J = 12.2 Hz, 1H), 4.28 (d, J = 3.0 Hz, 2H), 3.89 (t, J = 5.5 Hz, 2H), 3.73 (q, J = 13.1 Hz, 2H), 3.46 (d, J = 12.5 Hz, 1H), 3.19 (t, J = 12.3 Hz, 1H), 2.93 (t, J = 12.2 Hz, 3H), 2.71 (s, 3H), 2.44 (s, 3H), 2.00 (q, J = 6.9, 6.3 Hz, 1H), 1.17 (t, J = 7.4 Hz, 3H).

[0320] Example 6: Synthesis of compound 6

[0321]

[0322] Step 1: Preparation of compound 6-1

[0323] Intermediate 5-7 (160 mg, 345.83 μmol) and 3-(benzyloxy)picolinic acid (87.20 mg, 380.41 μmol) were dissolved in N,N-dimethylformamide (1 mL), 2-(7-azabenzotriazol-1- yl)-1,1,3,3-hexafluorophosphonic 6-oxide hexahydrate (157.79 mg, 414.99 μmol) and N,N-diisopropyl ethylamine (134.09 mg, 1.04 mmol, 180.71 μL) were added successively, and the resulting reaction system was stirred at 25 °C for 1 h. LC-MS showed that the reaction was completed. The reaction solution was directly concentrated under reduced pressure, and the resulting residue was purified by silica gel column (DCM / MeOH = 5%) to give compound 6-1 (210 mg) with a yield of 90.11%. LC-MS (ESI) [M+H] + : 675.2.

[0324] Step 2: Preparation of compound 6-2

[0325] Compound 6-1 (210.31 mg, 311.63 μmol) was dissolved in a solution of HCl in dioxane (4 M, 4.00 mmol, 1 mL), and the resulting reaction system was stirred at 25 °C for 1 h. LC-MS showed that the reaction was completed. The reaction solution was cooled to 25 °C, the pH was adjusted to neutral with saturated sodium bicarbonate, extracted with ethyl acetate (5 mL x 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude 6-2 (150 mg) with a yield of 75.13%. LC-MS (ESI) [M+H] + : 545.2.

[0326] Step 3: Preparation of compound 6-3

[0327] Compound 6-2 (67 mg, 123.03 μmol) was dissolved in dimethyl sulfoxide (0.5 mL), 3,6-dihydro-2H-pyran-4-boronic acid (23.61 mg, 184.54 μmol) and PhenCuPPhBr2 (3.49 mg, 6.15 μmol) were added, and the resulting reaction solution was stirred under air for about 12 h until the raw material was completely reacted. The reaction solution was quenched with water (5 mL), extracted with ethyl acetate (5 mL x 2), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by silica gel column (MeOH / DCM = 3%) to give compound 6-3 (43 mg) with a yield of 52.98%. LC-MS (ESI) [M+H] + : 627.2.

[0328] Step 4: Preparation of compound 6-4

[0329] Compound 6-3 (41 mg, 62.15 μmol) was dissolved in methanol (0.5 mL), tetrahydrofuran (0.5 mL), and a solution of lithium hydroxide (4.47 mg, 186.45 μmol) in water (0.2 mL) was added. The resulting reaction system was stirred at room temperature for 5 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to remove the organic solvent, and the remaining aqueous phase was adjusted to neutral pH with 1M dilute hydrochloric acid, extracted with ethyl acetate (5 mL x 2), and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude 6-4 (35 mg) which was directly used in the next step. The crude yield was 83.72%. LC-MS (ESI) [M+H] + : 599.2.

[0330] Step 5: Preparation of compound 6-5

[0331] Intermediate 6-4 (25 mg, 37.17 μmol), 3-chloro-4-aminobenzotrifluoride (10.90 mg, 55.75 μmol), and pyridine (14.70 mg, 185.84 μmol, 14.97 μL) were dissolved in dichloromethane (0.5 mL), and phosphorus oxychloride (17.10 mg, 111.50 μmol) was added. The resulting reaction system was stirred at 25°C for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column (DCM / MeOH = 50:1) to obtain compound 6-5 (23 mg) with a yield of 70.64%. LC-MS (ESI) [M+H] + : 776.4.

[0332] Step 6: Preparation of compound 6

[0333] Intermediate 6-5 (25 mg, 28.54 μmol) was dissolved in dichloromethane (0.5 mL), and boron trichloride (16.72 mg, 142.68 μmol) was added. The resulting reaction system was stirred at 25°C for 1 hour, and LC-MS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by HPLC preparation to obtain compound 6 (2.16 mg) with a yield of 11.03%. LC-MS (ESI) [M+H] + : 686.2.

[0334] 1H NMR (400 MHz, DMSO-d6) δ 10.18 (s, 1H), 8.63 (s, 1H), 8.15 (d, J = 8.8 Hz, 1H), 8.07 - 8.04 (m, 1H), 7.97 (d, J = 1.9 Hz, 1H), 7.72 (d, J = 9.0 Hz, 1H), 7.28 (d, J = 3.8 Hz, 2H), 6.41 (s, 1H), 5.16 (s, 2H), 4.52 (d, J = 12.3 Hz, 1H), 4.28 (d, J = 2.9 Hz, 2H), 3.89 (t, J = 5.4 Hz, 2H), 3.73 (t, J = 14.6 Hz, 2H), 3.18 (q, J = 12.7 Hz, 3H), 2.91 (q, J = 12.0, 9.3 Hz, 3H), 2.71 (tt, J = 6.6, 2.9 Hz, 4H), 1.17 (t, J = 7.4 Hz, 3H).

[0335] Example 7: Synthesis of compound 7

[0336]

[0337] Step 1: Preparation of compound 7-2

[0338] Compound 7-1 (23 g, 150 mmol) was dissolved in dimethyl sulfoxide (200 mL) at room temperature, sodium methoxide (135 g, 750 mmol) was added, the reaction was stirred at 120 °C for 16 hours under nitrogen protection, water was added and extracted with ethyl acetate for several times, the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was slurried with pure ethyl acetate to obtain the target compound 7-2 (11.9 g) with a yield of 43.28%. LC-MS (ESI) [M+H] + : 150.1.

[0339] Step 2: Preparation of compound 7-3

[0340] Intermediate 7-2 (11.9 g, 80 mmol) was dissolved in N-methyl pyrrolidone (240 mL) at room temperature, and m-chloroperbenzoic acid (32 g, 184 mmol) was added. The reaction was carried out at room temperature for 16 hours until the reaction was complete, methyl tert-butyl ether solution (60 mL) was added, and after stirring for ten minutes, the liquid was filtered off, washed with ethyl acetate, and the filter cake was collected. The filter cake was dissolved in methyl tert-butyl ether solution (200 mL), stirred for 30 minutes, and then the liquid was filtered off, washed with ethyl acetate, and the obtained solid was dried to obtain compound 7-3 (11.9 g) with a yield of 100%. LC-MS (ESI) [M+H] + : 166.1.

[0341] Step 3: Preparation of compound 7-4

[0342] Intermediate 7-3 (10.9 g, 66 mmol) was dissolved in phosphorus oxychloride (100 mL) at room temperature and reacted at 50 °C for 16 hours until the reaction was completed. The solvent was distilled off, the solution was adjusted to be alkaline with sodium hydroxide in an ice bath, extracted with ethyl acetate and water for several times, the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was separated and purified by column chromatography (silica, petroleum ether / ethyl acetate = 2 / 1) to obtain compound 7-4 (11.0 g) with a yield of 90.78%. LC-MS (ESI) [M+H] + : 183.9.

[0343] Step 4: Preparation of compound 7-5

[0344] Intermediate 7-4 (11 g, 60 mmol) was dissolved in tetrahydrofuran (110 mL) at room temperature, and then 3,4-dihydro-2H-pyran (7.6 g, 90 mmol) and p-toluenesulfonic acid monohydrate (1.1 g, 6 mmol) were added. The obtained reaction solution was reacted at room temperature for 16 hours until the reaction was completed, diluted with water (100 mL), extracted with ethyl acetate (200 mL x 3), and the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was separated and purified by column chromatography (silica, petroleum ether / ethyl acetate = 2 / 1) to obtain compound 7-5 (14.8 g) with a yield of 92.27%. LC-MS (ESI) [M+H] + : 267.9.

[0345] Step 5: Preparation of compound 7-6

[0346] Intermediate 7-5 (14 g, 52 mmol) and pinacol vinylboronate (12 g, 78 mmol) were dissolved in a mixed solution of 1,4-dioxane (70 mL) and water (35 mL) at room temperature, and then methyl sulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl) (2-amino-1,1'-biphenyl-2-yl) palladium (II) (1.3 g, 1.56 mmol) and cesium fluoride (20 g, 130 mmol) were added. After the reaction mixture was stirred at 110 °C for 16 hours, LC-MS showed that the reaction was completed. Water was added for dilution (50 mL), extracted with ethyl acetate (200 mL x 2), and the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (silica, petroleum ether / ethyl acetate = 5 / 1) to obtain compound 7-6 (13.2 g) with a yield of 98%. LC-MS (ESI) [M+H] + : 260.0.

[0347] Step 6: Preparation of compound 7-7

[0348] Intermediate 7-6 (13.2 g, 51 mmol) was dissolved in methanol (130 mL) at room temperature, and 10% palladium carbon catalyst (5 g) was added. The reaction was stirred at room temperature for 16 hours under hydrogen atmosphere until the reaction was completed. The reaction was concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (silica, petroleum ether / ethyl acetate = 2 / 1) to obtain compound 7-7 (10.9 g) with a yield of 81.9%. LC-MS (ESI) [M+H] + : 262.1.

[0349] Step 7: Preparation of compound 7-8

[0350] Compound 7-7 (3 g, 11 mmol) was dissolved in acetonitrile (30 mL) at room temperature, and ethyl bromoacetate (19 g, 15 mmol) was added. The reaction was carried out at 70°C for 16 hours under nitrogen protection until the reaction was completed. The solvent was removed by rotary evaporation under reduced pressure, and the residue was separated and purified by reverse phase column chromatography (A: formic acid / water, B: acetonitrile) to obtain the target compound 7-8 (1.2 g) with a yield of 31.20%. LC-MS (ESI) [M+H] + : 333.9.

