Hydrazine-structured compound, pharmaceutical composition thereof and use thereof

Abstract

Disclosed in the present invention are a hydrazine-structured compound, a pharmaceutical composition thereof and the use thereof. The hydrazine-structured compound of the present invention has a structure as shown in formula I. The hydrazine-structured compound as shown in formula I of the present invention has a good inhibitory effect on various mutations such as KRAS G12C, KRAS G12D, KRAS G12V and KRAS Q61H, and KRAS-amplified wild-type cells, and is expected to be used in the treatment and / or prevention of various Ras-related diseases.

Description

A hydrazine-based compound, its pharmaceutical composition and applications Technical Field

[0001] This invention relates to a hydrazine-based compound, its pharmaceutical composition, and its applications. Background Technology

[0002] Ras (Rat sarcoma viral oncogene) was first discovered in rat sarcoma. The mammalian ras gene family has three members: H-ras (HRAS), K-ras (KRAS), and N-ras (NRAS). K-ras has two variants, A and B, in its fourth exon. Ras genes are widely found in various eukaryotes such as mammals, fruit flies, fungi, nematodes, and yeast, with varying expression levels in different tissues. H-Ras is mainly expressed in skin and skeletal muscle, K-Ras is mainly expressed in the colon and thymus, and N-Ras is highly expressed in the testes. Ras proteins act as molecular switches in cell signal transduction, regulating signal transduction by binding to GTP / GDP, thereby modulating life processes such as cell proliferation, differentiation, aging, and apoptosis.

[0003] Ras mutations are closely related to the occurrence and development of human tumors, with approximately 30% of human tumors containing Ras mutations. KRAS mutations are the most common, accounting for about 85%, while NRAS and HRAS account for 12% and 3%, respectively. KRAS mutations are most prevalent in pancreatic cancer, colorectal cancer, and lung cancer; NRAS mutations are common in melanoma and acute myeloid leukemia; and HRAS mutations are common in bladder cancer and head and neck cancer. Ras proto-oncogene mutations mainly occur through point mutations. More than 150 different Ras point mutations have been identified, with mutations at glycine positions 12 and 13 and at glutamine position 61 being the most common.

[0004] For decades, researchers have focused on developing small-molecule inhibitors that directly target RAS. Scientists have long hoped to develop GTP-competitive inhibitors that directly act on the RAS protein. However, this has been unsuccessful due to the extremely strong affinity between GTP and RAS (pmol / L levels), the high concentration of GTP in cells (0.5 mM), and the lack of pockets in the RAS protein structure conducive to small molecule binding. In recent years, researchers have utilized KRAS... G12C Significant progress has been made in drug development using allosteric sites of mutants. In 2013, a research group reported on KRAS... G12C The discovery of small molecule inhibitors (Nature, 2013, 503, 548-551). They derived their inhibitors from KRAS. G12CA novel binding pocket located below the molecular switch II region was identified in the mutant. These inhibitors bind to this allosteric pocket and covalently bind to the nearby cysteine ​​residue Cys12, thereby selectively inhibiting KRAS. G12C Activation. Other researchers have reported KRAS with cellular activity. G12C Inhibitor (Science, 2016, 351, 604-608).

[0005] As one of the most important cancer driver genes, KRAS mutations in its G12C have made it possible to design drugs using irreversible covalent small molecules. Breakthroughs in related research have quickly made the development of corresponding drugs a hot topic in global anti-tumor drug research. In May 2021, Amgen's KRAS G12C inhibitor, Lumakras, received accelerated approval from the US FDA, achieving a major breakthrough in addressing the "undruggable" nature of KRAS.

[0006] Despite significant breakthroughs in KRAS G12C inhibitor research, current clinical trials show that these inhibitors suffer from short-lived efficacy and a high risk of developing drug resistance. Meanwhile, numerous other mutations, including those carrying KRAS G12D, G12V, G13D, G12R, G12S, and G12A mutations, as well as cancers with KRAS wild-type amplification, lack effective drugs. Therefore, there is an urgent need to find drugs that target KRAS mutations.

[0007] In summary, after decades of unremitting efforts, people have gradually deepened their understanding of Ras. To date, drugs targeting KRAS G12C mutations have been marketed. However, there are no particularly effective treatments for other different mutations. Finding compounds that have a good inhibitory effect on Ras remains a research hotspot and challenge in the field of new drug development. Summary of the Invention

[0008] The technical problem to be solved by the present invention is the lack of effective drugs for treating KRAS mutation diseases in the prior art, and provides a hydrazine-based compound, its pharmaceutical composition and application.

[0009] This invention provides a hydrazine-based compound as shown in Formula I, its stereoisomers, its pharmaceutically acceptable salts, its stereoisomers, pharmaceutically acceptable salts, its prodrugs, its deuterated compounds, or their PROTAC molecules:

[0010] Among them, R 1 for

[0011] X 1a For N or CRb1 R b1 For CN; X 1b and X 1c Independently, it can be H, F, or CH3;

[0012] X 2a For N or CH; X 2b For H or F; X 2c For H or F; X 2d For H, F,

[0013] X 3a and X 3b Independently N or CR b2 R b2 For H, CH3, or F; X 3c For H, F, Cl, CH3, CF3, CN, X 3d For CF3, I or

[0014] X 4a For CH3, Cl, CF3, -OCF2Cl or -OCHF2; X 4b For H, F, Cl or CH3; X 4c For H or CN;

[0015] It can be a single or double bond; X 5a CH3,

[0016] X 6a for X 6b For H or F;

[0017] X 7a For H or F;

[0018] X 8a For F, X 8b For H or F;

[0019] Y is O, S, NH, CH2 or

[0020] m1, m2 and n are independently 0, 1, 2, 3 or 4;

[0021] R 2 For hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups, C atoms substituted with one or more deuterium atoms 1-6Alkyl or -C(=O)R 21 ;

[0022] R 21 For hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl;

[0023] R 4 Independently deuterium, halogen, cyano, hydroxyl, C 1-6 Alkyl, with one or more R 4-1 Replacement C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 1-6 Alkyl-O-, with one or more R 4-2 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 4-3 Replacement C 1-6 Alkyl-S-, O=, C 3-10 Cycloalkyl-O-,-C(=O)R 4a -NR 4b1 R 4b2 -C(=O)OR 4c -C(=O)NR 4d1 R 4d2 or Alternatively, when n is 2, 3, or 4, any two R values ​​can be chosen. 4 Connected, independently forming 3- to 8-membered carbon rings with the atoms they are connected to, and bounded by one or more R... 4-4 Substituted 3- to 8-membered carbon rings, "4- to 8-membered heterocycles containing 1 to 3 heteroatoms independently selected from O, S, and N", or substituted by one or more R 4-5 The substituted "4- to 8-membered heterocycles containing 1 to 3 heteroatoms, which are independently selected from O, S, and N";

[0024] R 5 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 51 Replacement C 1-6 Alkyl, C 3-10 Cycloalkyl, hydroxyl, C 1-6 Alkyl-O-, with one or more R 52 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 53 Replacement C 1-6Alkyl-S-, amino, C 1-6 Alkyl-NH-, with one or more R 54 Replacement C 1-6 Alkyl-NH-, C 2-6 alkenyl, C 2-6 alkynyl or C 3-10 cycloalkyl-O-;

[0025] R 6 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 61 Replacement C 1-6 Alkyl, C 3-10 Cycloalkyl, hydroxyl, C 1-6 Alkyl-O-, with one or more R 62 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 63 Replacement C 1-6 Alkyl-S-, C 2-6 alkenyl, C 2-6 alkynyl, amino, C 1-6 Alkyl-NH-, with one or more R 64 Replacement C 1-6 Alkyl-NH- or C 3-10 cycloalkyl-O-;

[0026] T is N or CR 7 ;R 7 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 71 Replacement C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 Cycloalkyl, nitro, hydroxyl, C 1-6 Alkyl-O-, with one or more R 72 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 73 Replacement C 1-6 Alkyl-S-, C 1-6 Alkyl-NH-, with one or more R 74 Replacement C 1-6 Alkyl-NH- or C 3-10 cycloalkyl-O-;

[0027] R 51 R 52 R 53 R 54 R61 R 62 R 63 R 64 R 71 R 72 R 73 and R 74 Independently deuterium, halogen, cyano, C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C 2-6 Alkenyl, hydroxyl, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, -C(=O)R 51a -NR 51b1 R 51b2 -C(=O)OR 51c or -C(=O)NR 51d1 R 51d2 ;R 51a R 51b1 R 51b2 R 51c R 51d1 and R 51d2 Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl;

[0028] Or, any R 2 and R 6 Optional R 4 and R 6 Each of these compounds forms a 5-12 membered heterocyclic alkenyl group with the atoms to which it is attached, consisting of 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from O, S, and N, or is bounded by one or more R atoms. a The substituted "contains 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S and N";

[0029] R 4-1 R 4-2 R 4-3 R 4-4 R 4-5 and R a Independent of halogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6Alkyl groups, C atoms substituted with one or more deuterium atoms 1-6 Alkyl, C 2-6 alkenyl, cyano, hydroxyl, O=, C 1-6 Alkyl-O-, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, -NR 4f1 R 4f2 -C(=O)OR 4g or -C(=O)NR 4h1 R 4h2 ;

[0030] R 4a R 4b1 R 4b2 R 4c R 4d1 R 4d2 R 4e1 R 4e2 R 4f1 R 4f2 R 4g R 4h1 and R 4h2 Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl;

[0031] L is C 1-6 Alkylene, -OR L-1 -、-SR L-2 -or-NR L-3 (R L-4 )-;R L-1 R L-2 and R L-4 Independently for C 1-6 Alkylene or C substituted with one or more deuterium atoms 1-6 Alkylene; R L-3 For hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl;

[0032] R 3 C 3-12 cycloalkyl, with one or more R 3-1 Replacement C 3-12Cycloalkyl, "a 4- to 12-membered heterocycloalkyl group containing 1 to 3 heteroatoms independently selected from O, S, and N", and surrounded by one or more R 3-2 The substituted "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S, and N in a 4 to 12-membered heterocyclic alkyl group", and is subjected to one or more R 3-3 Replacement C 2-6 alkenyl or -NR 3-41 R 3-42 When there are multiple substituents, they may be the same or different.

[0033] R 3-41 and R 3-42 Independently hydrogen, C 1-6 Alkyl, C 3-10 cycloalkyl, C 1-6 Alkyl-O- or "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S and N in 4 to 12-membered heterocyclic alkyl groups";

[0034] R 3-1 R 3-2 and R 3-3 Independently for C 1-6 Alkyl, with one or more R 3-1-1 Replacement C 1-6 Alkyl, C 1-6 Alkyl-O-, halogen, aryl, with one or more R 3-1-8 Substituted aryl group, "5-12 membered heteroaryl group containing 1 to 4 heteroatoms independently selected from O, S, and N", or substituted by one or more R 3-1-9 Substituted "containing 1 to 4 heteroatoms, the heteroatoms being independently selected from O, S, and N 5-12 membered heteroaryl groups" or Or, any two Rs 3-1 Two Rs can be selected at random 3-2 Connected, independently forming 3- to 8-membered carbon rings with the atoms they are connected to, and bounded by one or more R... 3-1-2 Substituted 3- to 8-membered carbon rings, "4- to 8-membered heterocycles containing 1 to 3 heteroatoms independently selected from O, S, and N", or substituted by one or more R 3-1-3 Substituted "4-8 membered heterocycles containing 1-3 heteroatoms, the heteroatoms being independently selected from O, S, and N", C 6-20 aryl, with one or more R 3-1-4 Replacement C 6-20 Aryl, "a 5-12 membered heteroaryl group containing 1-4 heteroatoms independently selected from O, S, and N", and bonded by one or more R 3-1-5 Substituted "5-12 membered heteroaryl groups containing 1-4 heteroatoms, the heteroatoms being independently selected from O, S, and N", C 5-7 Cycloalkenyl, with one or more R 3-1-6 Replacement C5-7 Cycloalkenyl, "a 5-7 membered heterocyclic alkenyl group containing 1-3 heteroatoms independently selected from O, S, and N", or consisting of one or more R... 3-1-7 The substituents are “5-7 membered heterocyclic alkenyl groups containing 1-3 heteroatoms, which are independently selected from O, S, and N”; when there are multiple substituents, they may be the same or different;

[0035] R 3-1-1 Halogen, -NR 3b1 R 3b2 -OC(=O)NR 3c1 R 3c2 -OR 3d "Contains 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N" or is composed of one or more R 3-1-1-1 The substituted "contains 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N";

[0036] R 3d C 6-20 Aryl, "a 5-12 membered heteroaryl group containing 1 to 4 heteroatoms independently selected from O, S, and N", and bonded by one or more R 3d1 Replacement C 6-20 aryl, or, by one or more R 3d2 The substituted "contains 1 to 4 heteroatoms, the heteroatoms being independently selected from 5 to 12-membered heteroaryl groups of O, S and N";

[0037] R 3-1-1-1 for Halogen or C 1-6 alkyl;

[0038] R 3a1 R 3a2 R 3d1 and R 3d2 Independently hydrogen, halogen, cyano, C 1-6 Alkyl or with one or more R 3a11 Replacement C 1-6 Alkyl; R 3a11 Halogen or C 1-6 Alkyl-O-;

[0039] R 3-1-2 R 3-1-3 R 3-1-4 R 3-1-5 R 3-1-6 R 3-1-7 R 3-1-8 and R 3-1-9 Independently for C 1-6 Alkyl groups, C substituted with one or more halogens 1-6Alkyl groups, C atoms substituted with one or more deuterium atoms 1-6 Alkyl, halogen, cyano, C 2-6 alkenyl, C 2-6 alkynyl, hydroxyl, -C(=O)R 31a -NR 31b1 R 31b2 -C(=O)OR 31c -C(=O)NR 31d1 R 31d2 -S(O)2NR 31e1 R 31e2 -S(O)2R 31f -OC 1-6 Alkyl groups, -OC groups substituted with one or more halogens 1-6 Alkyl groups, -OC groups substituted with one or more deuterium atoms 1-6 Alkyl, -SC 1-6 Alkyl groups, -SC groups substituted with one or more halogens 1-6 Alkyl groups or -SC groups substituted with one or more deuterium groups 1-6 Alkyl group; when there are multiple substituents, they may be the same or different;

[0040] R 3b1 R 3b2 R 3c1 R 3c2 R 31a R 31b1 R 31b2 R 31c R 31d1 R 31d2 R 31e1 R 31e2 and R 31f Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 Alkyl group; when there are multiple substituents, they may be the same or different;