[0351] Step 8: Preparation of compound 7-9

[0352] Intermediate 7-8 (1.2 g, 3.6 mmol) was dissolved in N,N-dimethylformamide (10 mL) at room temperature, and N-bromosuccinimide (0.6 g, 3.6 mmol) was added. The reaction was carried out at room temperature for 16 hours until the reaction was completed. The reaction was diluted with water (50 mL) and extracted with ethyl acetate (100 mL x 3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was separated and purified by silica gel column chromatography (dichloromethane / (methanol:dichloromethane = 1:1) = 10 / 1) to obtain the target compound 7-9 (0.4 g) with a yield of 31.56%. LC-MS (ESI) [M+H] + : 411.8. Step 9: Preparation of compound 7-10

[0353] Intermediate 7-9 (0.4 g, 1.1 mmol) and N-Boc-1,2,5,6-tetrahydropyridine-4- boronic acid pinacol ester (0.7 g, 2.2 mmol) were dissolved in a mixed solvent of 1,4- dioxane (20 mL) and water (2 mL) at room temperature, and then chloro(2- dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)] palladium(II) (0.089 g, 0.1 mmol), potassium phosphate (0.7 g, 3.4 mmol) and 2- dicyclohexylphosphino-2',4',6'-triisopropyl biphenyl (0.1 g, 0.3 mmol) were added. After replacing nitrogen, the reaction mixture was stirred at 100 °C for 16 hours until the reaction was completed. Water (20 mL) was added for dilution, and extracted with ethyl acetate (100 mL x 3), and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the obtained residue was separated and purified by silica gel column chromatography (silica, dichloromethane / methanol = 10 / 1) to obtain the target compound 7-10 (0.3 g) with a yield of 57.8%. LC-MS (ESI) [M+H] + : 515.0.

[0354] Step 10: Preparation of compound 7-11

[0355] Compound 7-10 (0.3 g, 0.6 mmol) was dissolved in a mixed solution of tetrahydrofuran (2 mL) and ethanol (2 mL) at room temperature, and then Raney nickel catalyst (0.5 g) was added, and hydrogen was replaced. The reaction was carried out at room temperature for 16 hours until the reaction was completed, and then the filtrate was filtered and concentrated under reduced pressure. The obtained residue was purified and separated by C18 column chromatography (A: formic acid / water, B: acetonitrile) to obtain compound 7-11 (0.15 g) with a yield of 49.80%. LC-MS (ESI) [M+H] + : 417.9.

[0356] Step 11: Preparation of compound 7-12

[0357] Intermediate 7-11 (0.15 g, 0.29 mmol) was dissolved in tetrahydrofuran (6 mL) at room temperature, and then hydrochloric acid dioxane solution (2 ml, 1 mol / L) was added. The reaction was carried out at room temperature for 16 hours until the reaction was completed, and then dried by concentration under reduced pressure to obtain compound 7-12 (0.15 g) with a yield of 100%. LC-MS (ESI) [M+H] + : 333.1.

[0358] Step 12: Preparation of compound 7-13

[0359] Intermediate 7-12 (0.13 g, 0.25 mmol) was dissolved in N,N-dimethylformamide (1.5 mL) at room temperature, and then 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (0.067 g, 0.27 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.14 g, 0.37 mmol), and N,N-diisopropylethylamine (0.16 g, 1.26 mmol) were added. The reaction was allowed to proceed at room temperature for 16 hours until the starting material was completely consumed. Water (10 mL) was added to dilute the solution, and extraction was performed with ethyl acetate (20 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was separated and purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 7-13 (0.10 g) with a yield of 47.14%. LC-MS (ESI) [M+H] + : 559.2.

[0360] Step 13: Preparation of compound 7-14

[0361] Intermediate 7-13 (0.083 g, 0.15 mmol) was dissolved in dichloromethane (5 mL) at room temperature, and then 3,6-dihydro-2H-pyran-4-boronic acid (0.019 g, 0.15 mmol) and copper acetate monohydrate (0.045 g, 0.22 mmol) were added, followed by pyridine (0.047 g, 0.59 mmol). The reaction was allowed to proceed at room temperature for 16 hours until the reaction was completed. Water (10 mL) was added to dilute the solution, and extraction was performed with dichloromethane (20 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was separated and purified by C18 column chromatography (A: formic acid / water, B: acetonitrile) to obtain compound 7-14 (0.076 g) with a yield of 79.83%. LC-MS (ESI) [M+H] + : 641.1.

[0362] Step 14: Preparation of compound 7-15

[0363] Intermediate 7-14 (0.076 g, 0.12 mmol) was dissolved in a mixed solution of tetrahydrofuran (5 mL) and water (1 mL) at room temperature, and then lithium hydroxide (0.02 g, 0.47 mmol) was added. The reaction was allowed to proceed at room temperature for 16 hours until the reaction was completed. The solvent was removed by rotary evaporation under reduced pressure, water (10 mL) was added to dilute the solution, and the pH of the solution was adjusted to < 6 with dilute hydrochloric acid. Extraction was performed with ethyl acetate (20 mL x 2), and the combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Vacuum freeze-drying was performed to obtain compound 7-15 (0.076 g) with a yield of 100%. LC-MS (ESI) [M+H] + : 613.3.

[0364] Step 15: Preparation of compound 7-16

[0365] Intermediate 7-15 (0.056 g, 0.09 mmol), N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate (0.030 g, 0.108 mmol), 1-methylimidazole (0.011 g, 0.135 mmol) and 3-chloro-4-aminobenzotrifluoride (0.017 g, 0.09 mmol) were dissolved in acetonitrile (3 mL) at room temperature. After 1 hour of reaction at room temperature, LC-MS showed that the reaction was completed. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was purified by C18 column chromatography (A: formic acid / water, B: acetonitrile) to give compound 7-16 (0.092 g) with a yield of 100%. LC-MS (ESI) [M+H] + : 789.9.

[0366] Step 16: Preparation of compound 7

[0367] Compound 7-16 (0.09 g, 0.11 mmol) was dissolved in dichloromethane (5 mL) at room temperature, and boron trichloride (0.1 ml) was added. After replacement of nitrogen, the reaction was carried out at room temperature for 16 hours until the reaction was completed. The reaction solution was quenched by adding methanol (5 mL), and the solvent was removed by concentration under reduced pressure. Compound 7 (24.75 mg) was obtained by HPLC preparation purification with a yield of 30.1%. LC-MS (ESI) [M+H] + : 700.2.

[0368] 1 HNMR (400MHz, MeOH-d4) δ 8.60-8.54 (m, 1H), 8.42-8.33 (m, 1H), 8.21-8.15 (m, 1H), 7.82-7.76 (m, 1H), 7.66-7.58 (m, 1H), 6.59-6.48 (m, 1H), 5.38-5.31 (m, 2H), 4.85-4.78 (m, 1H), 4.36-4.31 (m, 2H), 4.20-4.09 (m, 1H), 4.00-3.94 (m, 2H), 3.31-3.22 (m, 1H), 3.11-2.79 (m, 6H), 2.77-2.72 (m, 2H), 2.55-2.50 (m, 3H), 1.74-1.50 (m, 2H), 1.37-1.31 (m, 3H).

[0369] Example 8: Synthesis of compound 8

[0370] HNMR (400MHz, MeOH-d4) δ 8.60-8.54 (m, 1H), 8.42-8.33 (m, 1H), 8.21-8.15 (m, 1H), 7.82-7.76 (m, 1H), 7.66-7.58 (m, 1H), 6.59-6.48 (m, 1H), 5.38-5.31 (m, 2H), 4.85-4.78 (m, 1H), 4.36-4.31 (m, 2H), 4.20-4.09 (m, 1H), 4.00-3.94 (m, 2H), 3.31-3.22 (m, 1H), 3.11-2.79 (m, 6H), 2.77-2.72 (m, 2H), 2.55-2.50 (m, 3H), 1.74-1.50 (m, 2H), 1.37-1.31 (m, 3H).

[0371] Step 1: Preparation of compound 8-1

[0372] Compound 4-11 (21 mg, 34.17 μmol), 2-methyl-4-trifluoromethylaniline (17.95 mg, 102.50 μmol), pyridine (27.03 mg, 341.66 μmol, 27.52 μL) were dissolved in DCM (1.5 mL), and phosphorus oxychloride (26.19 mg, 170.83 μmol) was added at room temperature. After the addition was completed, the system was stirred at room temperature for 1 hour until the reaction was complete. The reaction solution was quenched with water (5 mL), and the product was extracted with ethyl acetate (10 mL x 2). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 19:1) to obtain compound 8-1 (26 mg) with a yield of 65.0%. LC-MS (ESI) [M+H] + : 772.3.

[0373] Step 2: Preparation of compound 8

[0374] Compound 8-1 (20 mg, 25.91 μmol) was dissolved in DCM (2 mL), and a solution of boron trichloride in hexane (0.2 mL, 1 M) was added at room temperature. After stirring at room temperature for 1 hour until the reaction was complete, methanol (5 mL) was added to quench the reaction. The solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by HPLC preparation to obtain compound 8 crude product (12 mg, containing isomers). The crude product was further purified by SFC preparation (column: ChiralPak AD, 250 x 30 mm I.D., 10 μm, mobile phase: A: CO2; B: isopropanol (0.1% NH3H2O), component: B 40%, flow rate: 150 mL / min, back pressure: 100 bar, column temperature: 38°C, wavelength: 220 nm, cycle time: ~ 5 min) to obtain compound 8 (4.33 mg, yield 24.5%). The analysis method of the target product is as follows: column: ChiralPak AD, 50 x 4.6 mm I.D., 3 μm, mobile phase: A: CO2, B: isopropanol (0.05% DEA), component: B 40%, flow rate: 3 mL / min, back pressure: 100 bar, column temperature: 35°C, wavelength: 220 nm, LC-MS (ESI) [M+H] + : 682.3.

[0375] 1H NMR (400 MHz, MeOD-d4) d 8.41 (s, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.51 (s, 1H), 7.42 (d, J = 8.6 Hz, 1H), 6.81 (s, 1H), 5.28 (s, 2H), 4.33 (s, 2H), 4.00 - 3.74 (m, 4H), 3.15 - 3.02 (m, 4H), 2.86 (s, 3H), 2.76 - 2.65 (m, 1H), 2.44 (s, 3H), 2.33 (s, 3H), 2.19 - 2.09 (m, 1H), 2.03 - 1.93 (m, 1H), 0.84 (t, J = 6.8 Hz, 3H).

[0376] Example 9: Synthesis of compound 9

[0377]

[0378] Step 1: Preparation of compound 9-1

[0379] Compound 4-11 (30 mg, 48.81 μmol), 2-fluoro-4-trifluorophenylamine (34.97 mg, 195.23 μmol), pyridine (38.61 mg, 488.08 μmol, 39.32 μL) were dissolved in dichloromethane (1 mL), and phosphorus oxychloride (37.42 mg, 244.04 μmol) was added at room temperature. The reaction was stirred at room temperature for 1 hour until the reaction was complete. The reaction was quenched by pouring into water (5 mL), extracted with ethyl acetate (10 mL x 2), and the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated to dryness in vacuo. The residue was purified by preparative silica gel column chromatography (DCM:MeOH = 19:1) to obtain compound 9-1 (27.7 mg) with a yield of 73.28%. LC-MS (ESI) [M+H] + 776.2.