[0041] Or, R 4b1 and R 4b2 R 4d1 and R 4d2 R 4e1 and R 4e2 R 51b1 and R 51b2 R 51d1 and R 51d2 R 4f1 and R 4f2 R 4h1 and R 4h2 R 3a1 and R 3a2 R3b1 and R 3b2 R 3c1 and R 3c2 R 31b1 and R 31b2 R 31d1 and R 31d2 R 31e1 and R 31e2 Each of these atoms forms a 4-12 membered heterocyclic alkyl group with the N atom it is attached to, consisting of 1-3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from O, S, and N. b The substituted form "contains 1 to 3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N"; R b For deuterium, halogens, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups, C atoms substituted with one or more deuterium atoms 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 Cycloalkyl, nitro, hydroxyl, C 1-6 Alkyl-O-, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, amino, C 1-6 Alkyl-NH-, C substituted with one or more halogens 1-6 Alkyl-NH- or C-substituted with one or more deuterium groups 1-6 Alkyl-NH-;

[0042] The hydrazine compound with the structure shown in Formula I is not one of the following compounds:

[0043] In one scheme, in a hydrazine-based compound as shown in Formula I, R 1 for

[0044] X 1a For N or CR b1 R b1 For CN; X 1b and X 1c Independently H or F;

[0045] X 2aFor CH; X 2b H; X 2c For F; X 2d for

[0046] For single or double bonds, X 5a CH3 or

[0047] X 8a for X 8b For F;

[0048] Y is either O or CH2;

[0049] m1 and m2 are independently 1;

[0050] n is 0;

[0051] R 2 It is hydrogen or C 1-6 alkyl;

[0052] R 5 It is a halogen;

[0053] R 6 It is hydrogen, hydroxyl or C 1-6 Alkyl-O-;

[0054] T is N or CR 7 ;R 7 Halogen, cyano, C 1-6 Alkyl, C 3-10 cycloalkyl or with one or more R 71 Replacement C 1-6 alkyl;

[0055] R 71 It can be halogenated or cyano-based independently;

[0056] Or, R 2 and R 6 Each of them forms a 5-12 membered heterocyclic alkenyl group with the atoms to which they are attached, consisting of 2 to 4 heteroatoms, two of which are N and the other heteroatoms are independently selected from O, S and N.

[0057] L is -OR L-1 -;R L-1 C 1-6 Alkylene;

[0058] R 3 For one or more R 3-1 Replacement C 3-12 cycloalkyl, with one or more R 3-2The substituted group is defined as "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N"; when there are multiple substituents, they may be the same or different.

[0059] R 3-1 For one or more R 3-1-1 Replacement C 1-6 alkyl;

[0060] R 3-1-1 It is defined as "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S, and N in 4- to 12-membered heterocyclic alkyl groups" or being surrounded by one or more R groups. 3- 1-1-1 The substituted "contains 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N";

[0061] R 3-1-1-1 It is a halogen;

[0062] R 3-2 Independently halogen or

[0063] R 3a1 and R 3a2 It is a halogen.

[0064] In a certain scheme, the arbitrarily chosen R 2 and R 6 It forms a 5-12 membered heterocyclic alkenyl group with 2 to 4 heteroatoms, two of which are N and the other heteroatoms are independently selected from O, S and N, or is bonded by one or more R atoms. a The alternative is "containing 2 to 4 heteroatoms, two of which are N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S, and N". The heteroatom combination in "containing 2 to 4 heteroatoms, two of which are N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S, and N" is preferably (N, N), (N, N, and O), (N, N, and S), or (N, N, O, and S).

[0065] In one particular scheme, R 1 for

[0066] In one of the solutions, for

[0067] In one of the solutions, for

[0068] In one of the solutions, for

[0069] In one particular scheme, R 6 For H, hydroxyl, Me, -OMe, -SMe, Br, F, or -NHMe, or R 6 With R 2 form

[0070] In one embodiment, the hydrazine compound with the structure shown in Formula I is any of the following compounds:

[0071] In one embodiment, the hydrazine compound with the structure shown in Formula I is any of the following compounds:

[0072] In one embodiment, the hydrazine compound with the structure shown in Formula I is any of the following compounds:

[0073] The present invention also provides a pharmaceutical composition comprising substance A and pharmaceutical excipients; wherein substance A is a therapeutically effective amount of the above-described hydrazine compound of Formula I, its stereoisomer, its pharmaceutically acceptable salt, its stereoisomer pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule.

[0074] The present invention also provides the application of substance A in the preparation of RAS inhibitors, wherein substance A is a hydrazine-based compound as shown in Formula I, its stereoisomer, its pharmaceutically acceptable salt, its stereoisomer pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule.

[0075] In the application of substance A in the preparation of RAS inhibitors, the RAS refers to wild-type RAS and mutant RAS. The wild-type RAS may be wild-type amplified KRAS. The mutant RAS can be, for example, mutant KRAS, mutant HRAS, or mutant NRAS. The mutant KRAS can be mutant G12, G13, or Q61, such as KRAS G12C, KRAS G12D, KRAS G12S, KRAS G12A, KRAS G12V, or KRAS G13D, or even more specifically, KRAS G12C, KRAS G12D, or KRAS G12V. The mutant HRAS can be G12, G13, or Q61, such as mutant HRAS G12C, HRAS G12D, HRAS G12S, HRAS G12A, HRAS G12V, or HRAS G13D. The mutant NRAS can be mutant G12, G13, or Q61, such as NRAS G12C, NRAS G12D, NRAS G12S, NRAS G12A, NRAS G12V, or NRAS G12D. G13D.

[0076] The present invention also provides the application of substance A in the preparation of a drug for treating or preventing RAS-related diseases; wherein substance A is a hydrazine compound with a structure as shown in Formula I, its stereoisomer, its pharmaceutically acceptable salt, its stereoisomer pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule.

[0077] In the application of substance A in the preparation of drugs, the RAS can be wild-type RAS or mutant RAS. The wild-type RAS can be wild-type amplified KRAS. The mutant RAS can be, for example, mutant KRAS, mutant HRAS, or mutant NRAS. The mutant KRAS can be mutant G12, G13, or Q61, such as KRAS G12C, KRAS G12D, KRAS G12S, KRAS G12A, KRAS G12V, or KRAS G13D, or even more specifically, KRAS G12C, KRAS G12D, or KRAS G12V. The mutant HRAS can be G12, G13, or Q61, such as mutant HRAS G12C, HRAS G12D, HRAS G12S, HRAS G12A, HRAS G12V, or HRAS G13D. The mutant NRAS can be mutant G12, G13, or Q61, such as NRAS G12C, NRAS G12D, NRAS G12S, NRAS G12A, NRAS G12V, or NRAS G12D. G13D.

[0078] The application of substance A in the preparation of a pharmaceutical product, specifically for RAS-related diseases such as cancer. These cancers include, but are not limited to, one or more of the following: colon cancer, appendix cancer, pancreatic cancer, MYH-related polyposis, blood cancers, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophageal cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia, and melanoma.

[0079] This invention also provides the use of substance A in the preparation of a medicament for treating or preventing cancer; substance A is a hydrazine compound with a structure as shown in Formula I, its stereoisomer, its pharmaceutically acceptable salt, its pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule. The cancers include, for example, colon cancer, appendix cancer, pancreatic cancer, MYH-related polyposis, blood cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophageal cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia, and melanoma, or one or more of these.

[0080] The term "multiple" refers to 2, 3, 4, or 5.

[0081] The term "pharmaceutically acceptable salt" refers to a salt prepared from the compounds of the present invention with a relatively non-toxic, pharmaceutically acceptable acid or base. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting a sufficient amount of a pharmaceutically acceptable base with the neutral form of such compounds in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, lithium salts, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, zinc salts, bismuth salts, ammonium salts, and diethanolamine salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting a sufficient amount of a pharmaceutically acceptable acid with the neutral form of such compounds in a pure solution or a suitable inert solvent. Pharmaceutically acceptable acids include inorganic acids, including but not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, phosphoric acid, phosphorous acid, and sulfuric acid. The pharmaceutically acceptable acids include organic acids, including but not limited to: acetic acid, propionic acid, oxalic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, salicylic acid, tartaric acid, methanesulfonic acid, isonicotinic acid, acidic citric acid, oleic acid, tannic acid, pantothenic acid, hydrogen tartrate, ascorbic acid, gentian acid, fumaric acid, gluconic acid, succinic acid, formic acid, ethanesulfonic acid, dihydroxynaphthyl acid (i.e., 4,4'-methylene-bis(3-hydroxy-2-naphthylcarboxylic acid)), amino acids (e.g., glutamic acid, arginine), etc. When the compounds of the present invention contain relatively acidic and relatively basic functional groups, they can be converted into base addition salts or acid addition salts. For details, see Berge et al., "Pharmaceutical Salts", Journal of Pharmaceutical Science 66: 1-19 (1977) or Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl and Camille G. Wermuth, ed., Wiley-VCH, 2002).

[0082] The term "stereoisomer" refers to isomers of molecules that have the same order of interconnection of atoms or groups of atoms but different spatial arrangements, such as cis-trans isomers, optical isomers, or trans-blocked isomers. These stereoisomers can be separated, purified, and enriched by asymmetric synthesis methods or chiral separation methods (including but not limited to thin-layer chromatography, rotational chromatography, column chromatography, gas chromatography, high-performance liquid chromatography, etc.). They can also be obtained through chiral resolution by bonding (chemical bonding, etc.) or salt formation (physical bonding, etc.) with other chiral compounds.

[0083] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0084] The term "alkyl" refers to a straight-chain or branched alkyl group having a specified number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and similar alkyl groups.

[0085] The term "alkylene" refers to a linking group between two other classes. It can be straight or branched, and examples include, but are not limited to, -CH2-, -CH2CH2-, -CH2CH2CH2CH(CH3)-, and -CH2CH(CH2CH3)CH2-.

[0086] The term "alkoxy group" refers to the group -OR X , where R X It is an alkyl group as defined above.

[0087] The term "cycloalkyl" refers to a saturated cyclic group consisting only of carbon atoms, having a specified number of carbon atoms (e.g., C3 to C6), and can be monocyclic, bridged, or spirocyclic. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0088] The term "aryl" refers to an aromatic group consisting of carbon atoms, with each ring possessing aromaticity. Examples include phenyl or naphthyl groups.

[0089] The term "heteroaryl" refers to a cyclic group having a specified number of ring atoms (e.g., 5 to 12), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S). It can be monocyclic or polycyclic, and at least one ring is aromatic (conforming to Hückel's rule). Heteroaryl groups are linked to other segments of a molecule through either an aromatic or non-aromatic ring. Heteroaryl groups include, but are not limited to, furanyl, pyrroleyl, thiopheneyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, and indoleyl.

[0090] The term "heterocyclic alkyl" refers to a cyclic group having a specified number of ring atoms (e.g., 3 to 8), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S), which can be monocyclic, bridged, or spirocyclic, and each ring is saturated. Heterocyclic alkyl groups include, but are not limited to, aza-butanediol, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, piperidinyl, etc.

[0091] The term "hydroxyl group" refers to a -OH group.

[0092] The term "cyano" refers to a -CN group.

[0093] The term "C" refers to the determination of the carbon number range.x1 -C y1 Substituents such as "C" (where x1 and y1 are integers) x1 -C y1 "alkyl", "C" x1 -C y1 "Cycloalkyl, "C x1 -C y1 "Cycloalkenyl, "C x1 -C y1 "alkoxy group", "C" x1 -C y1 "Alkenyl, "C x1 -C y1 "Alkyne group, "C x1 -C y1 "Aryl, "C x1 -C y1 "Heteroaryl groups all indicate the number of carbons without substituents. For example, C1-C6 alkyl groups indicate C1-C6 alkyl groups without substituents."

[0094] In this invention, when R 4 and R 6 It forms a 5-12 membered heterocyclic alkenyl group with 1 to 4 heteroatoms, one of which is N and the other heteroatoms are independently selected from O, S and N, or is bonded by one or more R a2 When replacing "containing 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S and N", the heteroatom combination of "containing 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S and N" is (N), (N and O), (N and S) or (N, O and S).

[0095] In this invention, when R 4b1 and R 4b2 R 4d1 and R 4d2 R 4e1 and R 4e2 R 51b1 and R 51b2 R 51d1 and R 51d2 R 4f1 and R 4f2 R 4h1 and R 4h2 R 3a1 and R 3a2 R 3b1 and R 3b2 R 3c1 and R 3c2 R 31b1 and R 31b2 R31d1 and R 31d2 R 31e1 and R 31e2 Each of these compounds forms a 4-12 membered heterocyclic alkyl group with one to three heteroatoms, one of which is N, and the other heteroatoms are independently selected from O, S, and N, or is bonded to one or more R atoms. b When replacing "containing 1 to 3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N", the heteroatom combination of "containing 1 to 3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N" is (N), (N and O), (N and S) or (N, O and S).

[0096] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0097] The reagents and raw materials used in this invention are all commercially available.

[0098] The positive and progressive effects of this invention are as follows: the hydrazine-based compounds of Formula I in this invention have a good inhibitory effect on various mutations such as KRAS G12C, KRAS G12D, KRAS G12V, and KRAS Q61H, as well as wild-type cells with KRAS amplification, and are expected to treat and / or prevent various diseases related to Ras. Detailed Implementation

[0099] The present invention is further illustrated below by way of examples, but these examples do not limit the invention to the scope of the embodiments described. Experimental methods in the following examples, unless otherwise specified, were performed according to conventional methods and conditions, or as selected in the product instructions. All solvents used in the following examples are of analytical grade or chromatographic grade. When the solvents used in the following examples are mixed solvents, unless otherwise stated, all are volume ratios. In this invention, room temperature refers to ambient temperature, which is 10°C-35°C. Overnight refers to 8-15 hours. Reflux refers to the solvent reflux temperature at normal pressure.