[0380] Step 2: Preparation of compound 9

[0381] Compound 9-1 (30 mg, 38.67 pmol) was dissolved in DCM (3 mL), and boron trichloride (0.3 mL, 1 M) in n-hexane was added at room temperature. The reaction was stirred at room temperature for 0.6 hours until the reaction was complete, and then methanol (5 mL) was added to quench the reaction. The solvent was evaporated to dryness, and the residue was purified by HPLC to obtain the crude compound 9 (15 mg, containing isomers), which was further purified by SFC (column ChiralPak AD, 250 x 30 mm I.D., 10 pm; mobile phase [A: carbon dioxide, B: ethanol (containing 0.1% ammonia water)]; component B%: 30%, flow rate: 150 mL / min, column temperature: 38 °C, wavelength: 220 nm, cycle time: ~ 9 min) to obtain compound 9 (5.08 mg) with a yield of 33.8%. (The product peak had a retention time of 0.637 minutes, and the SFC analytical method was determined as follows: column: ChiralPak AD, 50 x 4.6 mm I.D., 3 pm; mobile phase [A: carbon dioxide, B: ethanol (containing 0.05% DEA)], 5-40% B; flow rate: 3 mL / min; column temperature: 35 °C). LC-MS (ESI) [M+H] + : 686.2.

[0382] 1 HNMR (400 MHz, Methanol-d4) δ 8.42-8.20 (m, 2H), 7.57 (d, J = 12.0 Hz, 1H), 7.47 (d, J = 8.4 Hz, 1H), 6.88-6.79 (m, 1H), 5.34 (s, 2H), 4.36 (s, 2H), 4.09-3.81 (m, 4H), 3.60-3.40 (m, 1H), 3.22-3.06 (m, 3H), 2.89 (s, 3H), 2.84-2.68 (m, 1H), 2.47 (s, 3H), 2.24-2.14 (m, 1H), 2.10-1.98 (m, 1H), 1.33-1.28 (m, 3H).

[0383] Example 10: Synthesis of compound 10

[0384]

[0385] Step 1: Preparation of compound 10-1

[0386] Intermediate 7-10 (200 mg, 0.39 mmol) was dissolved in tetrahydrofuran (6 mL) at room temperature, and hydrochloric acid dioxane solution (2 mL, 1 M) was added. After the reaction was completed at room temperature for 2 hours, LC-MS showed that the reaction was complete. The solvent was removed by rotary evaporation under reduced pressure to obtain compound 10-1 (200 mg) with a yield of 100%. LC-MS (ESI) [M+H]+ :331.1.

[0387] Step 2: Preparation of compound 10-2

[0388] Intermediate 10-1 (200 mg, 0.39 mmol) was dissolved in N,N-dimethylformamide (1.5 mL) at room temperature, then 5-(benzyloxy)-6-methylpyrimidine-4-carboxylic acid (95.2 mg, 0.39 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (222.2 mg, 0.585 mmol) and N,N-diisopropylethylamine (251.6 mg, 1.95 mmol) were added. After 1 hour of reaction at room temperature, LC-MS showed that the reaction was completed. Diluted with water (10 mL), extracted with ethyl acetate (10 mL x 2), the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (silica, dichloromethane / methanol = 10 / 1) to give compound 10-2 (129 mg) with a yield of 59.4%. LC-MS (ESI) [M+H] + :557.1.

[0389] Step 3: Preparation of compound 10-3

[0390] Intermediate 10-2 (69 mg, 0.124 mmol) was dissolved in dichloromethane (5 mL) at room temperature, then 3,6-dihydro-2H-pyran-4-boronic acid (23.8 mg, 0.186 mmol) and copper acetate monohydrate (49.9 mg, 0.25 mmol) were added, and triethylamine (37.6 mg, 0.372 mmol) was added. After 16 hours of reaction at room temperature, LC-MS showed that the reaction was completed. Diluted with water (10 mL), extracted with dichloromethane (20 mL x 2), the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was separated and purified by C18 column chromatography (acetonitrile / water = 45%, formic acid system) to give compound 10-3 (50 mg) with a yield of 63.1%. LC-MS (ESI) [M+H] + :639.1.

[0391] Step 4: Preparation of compound 10-4

[0392] Intermediate 10-3 (50 mg, 0.078 mmol) was dissolved in a mixture of tetrahydrofuran (5 mL) and water (1 mL) at room temperature, and lithium hydroxide monohydrate (13.1 mg, 0.31 mmol) was added. After the reaction was carried out at room temperature for 0.5 h, LC-MS showed that the reaction was complete. The solution was adjusted to pH 3 with dilute hydrochloric acid (1 M), extracted with ethyl acetate (10 mL x 2), and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and freeze-dried in vacuum to obtain compound 10-4 (46 mg) with a yield of 96.6%. LC-MS (ESI) [M+H] + : 611.2.

[0393] Step 5: Preparation of compound 10-5

[0394] Intermediate 10-4 (40 mg, 0.0655 mmol) and 3-chloro-4-aminobenzotrifluoride (38.4 mg, 0.196 mmol) were dissolved in pyridine (1 mL) at room temperature, and 4 drops of phosphorus oxychloride were added dropwise. The reaction was carried out at room temperature for 0.5 h until the reaction was complete, diluted with water (5 mL), extracted with ethyl acetate (10 mL x 2), and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by C18 column chromatography (acetonitrile / water = 68%, formic acid system) to obtain the target compound 10-5 (20 mg) with a yield of 38.7%. LC-MS (ESI) [M+H] + : 788.2.

[0395] Step 6: Preparation of compound 10

[0396] Compound 10-5 (20 mg, 0.025 mmol) was dissolved in dichloromethane (5 mL) at room temperature, and boron trichloride (0.1 mL, 1 M) in dichloromethane was added. After replacement of nitrogen and reaction at room temperature for 2 h, LC-MS showed that the reaction was complete. Methanol (2 mL) was added to quench, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by HPLC preparation to obtain compound 10 (5.02 mg) with a yield of 28.8%. LC-MS (ESI) [M+H] + : 698.2.

[0397] 1H NMR (400 MHz, DMSO-d6) δ 10.28 (s, 2H), 8.61 (d, J = 5.8 Hz, 1H), 8.53 (s, 1H), 8.07 (d, J = 8.6 Hz, 1H), 7.95 (s, 1H), 7.71 (d, J = 8.8 Hz, 1H), 6.46 (s, 1H), 5.52 (d, J = 66.2 Hz, 1H), 5.31 - 5.08 (m, 2H), 4.40 - 3.94 (m, 4H), 3.85 (t, J = 5.4 Hz, 2H), 3.72 - 3.39 (m, 1H), 2.82 - 2.60 (m, 3H), 2.45 - 2.35 (m, 5H), 2.16 - 1.94 (m, 2H), 1.17 (t, J = 7.5 Hz, 3H).

[0398] Example 11: Synthesis of compound 11

[0399]

[0400] Step 1: Preparation of compound 11-1

[0401] Compound 5-8 (700 mg, 1.01 mmol) was dissolved in a mixture solvent of MeOH (3 mL), THF (9 mL), water (3 mL), and then LiOH (85.05 mg, 3.55 mmol) was added. The resulting mixture was stirred at 25 °C for about 1 hour until the reaction was completed. The mixture was adjusted to pH < 4 with dilute hydrochloric acid, extracted with ethyl acetate (20 mL x 2), the organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give compound 11-1 (650 mg) with a yield of 96.8%. LC-MS (ESI) [M+H] + : 662.4.

[0402] Step 2: Preparation of compound 11-2

[0403] Compound 11-1 (350 mg, 528.84 μmol), 2-chloro-4-trifluoromethylaniline (155.14 mg, 793.27 μmol), and pyridine (209.16 mg, 2.64 mmol, 213.01 μL) were dissolved in DCM (5 mL), and phosphorus oxychloride (121.63 mg, 793.27 μmol, 73.94 μL) was added. After addition, the resulting reaction solution was stirred at 25 °C for 1 hour until the raw material was completely reacted. The solvent was removed by rotary evaporation under reduced pressure, and the resulting residue was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 11-2 (310 mg) with a yield of 69.8%. LC-MS (ESI) [M+H] + : 839.2.

[0404] Step 3: Synthesis of compound 11-3

[0405] Compound 11-2 (310 mg, 369.32 μmol) was dissolved in HCl methanol solution (4 M, 40.00 mmol, 10 mL), the mixture was reacted at 25 °C for 1 h until the raw material was completely reacted, the reaction solution was slowly added to saturated sodium bicarbonate solution (40 mL), extracted with ethyl acetate (40 mL x 2), the organic phase was combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure, the residue was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 11-3 (183 mg) with a yield of 69.9%. LC-MS (ESI) [M+H] + : 709.2.

[0406] Step 4: Synthesis of compound 11-4

[0407] Compound 11-3 (183 mg, 258.07 μmol) was dissolved in DMSO (4 mL), (1-(tert- butyloxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)boronic acid (117.20 mg, 516.14 μmol), PhenCuPPhBr2 (87.95 mg, 154.84 μmol) were added, the resulting reaction mixture was reacted at 90 °C under air atmosphere for 16 h until the raw material was completely reacted, the solvent was removed by rotary evaporation under reduced pressure, the residue was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to give compound 11-4 (200 mg) with a yield of 87.04%. LC-MS (ESI) [M+H] + : 890.4.

[0408] Step 12: Synthesis of compound 11-5

[0409] Compound 11-4 (200 mg, 146.01 μmol) was dissolved in DCM (3 mL), TFA (1.49 g, 13.06 mmol, 1 mL) was added, the resulting mixture was reacted at 25 °C for about 2 h until the raw material was completely reacted, the reaction solution was slowly poured into saturated sodium bicarbonate solution (20 mL), extracted with ethyl acetate (20 mL x 2), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure, the residue was separated by silica gel column chromatography (DCM / MeOH = 5 / 1) to give compound 11-5 (110 mg) with a yield of 95.34%. LC-MS (ESI) [M+H] + : 790.2.

[0410] Step 13: Synthesis of compound 11-6

[0411] Compound 11-5 (100 mg, 126.55 μmol) was dissolved in DCM (4 mL), TEA (76.83 mg, 759.27 μmol, 105.90 μL) was added, and cyclopropylsulfonyl chloride (88.95 mg, 632.73 μmol) was slowly added dropwise at 25 °C. The reaction was carried out at room temperature for about 2 hours until the raw material was completely reacted. The mixture was poured into a saturated sodium bicarbonate solution (10 mL), and then extracted with ethyl acetate (10 mL x 2). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained residue was subjected to silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 11-6 (100 mg) with a yield of 88.4%. LC-MS (ESI) [M+H] + : 894.2.

[0412] Step 14: Synthesis of compound 11

[0413] Compound 11-6 (100 mg, 111.81 μmol) was dissolved in DCM (4 mL), and a solution of boron trichloride (1 M, 1 mL) in dichloromethane was added. The obtained mixture was reacted at 25 °C for 1 hour until the raw material was completely reacted. The solvent was removed by concentration under reduced pressure, and the residue was subjected to HPLC preparation separation to obtain compound 11 (5.05 mg) with a yield of 5.62%. LC-MS (ESI) [M+H] + : 804.2.

[0414] 1 H NMR (400 MHz, DMSO-d6) δ 10.19 (s, 1H), 8.65 (s, 1H), 8.56 (s, 1H), 8.16 (s, 1H), 7.97 (s, 1H), 7.72 (s, 1H), 6.41 (s, 1H), 5.16 (s, 2H), 4.49 (d, J = 12.5 Hz, 1H), 4.02 (s, 2H), 3.73 (d, J = 12.0 Hz, 2H), 3.55 (s, 2H), 3.46 (d, J = 6.5 Hz, 3H), 3.19 (s, 4H), 2.98 - 2.81 (m, 5H), 2.71 (d, J = 10.8 Hz, 2H), 2.44 (s, 3H), 1.17 (s, 3H).

[0415] Example 12: Synthesis of compound 12

[0416]

[0417] Step 1: Synthesis of compound 12-1

[0418] Compound 5-11 (100 mg, 162.96 μmol), 4-sulfenyl aniline (42.86 mg, 195.55 μmol), pyridine (64.45 mg, 814.78 μmol, 65.64 μL) were dissolved in dichloromethane (3 mL), phosphorus oxychloride (37.48 mg, 244.43 μmol) was added, the resulting reaction system was stirred at 25 °C for about 1 h until the reaction was completed, the reaction liquid was concentrated under reduced pressure, the obtained residue was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 12-1 (116 mg, 142.36 μmol), the yield was 87.36 %. LC-MS (ESI) [M+H] + : 815.2.

[0419] Step 2: Synthesis of compound 12

[0420] Compound 12-1 (106 mg, 130.09 μmol) was dissolved in DCM (3 mL), a solution of boron trichloride (1 M, 2 mL) in dichloromethane was added, and the reaction was stirred at 25 °C for about 1 h until the reaction was completed. The reaction liquid was concentrated under reduced pressure, and the obtained residue was separated by HPLC preparation purification to obtain compound 12 (4.5 mg, 6.21 μmol), the yield was 4.77 %. LCMS (ESI) [M+H] + : 725.2.

[0421] 1 H NMR (400 MHz, DMSO-d6) δ 10.90 (s, 1H), 8.63 (s, 1H), 8.55 (s, 1H), 7.88 (d, J = 9.3 Hz, 2H), 7.78 (d, J = 9.0 Hz, 2H), 6.41 (s, 1H), 4.99 (s, 2H), 4.49 (d, J = 11.3 Hz, 1H), 4.27 (s, 2H), 3.89 (s, 2H), 3.75 (d, J = 30.6 Hz, 2H), 3.51 (s, 1H), 3.19 (s, 2H), 2.90 (s, 3H), 2.71 (s, 3H), 2.43 (s, 3H), 1.15 (s, 3H).

[0422] Example 13: Synthesis of compound 13

[0423]

[0424] Step 1: Synthesis of compound 13-1

[0425] Compound 4-11 (100 mg, 162.7 μmol), 3-chloro-5-(trifluoromethyl)pyridin-2-amine (64 mg, 325.4 μmol), pyridine (64.4 mg, 813.5 μmol) were dissolved in dichloromethane (5 mL) under nitrogen protection, and phosphorus oxychloride (49.9 mg, 325.4 μmol) was added. The mixture was stirred at 25 °C for half an hour until the reaction was completed. The reaction solution was concentrated under reduced pressure, and the obtained residue was separated by silica gel column chromatography (DCM / MeOH = 10 / 1) to obtain compound 13-1 (40 mg) with a yield of 31%. LC-MS (ESI) [M+H] + : 793.2.

[0426] Step 2: Synthesis of compound 13

[0427] Compound 13-1 (16 mg, 20.2 μmol) was added to trifluoroacetic acid (1 mL) under nitrogen protection, and heated to 90 °C for 0.5 hours until the reaction was completed. The reaction solution was concentrated under reduced pressure, and the obtained residue was separated by HPLC preparation purification to obtain compound 13 (3.0 mg) with a yield of 21%. LC-MS (ESI) [M+H] + : 703.2.

[0428] 1 H NMR (400 MHz, Methanol-d4) δ 8.70 (s, 1H), 8.56 (s, 1H), 8.35 (s, 1H), 6.89 (s, 1H), 5.54 (s, 2H), 4.39 (s, 2H), 4.12 (s, 1H), 4.00 (s, 2H), 3.92 (s, 2H), 3.50 (s, 3H), 3.15 (s, 2H), 2.93 (s, 3H), 2.74 (s, 1H), 2.54 (s, 3H), 1.34 (s, 3H).

[0429] Example 14: Synthesis of compound 14

[0430]

[0431] Step 1: Synthesis of compound 14-1

[0432] Compound 4-11 (100 mg, 162.7 μmol), 5-chloro-2-methyl-4- (trifluoromethyl) aniline (37.1 mg, 177.2 μmol) and pyridine (63.7 mg, 805.3 μmol) were dissolved in dichloromethane (6 mL) under nitrogen protection, and a solution of phosphorus oxychloride (27.2 mg, 177.2 μmol) in dichloromethane (0.2 mL) was added. The mixture was stirred at 25 °C for 0.5 hours until the reaction was completed. The reaction solution was poured into water, extracted with dichloromethane (2 x 5 mL), and the organic phase was dried over anhydrous sodium sulfate and rotary evaporated under reduced pressure. The residue was separated and purified by silica gel column chromatography (dichloromethane:methanol = 93:7) to obtain compound 14-1 (110 mg) with a yield of 84%. LC-MS (ESI) [M+H] + : 806.4.

[0433] Step 2: Synthesis of compound 14

[0434] Compound 14-1 (150 mg, 184.2 μmol) was added to trifluoroacetic acid (5 mL) under nitrogen protection, and the reaction was carried out at 90 °C for half an hour until the reaction was completed. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by HPLC preparation to obtain compound 14 (99.7 mg) with a yield of 76%. LC-MS (ESI) [M+H] + : 716.3.

[0435] 1 H NMR (400 MHz, DMSO-d6) δ 10.24 (s, 1H), 10.11 (s, 1H), 8.58 (s, 1H), 7.99 (s, 1H), 7.76 (s, 1H), 6.79 - 6.72 (m, 1H), 5.27 (s, 2H), 4.51 (d, J = 12.4 Hz, 1H), 4.33 (q, J = 2.9 Hz, 2H), 3.91 (t, J = 5.5 Hz, 2H), 3.70 (d, J = 11.4 Hz, 2H), 3.49 (d, J = 12.6 Hz, 1H), 3.23 (s, 1H), 3.08 - 2.91 (m, 3H), 2.78 (d, J = 23.3 Hz, 3H), 2.59 (d, J = 11.4 Hz, 1H), 2.45 (s, 3H), 2.37 (s, 3H), 1.20 (t, J = 7.4 Hz, 3H).

[0436] Example 15: Synthesis of compound 15

[0437]

[0438] Step 1: Synthesis of compound 15-1

[0439] Compound 4-11 (100 mg, 162.7 μmol), 2-chloro-5-fluoro-4- (trifluoromethyl) aniline (37.8 mg, 177.2 μmol), pyridine (63.7 mg, 805.3 μmol) were dissolved in dichloromethane (6 mL) under nitrogen protection, and phosphorus oxychloride (27.2 mg, 177.2 μmol) was added. The mixture was stirred at 25 °C for half an hour until the reaction was completed. The reaction solution was poured into water, extracted with dichloromethane (2 x 5 mL), and the organic phase was dried over anhydrous sodium sulfate and rotary evaporated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane:methanol = 93:7) to obtain compound 15-1 (120 mg) with a yield of 87%. LC-MS (ESI) [M+H] + : 810.3.

[0440] Step 2: Synthesis of compound 15

[0441] Compound 15-1 (100 mg, 117.3 μmol) was added to trifluoroacetic acid (5 mL) under nitrogen protection, and heated to 90 °C for half an hour until the reaction was completed. The reaction solution was concentrated under reduced pressure, and the residue was purified by HPLC to obtain compound 15 (70.2 mg) with a yield of 83%. LC-MS (ESI) [M+H] + : 720.2.

[0442] 1 H NMR (400 MHz, DMSO-d6) δ 10.47 (s, 1H), 10.24 (s, 1H), 8.58 (s, 1H), 8.12 (d, J = 12.8 Hz, 1H), 8.02 (d, J = 7.3 Hz, 1H), 6.74 (t, J = 1.5 Hz, 1H), 5.35 (s, 2H), 4.51 (d, J = 12.5 Hz, 1H), 4.31 (d, J = 3.0 Hz, 2H), 3.90 (t, J = 5.5 Hz, 2H), 3.77 - 3.62 (m, 2H), 3.49 (d, J = 12.6 Hz, 1H), 3.22 (t, J = 12.0 Hz, 1H), 2.97 (dd, J = 11.3, 5.0 Hz, 3H), 2.77 (d, J = 22.1 Hz, 3H), 2.58 (d, J = 11.5 Hz, 1H), 2.44 (s, 3H), 1.18 (t, J = 7.4 Hz, 3H).

[0443] Example 16: Synthesis of compound 16

[0444]

[0445] Step 1: Synthesis of compound 16-1

[0446] Compound 4-11 (100 mg, 162.7 μmol), bicyclo[4.2.0]octa-1(6),2,4-triene-3-amine (28.7 mg, 241.1 μmol) (synthesis reference WO2023284837), pyridine (63.6 mg, 803.7 μmol, 64.75 μL) were dissolved in dichloromethane (5 mL), phosphorus oxychloride (29.6 mg, 192.89 μmol) was added, the mixture was stirred at 25 °C for 1 hour until the reaction was completed, the reaction solution was poured into saturated aqueous sodium bicarbonate solution (3 mL), extracted with dichloromethane (2 x 5 mL), the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure, the residue was separated by silica gel column chromatography (dichloromethane:methanol = 15:1) to obtain compound 16-1 (75 mg) with a yield of 61%. LC-MS (ESI) [M+H] + : 716.4.