[0100] The following is a list of abbreviations used in the examples: DMF N,N-dimethylformamide ACN Acetonitrile DIPEA Diisopropylethylamine TIPS Triisopropylsilyl THF Tetrahydrofuran DCM Dichloromethane EA Ethyl acetate DMSO Dimethyl sulfoxide PE Petroleum ether MeOH Methanol cataCXium A Pd G3 Methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) Pd(DPEphos)Cl2 Bis(diphenylphosphine ether)palladium(II) dichloride DABCO 1,4-diazabicyclo[2.2.2]octane TFA Trifluoroacetic acid Boc Tert-butoxycarbonyl m-CPBA m-chloroperoxybenzoic acid Bn Benzyl NIS N-iodosuccinimide THP 2-Tetrahydropyran DHP 3,4-dihydro-2H-pyran TBAF Tetrabutylammonium fluoride

[0101] Synthetic routes of compounds 1 and 2

[0102] Synthesis of compound 1-c

[0103] 7-Bromo-2,4-dichloro-6,8-difluoroquinazoline (400 mg, 1.27 mmol), tetrahydrofuran (100 mL), DIPEA (444 mg, 3.44 mmol), and tert-butyl 1,2-diazaphen-1-carboxylic acid (383 mg, 1.91 mmol) were added to the reaction flask, and the mixture was stirred at this temperature for 12 hours. The reaction solution was concentrated at room temperature, and the residue was extracted with dichloromethane (100 mL x 3) by adding saturated sodium bicarbonate solution. The organic phase was concentrated and purified by column chromatography (PE / EA = 10 / 1) to give compound 1-c (396 mg, 65%). LC-MS (ESI): m / z = 477.6 (M+H) + .

[0104] Synthesis of compound 1-b

[0105] Compound 1-c (400 mg, 0.84 mmol), tetrahydrofuran (15 mL), DABCO (376 mg, 3.35 mmol), DMF (15 mL), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (400 mg, 2.51 mmol), and cesium carbonate (818 mg, 2.51 mmol) were added to the reaction flask, and the mixture was stirred overnight under nitrogen. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was concentrated and purified by column chromatography (MeOH / DCM = 1 / 10) to give compound 1-b (380 mg, 76%). LC-MS (ESI): m / z = 600.1 (M+H) + .

[0106] Synthesis of compound 1-a

[0107] 1-b (231 mg, 0.39 mmol), toluene (5 mL), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboronyl-2-yl)-7-fluorobenzo[b]thiophene-2-yl) tert-butyl carbamate (467 mg, 1.15 mmol), cesium carbonate (627 mg, 1.92 mmol), and PdCl2 (DPEphos) (28 mg, 0.04 mmol) were added to a microwave-safe tube. The tube was purged three times with nitrogen and then stirred in an oil bath preheated to 100 °C for 1.5 hours. The reaction mixture was directly evaporated to dryness and purified by column chromatography (mobile phase: dichloromethane / methanol = 100 / 0; 100 / 6) to give compound 1-a (243 mg, 78%). LC-MS (ESI): m / z = 812.8 (M+H) + .

[0108] Synthesis of compounds 1 and 2

[0109] 1-a (236 mg, 0.29 mmol), dichloromethane (9 mL), and trifluoroacetic acid (3 mL) were added to the reaction flask and stirred at room temperature for 3 hours. The reaction solution was evaporated to dryness, saturated sodium bicarbonate (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The extract was concentrated and chirally resolved to give compound 1 (31 mg, 17%) and compound 2 (27 mg, 15%).

[0110] Compound 1: LC-MS (ESI): m / z = 612.7 (M+H) + ; 1H NMR (400MHz, DMSO_d6): δ9.12 (1H, dd, J1=12.8Hz, J2=1.6Hz), 8.11 (2H, s), 7. 33(1H,dd,J1=8.4Hz, J2=5.2Hz), 7.15(1H,t,J=8.8Hz), 5.70(1H,t,J=6.4Hz), 5.28(1H,d,J=53.6Hz),4.15-3.85(4H,m),3.18-2.89(6H,m),2.88-2.76(1H,m ),2.08-1.96(2H,m),1.94-1.80(3H,m),1.79-1.72(2H,m),1.73-1.55(4H,m).

[0111] Compound 2: LC-MS (ESI): m / z = 612.7 (M+H) + ; 1 H NMR (400MHz, DMSO_d6): δ9.12 (1H, dd, J1=12.8Hz, J2=1.6Hz), 8.12 (2H, s), 7. 33(1H,dd,J1=8.4Hz, J2=5.6Hz), 7.15(1H,t,J=8.4Hz), 5.70(1H,t,J=6.0Hz), 5.28(1H,d,J=53.2Hz),4.15-3.85(4H,m),3.18-2.89(6H,m),2.88-2.76(1H,m ),2.18-1.96(2H,m),1.94-1.80(3H,m),1.79-1.72(2H,m),1.73-1.55(4H,m).

[0112] Synthetic routes of compounds 3 and 4

[0113] Synthesis of compound 3-d

[0114] To a reaction flask, add 200 mg (0.66 mmol) of 1,4,5-oxadiazine-heptane-4,5-dicarboxylic acid di-tert-butyl ester, 10 mL of dichloromethane, and 3 mL of trifluoroacetic acid. Stir at room temperature under nitrogen for 3 hours. The reaction mixture was evaporated to dryness to give compound 3-d (67 mg, 99%), which was used directly in the next reaction without purification. LC-MS (ESI): m / z 103.4 (M+H) + .

[0115] Synthesis of compound 3-c

[0116] Compound 3-d (67 mg, 0.66 mmol), tetrahydrofuran (10 mL), and N,N-diisopropylethylamine (0.54 mL, 3.28 mmol) were added to the reaction flask. 7-Bromo-2,4-dichloro-6,8-difluoroquinazoline (206 mg, 0.66 mmol) was added under ice-water bath conditions. After addition, the mixture was allowed to warm to room temperature and stirred overnight. The next day, water was added to the reaction mixture, and the solution was extracted eight times with ethyl acetate. The combined organic phase suspension was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–30%) to give compound 3-c (204 mg, 82%). LC-MS (ESI): m / z 379.1 (M+H) + .

[0117] Synthesis of compound 3-b

[0118] Compound 3-c (204 mg, 0.54 mmol) was added to a reaction flask, followed by 10 mL of dried tetrahydrofuran, 10 mL of dried N,N-dimethylformamide, 257 mg (1.61 mmol) of ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol, 525 mg (1.61 mmol) of cesium carbonate, and 241 mg (2.15 mmol) of triethylenediamine. The mixture was stirred overnight at room temperature under nitrogen atmosphere. The next day, water was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate. The combined organic phases were washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 3-b (140 mg, 52%). LC-MS (ESI): m / z 502.2 (M+H) + .

[0119] Synthesis of compound 3-a

[0120] Compound 3-b (140 mg, 0.28 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (225 mg, 0.56 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (20 mg, 0.028 mmol), cesium carbonate (272 mg, 0.84 mmol), and toluene (10 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 3-a (169 mg, 85%). LC-MS (ESI): m / z 714.7 (M+H) + .

[0121] Synthesis of Compounds 3 and 4

[0122] Trifluoroacetic acid (2 mL) was added to a 6 mL solution of compound 3-a (169 mg, 0.24 mmol) in dichloromethane at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, methanol was added, and the pH was adjusted to 7 with saturated sodium bicarbonate solution. The solution was evaporated to dryness, purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%), and further chiral resolution was performed to give compound 3 (39 mg, 27%) and compound 4 (31 mg, 21%).

[0123] Compound 3: LC-MS (ESI): m / z 614.4 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ9.12 (1H, d, J = 11.6Hz), 8.13 (2H, s), 7.40-7.28 (1H, m),7.16(1H,t,J=8.8Hz),5.98(1H,t,J=6.0Hz),5.28(1H,d,J=54.0Hz),4. 29-4.17(1H,m),4.17-4.03(2H,m),4.02-3.90(3H,m),3.78-3.68(2H,m),3 .16-2.96(5H,m),2.89-2.76(1H,m),2.19-1.94(3H,m),1.91-1.68(3H,m);

[0124] Compound 4: LC-MS (ESI): m / z 614.4 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ9.12 (1H, d, J = 12.4Hz), 8.13 (2H, s), 7.40-7.28 (1H, m),7.16(1H,t,J=8.8Hz),5.98(1H,t,J=6.0Hz),5.28(1H,d,J=54.0Hz),4. 29-4.17(1H,m),4.17-4.03(2H,m),4.02-3.90(3H,m),3.78-3.68(2H,m),3 .16-2.96(5H,m),2.89-2.76(1H,m),2.19-1.94(3H,m),1.91-1.68(3H,m).

[0125] Synthetic route of compound 5

[0126] Synthesis of compound 5-g

[0127] In a reaction flask, add 4-bromo-2,3,5,6-tetrafluorobenzoic acid (5 g, 18.32 mmol) and dichloromethane (60 mL). Add 3 drops of DMF at room temperature. After stirring in an ice bath for 10 minutes, add 6.2 mL of oxaloyl chloride (73.28 mmol). Stir the reaction mixture at room temperature for 1 hour. Remove the solvent by evaporation. Dilute the residue with anhydrous tetrahydrofuran (50 mL) to prepare solution A. In another reaction flask, add sodium hydroxide (3.35 g, 83.75 mmol) and pure water (120 mL). After stirring in an ice bath for 5 minutes, add S-methylisothiourea hemisulfate (15.30 g, 54.95 mmol). After stirring in an ice bath for 5 minutes, add tetrahydrofuran (20 mL) and solution A. Stir the reaction mixture in an ice bath for 30 minutes. Extracted with ethyl acetate / petroleum ether (1:1), washed twice with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude product. Anhydrous DMF (60 mL) was added. After purging with nitrogen three times, the reaction mixture was heated to 120 °C and stirred for 2 hours. Cooled to room temperature, the mixture was added dropwise to ice water (containing 12 mL of saturated sodium bicarbonate aqueous solution), filtered, the filter cake washed with water and petroleum ether, and air-dried overnight. The solid was slurried with ethyl acetate / petroleum ether (1:1) to give 5-g (5.0 g, yield 84%) of the compound. LC-MS (ESI): m / z 325.0 (M+H) + .

[0128] Synthesis of compound 5-f

[0129] Under nitrogen atmosphere in an ice-water bath, NaH (388 mg, 0.13 mmol) was suspended in 10 mL of anhydrous DMF, followed by the dropwise addition of anhydrous methanol (336 μL, 8.31 mmol). The reaction mixture was stirred in an ice-water bath for 1 hour. Then, a 10 mL solution of DMF containing compound 5-g (1.0 g, 2.77 mmol, 90%) was added dropwise. After the addition was complete, the reaction mixture was heated to room temperature and stirred for 2 hours. The reaction mixture was quenched with saturated ammonium chloride solution (50 mL), resulting in the precipitation of a large amount of solid. The solid was filtered, washed with water (50 mL), collected, and dried under vacuum overnight. Compound 5-f (1.15 g, crude product) was obtained. The crude product was used directly in the next reaction without purification. LC-MS (ESI): m / z = 337.1 [M+H] + .

[0130] Synthesis of compound 5-e

[0131] Compound 5-f (1.15 g, 3.41 mmol) was dissolved in 30 mL of anhydrous acetonitrile at room temperature. DIPEA (4.23 mL, 25.58 mmol) and phosphorus oxychloride (4.77 mL, 51.17 mmol) were added, and the reaction mixture was stirred at 80 °C under nitrogen for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (EA / PE = 0–30%) to give compound 5-e (600 mg, 49%). LC-MS (ESI): m / z = 354.9 [M+H] + .

[0132] Synthesis of compound 5-d

[0133] Compound 5-e (600 mg, 1.69 mmol) was dissolved in 50 mL of anhydrous THF at room temperature. DIPEA (1.39 mL, 8.44 mmol) and tert-butyl 1,2-diazacycloheptan-1-carboxylate (338 mg, 1.69 mmol) were added in an ice-water bath. The reaction mixture was brought to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (EA / PE = 0–20%) to give compound 5-d (649 mg, 74%). LC-MS (ESI): m / z = 519.1 [M+H] + .

[0134] Synthesis of compound 5-c

[0135] Compound 5-d (649 mg, 1.25 mmol) was dissolved in 50 mL of ethyl acetate at room temperature. m-CPBA (761 mg, 3.75 mmol, 85%) was added in an ice-water bath. The reaction mixture was brought to room temperature and stirred for 2 hours. The reaction solution was quenched with saturated sodium sulfite solution (50 mL), extracted with ethyl acetate (50 mL x 2), and the organic phase was washed successively with saturated sodium bicarbonate solution (100 mL) and saturated brine (100 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid column chromatography (EA / PE = 0–50%) to give compound 5-c (588 mg, 85%). LC-MS (ESI): m / z = 551.3 [M+H] + .

[0136] Synthesis of compound 5-b

[0137] Compound 5-c (580 mg, 1.05 mmol) was dissolved in 40 mL of anhydrous dichloromethane at room temperature. ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (502 mg, 3.16 mmol) was added, followed by sodium tert-butoxide (354 mg, 3.68 mmol) in an ice-water bath. The reaction mixture was brought to room temperature and stirred for 1 hour. The reaction solution was cooled in an ice-water bath, quenched with water (50 mL), extracted with DCM (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid separation column chromatography (MeOH / DCM = 0–10%) to give compound 5-b (565 mg, 85%). LC-MS (ESI): m / z = 630.2 [M+H] + .

[0138] Synthesis of compound 5-a

[0139] Compound 5-b (150 mg, 0.24 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboronyl-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (192 mg, 0.48 mmol), cesium carbonate (233 mg, 0.71 mmol), dichloro[bis(diphenylphosphine)ether]palladium(II) (17 mg, 0.024 mmol), and anhydrous toluene (10 mL) were added to a microwave-safe tube at room temperature. The mixture was purged with nitrogen three times, and then stirred at 100 °C for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (MeOH / DCM = 0–10%) to give compound 5-a (200 mg, 99%). LC-MS (ESI): m / z = 842.3 [M+H] + .

[0140] Synthesis of Compound 5

[0141] Compound 5-a (200 mg, 0.24 mmol) was dissolved in 20 mL of dichloromethane at room temperature. Trifluoroacetic acid (6 mL) was added under ice-water bath conditions, and the reaction mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (alkaline method, ammonium bicarbonate system). The residue was then lyophilized to give compound 5 (100 mg, 65%). LC-MS (ESI): m / z = 642.2 [M+H] + ; 1H NMR(DMSO-d6,400MHz): δ8.16(2H,s),7.41-7.35(1H,m),7.16(1H,t,J=8.8Hz) ,6.57(1H,s),5.30(1H,d,J=54.0Hz),4.14(1H,d,J=10.0Hz),4.06(1H,d,J=10 .4Hz),3.85(3H,s),3.75(2H,t,J=5.6Hz),3.23-3.05(3H,m),2.93-2.72(3H,m ),2.21-2.01(3H,m),1.95-1.75(5H,m),1.73-1.64(2H,m),1.62-1.51(2H,m).