[0447] Step 2: Synthesis of compound 16

[0448] Compound 16-1 (75 mg, 98.2 μmol) was added to trifluoroacetic acid (1.5 mL) under nitrogen protection, heated to 90 °C for half an hour until the reaction was completed, the reaction solution was concentrated under reduced pressure, the residue was separated by HPLC preparation purification to obtain compound 16 (29.5 mg) with a yield of 48%. LC-MS (ESI) [M+H] + : 626.4.

[0449] 1 H NMR (400 MHz, DMSO-d6) δ 10.36 (s, 1H), 10.23 (s, 1H), 8.57 (s, 1H), 7.37 (s, 1H), 7.28 (dd, J = 8.1, 1.5 Hz, 1H), 7.02 (d, J = 8.0 Hz, 1H), 6.73 (s, 1H), 5.10 (s, 2H), 4.50 (d, J = 12.5 Hz, 1H), 4.31 (d, J = 3.0 Hz, 2H), 3.89 (t, J = 5.5 Hz, 2H), 3.74 - 3.65 (m, 2H), 3.48 (d, J = 12.8 Hz, 1H), 3.23 (d, J = 12.0 Hz, 1H), 3.08 (s, 4H), 2.97 (d, J = 8.8 Hz, 3H), 2.76 (d, J = 21.6 Hz, 3H), 2.57 (d, J = 11.5 Hz, 1H), 2.44 (s, 3H), 1.17 (t, J = 7.4 Hz, 3H).

[0450] Example 17: Synthesis of compound 17

[0451]

[0452] Step 1: synthesis of compound 17-1

[0453] Compound 4-11 (120 mg, 195.2 μmol), 2,3-dihydro-1H-inden-5-amine (38.5 mg, 289.3 μmol), pyridine (76.3 mg, 964.5 μmol) were dissolved in dichloromethane (5 mL) under nitrogen protection, phosphorus oxychloride (35.5 mg, 231.5 μmol) was added, the resulting mixture was stirred at 25 °C for 1 hour until the reaction was completed, the reaction solution was poured into saturated aqueous sodium bicarbonate solution (3 mL) to quench, extracted with dichloromethane (2 x 5 mL), the organic phase was dried over anhydrous sodium sulfate, rotary evaporation under reduced pressure, the resulting residue was separated by silica gel column chromatography (dichloromethane:methanol = 15:1) to give compound 17-1 (120 mg) with a yield of 81%. LC-MS (ESI) [M+H] + :730.2.

[0454] Step 2: synthesis of compound 17

[0455] Compound 17-1 (120 mg, 155.9 μmol) was added to trifluoroacetic acid (1.0 mL) under nitrogen protection, the reaction was stirred at 90 °C for 1 hour until the reaction was completed, the reaction solution was concentrated under reduced pressure, the resulting residue was separated by HPLC preparation purification to give compound 17 (67.6 mg) with a yield of 68%. LC-MS (ESI) [M+H] + :640.4.

[0456] 1 H NMR (400 MHz, DMSO-d6) δ 10.38 (s, 1H), 8.53 (s, 1H), 7.49 (d, J = 1.6 Hz, 1H), 7.25 (dd, J = 8.1, 2.0 Hz, 1H), 7.15 (d, J = 8.1 Hz, 1H), 6.75 - 6.71 (m, 1H), 5.10 (s, 2H), 4.50 (d, J = 12.8 Hz, 1H), 4.31 (q, J = 2.9 Hz, 2H), 3.89 (t, J = 5.5 Hz, 2H), 3.76 - 3.65 (m, 2H), 3.50 (d, J = 12.8 Hz, 1H), 3.22 (t, J = 12.3 Hz, 2H), 2.97 (d, J = 8.9 Hz, 3H), 2.80 (q, J = 7.0 Hz, 7H), 2.57 (d, J = 10.9 Hz, 1H), 2.43 (s, 3H), 2.02 - 1.95 (m, 2H), 1.17 (t, J = 7.4 Hz, 3H).

[0457] Example 18: synthesis of compound 18

[0458]

[0459] Step 1: Synthesis of compound 18-1

[0460] Compound 4-6 (9 g, 15.4 mmol) was added to ethanol (400 mL), and then palladium on carbon (5%, 1.6 g) and palladium hydroxide on carbon (5%, 2.2 g) were added. The reaction was heated to reflux under hydrogen atmosphere for 16 hours until the reaction was completed. The reaction solution was filtered and concentrated under reduced pressure. The obtained crude product was purified by slurry with mixed solvents (petroleum ether: ethyl acetate = 5:1) to obtain compound 18-1 (6 g) with a yield of 85%. LC-MS (ESI) [M+H] + : 435.4.

[0461] Step 2: Synthesis of compound 18-2

[0462] Compound 18-1 (6.75 g, 14.43 mmol), (3,6-dihydro-2H-pyran-4-yl)boronic acid (3.69 g, 28.86 mmol) and pyridine (11.41 g, 144.28 mmol, 11.62 mL) were dissolved in dichloromethane (140 mL) and ultrasonically dissolved. After the solution was clear, copper acetate (7.86 g, 43.28 mmol) was added. The reaction was carried out at room temperature under oxygen atmosphere for 2 hours until the reaction was completed. The reaction solution was filtered through diatomite. The filtrate was washed with water (3 x 30 mL), saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was separated and purified by silica gel column (dichloromethane:methanol = 20:1) to obtain compound 18-2 (7.43 g) with a yield of 92%. LC-MS (ESI) [M+H] + : 517.3.

[0463] Step 3: Synthesis of compound 18-3

[0464] Compound 18-2 (7.43 g, 13.31 mmol) was dissolved in hydrochloric acid dioxane solution (4 mol / L, 75 mL) under nitrogen protection. The reaction was stirred at 25°C for half an hour until the reaction was completed. The reaction solution was concentrated under reduced pressure to remove the solvent. Water (100 mL) was added. The pH was adjusted to 8 with saturated sodium carbonate solution. The reaction solution was extracted with dichloromethane (3 x 50 mL). The organic phase was combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 18-3 (5.4 g) with a yield of 93%. LC-MS (ESI) [M+H] + : 417.4.

[0465] Step 4: Synthesis of compound 18-4

[0466] Compound 18-3 (1.72 g, 4.13 mmol) was dissolved in anhydrous N,N- dimethylformamide (20 mL) under nitrogen protection, 5-methoxypyrimidine-4- carboxylic acid (0.76 g, 4.96 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.14 g, 8.26 mmol) and N,N-diisopropyl ethylamine (0.53 g, 4.13 mmol) were added, and the reaction was stirred at 25 °C for 16 hours until the reaction was completed. Water (40 mL) was added for dilution, and the reaction was extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (methanol: dichloromethane = 1:9) to obtain compound 18-4 (2.0 g) with a yield of 88%. LC-MS (ESI) [M+H]: 553.4. + : 553.4.

[0467] Step 5: Synthesis of compound 18-5

[0468] Compound 18-4 (2.5 g, 4.5 mmol) was dissolved in tetrahydrofuran / water (16 mL / 4 mL) under nitrogen protection, and lithium hydroxide monohydrate (0.76 g, 18.1 mmol) was added. The reaction was stirred at 25 °C for 1 hour until the reaction was completed. Dilute hydrochloric acid (2 mol / L) was added dropwise to adjust pH = 2, water (20 mL) was added, and the reaction was extracted with ethyl acetate (20 mL x 3). The combined organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 18-5 (2.0 g) with a yield of 84%. LC-MS (ESI) [M+H]: 525.2. + : 525.2.

[0469] Step 6: Synthesis of compound 18-6

[0470] Compound 18-5 (0.2 g, 0.38 mmol) was dissolved in dichloromethane (10 mL) under nitrogen protection, 2-chloro-4-trifluoromethylaniline (0.089 g, 0.42 mmol) and pyridine (0.30 g, 3.8 mmol) were added, and a solution of phosphorus oxychloride (0.07 g, 0.46 mmol) in dichloromethane (3 mL) was added. The reaction was stirred at 25 °C for 15 minutes until the reaction was completed. Water (10 mL) was added for dilution, and the reaction was extracted with dichloromethane (10 mL x 3). The combined organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (methanol / dichloromethane = 5%) to obtain compound 18-6 (0.1 g) with a yield of 37%. LC-MS (ESI) [M+H]: 702.2. + : 702.2.

[0471] Step 7: Synthesis of compound 18

[0472] Compound 18-6 (0.15 g, 0.21 mmol) was dissolved in dry N-N dimethylformamide (4 mL) under nitrogen protection, and aluminum chloride (0.28 g, 2.1 mmol) was added. The reaction was carried out in a microwave reactor at 150 °C for 1 hour. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was purified by HPLC to obtain compound 18 (20 mg), with a yield of 14%. LC-MS (ESI) [M+H] + : 688.4.

[0473] 1 H NMR (400 MHz, MeOD-d4) d 8.65 (s, 1H), 8.41 (s, 1H), 8.16 (d, J = 8.5 Hz, 1H), 7.83 (d, J = 1.6 Hz, 1H), 7.62 (dd, J = 8.6, 1.5 Hz, 1H), 6.88 (s, 1H), 5.44 - 5.36 (m, 2H), 4.69 (d, J = 12.6 Hz, 1H), 4.39 (d, J = 2.8 Hz, 2H), 4.01 - 3.95 (m, 2H), 3.95 - 3.83 (m, 2H), 3.64 - 3.54 (m, 1H), 3.51 - 3.39 (m, 1H), 3.19 - 3.10 (m, 3H), 2.97 - 2.85 (m, 3H), 2.75 (d, J = 11.3 Hz, 1H), 1.33 (t, J = 5.0 Hz, 3H).

[0474] Example 19: Synthesis of compound 19

[0475]

[0476] Step 1: Synthesis of compound 19-1

[0477] Compound 18-3 (0.26 g, 0.62 mmol) was dissolved in N,N-dimethylformamide (3 mL) under nitrogen protection, and then 3-(benzyloxy)picolinic acid (0.17 g, 0.74 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.28 g, 0.74 mmol) and N,N-diisopropyl ethylamine (0.4 g, 3.1 mmol) were added. The reaction was stirred at 25 °C for 1 hour until completion. Water (6 mL) was added for dilution, and then extracted with ethyl acetate (5 mL x 3). The organic phase was combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (methanol / dichloromethane = 6%) to obtain compound 19-1 (0.35 g) with a yield of 89%. LC-MS (ESI) [M+H] + : 628.3.