[0142] Synthetic route of compound 6

[0143] Synthesis of compound 6-c

[0144] Add 3-d (67 mg, 0.66 mmol), tetrahydrofuran (10 mL), and N,N-diisopropylethylamine (0.54 mL, 3.28 mmol) to the reaction flask. Then add 7-bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (239 mg, 0.66 mmol) under ice-water bath conditions. Allow the mixture to cool to room temperature and stir for 2 hours. The reaction solution was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–50%) to give compound 6-c (253 mg, 90%). LC-MS (ESI): m / z 429.0 (M+H) + .

[0145] Synthesis of compound 6-b

[0146] Add 6-c (253 mg, 0.59 mmol), dried tetrahydrofuran (10 mL), dried N,N-dimethylformamide (10 mL), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (281 mg, 1.77 mmol), cesium carbonate (576 mg, 1.77 mmol), and triethylenediamine (264 mg, 2.36 mmol) to the reaction flask. Stir the mixture overnight at room temperature under nitrogen. The next day, add water to the reaction mixture, extract twice with ethyl acetate, wash the combined organic phases three times with saturated brine, dry to anhydrous sodium sulfate, filter, evaporate to dryness, and purify the crude product by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 6-b (49 mg, 15%). LC-MS (ESI): m / z 552.0 (M+H) + .

[0147] Synthesis of compound 6-a

[0148] 6-b (49 mg, 0.089 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (72 mg, 0.18 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (9 mg, 0.009 mmol), cesium carbonate (145 mg, 0.44 mmol), and toluene (5 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 6-a (40 mg, 59%). LC-MS (ESI): m / z 764.6 (M+H) + .

[0149] Synthesis of Compound 6

[0150] Trifluoroacetic acid (2 mL) was added to a solution of 6-a (40 mg, 0.052 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 6 (5 mg, 14%). LC-MS (ESI): m / z 664.2 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ9.83(1H,s),8.06(2H,s),7.28-7.20(1H,m),7.17-7.07(1H,m),6.15(1H,t,J=6.4Hz),5.27 (1H,d,J=54.0Hz),4.32-3.90(6H,m),3.78-3.68(2H,m),3.14-2.96(5H,m),2.87-2.76(1H,m),2.16-1.61(6H,m).

[0151] Synthetic route of compound 8

[0152] Synthesis of compound 8-h

[0153] Under ice-water bath conditions, benzyl alcohol (859 μL, 8.31 mmol), N,N-dimethylformamide (10 mL), and sodium hydride (221 mg, 5.54 mmol) were added to the reaction flask. The mixture was stirred under nitrogen atmosphere for 1 hour at this temperature, followed by the addition of 5-g of the compound (900 mg, 2.77 mmol). After the addition was complete, the mixture was allowed to warm to room temperature and stirred for 3 hours. Sodium hydride (221 mg, 5.54 mmol) was then added under ice-water bath conditions, and the mixture was stirred for 1 hour at this temperature. Under ice-water bath conditions, 10 mL of saturated ammonium chloride solution was added dropwise to the reaction mixture, resulting in the precipitation of a large amount of solid. The solid was filtered, washed with water, and then removed with ethanol. The mixture was then dried under a pump to obtain compound 8-h (1130 mg, 99%), which was used directly in the next reaction without purification. LC-MS (ESI): m / z 413.0 (M+H) + .

[0154] Synthesis of compound 8-g

[0155] Compound 8-h (1130 mg, 2.74 mmol), acetonitrile (30 mL), N,N-diisopropylethylamine (3.39 mL, 20.51 mmol), and phosphorus oxychloride (3.82 mL, 41.02 mmol) were added to the reaction flask, and the mixture was stirred at 80 °C for 2 hours under nitrogen atmosphere. The mixture was evaporated to dryness, rinsed twice with dichloromethane, and dried under vacuum to give crude compound 8-g (1180 mg, 100%), which was used directly in the next reaction without purification. LC-MS (ESI): m / z 431.1 (M+H) + .

[0156] Synthesis of compound 8-f

[0157] Compound 8-g (1180 mg, 2.73 mmol), tetrahydrofuran (30 mL), N,N-diisopropylethylamine (2.26 mL, 13.67 mmol), and tert-butyl 1,2-diazaphen-1-carboxylate (547 mg, 2.73 mmol) were added to the reaction flask under ice-water bath conditions. The mixture was allowed to warm to room temperature and stirred overnight. The next day, the solution was quenched with water, extracted twice with ethyl acetate, washed with saturated brine, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–5%) to give compound 8-f (811 mg, 50%). LC-MS (ESI): m / z 595.0 (M+H) + .

[0158] Synthesis of compound 8-e

[0159] Compound 8-f (811 mg, 1.36 mmol), ethyl acetate (30 mL), and m-chloroperoxybenzoic acid (829 mg, 4.09 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 1 hour. A saturated sodium sulfite solution was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution, then with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–30%) to give compound 8-e (800 mg, 94%). LC-MS (ESI): m / z 627.4 (M+H) + .

[0160] Synthesis of compound 8-d

[0161] Compound 8-e (800 mg, 1.28 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (609 mg, 3.83 mmol), dry dichloromethane (40 mL), and sodium tert-butoxide (429 mg, 4.46 mmol) were added to the reaction flask under ice-water bath conditions. The mixture was slowly heated to room temperature and stirred for 1 hour. The reaction solution was then directly purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 8-d (655 mg, 73%). LC-MS (ESI): m / z 706.4 (M+H) + .

[0162] Synthesis of compound 8-c

[0163] Compound 8-d (655 mg, 0.93 mmol), dichloromethane (9 mL), and trifluoroacetic acid (3 mL) were added to a reaction flask and stirred at room temperature for 2 hours. The mixture was evaporated to dryness, extracted twice with saturated sodium bicarbonate solution and ethyl acetate, evaporated to dryness again, and purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 8-c (466 mg, 97%). LC-MS (ESI): m / z 516.1 (M+H) + .

[0164] Synthesis of compound 8-b

[0165] Compound 8-c (460 mg, 0.89 mmol), ethanol (30 mL), and paraformaldehyde (460 mg, 15.32 mmol) were added to a reaction flask, and the mixture was stirred overnight at 100 °C under nitrogen. The next day, the reaction solution was evaporated to dryness and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 8-b (150 mg, 32%). LC-MS (ESI): m / z 528.1 (M+H) + .

[0166] Synthesis of compound 8-a

[0167] Compound 8-b (50 mg, 0.095 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoborhexane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (77 mg, 0.19 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (7 mg, 0.009 mmol), cesium carbonate (154 mg, 0.47 mmol), and toluene (10 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 8-a (63 mg, 90%). LC-MS (ESI): m / z 740.8 (M+H) + .

[0168] Synthesis of Compound 8

[0169] Trifluoroacetic acid (2 mL) was added to a solution of compound 8-a (63 mg, 0.085 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 8 (13 mg, 24%). LC-MS (ESI): m / z 640.2 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ8.13(2H,s),7.41-7.32(1H,m),7.16(1H,t,J=8.8Hz),5.50-5.17(3H,m), 4.80-4.64(1H,m),4.18-3.92(2H,m),3.21-2.98(6H,m),2.91-2.78(1H,m),2.22-1.48(12H,m).

[0170] Synthetic route of compound 9

[0171] Synthesis of compound 9-c

[0172] 7-Bromo-2,4,6-trichloro-8-fluoroquinazoline (300 mg, 0.91 mmol) and THF (20 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement, and 3-d (102 mg, 1.00 mmol) and DIPEA (469 mg, 3.63 mmol) were added under ice bath conditions. The reaction mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 0 / 10) to give compound 9-c (320 mg, 89%).

[0173] Synthesis of compound 9-b

[0174] At room temperature, 9-c (320 mg, 0.81 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (257 mg, 1.62 mmol), THF (10 mL), DMF (10 mL), triethylenediamine (363 mg, 3.23 mmol), and cesium carbonate (790 mg, 2.42 mmol) were added sequentially to a reaction flask. The mixture was protected by N2 displacement and reacted overnight at room temperature. Water was added to the reaction solution, and the mixture was extracted with EA. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 9-b (100 mg, 24%). LC-MS (ESI): m / z 519.8 (M+H) + .

[0175] Synthesis of compound 9-a

[0176] At room temperature, 9-b (100 mg, 0.19 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (156 mg, 0.39 mmol), toluene (5 mL), bis(diphenylphosphine ether)palladium dichloride (14 mg, 0.02 mmol), and cesium carbonate (314 mg, 0.96 mmol) were added sequentially to a microwave-safe mixture. The mixture was protected with N2 displacement and heated to 100 °C for 3 hours. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 9-a (80 mg, 57%). LC-MS (ESI): m / z 730.6 (M+H) + .

[0177] Synthesis of Compound 9

[0178] 9-a (80 mg, 0.11 mmol), DCM (4 mL), and trifluoroacetic acid (1.5 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to give compound 9 (23 mg, 33%). LC-MS (ESI): m / z 630.4 (M+H) + ; 1HNMR(DMSO-d6,400MHz): δ9.45(1H,s),8.10(2H,s),7.41-6.98(2H,m),6.11-5.96(1H,m),5.45-5.09(1H ,d,J=54.0Hz),4.36-3.83(6H,m),3.73(2H,s),3.18-2.92(5H,m),2.89-2.74(1H,m),2.23-1.60(6H,m).

[0179] Synthetic route of compound 10

[0180] Synthesis of compound 10-c

[0181] Compound 3-d (338 mg, 3.31 mmol) and N,N-diisopropylethylamine (2139 mg, 16.55 mmol) were added to a solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (835 mg, 3.31 mmol) in dichloromethane (20 mL) under dry ice and ethyl acetate bath. The mixture was reacted at this temperature for 2 hours, resulting in the precipitation of a large amount of insoluble matter. The mixture was filtered, and the filter cake was collected. Water was added to the filtrate, and the extract was obtained with dichloromethane (60 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The evaporated solid was combined with the filter cake to give compound 10-c (1.05 g, 100%). LC-MS (ESI): m / z 318.0 (M+H) + .

[0182] Synthesis of compound 10-b

[0183] Under ice-water bath conditions, a solution of compound 10-c (500 mg, 1.57 mmol) in dichloromethane (150 mL) was reacted with ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (275 mg, 1.73 mmol), followed by the slow addition of sodium tert-butoxide (302 mg, 3.14 mmol). The mixture was reacted at this temperature for 2 hours. Water was added to the reaction solution, and the mixture was extracted with dichloromethane (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by rapid column chromatography (mobile phase: DCM:MeOH = 10:1 / DCM, 0-100%) to give compound 10-b (540 mg, 78%). LC-MS (ESI): m / z 441.3 (M+H) + .

[0184] Synthesis of compound 10-a

[0185] Compound 10-b (200 mg, 0.45 mmol), 6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-((triisopropylsilyl)ethynyl)naphthyl-2-amine (318 mg, 0.68 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II)cataCXium A Pd G3 (33 mg, 0.045 mmol), potassium phosphate (289 mg, 1.36 mmol), tetrahydrofuran (12 mL), and water (2 mL) were added to a 30 mL microwave tube. The microwave tube was sealed, purged with nitrogen several times, and heated to 65 °C overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL * 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give compound 10-a (338 mg, 100%). The crude product was used directly in the next reaction without further purification.

[0186] Synthesis of Compound 10

[0187] Cesium fluoride (688 mg, 4.53 mmol) was added to a solution of compound 10-a (338 mg, 0.45 mmol) in N,N-dimethylformamide (15 mL), and the reaction mixture was reacted at room temperature for 1 hour. A large amount of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by Prep-HPLC to give compound 10 (36 mg, 14%). LC-MS (ESI): m / z 590.3 (M+H) + .

[0188] Synthetic route of compound 11

[0189] Synthesis of compound 11-a

[0190] Compound 10-b (100 mg, 0.23 mmol), ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)naphth-1-yl)ethynyl)triisopropylsilane (154 mg, 0.34 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II)cataCXium A Pd G3 (17 mg, 0.023 mmol), potassium phosphate (144 mg, 0.68 mmol), tetrahydrofuran (3 mL), and water (0.5 mL) were added to a 10 mL microwave tube. The microwave tube was sealed, purged with nitrogen several times, and heated to 65 °C overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL * 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give compound 11-a (165 mg, 100%). The crude product was used directly in the next reaction without purification. LC-MS (ESI): m / z 731.5 (M + H) + .

[0191] Synthesis of Compound 11

[0192] Cesium fluoride (343 mg, 2.26 mmol) was added to a solution of compound 11-a (165 mg, 0.23 mmol) in N,N-dimethylformamide (10 mL), and the reaction mixture was reacted at room temperature for 1 hour. A large amount of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by Prep-HPLC to give compound 11 (20 mg, 15%). LC-MS (ESI): m / z 575.3 (M+H) + .

[0193] Synthetic route of compound 12

[0194] Synthesis of Compound 12

[0195] To a solution of compound 10 (95 mg, 0.16 mmol) in ethyl acetate (15 mL) and methanol (3 mL), 10% palladium on carbon (171 mg, 1.61 mmol) was added. The reaction solution was purged with hydrogen several times, and the reaction was carried out at room temperature for 3 hours. The reaction was completed by LC-MS. The reaction solution was filtered, the filtrate was evaporated to dryness, and the crude product was purified by Prep-HPLC to give compound 12 (26 mg, 27%). LC-MS (ESI): m / z 594.3 (M+H) + .

[0196] Synthetic route of compound 13

[0197] Synthesis of compound 13-d

[0198] Compound 7-bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (1000 mg, 2.37 mmol) was dissolved in 50 mL of anhydrous THF at room temperature. DIPEA (1.18 mL, 7.11 mmol) and tert-butyl 1,2-diazacycloheptan-1-carboxylate (475 mg, 2.37 mmol) were added in an ice-water bath, and the reaction mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (EA / PE = 0–30%) to give compound 13-d (770 mg, 55%). LC-MS (ESI): m / z = 585.0 [M+H] + .

[0199] Synthesis of compound 13-c

[0200] Compound 13-d (770 mg, 1.31 mmol) was dissolved in 10 mL of anhydrous DMF at room temperature. Anhydrous THF (10 mL) was added, followed by ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (628 mg, 3.94 mmol), cesium carbonate (1.29 g, 3.94 mmol), and 1,4-diazabicyclo[2,2,2]octane (590 mg, 5.26 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled in an ice-water bath, quenched with water (100 mL), and extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (100 mL x 5), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (MeOH / DCM = 0–10%) to give compound 13-c (928 mg, 100%). LC-MS (ESI): m / z = 708.2 [M+H] + .