[0478] Step 2: Synthesis of compound 19-2

[0479] Compound 19-1 (0.28 g, 0.45 mmol) was dissolved in tetrahydrofuran / water (5 mL / 1 mL) under nitrogen protection, and then lithium hydroxide monohydrate (57 mg, 1.35 mmol) was added. The reaction was stirred at 25 °C for 1 hour until completion. The reaction solution was adjusted to pH = 2 with dilute hydrochloric acid (2 mol / L), and then diluted with water (10 mL). Extraction was performed with ethyl acetate (10 mL x 3), and then the organic phase was combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 19-2 (0.24 g) with a yield of 90%. LC-MS (ESI) [M+H] + : 600.3.

[0480] Step 3: Synthesis of compound 19-3

[0481] Compound 19-2 (100 mg, 0.17 mmol) was dissolved in dichloromethane (5 mL) under nitrogen protection, and then intermediate 2-chloro-5-fluoro-4-(trifluoromethyl)aniline (44 mg, 0.20 mmol) and pyridine (130 mg, 1.70 mmol) were added. A solution of phosphorus oxychloride (31 mg, 0.20 mmol) in dichloromethane (3 mL) was added, and then the reaction was stirred at room temperature for 15 minutes until completion. Water (6 mL) was added for dilution, and then extraction was performed with dichloromethane (5 mL x 3). The organic phase was combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (methanol / dichloromethane = 5%) to obtain compound 19-3 (66 mg) with a yield of 50%. LC-MS (ESI) [M+H] + : 795.3.

[0482] Step 4: Synthesis of compound 19

[0483] Compound 19-3 (56 mg, 0.70 mmol) was dissolved in trifluoroacetic acid (3 mL) under nitrogen protection, heated to 90 °C and stirred for 3 hours until the reaction was completed. The reaction was concentrated under reduced pressure, the residue was diluted with water (6 mL), extracted with dichloromethane (5 mL x 3), the organic phases were combined, washed with water (5 mL x 6) and saturated sodium chloride solution (5 mL) successively, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by HPLC to obtain compound 19 (7.0 mg) with a yield of 14%. LC-MS (ESI) [M+H] + : 705.3.

[0484] 1 H NMR (400 MHz, MeOD-d4) δ 8.17 (d, J = 12.6 Hz, 1H), 8.09 (s, 1H), 7.85 (d, J = 7.1 Hz, 1H), 7.36 (d, J = 2.8 Hz, 2H), 6.87 (s, 1H), 5.43 (s, 2H), 4.77 - 4.66 (m, 1H), 4.42 - 4.33 (m, 2H), 3.99 (t, J = 5.4 Hz, 2H), 3.95 - 3.83 (m, 2H), 3.76 - 3.60 (m, 1H), 3.38 (d, J = 22.3 Hz, 1H), 3.18 - 3.09 (m, 3H), 2.96 - 2.84 (m, 3H), 2.79 - 2.68 (m, 1H), 1.32 (t, J = 7.4 Hz, 3H).

[0485] Example 20: Synthesis of compound 20

[0486]

[0487] Step 1: Synthesis of compound 20-1

[0488] Compound 19-2 (100 mg, 0.17 mmol), 2-chloro-4-trifluoromethylaniline (48.92 mg, 0.25 mmol) and phosphorus oxychloride (76.71 mg, 500.3 μmol) were dissolved in DCM (6 mL) under nitrogen protection, and pyridine (79.15 mg, 1.0 mmol) was added. The mixture was stirred at 25 °C for half an hour until the reaction was completed. Water (6 mL) was added to dilute, extracted with dichloromethane (5 mL x 3), the organic phases were combined, washed with saturated brine (5 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / dichloromethane = 10 / 1) to obtain compound 20-1 (122 mg) with a yield of 94%. LC-MS (ESI) [M+H] +:777.2.

[0489] Step 2: Synthesis of compound 20

[0490] Compound 20-1 (122 mg, 0.16 mmol) was dissolved in trifluoroacetic acid (1 mL) under nitrogen protection, heated to 90 °C and stirred for half an hour until the reaction was completed. The reaction was concentrated under reduced pressure and dried. The residue was purified by HPLC to obtain compound 20 (7.2 mg) with a yield of 6.7%. LC-MS (ESI) [M+H] + :687.2.

[0491] 1 H NMR (400 MHz, MeOD-d4) δ 8.15 (s, 1H), 8.07 (s, 1H), 7.81 (s, 1H), 7.60 (d, J = 10.1 Hz, 1H), 7.34 (s, 2H), 6.86 (s, 1H), 5.38 (s, 2H), 4.36 (s, 2H), 3.94 (d, J = 36.8 Hz, 4H), 3.64 (s, 1H), 3.45 (s, 2H), 3.15 (s, 3H), 2.90 (s, 3H), 2.69 (s, 1H), 1.33 (s, 3H).

[0492] Example 21: Synthesis of compound 21

[0493]

[0494] Step 1: Synthesis of compound 21-1

[0495] Compound 4-11 (33.6 mg, 54.7 μmol), 4-(pentafluorothio)aniline (11.98 mg, 54.7 μmol), pyridine (21.6 mg, 273.3 μmol) were dissolved in dichloromethane (5 mL) under nitrogen protection, and a solution of phosphorus oxychloride (25.2 mg, 164.0 μmol) in dichloromethane (0.5 mL) was added. The mixture was stirred at 25 °C for 1 hour until the reaction was completed. The reaction was quenched by pouring into saturated aqueous sodium bicarbonate solution (3 mL), and extracted with dichloromethane (2 x 5 mL). The organic phase was dried over anhydrous sodium sulfate and rotary evaporated under reduced pressure. The obtained crude product was separated by silica gel column chromatography (dichloromethane:methanol = 9:1) to obtain compound 21-1 (42 mg) with a yield of 94%. LC-MS (ESI) [M+H] + :816.2.

[0496] Step 2: Synthesis of compound 21

[0497] Compound 21-1 (42 mg, 51.5 μmol) was dissolved in dichloromethane (5 mL) under nitrogen protection, then boron trichloride (25.13 mg, 214.51 μmol, 0.2 mL) was added, and the reaction was stirred at 25 °C for 1 h until completion. Methanol (1 mL) was added to quench the reaction, and the obtained crude product was concentrated under reduced pressure and purified by HPLC to obtain compound 21 (12 mg) with a yield of 39%. LC-MS (ESI) [M+H] + = 726.2.

[0498] 1 H NMR (400 MHz, DMSO-d6) δ 11.04 (s, 1H), 10.10 (s, 1H), 8.56 (s, 1H), 7.88 (d, J = 9.3 Hz, 2H), 7.77 (d, J = 8.9 Hz, 2H), 6.70 - 6.52 (m, 1H), 5.36 (s, 2H), 4.50 (d, J = 12.4 Hz, 1H), 4.33 (q, J = 2.8 Hz, 2H), 3.93 (t, J = 5.4 Hz, 2H), 3.70 (t, J = 12.0 Hz, 2H), 3.47 (d, J = 12.7 Hz, 1H), 3.20 (d, J = 11.1 Hz, 2H), 3.09 - 2.91 (m, 3H), 2.76 (d, J = 24.2 Hz, 3H), 2.57 (d, J = 11.4 Hz, 1H), 2.44 (s, 3H), 1.17 (t, J = 7.4 Hz, 3H).

[0499] Example 22: Synthesis of compound 22

[0500]

[0501] Step 1: Synthesis of compound 22-1

[0502] Compound 4-11 (120 mg, 195.2 μmol), 2,4-dichloro-5-fluoroaniline (52.7 mg, 292.9 μmol), pyridine (77.2 mg, 976.2 μmol) were dissolved in dichloromethane (3 mL) under nitrogen protection, then phosphorus oxychloride (44.9 mg, 292.9 μmol) was added, and the obtained mixture was stirred at 25 °C for 1 h until completion. Water (3 mL) was added to quench the reaction, and the reaction was extracted with dichloromethane (2 x 5 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was separated by silica gel column chromatography (dichloromethane:methanol = 19:1) to obtain compound 22-1 (117 mg) with a yield of 87%. LC-MS (ESI) [M+H] + : 776.4.

[0503] Step 2: Synthesis of compound 22

[0504] Compound 22-1 (117 mg, 150.7 μmol) was dissolved in trifluoroacetic acid (2.5 mL) under nitrogen protection, and the mixture was stirred at 90 °C for 0.5 h until the reaction was completed. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (20 mL). The solution was washed with water (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by slurry with petroleum ether / ethyl acetate (4:1) to give compound 22 (51.7 mg) in 50% yield. LC-MS (ESI) [M+H] + : 686.2.

[0505] 1 H NMR (400 MHz, DMSO-d6) δ 10.28 (s, 1H), 8.58 (s, 1H), 8.00-7.90 (m, 2H), 6.82-6.70 (m, 1H), 5.28 (s, 2H), 4.51 (d, J = 12.0 Hz, 1H), 4.39-4.29 (m, 2H), 3.95-3.86 (m, 2H), 3.80-3.64 (m, 2H), 3.54-3.45 (m, 1H), 3.28-3.16 (m, 1H), 3.09-2.88 (m, 3H), 2.86-2.73 (m, 3H), 2.66-2.55 (m, 1H), 2.44 (s, 3H), 1.19 (t, J = 8.0 Hz, 3H).

[0506] Example 23: Synthesis of compound 23

[0507]

[0508] Step 1: Synthesis of compound 23-1

[0509] Compound 4-11 (100 mg, 161.1 μmol), 4-amino-2,5-difluorobenzotrifluoride (34.9 mg, 177.2 μmol), and pyridine (63.7 mg, 805.3 μmol) were dissolved in dichloromethane (6 mL) under nitrogen protection, and a solution of phosphorus oxychloride (27.2 mg, 177.2 μmol) in dichloromethane (0.2 mL) was added. The mixture was stirred at 25 °C for 0.5 h until the reaction was completed. The reaction mixture was quenched with water (3 mL) and extracted with dichloromethane (2 x 5 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (dichloromethane:methanol = 93:7) to give compound 23-1 (120 mg) in 89% yield. LC-MS (ESI) [M+H] + : 794.4.

[0510] Step 2: Synthesis of compound 23

[0511] Compound 23-1 (110 mg, 119.7 μmol) was dissolved in trifluoroacetic acid (5 mL) under nitrogen protection, and the reaction was stirred at 90 °C for 0.5 hours until the reaction was complete. After concentration under reduced pressure, water (10 mL) was added, and dichloromethane (20 mL x 2) was used for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was subjected to silica gel column chromatography (dichloromethane:methanol = 90:10) to obtain compound 23 (63.8 mg) with a yield of 75%. LC-MS (ESI) [M+H] + : 704.3.