[0201] Synthesis of compound 13-b

[0202] Compound 13-c (100 mg, 0.14 mmol), cuprous cyanide (63 mg, 0.71 mmol), and anhydrous DMSO (1 mL) were added to a microwave-safe tube at room temperature. The mixture was purged with nitrogen three times and then stirred at 80 °C for 16 hours. The reaction mixture was cooled to room temperature, quenched with water, and extracted with ethyl acetate (100 mL). The aqueous phase was then extracted with DCM (100 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in 3 mL of DMF, and 10 mL of water was added. A solid precipitated, which was filtered, the filter cake was washed with water, and the solid was collected. It was dissolved in 20 mL of methanol, concentrated under reduced pressure, and the residue was dried under vacuum for 2 hours to give crude compound 13-b (54 mg, 64%). The crude product was used directly in the next reaction without purification. LC-MS (ESI): m / z = 607.1 [M+H] + .

[0203] Synthesis of compound 13-a

[0204] Compound 13-b (54 mg, 0.089 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboronyl-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (72 mg, 0.18 mmol), cesium carbonate (87 mg, 0.27 mmol), dichloro[bis(diphenylphosphine)ether]palladium(II) (6.4 mg, 0.009 mmol), and anhydrous toluene (3 mL) were added to a microwave-safe tube at room temperature. The mixture was purged with nitrogen three times, and then stirred at 100 °C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (MeOH / DCM = 0–10%) to give compound 13-a (50 mg, 68%). LC-MS (ESI): m / z = 819.5 [M+H] + .

[0205] Synthesis of Compound 13

[0206] Compound 13-a (50 mg, 0.061 mmol) was dissolved in 6 mL of dichloromethane at room temperature. Trifluoroacetic acid (3 mL) was added under ice-water bath conditions, and the reaction mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (alkaline method, ammonium bicarbonate system). The residue was then lyophilized to give compound 13 (1.1 mg, 2.9%). LC-MS (ESI): m / z = 619.2 [M+H] + .

[0207] Synthetic route of compound 14

[0208] Synthesis of compound 14-b

[0209] Add 13-c (100 mg, 0.14 mmol), methylboric acid (17 mg, 0.28 mmol), potassium carbonate (59 mg, 0.42 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloromethane complex (12 mg, 0.014 mmol), 1,4-dioxane (10 mL), and water (1 mL) to a microwave tube. Seal the tube, purge with nitrogen three times, stir overnight at 50 °C, and then continue stirring overnight at 60 °C. Reduce the reaction solution to dryness by rotary evaporation and purify by column chromatography (methanol:dichloromethane = 0–10%) to give compound 14-b (50 mg, 59%). LC-MS (ESI): m / z 596.2 (M+H) + .

[0210] Synthesis of compound 14-a

[0211] Compound 14-b (50 mg, 0.084 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (102 mg, 0.25 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (6 mg, 0.008 mmol), cesium carbonate (137 mg, 0.42 mmol), and toluene (10 mL) were added to a microwave tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 4 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 14-a (68 mg, 100%). LC-MS (ESI): m / z 808.5 (M+H) + .

[0212] Synthesis of Compound 14

[0213] Trifluoroacetic acid (2 mL) was added to a solution of compound 14-a (68 mg, 0.084 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 14 (15 mg, 29%). LC-MS (ESI): m / z 608.3 (M+H) + ; 1H NMR(400M,DMSO-d6)δ9.11(1H,s),8.03(2H,s),7.22-7.08(2H,m),5.71-5.60(1H,m),5.32(1H,d,J=54 .4Hz),4.23-3.84(4H,m),3.29-2.83(6H,m),2.27-2.00(6H,m),1.98-1.75(5H,m),1.74-1.54(4H,m).

[0214] Synthetic route of compound 15

[0215] Synthesis of compound 15-a

[0216] Compound 1-b (70 mg, 0.12 mmol), toluene (5 mL), (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)benzo[d]thiazolyl) tert-butyl carbamate (73 mg, 0.23 mmol), cesium carbonate (190 mg, 0.58 mmol), and PdCl2 (DPEphos) (8 mg, 0.01 mmol) were added to a microwave-safe tube. The tube was purged with nitrogen three times and then placed in an oil bath preheated to 100 °C with stirring for 1.5 hours. The reaction mixture was directly evaporated to dryness and subjected to column chromatography (mobile phase: dichloromethane / methanol = 100 / 0; 100 / 6) to give compound 15-a (79 mg, 86%). LC-MS (ESI): m / z = 788.7 (M+H) + .

[0217] Synthesis of Compound 15

[0218] 15-a (79 mg, 0.10 mmol), dichloromethane (6 mL), and trifluoroacetic acid (2 mL) were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction solution was evaporated to dryness, and the reaction was quenched with saturated sodium bicarbonate (10 mL). The mixture was extracted with dichloromethane (20 mL * 3), concentrated, and purified by HPLC to obtain compound 15 (39 mg, 67%). LC-MS (ESI): m / z = 588.2 (M + H) + .

[0219] Synthetic route of compound 16

[0220] Synthesis of compound 16-l

[0221] To a solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (300 mg, 1.19 mmol) in dichloromethane (30 mL), tert-butyl 1,2-diazaphen-1-carboxylic acid (238 mg, 1.19 mmol) and N,N-diisopropylethylamine (461 mg, 3.56 mmol) were added, and the mixture was reacted at this temperature for 1 hour. The reaction mixture was then brought to room temperature and reacted for another hour. Water was added, and the mixture was extracted with dichloromethane (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give 16-l (490 mg, 99%). LC-MS (ESI): m / z 416.0 (M+H) + .

[0222] Synthesis of compound 16-k

[0223] Under ice-water bath conditions, 20 mL of dichloromethane ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (281 mg, 1.77 mmol) was added to a solution of 16-l (490 mg, 1.18 mmol) in (20 mL) of dichloromethane. Sodium tert-butoxide (226 mg, 2.35 mmol) was then slowly added, and the mixture was reacted at this temperature for 2 hours. Water was then added, and the mixture was extracted with dichloromethane (50 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by rapid column chromatography (mobile phase: dichloromethane:methanol = 10:1 / dichloromethane, 0-50%) to give 16-k (561 mg, 88%). LC-MS (ESI): m / z 539.4 (M+H) + .

[0224] Synthesis of compound 16-j

[0225] 3-Bromo-2-methylaniline (18.20 g, 97.82 mmol) and dimethyl sulfoxide (100 mL) were added to a reaction flask and stirred in an ice-water bath for 10 minutes. Then, NIS (18.61 g, 107.60 mmol) was slowly added to the reaction mixture, and the mixture was stirred at room temperature for 0.5 hours. Water (500 mL) was added to the reaction mixture, and the mixture was filtered. The filter cake was washed with water (200 mL x 3). The filter cake was dried under vacuum to obtain compound 16-j (28.89 g, 95%). LC-MS (ESI): m / z = 312.0 (M + H) + .

[0226] Synthesis of compound 16-i

[0227] 16-J (28.89 g, 92.61 mmol) and acetic acid (300 mL) were added to the reaction flask, and the mixture was stirred for 10 minutes in an ice-water bath. A solution of sodium nitrite (14.70 g, 213.01 mmol) in water (40 mL) was slowly added dropwise to the mixture. The reaction mixture was slowly heated to 35 °C and stirred for 48 hours. The reaction mixture was poured into water (1500 mL), filtered, and the filter cake was washed with water (100 mL x 3). The filter cake was dried under vacuum to give compound 16-i (17.61 g, 59%). LC-MS (ESI): m / z = 323.0 (M + H) + .

[0228] Synthesis of compound 16-h

[0229] 3,4-Dihydro-2H-pyran (17.82 g, 208.07 mmol), p-toluenesulfonic acid (7.42 g, 39.01 mmol), 16-i (16.80 g, 52.02 mmol), and tetrahydrofuran (300 mL) were added to the reaction flask, and the mixture was stirred at room temperature for 4 hours. Saturated sodium bicarbonate (300 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (300 mL x 2). The organic phase was concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 20 / 1; 10 / 1) to give compound 16-h (15.98 g, 75%). LC-MS (ESI): m / z = 407.0 (M+H) + .

[0230] Synthesis of compound 16-g

[0231] Add 16-h (15.60 g, 38.33 mmol), tetrahydrofuran (300 mL), and cyclobutanone (15.00 g, 214.01 mmol) to the reaction flask, and stir at -78 °C for 10 min under nitrogen protection. Add n-butyllithium n-hexane solution (2.5 M, 45.62 mL, 114.05 mmol) slowly dropwise to the reaction solution over 50 min. Continue stirring at -78 °C for 3 h. Quench the reaction with saturated ammonium chloride (200 mL) at low temperature, and extract with ethyl acetate (200 mL x 3). Concentrate the organic phase and purify by column chromatography (petroleum ether / ethyl acetate = 20 / 1; 10 / 3) to give compound 16-g (6.43 g, 48%). LC-MS (ESI): m / z = 351.3 (M + H). + .

[0232] Synthesis of compound 16-f

[0233] 16-g (3.50 g, 9.97 mmol), dichloromethane (350 mL), water (200 mL), silver nitrate (0.87 g, 5.12 mmol), and potassium persulfate (10.00 g, 37.00 mmol) were slowly added to a mixed solution in a reaction flask, followed by nitrogen protection. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was then poured into a saturated aqueous solution of sodium sulfite (100 mL) and extracted with dichloromethane (100 mL x 3). The organic phase was concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 10 / 1; 3 / 1) to give compound 16-f (0.93 g, 27%). LC-MS (ESI): m / z = 349.1 (M+H) + .

[0234] Synthesis of compound 16-e

[0235] Triisopropylsilylacetylene (1.15 g, 6.29 mmol) and toluene (30 mL) were added to the reaction flask and stirred in an ice-water bath for 10 minutes. A 2.5 M, 2 mL, 5.00 mmol solution of n-butyllithium in hexane was slowly added dropwise to the solution, and the mixture was stirred in an ice-water bath under nitrogen protection for 2 hours. A 30 mL solution of 16-f (0.45 g, 1.29 mmol) in toluene was slowly added dropwise to the previous reaction mixture in an ice-water bath, and stirring continued for 3 hours. The reaction was quenched by adding 50 mL of saturated ammonium chloride aqueous solution, extracted with ethyl acetate (50 mL x 3), the organic phase was concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 100 / 0; 3 / 1) to give compound 16-e (0.61 g, 89%). LC-MS (ESI): m / z = 531.3 (M + H). + .

[0236] Synthesis of compound 16-d

[0237] Add 16-e (250 mg, 0.47 mmol), dichloromethane (20 mL), and triethylsilane (137 mg, 1.18 mmol) to the reaction flask and stir in an ice-water bath for 10 minutes. Add trifluoroacetic acid (214 mg, 1.88 mmol) dropwise to the reaction solution. Slowly raise the reaction solution to room temperature and stir for 3 hours. Quench the reaction with saturated sodium bicarbonate (20 mL) and extract with dichloromethane (30 mL x 3). Concentrate the organic phase and purify by column chromatography (petroleum ether / ethyl acetate = 10 / 1; 5 / 1) to give compound 16-d (180 mg, 75%). LC-MS (ESI): m / z = 513.1 (M+H) + .

[0238] Synthesis of compound 16-c

[0239] 16-d (180 mg, 0.35 mmol), methanol (30 mL), tetrahydroxydiboron (94 g, 1.05 mmol), cataCXium A PdG3 (51 mg, 0.07 mmol), and triethylamine (35 mg, 0.35 mmol) were added to a reaction flask under nitrogen protection. The reaction mixture was slowly heated to 50 °C and stirred for 2 hours. The reaction mixture was concentrated, and the solid was washed with ethyl acetate (10 mL). Solid compound 16-c (140 mg, 83%) was given. LC-MS (ESI): m / z = 479.2 (M+H) + .

[0240] Synthesis of compound 16-b

[0241] 16-c (107 mg, 0.22 mmol), tetrahydrofuran (6 mL), water (0.8 mL), 16-k (120 mg, 0.22 mmol), potassium phosphate (189 mg, 0.89 mmol), and cataCXium A Pd G3 (49 mg, 0.07 mmol) were added to a microwave-safe tube. The tube was purged with nitrogen three times and then stirred in an oil bath preheated to 65 °C for 12 hours. The reaction solution was concentrated and purified by column chromatography (DCM / MeOH = 10 / 1) to give compound 16-b (86 mg, 41%). LC-MS (ESI): m / z = 938.2 (M+H) + .

[0242] Synthesis of compound 16-a

[0243] 16-b (86 mg, 0.09 mmol), dichloromethane (3 mL), and trifluoroacetic acid (1 mL) were added to the reaction flask, and the mixture was stirred at room temperature for 3 hours. The mixture was then neutralized with saturated sodium bicarbonate aqueous solution (10 mL), extracted with dichloromethane (20 mL x 3), and the organic phase was concentrated to give compound 16-a (43 mg, 62%). LC-MS (ESI): m / z = 753.7 (M+H) + .

[0244] Synthesis of Compound 16

[0245] 16-a (41 mg, 0.05 mmol), cesium fluoride (124 mg, 0.82 mmol), and DMF (3 mL) were added to a reaction flask at room temperature. The reaction mixture was stirred at room temperature for 4 hours. After filtration, the solution was purified by preparative HPLC to give compound 16 (2.7 mg, 8%). LC-MS (ESI): m / z = 597.3 (M+H) + .

[0246] Synthetic route of compound 17

[0247] Synthesis of compound 17-d

[0248] Add 16-e (210 mg, 0.40 mmol), dichloromethane (20 mL), and triethylsilane (200 mg, 1.72 mmol) to the reaction flask and stir at -78 °C for 10 minutes. Add trifluoroacetic acid (380 mg, 3.33 mmol) dropwise to the reaction solution. Continue stirring at -78 °C for 3 hours. Quench the reaction with saturated sodium bicarbonate (50 mL) and extract with dichloromethane (100 mL x 2). Concentrate the organic phase and purify by column chromatography (petroleum ether / ethyl acetate = 20 / 1; 5 / 1) to give compound 17-d (200 mg, 98%). LC-MS (ESI): m / z = 515.2 (M+H) + .

[0249] Synthesis of compound 17-c

[0250] 17-d (210 mg, 0.41 mmol), cataCXium A PdG3 (59 mg, 0.08 mmol), tetrahydroxydiboron (110 mg, 1.22 mmol), triethylamine (41 mg, 0.41 mmol), and methanol (30 mL) were added to the reaction flask under nitrogen protection. The reaction mixture was slowly heated to 50 °C and stirred for 2 hours. The reaction mixture was concentrated to obtain solid compound 17-c (200 mg), which was then added to the next reaction step. LC-MS (ESI): m / z = 481.3 (M+H) + .

[0251] Synthesis of compound 17-b

[0252] 17-c (107 mg, 0.22 mmol), tetrahydrofuran (6 mL), water (0.8 mL), 16-k (120 mg, 0.22 mmol), potassium phosphate (189 mg, 0.89 mmol), and cataCXium A Pd G3 (49 mg, 0.07 mmol) were added to a microwave-safe tube. The tube was purged with nitrogen three times and then stirred in an oil bath preheated to 65 °C for 12 hours. The reaction solution was concentrated and purified by column chromatography (DCM / MeOH = 10 / 1) to give compound 17-b (110 mg, 53%). LC-MS (ESI): m / z = 940.1 (M+H) + .

[0253] Synthesis of compound 17-a

[0254] 17-b (110 mg, 0.12 mmol), dichloromethane (2 mL), and trifluoroacetic acid (1 mL) were added to the reaction flask and stirred at room temperature for 2 hours. The mixture was then neutralized with saturated sodium bicarbonate aqueous solution (10 mL), extracted with dichloromethane (20 mL x 3), and the organic phase was concentrated to give compound 17-a (76 mg, 86%). LC-MS (ESI): m / z = 756.1 (M+H). + .

[0255] Synthesis of Compound 17

[0256] 17-a (76 mg, 0.10 mmol), tetrabutylammonium fluoride tetrahydrofuran solution (1 M, 15 mL, 15.00 mmol), and tetrahydrofuran (20 mL) were added to a reaction flask at room temperature. The reaction mixture was stirred at room temperature for 8 hours. The solution was concentrated, and saturated sodium bicarbonate aqueous solution (60 mL) was added. Extraction was performed with ethyl acetate (100 mL * 3). The organic phase was concentrated and purified by HPLC to give compound 17 (23 mg, 38%). LC-MS (ESI): m / z = 599.8 (M + H). + .

[0257] Synthetic route of compound 18

[0258] Synthesis of compound 18-c

[0259] 1,2-diazaphen-1-carboxylic acid tert-butyl ester (606 mg, 3.03 mmol) was added to a tetrahydrofuran (50 mL) solution of 7-bromo-2,4,6-trichloro-8-fluoroquinazoline (1 g, 3.03 mmol) and N,N-diisopropylethylamine (1173 mg, 9.08 mmol) under ice-water bath conditions. The mixture was allowed to warm to room temperature and stirred overnight. The next day, water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–10%) to give 18-c (1350 mg, 90%). LC-MS (ESI): m / z 493.1 (M+H) + .

[0260] Synthesis of compound 18-b

[0261] At room temperature, 18-c (1000 mg, 2.02 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (644 mg, 4.05 mmol), THF (15 mL), DMF (15 mL), triethylenediamine (908 mg, 8.09 mmol), and cesium carbonate (1978 mg, 6.07 mmol) were added sequentially to a reaction flask. The mixture was protected by N2 displacement and reacted overnight at room temperature. Water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL * 2). The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 0 / 10) to give compound 18-b (500 mg, 40%).

[0262] Synthesis of compound 18-a

[0263] At room temperature, 18-b (80 mg, 0.13 mmol), (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)benzo[d]thiazolyl) tert-butyl carbamate (81 mg, 0.26 mmol), toluene (4 mL), bis(diphenylphosphine ether) palladium dichloride (9 mg, 0.01 mmol), and cesium carbonate (211 mg, 0.65 mmol) were added sequentially to a microwave-safe mixture. The mixture was protected by N2 displacement and heated to 100 °C for 3 hours. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 18-a (100 mg, 96%).

[0264] Synthesis of Compound 18

[0265] 18-a (100 mg, 0.12 mmol), DCM (6 mL), and trifluoroacetic acid (2 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to give compound 18 (25 mg, 33%). LC-MS (ESI): m / z 604.3 (M+H) + ; 1 HNMR(DMSO-d6,400MHz): δ9.42(1H,s),7.92(2H,s),7.30-7.17(1H,m),7.12-6.98(2H,t,J=8.8Hz),5.81-5.62(1H,t, J=6.0Hz),5.40-5.14(1H,d,J=53.6Hz),4.16-3.89(4H,m),3.18-2.91(5H,m),2.89-2.74(1H,m),2.20-1.53(11H,m).

[0266] Synthetic route of compound 19

[0267] Synthesis of compound 19-d

[0268] 16-f (450 mg, 1.29 mmol) and THF (10 mL) were added sequentially to the reaction flask at room temperature. The mixture was protected by N2 displacement, and a tetrahydrofuran solution of methyl magnesium bromide (1 M, 6.44 mL, 6.44 mmol) was added dropwise under ice bath. After the addition was complete, the reaction was allowed to proceed overnight at room temperature. The reaction solution was quenched with saturated ammonium chloride aqueous solution and extracted with ethyl acetate. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. DCM (10 mL) was added to dissolve the filtrate, and the mixture was protected by N2 displacement. Triethylsilane (1098 mg, 5.15 mmol) and TFA (588 mg, 5.15 mmol) were added dropwise under dry ice and acetone bath. The reaction was maintained at low temperature for 2 hours. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 8 / 2) to obtain mixture 19-d (230 mg). LC-MS(ESI):m / z=347.1 / 349.1[M+H] + .

[0269] Synthesis of compound 19-c

[0270] 19-d (230 mg), EA (4 mL), and 5% rhodium on carbon (290 mg, 2.82 mmol) were added sequentially to a reaction flask at room temperature. The mixture was purged with hydrogen for protection and reacted at 40 °C for 2 hours. The reaction solution was filtered, and the filtrate was concentrated to give compound 19-c (220 mg, 49%). LC-MS (ESI): m / z = 349.1 [M+H] + .

[0271] Synthesis of compound 19-b

[0272] At room temperature, 19-c (200 mg, 0.57 mmol), methanol (4 mL), tetrahydroxydiboron (205 mg, 2.29 mmol), cataCXium A Pd G3 (20 mg, 0.03 mmol), and triethylamine (232 mg, 2.29 mmol) were added sequentially to a reaction flask. The mixture was protected with N2 displacement and reacted at 50 °C for 1 hour. The reaction solution was concentrated to give crude compound 19-b (400 mg). LC-MS (ESI): m / z = 315.0 [M+H] + .

[0273] Synthesis of compound 19-a

[0274] At room temperature, crude 19-b (200 mg, 0.29 mmol), 16-k (343 mg, 0.64 mmol), cataCXium A Pd G3 (23 mg, 0.03 mmol), potassium phosphate (203 mg, 0.96 mmol), THF (10 mL), and water (2 mL) were added sequentially to a microwave-safe tube. The mixture was protected with nitrogen purging and heated to 65 °C overnight. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give crude compound 19-a (420 mg). LC-MS (ESI): m / z 774.0 (M+H) + .

[0275] Synthesis of Compound 19

[0276] Crude 19-a (420 mg), DCM (20 mL), and trifluoroacetic acid (7 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to obtain compound 19 (92 mg, 55%). LC-MS (ESI): m / z 589.5 (M+H) + ; 1 HNMR(DMSO_d6,400MHz): δ12.92(1H,m),10.40-10.20(1H,m),7.59-7.40(1H,m),7.34(1H,s),5.94-5.84(1H,m),5.40- 5.17(1H,d,J=55.2Hz),4.23-3.90(4H,m),3.21-2.91(7H,m),2.89-2.74(1H,m),2.20-1.55(17H,m),1.00-0.74(3H,m).

[0277] Synthesis of compounds 20 and 21

[0278] Compound 6 (345 mg) was synthesized using the same method as compound 6, and was purified by chiral resolution to give compound 20 (129 mg, 37%) and compound 21 (145 mg, 42%).

[0279] Compound 20: LC-MS (ESI): m / z = 664.6 (M+H) + ; 1H NMR (400MHz, CDCl3): δ9.66(1H,s),7.17(1H,dd,J1=8.0Hz,J2=4.4Hz),6.96(1H,t,J=8.8Hz),5.82(2H,s),5.29(1H,d,J=52.0Hz),4.45-4.25( 3H,m),4.24-4.09(2H,m),4.04-3.92(2H,m),3.83(2H,t,J=4.4Hz),3.4 6-3.15(5H,m),3.06-2.94(1H,m),2.24-2.12(2H,m),2.08-1.80(4H,m).

[0280] Compound 21: LC-MS (ESI): m / z = 664.6 (M+H) + ; 1 H NMR (400MHz, CDCl3): δ9.65(1H,s),7.19-7.12(1H,m),6.95(1H,t,J=8.4Hz),6.26(2H,s),5.22(1H,d,J=53.2Hz),4.40- 4.12(4H,m),4.09-3.86(3H,m),3.81(2H,s),3.49-3.15(5H,m),3.06-2.94(1H,m),2.40-2.09(3H,m),2.08-1.80(3H,m).

[0281] Synthetic routes of compounds 22 and 23

[0282] Synthesis of compound 22-g

[0283] Under ice-water bath conditions, benzyl alcohol (1242 μL, 12.01 mmol), DMF (20 mL), and sodium hydroxide (60%, 320 mg, 8.00 mmol) were added to the reaction flask. The mixture was stirred under nitrogen atmosphere for 1 hour at this temperature. Then, 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(1H)-one (1121 mg, 4.00 mmol) was added. After the addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 3 hours. While the reaction mixture was cooled in an ice-water bath, 100 mL of saturated ammonium chloride solution was added dropwise, resulting in the precipitation of a large amount of solid, which was filtered. The solid was washed with water, removing water with ethanol, and then dried using an oil pump to obtain 22 g of the compound, which was used directly in the next reaction without purification. LC-MS (ESI): m / z = 352.0 (M+H) + .

[0284] Synthesis of compound 22-f

[0285] 22-g (704 mg, 2.00 mmol), dimethyl sulfoxide (10 mL), N,N-diisopropylethylamine (776 mg, 6.00 mmol), and N-phenylbis(trifluoromethanesulfonyl)imide (2145 mg, 6.00 mmol) were added to the reaction flask and stirred at room temperature for 2 hours. 1,2-diazepine (401 mg, 4.00 mmol) and N,N-diisopropylethylamine (776 mg, 6.00 mmol) were added to the reaction mixture, and the mixture was stirred at 50°C for 1 hour under nitrogen protection. Saturated sodium bicarbonate (30 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 2). The organic phase was concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1; 1 / 1) to give compound 22-f (410 mg, 47%). LC-MS (ESI): m / z = 434.2 (M+H) + .

[0286] Synthesis of compound 22-e

[0287] 22-f (410 mg, 0.95 mmol), dichloromethane (3 mL), and trifluoroacetic acid (1 mL) were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The reaction solution was neutralized with saturated sodium bicarbonate aqueous solution (10 mL), extracted with dichloromethane (20 mL x 3), the organic phase was concentrated, and purified by column chromatography (dichloromethane / methanol = 10 / 1) to give compound 22-e (131 mg, 40%). LC-MS (ESI): m / z = 344.0 (M+H) + .

[0288] Synthesis of compound 22-d

[0289] 22-e (131 mg, 0.38 mmol), methanol (20 mL), paraformaldehyde (100 mg, 3.33 mmol), and trifluoroacetic acid (0.06 mL, 0.76 mmol) were added to the reaction flask. The mixture was stirred at 80 °C for 3 hours under nitrogen protection. The reaction solution was concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 10 / 1; 3 / 1) to give compound 22-d (98 mg, 72%). LC-MS (ESI): m / z = 356.1 (M+H) + .

[0290] Synthesis of compound 22-c

[0291] 22-d (129 mg, 0.36 mmol) and ethyl acetate (20 mL) were added to the reaction flask, followed by the addition of 80% pure m-chloroperoxybenzoic acid (196 mg, 0.91 mmol) at room temperature. The mixture was stirred for 3 hours under nitrogen protection in an ice-water bath. The reaction solution was poured into a saturated sodium bicarbonate aqueous solution (180 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 10 / 1; 3 / 1) to give compound 22-c (98 mg, 70%). LC-MS (ESI): m / z = 388.1.1 (M+H) + .

[0292] Synthesis of compound 22-b

[0293] Add 22-c (98 mg, 0.25 mmol) and toluene (30 mL) to the reaction flask and stir in an ice-water bath for 10 minutes. Add ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (60 mg, 0.38 mmol) and sodium tert-butoxide (29 mg, 0.30 mmol) to the reaction solution and stir in an ice-water bath under nitrogen protection for 1.5 hours. Add saturated ammonium chloride aqueous solution (50 mL) to the reaction solution, extract with ethyl acetate (30 mL * 3), concentrate the organic phase, and purify by column chromatography (dichloromethane / methanol = 20 / 1) to give compound 22-b (62 mg, 53%). LC-MS (ESI): m / z = 467.3 (M+H) + .

[0294] Synthesis of compound 22-a

[0295] 22-b (63 mg, 0.14 mmol), tetrahydrofuran (3 mL), water (0.4 mL), 6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-((triisopropylsilyl)ethynyl)naphthyl-2-amine (189 mg, 0.41 mmol), potassium phosphate (143 mg, 0.68 mmol), and cataCXium A Pd G3 (29 mg, 0.04 mmol) were added to a microwave tube. The tube was purged with nitrogen three times and then stirred in an oil bath preheated to 65 °C for 12 hours. The reaction mixture was concentrated and purified by column chromatography (DCM / MeOH = 10 / 1) to give compound 22-a (42 mg, 40%). LC-MS (ESI): m / z = 772.9 (M+H) + .

[0296] Synthesis of Compounds 22 and 23

[0297] 22-a (42 mg, 0.05 mmol), cesium fluoride (83 mg, 0.54 mmol), and DMF (4 mL) were added to a reaction flask at room temperature. The reaction mixture was stirred at room temperature for 3 hours. After filtration, compound 22 (4.1 mg, 12%) was purified by preparative HPLC. LC-MS (ESI): m / z = 616.3 (M+H) + Compound 23 (6.4 mg, 19.5%), LC-MS (ESI): m / z = 604.3 (M+H) + .