[0512] 1 H NMR (400 MHz, DMSO-d6) δ 10.92 (s, 1H), 10.23 (s, 1H), 8.57 (s, 1H), 8.20 (dd, J = 12.6, 6.1 Hz, 1H), 7.88 (dd, J = 10.6, 6.6 Hz, 1H), 6.73 (p, J = 1.7 Hz, 1H), 5.30 (s, 2H), 4.51 (d, J = 12.4 Hz, 1H), 4.31 (q, J = 2.9 Hz, 2H), 3.89 (t, J = 5.5 Hz, 2H), 3.76 - 3.64 (m, 2H), 3.49 (d, J = 12.6 Hz, 1H), 3.22 (t, J = 11.8 Hz, 1H), 2.97 (d, J = 9.6 Hz, 3H), 2.76 (d, J = 19.9 Hz, 3H), 2.58 (d, J = 11.2 Hz, 1H), 2.44 (s, 3H), 1.17 (t, J = 7.5 Hz, 3H).

[0513] Comparative Example

[0514] Example 42 in WO2022249060A1 was used as the control compound HRO761, and the compound was prepared by referring to the method in the patent.

[0515] Experimental Example 1: Detection of WRN helicase activity

[0516] 1. Experimental instruments

[0517] The instrument information used in this experimental example is shown in Table 1.

[0518] Table 1

[0519] Instrument name Instrument manufacturer Model Centrifuge Eppendorf 5810R Microplate reader PerkinElmer EnVision-2015

[0520] 2. Experimental materials

[0521] The WRN enzyme used in the experiment has a His-TEV tag at the N-terminus and is expressed in eukaryotic cells with a purity of 90%. The two single-stranded DNAs used for detection are labeled with BHQ2 and Cy5, respectively. When the two fluorescent molecules are close to each other (when the DNA is in a double-stranded state), no fluorescence signal can be detected due to the quenching effect. WRN has helicase activity and can unwind double-stranded DNA into single-stranded DNA, thereby generating a fluorescence signal. The information of other reagents and consumables required for the experiment is shown in Table 2.

[0522] Table 2

[0523] Reagent Brand Catalogue number Bicine Sigma B8660 KCl Sigma P9541 MgCl2 Sigma M1028 F-127 Sigma P2443 Tween 20 Thermo Fisher 646547 Tris BBI Life Sciences G9391 NaCl BBI Life Sciences A600194-0500 TCEP Sigma BSG Sigma DNA A610476-001 Jinsirui NA ATP Sigma 96-well V-bottom plate A7699 Biunten FPT019 384-well plate Greiner Example 781209

[0524] 3. Experimental method

[0525] The two labeled single-stranded DNAs were annealed to form double-stranded DNA. Annealing buffer: 12 mM Tris (pH 8.0), 300 mM NaCl, 12 mM MgCl2, 2 mM DTT. Annealing program: 95°C, 5 min.

[0526] Use buffer (20 mM Bicine, 10 mM KCl, 1 mM MgCl2, 0.005% BSG, 1 mM TCEP, 0.1% F-127, pH 7.5) to configure 2x WRN enzyme (2 nM) and 2x substrate (100 nM double-stranded DNA, 1000 nM capture DNA, 100 μM ATP). The test compound is dissolved in DMSO to 10 mM, and is gradiently diluted with a 96-well V-bottom plate. Add 0.5 μL of the compound to 25 μL of 2x WRN enzyme, and incubate at room temperature for 30 min. Add 25 μL of 2x substrate, and react at room temperature for 30 min. Use an enzyme marker to detect, excitation light 620 nm, and emission light 685 nm.

[0527] 4. Data analysis

[0528] Use GraphPad Prism 8 software to fit the concentration-effect curve, and calculate the compound concentration IC 50 50% inhibition effect. First, calculate the percentage inhibition rate corresponding to each compound concentration, and then use the "log(inhibitor) vs. normalized response--Variable slope" equation of GraphPad Prism 8 software to fit the concentration-effect curve, thereby obtaining the IC 50 .

[0529] Inhibition rate (%) = (average fluorescence intensity of positive control well - fluorescence intensity of compound well) / (average fluorescence intensity of positive control well - average fluorescence intensity of negative control well) x 100

[0530] Positive control: 25 μL 2× WRN enzyme + 0.5 μL DMSO + 25 μL 2× substrate

[0531] Negative control: 25 μL 2× buffer + 0.5 μL DMSO + 25 μL 2× substrate

[0532] The experimental results are shown in Table 3.

[0533] Table 3

[0534] WRN helicase activity (IC50 nM) Example 4 Example 15 60.92 Example 22 70.15 Instrument name 72.12

[0535] The above tests show that the compound of the present invention has good WRN helicase activity and has the potential to be further developed into a WRN helicase inhibitor.

[0536] Experimental Example 2 WRN hydrolase activity detection experiment

[0537] 1. Experimental instruments

[0538] The instrument information used in this experiment is shown in Table 4.

[0539] Table 4

[0540] Instrument manufacturer Model Centrifuge Eppendorf Microplate reader 5810R PerkinElmer EnVision-2015 Reagent

[0541] 2. Experimental Materials

[0542] The WRN enzyme used in the experiment has a His-TEV tag at its N-terminus and is expressed in eukaryotic cells with a purity of 90%. The detection kit (ADP-Glo TM The Kinase Assay was purchased from Promega, catalog number V9101. Store in aliquots at -40°C. The kit can quantitatively detect the amount of ADP generated in the reaction. WRN hydrolyzes ATP to produce ADP. Adding ADP-Glo ​​reagent consumes excess ATP in the reaction system. Adding Kinase Detection Reagent converts the generated ADP into ATP, generating chemiluminescence. Detecting the chemiluminescent signal using a microplate reader reflects WRN enzyme activity. Other reagents and consumables required for the experiment are listed in Table 5.

[0543] Table 5

[0544] Brand Catalogue number Bicine Sigma KCl B8660 Sigma Sigma P9541 MgCl2 Tween 20 M1028 Thermo Fisher Tris 28320 BBI Life Sciences NaCl A600194-0500 BBI Life Sciences TCEP A610476-001 Sigma BSG 646547 Sigma DNA G9391 Jinsirui NA ATP Sigma 96-well V-bottom plate A7699 Biunten FPT019 384-well plate PE Example 6008280

[0545] 3. Experimental methods

[0546] Anneal single-stranded DNA to form double-stranded DNA. 5× Annealing Buffer: 50 mM Tris pH 8.0, 100 mM NaCl. Annealing Procedure: 95°C, 5 min.

[0547] The 2x WRN enzyme (1 nM) and 2x substrate (1 nM double-stranded DNA, 100 mM ATP) were configured using the buffer (20 mM Bicine (pH 7.5), 10 mM KCl, 10 mM MgCl2, 0.005% BSG, 0.002% Tween 20, 1 mM TCEP). The test compound was dissolved in DMSO to 10 mM and gradiently diluted with a 96-well V-bottom plate. First, 25 mL of 2x WRN enzyme and 0.5 mL of compound were added to a 96-well plate, and incubated at room temperature for 30 min. Then, 25 mL of 2x substrate was added, and reacted at room temperature for 60 min. 5 mL from the reaction plate was added to a 384-well plate for detection, 5 mL of ADP-Glo reagent was added, and incubated at room temperature for 60 min, 10 uL of Kinase Detection Reagent was added, and incubated at room temperature for 40 min. The chemiluminescence signal was detected by a microplate reader.

[0548] 4. Data analysis

[0549] The concentration-effect curve was fitted with GraphPad Prism 8 software, and the compound concentration IC 50 was calculated for 50% inhibition effect. First, the percentage inhibition rate corresponding to each compound concentration was calculated, and then the concentration-effect curve was fitted using the "log(inhibitor) vs. normalized response--Variable slope" equation of GraphPad Prism 8 software, so as to obtain the IC 50 .

[0550] Inhibition rate (%) = (average luminescence intensity of positive control well - luminescence intensity of compound well) / (average luminescence intensity of positive control well - average luminescence intensity of negative control well) x 100

[0551] Positive control: 25 mL of 2x WRN enzyme + 0.5 mL of DMSO + 25 mL of 2x substrate

[0552] Negative control: 25 mL of 2x buffer + 0.5 mL of DMSO + 25 mL of 2x substrate

[0553] The experimental results are shown in Table 6.

[0554] Table 6

[0555] WRN helicase activity (IC50 nM) Example 4 Example 5 88.53 Example 6 79.07 Example 7 134.5 Example 8 86.94 Example 9 118 Example 10 94.9 Example 11 237.8 Example 12 65.27 Example 13 85.98 Example 14 104 Example 15 30.8 Example 16 34.29 Example 17 42.59 Example 18 34.66 Example 19 64.26 Example 20 44.29 Example 21 35.5 Example 22 24.39 Example 23 46.77 Instrument name 41.94

[0556] The above test results show that the compound of the present application has good WRN hydrolytic enzyme activity.

[0557] Experimental Example 3 Cell Proliferation Experiment

[0558] 1. Experimental instruments

[0559] The instrument information used in this experimental example is shown in Table 7.

[0560] Table 7

[0561] Instrument manufacturer Model Biosafety cabinet Thermo 1300 SERIES A2 Thermo CO2 incubator Cell counter 371 Beckman Coulter Vi-CELL XR Centrifuge Eppendorf Microplate reader 5810R Molecular Devices SpectraMax i3x Instrument name

[0562] 2. Experimental materials

[0563] The information of other reagents and consumables required in the experiment is shown in Table 8.

[0564] Table 8

[0565]

[0566]

[0567] 3. Experimental method

[0568] SW48 cells were plated in a 96-well cell culture plate at a cell density of 1,500 cells / well, and LoVo, HCT116, and SW620 cells were plated at a cell density of 800 cells / well, and incubated in a 37°C incubator overnight. The test compound diluted with DMSO was added, and after 5 days of incubation, the cell viability was detected using the CellTiter-Glo kit. The CellTiter-Glo reagent was equilibrated to room temperature, and an appropriate amount was added to the cell plate, which was shaken at room temperature for 12 minutes. The luminescence signal was detected using a microplate reader.

[0569] 4. Data analysis

[0570] The luminescence signal values of the DMSO-treated wells and the cell-free wells were taken as the negative and positive controls, respectively, to calculate the inhibition rate of the compound on cell proliferation. The concentration-inhibition rate curve was fitted using the "log(inhibitor) vs. normalized response - Variable slope" equation of the GraphPad Prism 8 software, and the IC 50 .

[0571] Inhibition rate (%) = [1 - (luminescence intensity of compound well - average luminescence intensity of positive control well) / (average luminescence intensity of negative control well - average luminescence intensity of positive control well)] x 100

[0572] The experimental results are shown in Table 9.

[0573] Table 9

[0574]

[0575]

[0576] ND indicates not tested.