[0298] Synthetic route of compound 24

[0299] Synthesis of compound 24-k

[0300] 2-Amino-4-bromo-5-chloro-3,6-difluorobenzoic acid (2 g, 6.98 mmol), tetrahydrofuran (20 mL), and N,N'-carbonyldiimidazole (1.7 g, 10.47 mmol) were added to a reaction flask, and the mixture was stirred at 50 °C for half an hour under nitrogen atmosphere. The reaction solution was cooled to room temperature and slowly added dropwise to ammonia water at 0 °C (20 mL, 148.36 mmol). After the addition was complete, stirring was continued at room temperature for another half hour. The reaction solution was quenched with water and extracted twice with ethyl acetate. The organic phase was washed with saturated brine, evaporated to dryness, and purified by column chromatography (mobile phase: methanol / dichloromethane = 0–10%) to give compound 24-k (1.9 g, 95%). LC-MS (ESI): m / z 284.9 (M+H) + .

[0301] Synthesis of compound 24-j

[0302] Compound 24-k (2 g, 7.01 mmol) was added to a reaction flask, along with 50 mL of dried tetrahydrofuran. The mixture was heated to 40 °C, and sodium hydrogen hydride (0.7 g, 17.52 mmol) was added in portions. The mixture was stirred for 10 minutes, followed by the addition of N,N'-thiocarbonyldiimidazole (1.87 g, 10.51 mmol) in portions. The mixture was then stirred at 60 °C for 30 minutes. The reaction mixture was quenched with saturated ammonium chloride solution under ice-water bath cooling. The pH was adjusted to 5–6 with dilute hydrochloric acid, and the tetrahydrofuran was removed by vortexing, resulting in a large amount of solid. The mixture was filtered, the filter cake was washed with water, and dried in air to give compound 24-j (2.3 g, 100%). LC-MS (ESI): m / z 326.9 (M+H) + .

[0303] Synthesis of compound 24-i

[0304] Compound 24-j (2.3 g, 7.02 mmol), methanol (40 mL), sodium methoxide (0.57 g, 10.53 mmol), and methyl iodoform (0.87 mL, 14.05 mmol) were added to a reaction flask and stirred at room temperature for 1 hour under nitrogen atmosphere. 20 mL of water was slowly added to the reaction mixture, and the mixture was stirred at room temperature for 10 minutes. The mixture was then filtered. The filter cake was washed with water and air-dried to give compound 24-i (2.27 g, 95%). LC-MS (ESI): m / z 341.0 (M+H) + .

[0305] Synthesis of compound 24-h

[0306] Benzyl alcohol (908 μL, 8.78 mmol) was added to the reaction flask under an ice-water bath. N,N-dimethylformamide (10 mL) and sodium hydride (410 mg, 10.24 mmol) were dried. The mixture was stirred under nitrogen atmosphere for 1 hour at this temperature. Then, compound 24-i (1000 mg, 2.93 mmol) was added, and the mixture was stirred for another hour at this temperature. Under ice-water bath cooling, 10 mL of saturated ammonium chloride solution was added dropwise to the reaction mixture. A large amount of solid precipitated out and was filtered. The solid was washed with water, removing water with ethanol, and then dried under a pump to obtain compound 24-h (1720 mg), which was used directly in the next reaction without purification. LC-MS (ESI): m / z 429.0 (M+H) + .

[0307] Synthesis of compound 24-g

[0308] Compound 24-h (1720 mg), acetonitrile (40 mL), N,N-diisopropylethylamine (4.96 mL, 30.02 mmol), and phosphorus oxychloride (5.60 mL, 60.04 mmol) were added to a reaction flask, and the mixture was stirred at 80 °C for 2 hours under nitrogen atmosphere. The reaction solution was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–30%) to give compound 24-g (937 mg, 71% in two steps). LC-MS (ESI): m / z 446.9 (M+H) + .

[0309] Synthesis of compound 24-f

[0310] Compound 24-g (937 mg, 2.09 mmol), tetrahydrofuran (30 mL), N,N-diisopropylethylamine (1.73 mL, 10.46 mmol), and tert-butyl 1,2-diazaphen-1-carboxylate (419 mg, 2.09 mmol) were added to the reaction flask under ice-water bath conditions. The reaction mixture was allowed to warm to room temperature and stirred overnight. The next day, the reaction mixture was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–10%) to give compound 24-f (482 mg, 38%). LC-MS (ESI): m / z 611.1 (M+H) + .

[0311] Synthesis of compound 24-e

[0312] Compound 24-f (482 mg, 0.79 mmol), ethyl acetate (20 mL), and m-chloroperoxybenzoic acid (480 mg, 2.36 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 1 hour. The reaction mixture was quenched with saturated sodium sulfite solution and extracted twice with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution, then with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–30%) to give compound 24-e (422 mg, 83%). LC-MS (ESI): m / z 643.1 (M+H) + .

[0313] Synthesis of compound 24-d

[0314] Compound 24-e (422 mg, 0.66 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (313 mg, 1.97 mmol), dry dichloromethane (20 mL), and sodium tert-butoxide (220 mg, 2.29 mmol) were added to the reaction flask under ice-water bath conditions. The mixture was slowly heated to room temperature and stirred for 1 hour. The reaction solution was then directly purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 24-d (380 mg, 80%). LC-MS (ESI): m / z 722.1 (M+H) + .

[0315] Synthesis of compound 24-c

[0316] Compound 24-d (380 mg, 0.53 mmol), dichloromethane (9 mL), and trifluoroacetic acid (3 mL) were added to a reaction flask and stirred at room temperature for 2 hours. The reaction mixture was evaporated to dryness, and saturated sodium bicarbonate solution was added. The mixture was extracted twice with ethyl acetate. The organic phase was evaporated to dryness and purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 24-c (222 mg, 79%). LC-MS (ESI): m / z 532.1 (M+H) + .

[0317] Synthesis of compound 24-b

[0318] Compound 24-c (222 mg, 0.42 mmol), toluene (15 mL), paraformaldehyde (1000 mg, 33.30 mmol), and a few drops of trifluoroacetic acid were added to a reaction flask. The mixture was stirred at 100 °C for 30 minutes under nitrogen atmosphere. The reaction solution was evaporated to dryness, methanol was added, and the pH was adjusted to 7 with saturated sodium bicarbonate solution. The solution was evaporated to dryness again, and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 24-b (200 mg, 88%). LC-MS (ESI): m / z 543.9 (M+H) + .

[0319] Synthesis of compound 24-a

[0320] Compound 24-b (230 mg, 0.42 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (341 mg, 0.84 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (30 mg, 0.042 mmol), cesium carbonate (688 mg, 2.11 mmol), and toluene (15 mL) were added to a microwave tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol / dichloromethane = 0–5%) to give compound 24-a (310 mg, 97%). LC-MS (ESI): m / z 756.0 (M+H) + .

[0321] Synthesis of Compound 24

[0322] Trifluoroacetic acid (3 mL) was added to a 9 mL solution of compound 24-a (100 mg, 0.13 mmol) in dichloromethane at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 24 (40 mg, 46%). LC-MS (ESI): m / z 656.2 (M+H) + ;1 H NMR (400M, DMSO-d6): δ8.08(2H,s),7.33-7.20(1H,m),7.19-7.09(1H,m),5.55-5.15(3H,m),4.86-4.61 (1H,m),4.17-3.92(2H,m),3.20-2.95(5H,m),2.89-2.77(1H,m),2.18-1.94(3H,m),1.93-1.13(10H,m).

[0323] Synthetic route of compound 25

[0324] Synthesis of compound 25-f

[0325] Compound 24-g (1147 mg, 2.56 mmol), tetrahydrofuran (50 mL), N,N-diisopropylethylamine (2.11 mL, 12.80 mmol), and [1,4,5]oxadiazaporide (261 mg, 2.56 mmol) were added to the reaction flask under ice-water bath conditions. The mixture was allowed to warm to room temperature and stirred overnight. The next day, 112 mg of [1,4,5]oxadiazaporide was added to the reaction mixture, and stirring was continued for 1 hour. The reaction mixture was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–30%) to give compound 25-f (354 mg, 27%). LC-MS (ESI): m / z 513.1 (M+H) + .

[0326] Synthesis of compound 25-e

[0327] Compound 25-f (354 mg, 0.69 mmol), dichloromethane (6 mL), and trifluoroacetic acid (2 mL) were added to a reaction flask, and the mixture was stirred at room temperature for 1 hour. The solution was then evaporated to dryness to give compound 25-e (crude product). LC-MS (ESI): m / z 423.1 (M+H) + .

[0328] Synthesis of compound 25-d

[0329] Compound 25-e (292 mg, 0.69 mmol), toluene (10 mL), paraformaldehyde (500 mg, 16.65 mmol), and a few drops of trifluoroacetic acid were added to a reaction flask. The mixture was stirred at 100 °C for 30 minutes under nitrogen atmosphere. The reaction solution was evaporated to dryness, methanol was added, and the pH was adjusted to 7 with saturated sodium bicarbonate solution. The solution was evaporated to dryness again, and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–30%) to give compound 25-d (292 mg). LC-MS (ESI): m / z 435.0 (M+H) + .

[0330] Synthesis of compound 25-c

[0331] Compound 25-d (292 mg, 0.67 mmol), ethyl acetate (20 mL), and m-chloroperoxybenzoic acid (408 mg, 2.01 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 1 hour. A saturated sodium sulfite solution was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution, then with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–5%) to give compound 25-c (285 mg, 91%) (containing sulfones and sulfoxides). LC-MS (ESI): m / z 467.0 (M+H) + LC-MS (ESI): m / z 451.0 (M+H) + .

[0332] Synthesis of compound 25-b

[0333] Compound 25-c (285 mg, 0.61 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (291 mg, 1.83 mmol), dry dichloromethane (20 mL), and sodium tert-butoxide (205 mg, 2.13 mmol) were added to a reaction flask under ice-water bath conditions. The mixture was slowly heated to room temperature and stirred for 1 hour. The reaction solution was then directly purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 25-b (250 mg, 75%). LC-MS (ESI): m / z 546.0 (M+H) + .

[0334] Synthesis of compound 25-a

[0335] Compound 25-b (100 mg, 0.18 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (148 mg, 0.37 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (18 mg, 0.018 mmol), cesium carbonate (298 mg, 0.91 mmol), and toluene (15 mL) were added to a microwave tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–7%) to give compound 25-a (127 mg, 92%). LC-MS (ESI): m / z 758.3 (M+H) + .

[0336] Synthesis of Compound 25

[0337] Trifluoroacetic acid (2 mL) was added to a solution of compound 25-a (127 mg, 0.17 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 25 (48 mg, 44%). LC-MS (ESI): m / z 658.2 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ8.08(2H,s),7.31-7.21(1H,m),7.20-7.07(1H,m),5.59-5.15(3H,m),4.69(1H,s),4.14-4.05(1H,m),4.0 4-3.96(1H,m),3.96-3.86(2H,m),3.84-3.38(4H,m),3.21-2.94(4H,m),2.90-2.76(1H,m),2.17-1.95(3H,m),1.90-1.69(3H,m).

[0338] Synthetic route of compound 26

[0339] Synthesis of compound 26-d

[0340] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (2 g, 4.74 mmol) in dichloromethane (30 mL), tert-butyl 1,2-diazacycloheptan-1-carboxylate (0.95 g, 4.74 mmol) and N,N-diisopropylethylamine (2.48 mL, 14.22 mmol) were added, and the temperature was slowly raised to room temperature. The mixture was reacted overnight at this temperature. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane (60 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give compound 26-d (2.77 g). The crude product was used directly in the next reaction without purification. LC-MS (ESI): m / z 585.0 (M+H) + .

[0341] Synthesis of compound 26-c

[0342] Compound 26-d (2.77 g, 4.73 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (1.51 g, 9.46 mmol), tetrahydrofuran (20 mL), N,N-dimethylformamide (10 mL), cesium carbonate (4.62 g, 14.19 mmol), and 1,4-diazabicyclo[2,2,2]octane (2.65 g, 23.65 mmol) were added to the reaction flask and stirred overnight at room temperature under nitrogen. Water was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate (60 mL * 2). The organic phase was washed three times with saturated brine, evaporated to dryness, and purified by column flash chromatography (Biotage, 25g, Sillica gel, UV254, mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 26-C (2.35g, 70%). LC-MS (ESI): m / z 708.0 (M+H) + .

[0343] Synthesis of compound 26-b

[0344] Compound 26-c (200 mg, 0.28 mmol), cyclopropylboronic acid (73 mg, 0.85 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (21 mg, 0.028 mmol), potassium phosphate (195 mg, 0.85 mmol), and 1,4-dioxane (8 mL) were added to a 30 mL microwave-safe tube. The tube was sealed, purged with nitrogen several times, and heated to 80 °C overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL * 2). The combined organic phases were washed once with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The purified organic phases were then purified by column flash chromatography (mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 26-b (170 mg, 97%). LC-MS (ESI): m / z 622.3 (M+H) + .

[0345] Synthesis of compound 26-a

[0346] Compound 26-b (170 mg, 0.27 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (221 mg, 0.55 mmol), dichloro[bis(diphenylphosphine)ether]palladium(II) (20 mg, 0.027 mmol), cesium carbonate (267 mg, 0.82 mmol), and toluene (10 mL) were added to a 30 mL microwave-safe tube. The tube was sealed, purged with nitrogen several times, and heated to 100 °C overnight. The reaction mixture was evaporated to dryness, water was added, and the mixture was extracted with ethyl acetate (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column flash chromatography (mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 26-a (210 mg, 92%). LC-MS (ESI): m / z 835.0 (M+H) + .

[0347] Synthesis of Compound 26

[0348] Trifluoroacetic acid (3 mL) was added to a solution of compound 26-a (210 mg, 0.25 mmol) in dichloromethane (10 mL) at room temperature, and the mixture was reacted overnight at this temperature. The reaction solution was evaporated to dryness, and saturated aqueous sodium bicarbonate solution was added, followed by extraction with ethyl acetate (30 mL). The organic phase was evaporated to dryness, and the crude product was purified by preparative HPLC to give compound 26 (1.5 mg, 0.94%). LC-MS (ESI): m / z 634.3 (M+H) + .

[0349] Synthetic route of compound 27

[0350] Synthesis of compound 27-c

[0351] Compound 26-c (200 mg, 0.28 mmol), tris(o-methylphenyl)phosphine (17 mg, 0.056 mmol), palladium acetate (6 mg, 0.028 mmol), triethylamine (0.078 mL, 0.57 mmol), and DMF (3 mL) were added to a 10 mL microwave-safe tube. The tube was sealed and purged with nitrogen several times. Acrylonitrile (0.056 mL, 0.85 mmol) was then added. The mixture was heated to 80 °C and stirred overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give compound 27-c (178 mg), which was used directly in the next reaction. LC-MS (ESI): m / z 633.6 (M+H) + .