[0577] The above test results show that the compound of the present application has inhibitory effect on the proliferation of MSI-H colorectal cancer cell lines SW48 and HCT116 cells, but has no inhibitory effect on MSS SW620 cells.

[0578] Experimental Example 4 γH2AX induction experiment

[0579] 1. Experimental instruments

[0580] The instrument information used in this experimental example is shown in Table 10.

[0581] Table 10

[0582] Instrument manufacturer Model Biosafety cabinet Thermo 1300 SERIES A2 Thermo CO2 incubator Cell counter 371 Beckman Coulter Vi-CELL XR Centrifuge Eppendorf Shaker 5810R Thermo Microplate reader 13687716 PerkinElmer EnVision-2015 Compound number

[0583] 2. Experimental materials

[0584] The information of other reagents and consumables required in the experiment is shown in Table 11.

[0585] Table 11

[0586]

[0587] 3. Experimental method:

[0588] SW48, HCT116 and SW620 cells were plated in 96-well cell culture plates at a cell density of 15,000-25000 cells per well and incubated overnight in a 37°C incubator. Test compounds were added at a DMSO gradient, and incubated for 2-3 days. The level of γ-H2AX was detected using HTRF Phospho-H2AX (SER139) Detection Kits. 4x Lysis buffer and Detection buffer were equilibrated to room temperature and ready for use. 4x Lysis buffer was configured to 1x Lysis buffer using ddH2O, and 50 μL 1x Lysis buffer was added to each well after the medium was aspirated, and incubated at room temperature with shaking for 30 minutes. 16 μL of lysate was transferred to a 384-well plate, and 4 μL of diluted mixed antibody was added (Phospho-H2AX d2 antibody and Phospho-H2AX Eu Cryptate antibody were diluted 20-fold using Detection buffer, respectively, and then mixed evenly at a 1:1 volume ratio). The plate was incubated at room temperature for 2-24 hours, and the fluorescence signals at 655 nm and 615 nm were detected using a microplate reader. A parallel processing plate was set up for the experiment, and CellTiter-Glo kit was used to detect cell viability. The CellTiter-Glo reagent was equilibrated to room temperature, and an appropriate amount was added to the cell plate, which was shaken at room temperature for 12 minutes. The luminescence signal was detected using a microplate reader.

[0589] The fluorescence signal values of the DMSO-treated wells and the positive drug-treated wells were used as negative and positive controls, respectively, to calculate the level of γ-H2AX induced by the compound in cells. The concentration-induction rate curve was fitted using the "log (agonist) vs. response - Variable slope (four parameters)" equation of GraphPad Prism 8 software, and the EC50 value was obtained. 50 .

[0590] Cell survival rate (%) = luminescence intensity of compound well / average luminescence intensity of negative control well x 100

[0591] HTRF Ratio = 665 nm fluorescence signal value / 615 nm fluorescence signal value x 10 4

[0592] The experimental results are shown in Table 12.

[0593] Table 12

[0594]

[0595] The above test results show that the treatment of the compound of the present application can significantly induce DNA damage accumulation (increase of γ-H2AX level) of MSL-H colorectal cancer cell lines SW48 and HCT116, but has no inducing effect on MSS SW620 cells.

[0596] Experimental Example 5 Mouse pharmacokinetic test experiment

[0597] 1. Experimental purpose

[0598] CD-1 mice were used as test animals, and the mice were orally administered with Example 4, Example 22 and Comparative Example HRO761. The drug concentration in the plasma of the mice at different times was determined by LC-MS / MS method to study the pharmacokinetic characteristics of the compound of the present application in mice.

[0599] 2. Experimental scheme

[0600] 2.1 Experimental drugs and animals

[0601] Experimental drugs: Example 4, Example 22 and Comparative Example HRO761;

[0602] Animals: CD-1 mice, male, 24-25 g, purchased from Shanghai Jihui Experimental Animal Co., Ltd.

[0603] 2.2 Drug preparation

[0604] An appropriate amount of Example 4, Example 22 and Comparative Example HRO761 was weighed, and an appropriate amount of dimethyl sulfoxide, solutol and physiological saline (final solvent 5% DMSO + 10% solutol + 85% saline) was added in turn, vortexed and ultrasonically prepared into a 1.0 mg / mL drug solution.

[0605] 2.3 Drug administration

[0606] Three mice in the gavage group of Example 4, Example 22 and Comparative Example HRO761 were fasted overnight and then administered with the drugs (dose 10 mg / kg, administration volume 10 mL / kg), and the mice were fed 4 hours after administration.

[0607] 3. Experimental operation

[0608] The animals were collected with 0.04 mL of blood before administration and at 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours and 24 hours after administration, and the blood was anticoagulated with heparin sodium. The blood samples were placed on ice after collection, and the plasma was separated by centrifugation (centrifugation conditions: 8000 rpm, 5 minutes). The collected plasma was stored at -80°C before analysis.

[0609] The content of the test compound in the plasma of the mice after gavage administration was determined by LC-MS / MS method.

[0610] 4. Pharmacokinetic parameter results

[0611] The concentration of the compound in the plasma at different time points was calculated according to the calculation, and the pharmacokinetic parameters of Example 4, Example 22 and Comparative Example HR0761 in mice were calculated by using winnonlin software. The pharmacokinetic parameters of Example 4, Example 22 and Comparative Example HR0761 of the application are shown in Table 13.

[0612] Table 13

[0613] Example 4 <![CDATA[C max ,ng / mL]]> AUC 0-i , ng.h / mL Example 22 24773 51542 Comparative Example HRO761 24612 83414 Compound number 11206 28343

[0614] The above test results show that the compound of the application has good mouse pharmacokinetic characteristics.

[0615] Experimental Example 6 Rat Pharmacokinetic Test Experiment:

[0616] 1. Experimental purpose

[0617] SD rats were used as experimental animals, and Example 4, Example 15 and Comparative Example HR0761 were administered by gavage. The drug concentration in the rat plasma at different times was determined by LC-MS / MS method to study the pharmacokinetic characteristics of Example 4, Example 15 and Comparative Example HR0761 in rats.

[0618] 2. Experimental scheme

[0619] 2.1 Experimental drugs and animals

[0620] Experimental drugs: Example 4, Example 15 and Comparative Example HR0761;

[0621] Animals: SD rats, male, 240-260 g, purchased from Shanghai Jihui Experimental Animal Co., Ltd.

[0622] 2.2 Drug preparation

[0623] An appropriate amount of Example 4, Example 15 and Comparative Example HR0761 was weighed, and an appropriate amount of dimethyl sulfoxide, solutol and physiological saline (final solvent 5% DMSO + 10% solutol + 85% saline) was added. Vortex oscillation, ultrasonic preparation of 1.0 mg / mL drug solution.

[0624] 2.3 Drug administration

[0625] The rats in the gavage groups of Example 4, Example 15 and Comparative Example HR0761 (3 rats per group) were fasted overnight and then administered by gavage (dose 10 mg / kg, administration volume 10 mL / kg). Food was given 4 hours after administration.

[0626] 3. Experimental operation

[0627] Animals were bled 0.2 mL of blood before dosing and at 0.083 hour, 0.25 hour, 0.5 hour, 1 hour, 2 hour, 4 hour, 6 hour, 8 hour and 24 hour after dosing, respectively, and anticoagulated with sodium heparin. Blood samples were placed on ice after collection and centrifuged to separate plasma (centrifugation conditions: 8000 rpm, 5 minutes). The collected plasma was stored at -80°C before analysis.

[0628] Determination of the content of Example 4, Example 15 and Comparative Example HR0761 in rat plasma after oral administration by LC-MS / MS method

[0629] 4. Pharmacokinetic parameter results

[0630] According to the calculated concentration of the compound in plasma at different time points, the pharmacokinetic parameters of Example 4, Example 15 and Comparative Example HR0761 in rats were calculated by winnonlin software. The pharmacokinetic parameters of Example 4, Example 15 and Comparative Example HR0761 of the present application are shown in Table 14.

[0631] Table 14

[0632] Example 4 C max ,ng / mL]]> AUC 0-i , ng.h / mL Example 15 19180 170483 Comparative Example HRO761 9016 46423 ​ 6965 63602

[0633] The above test results show that the compound of the present application has good rat pharmacokinetic characteristics.

[0634] The above illustrates the exemplary embodiments of the present application. It should be understood that the protection scope of the present application is not limited to the above exemplary embodiments. Any modification, equivalent replacement, improvement, etc. made by those skilled in the art within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in, Structural unit Selected from Structural unit Selected from: Structural unit Selected from Ring C is selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl or pyrazinyl; Ring E is selected from phenyl, pyridyl, pyrimidinyl, pyridazinyl or pyrazinyl; L1 is selected from a single bond; L2 is selected from -C(=O)-; R3 and R4 are each independently selected from H; R5 are independently selected from H, F, Cl, Br, SF5 or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups; Alternatively, two R5s are connected together to form a C 3-6 Cycloalkyl, the C 3-6 Cycloalkyl is optionally substituted with 1, 2 or 3 R; R7 are independently selected from H, F, Cl, Br, OH or C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R groups; R8 is selected from H; m and q are independently selected from 0, 1, 2 or 3; R is independently selected from H, F, Cl, Br, I or C 1-6 Alkyl, the C 1-6 Alkyl is optionally substituted with 1, 2 or 3 R'; R' is selected from F, Cl, Br or I.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R is each independently selected from H, F, Cl, Br, CH3, CF3, CHF2, CH2F, CF2Cl, CF2Br or CF2I.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R5 are independently selected from H, F, Cl, Br, SF5, Me or The Me and Optionally substituted with 1, 2 or 3 R; Alternatively, two R5s are connected together to form described Optionally substituted with 1, 2 or 3 R.

4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R5 are independently selected from H, F, Cl, Br, SF5, Me, CF3, Alternatively, two R5s are connected together to form 5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 6. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein R7 are independently selected from H, F, Cl, Br, OH, Me or The Me and Optionally substituted with 1, 2 or 3 R.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein R7 are independently selected from H, F, Cl, Br, OH, Me, CF3, 8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein Structural unit Selected from 9. A compound of the following formula or a pharmaceutically acceptable salt thereof, selected from 10. A pharmaceutical composition, wherein Comprising the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof.

11. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 10, in the preparation of a medicament for treating tumor-related diseases.

12. The use according to claim 11, wherein: The tumor is a malignant tumor with high microsatellite instability, a malignant tumor with mismatch repair deficiency, or a malignant tumor with a large number of (TA) n Repeated sequence malignancies.

13. The use according to claim 11, wherein: The tumor-related diseases are one or more diseases related to solid tumors.

14. The use according to claim 11, wherein: The tumor-related diseases include one or more of colorectal cancer, gastric cancer, endometrial cancer and ovarian cancer.

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