[0352] Synthesis of compound 27-b

[0353] Under nitrogen protection and an ice-water bath, a solution of triethyllithium borohydride (superhydrogen) tetrahydrofuran (1 M, 2.81 mL, 2.81 mmol) was slowly added to a 15 mL solution of tetrahydrofuran containing 178 mg (0.28 mmol). The temperature was slowly raised to room temperature and stirred overnight. The reaction mixture was cooled to 0 °C, and methanol and water were added. Extraction was performed with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column flash chromatography (mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 27-b (135 mg, 75.6%). LC-MS (ESI): m / z 635.1 (M+H) + .

[0354] Synthesis of compound 27-a

[0355] Compound 27-b (135 mg, 0.21 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (172 mg, 0.43 mmol), dichloro[bis(diphenylphosphine)ether]palladium(II) (15 mg, 0.021 mmol), cesium carbonate (208 mg, 0.64 mmol), and toluene (5 mL) were added to a 30 mL microwave-safe tube. The tube was sealed, purged with nitrogen several times, and heated to 100 °C overnight. The reaction mixture was evaporated to dryness, water was added, and the mixture was extracted with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column flash chromatography (mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 27-a (154 mg, 85.60%). LC-MS (ESI): m / z 847.8 (M+H) + .

[0356] Synthesis of Compound 27

[0357] Trifluoroacetic acid (2 mL) was added to a solution of compound 27-a (154 mg, 0.18 mmol) in dichloromethane (15 mL) at room temperature, and the mixture was reacted at this temperature for 2 hours. The reaction solution was evaporated to dryness, and saturated sodium bicarbonate aqueous solution was added, followed by extraction with ethyl acetate (30 mL). The organic phase was evaporated to dryness, and the crude product was purified by preparative HPLC to give compound 27 (23 mg, 19.56%). LC-MS (ESI): m / z 647.3 (M+H) + ; 1H NMR (400MHz, CDCl3): δ9.13 (1H, s), 7.13 (1H, dd, J = 7.6, 5.2Hz), 6.98 (1H, t, J = 8. 8Hz),6.27(2H,d,J=17.6Hz),5.27(1H,d,J=53.2Hz),4.30–4.10(4H,m),3.99–3. 88(1H,m),3.37–3.11(5H,m),3.06–2.96(1H,m),2.92–2.82(1H,m),2.79–2.69(1 H,m),2.47–2.28(3H,m),2.26–2.15(3H,m),2.02–1.89(4H,m),1.83–1.69(4H,m).

[0358] Synthetic route of compound 28

[0359] Synthesis of compound 28-a

[0360] Compound 10-b (35 mg, 0.08 mmol), compound 19-b (25 mg, 0.08 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II)cataCXium A Pd G3 (6 mg, 0.008 mmol), K3PO4 (51 mg, 0.24 mmol), THF (5 mL), and water (1 mL) were added to a 30 mL microwave tube. The microwave tube was sealed, purged with nitrogen several times, and heated to 65 °C overnight. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column flash chromatography (mobile phase: dichloromethane / methanol = 10:1 / dichloromethane, 0-100%) to give compound 28-a (42 mg, 78%). LC-MS(ESI): m / z 676.0 (M+H) + .

[0361] Synthesis of Compound 28

[0362] Trifluoroacetic acid (2 mL) was added to a solution of compound 28-a (42 mg, 0.062 mmol) in dichloromethane (10 mL) at room temperature, and the mixture was reacted at this temperature for 3 hours. The reaction solution was evaporated to dryness, and saturated sodium bicarbonate aqueous solution was added, followed by extraction with ethyl acetate (30 mL). The organic phase was evaporated to dryness, and the crude product was purified by preparative HPLC to give compound 28 (3.2 mg, 8.7%). LC-MS (ESI): m / z 591.3 (M+H) + .

[0363] Synthetic route of compound 29

[0364] Synthesis of compound 29-p

[0365] 2-Amino-4-bromo-3,6-difluorobenzoic acid (2500 mg, 9.92 mmol), DMF (10 mL), and DCM (40 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement, and NIS (4464 mg, 19.84 mmol) was added. The reaction mixture was reacted at room temperature for 5 hours. The reaction solution was extracted twice with ethyl acetate after adding water. The organic phases were combined, washed twice with water and brine, dried over anhydrous sodium sulfate, and concentrated. The solution was stirred into silica gel and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 3 / 7) to give compound 29-p (2400 mg, 64%). LC-MS (ESI): m / z = 375.8 (MH) - .

[0366] Synthesis of compound 29-o

[0367] Compound 29-p (2400 mg, 6.35 mmol), THF (30 mL), and CDI (1545 mg, 9.53 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement and reacted at 50 °C for 30 minutes. Ammonia solution (25 mL) at 0 °C was slowly added dropwise. The reaction was continued at room temperature for 1 hour. The reaction mixture was extracted twice with ethyl acetate after adding water. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, and concentrated. The solution was then purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 9 / 1) using silica gel to give compound 29-o (2200 mg, 92%). LC-MS (ESI): m / z = 376.9 (M+H) + .

[0368] Synthesis of compound 29-n

[0369] Compound 29-o (2200 mg, 5.84 mmol) and THF (50 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by nitrogen displacement and heated to 40 °C. NaH (60%, 584 mg, 14.59 mmol) was added in portions. After stirring for 10 minutes, N,N-thiocarbazyldiimidazole (1560 mg, 8.75 mmol) was added in portions. The mixture was heated to 60 °C and reacted for half an hour. The reaction solution was cooled to 0 °C, quenched with saturated ammonium chloride solution, and adjusted to weakly acidic with dilute hydrochloric acid. THF was removed under reduced pressure, resulting in the precipitation of a solid. The solid was filtered, washed with water, and dried to obtain crude compound 29-n (2700 mg). LC-MS (ESI): m / z = 418.6 (M+H) + .

[0370] Synthesis of compound 29-m

[0371] Crude 29-n (2700 mg) and methanol (50 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by nitrogen displacer treatment, and sodium methoxide (522 mg, 9.67 mmol) and methyl iodide (1829 mg, 12.89 mmol) were added. The reaction was allowed to proceed at room temperature for 1 hour. The reaction was quenched by slowly adding 20 mL of water, resulting in the precipitation of a solid. The mixture was stirred for 10 minutes. The mixture was filtered, the filter cake was washed with water, and dried to give compound 29-m (2300 mg, 91%). LC-MS (ESI): m / z = 434.8 (M+H) + .

[0372] Synthesis of compound 29-l

[0373] Compound 29-m (2000 mg, 4.62 mmol), acetonitrile (50 mL), phosphorus oxychloride (10622 mg, 69.28 mmol), and DIPEA (11940 mg, 92.38 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected with nitrogen and heated to 80 °C for 3 hours. The reaction solution was concentrated. The solution was then purified by column chromatography (mobile phase: PE / EA 10 / 0 to 9 / 1) to give compound 29-l (1500 mg, 72%). LC-MS (ESI): m / z = 450.8 (M+H) + .

[0374] Synthesis of compound 29-k

[0375] Compound 29-l (1500 mg, 3.32 mmol) and THF (80 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement, and a methanol solution of sodium methoxide (30% wt, 598 mg, 3.32 mmol) was slowly added dropwise under dry ice and acetone bath. After the addition was complete, the reaction mixture was reacted for 1 hour. The reaction solution was quenched with water, and THF was removed under reduced pressure, resulting in a solid precipitation. The precipitate was filtered, washed with water, and dried to give compound 29-k (1380 mg, 93%). LC-MS (ESI): m / z = 446.8 (M+H) + .

[0376] Synthesis of compound 29-j

[0377] Compound 29-k (1380 mg, 3.09 mmol), DMAC (50 mL), and cuprous iodide (1764 mg, 9.26 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement, and methyl fluorosulfonyl difluoroacetate (1779 mg, 9.26 mmol) was added. The mixture was heated to 90 °C and reacted for 3 hours. The reaction solution was filtered, and the filtrate was extracted with water and EA. The organic phases were combined, washed with water and brine, dried, and concentrated. The solution was stirred into silica gel and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 9 / 1) to give compound 29-j (1000 mg, 83%). LC-MS (ESI): m / z = 389.0 (M+H) + .

[0378] Synthesis of compound 29-i

[0379] Compound 29-j (1000 mg, 2.57 mmol), acetic acid (15 mL), and sodium iodide (4000 mg, 26.69 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement and heated to 90 °C for 3 hours. The reaction solution was concentrated, adjusted to alkalinity with saturated sodium bicarbonate solution, and extracted with EA. The organic phases were combined, washed with water and brine, dried, and concentrated. The solution was stirred into silica gel and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 5 / 5) to give compound 29-i (600 mg, 62%). LC-MS (ESI): m / z = 374.9 (M+H) + .

[0380] Synthesis of compound 29-h

[0381] Compound 29-i (600 mg, 1.60 mmol), DMF (10 mL), and benzyl alcohol (346 mg, 3.20 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement, and NaH (60%, 224 mg, 5.60 mmol) was added in portions under ice bath conditions. After the addition was complete, the reaction mixture was reacted under ice bath conditions for 1 hour. The reaction solution was quenched with saturated ammonium chloride aqueous solution and extracted with EA. The organic phases were combined, washed with water and brine, dried, and concentrated. The solution was stirred into silica gel and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 5 / 5) to give compound 29-h (570 mg, 77%). LC-MS (ESI): m / z = 462.7 (M+H) + .

[0382] Synthesis of compound 29-g

[0383] Compound 29-h (570 mg, 1.23 mmol), acetonitrile (10 mL), phosphorus oxychloride (2830 mg, 18.46 mmol), and DIPEA (3181 mg, 24.61 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 and heated to 80 °C for 3 hours. The reaction solution was concentrated. The solution was then purified by column chromatography (mobile phase: PE / EA 10 / 0 to 9 / 1) to give compound 29-g (250 mg, 42%). LC-MS (ESI): m / z = 481.0 (M+H) + .

[0384] Synthesis of compound 29-f

[0385] Compound 29-g (250 mg, 0.52 mmol), THF (10 mL), [1,4,5]oxadiazazepoxide (106 mg, 1.04 mmol), and DIPEA (268 mg, 2.07 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement and reacted overnight at room temperature. The reaction solution was concentrated. The solution was then purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 29-f (230 mg, 81%). LC-MS (ESI): m / z = 547.0 (M+H) + .

[0386] Synthesis of compound 29-e

[0387] Compound 29-f (230 mg, 0.42 mmol), DCM (6 mL), and TFA (2 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement and reacted at room temperature for 2 hours. The reaction solution was concentrated to give crude compound 29-e (260 mg). LC-MS (ESI): m / z = 456.9 (M+H) + .

[0388] Synthesis of compound 29-d

[0389] At room temperature, crude 29-e (260 mg), toluene (10 mL), paraformaldehyde (400 mg, 13.32 mmol), and 5 drops of acetic acid were added sequentially to a reaction flask. The mixture was protected with nitrogen displacer and heated to 100 °C for half an hour. The reaction solution was concentrated. Methanol was added, and the solution was adjusted to alkalinity with saturated sodium bicarbonate aqueous solution and concentrated. Silica gel was added, and the solution was purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 29-d (170 mg, 86%). LC-MS (ESI): m / z = 469.0 (M+H) + .

[0390] Synthesis of compound 29-c

[0391] Compound 29-d (170 mg, 0.36 mmol), DCM (10 mL), and m-chloroperoxybenzoic acid (250 mg, 1.45 mmol) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement and reacted at room temperature for 3 hours. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 29-c (130 mg, 72%). LC-MS (ESI): m / z = 500.3 (M+H) + .

[0392] Synthesis of compound 29-b

[0393] Compound 29-c (130 mg, 0.26 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (54 mg, 0.34 mmol), and DCM (6 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement, and sodium tert-butoxide (50 mg, 0.52 mmol) was added under ice bath conditions. The reaction mixture was reacted at room temperature for 2 hours. Water was added to the reaction solution, and the mixture was extracted with DCM. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 29-b (90 mg, 60%). LC-MS (ESI): m / z 580.2 (M+H) + .

[0394] Synthesis of compound 29-a

[0395] Compound 29-b (90 mg, 0.16 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoborhexane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (188 mg, 0.47 mmol), toluene (4 mL), bis(diphenylphosphine ether)palladium dichloride (22 mg, 0.03 mmol), and cesium carbonate (253 mg, 0.78 mmol) were added sequentially to a microwave-safe mixture. The mixture was protected with N2 displacement and heated to 100 °C overnight. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 29-a (55 mg, 45%). LC-MS (ESI): m / z 792.6 (M+H) + .

[0396] Synthesis of Compound 29

[0397] Compound 29-a (55 mg, 0.07 mmol), DCM (6 mL), and trifluoroacetic acid (2 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to obtain compound 29 (3.3 mg, 7%). LC-MS (ESI): m / z 692.5 (M+H) + .

[0398] Synthetic route of compound 30

[0399] Synthesis of compound 30-a

[0400] Compound 7-b (118 mg, 0.18 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (26 mg, 0.04 mmol), 1,4-dioxane (6 mL), compound 19-b (114 mg, 0.36 mmol), and cesium carbonate (295 mg, 0.90 mmol) were added to a microwave tube. The tube was sealed, purged three times with nitrogen, and heated to 95 °C for 4 hours. The reaction mixture was concentrated and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 30-a (89 mg, 58%). LC-MS (ESI): m / z = 842.3 (M+H) + .

[0401] Synthesis of Compound 30

[0402] Trifluoroacetic acid (1 mL) was added to a solution of compound 30-a (89 mg, 0.11 mmol) in dichloromethane (3 mL) at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated and purified by preparative HPLC to give compound 30 (4.9 mg, 7%). LC-MS (ESI): m / z = 658.5 (M+H) + .

[0403] Synthetic route of compound 31

[0404] Synthesis of compound 31-j

[0405] 2-(tert-butyl)hydrazine-1,2-dicarboxylic acid-1-benzyl ester (5 g, 18.78 mmol), N,N-dimethylformamide (50 mL), 1,3-dibromopropane (1.91 mL, 18.78 mmol), and cesium carbonate (18.35 g, 56.33 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 3 hours. Water was added to the reaction solution, and the mixture was extracted twice with ethyl acetate. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to give compound 31-j. LC-MS (ESI): m / z 307.1 (M+H) + .

[0406] Synthesis of compound 31-i

[0407] Compound 31-j (1 g, 3.26 mmol), dichloromethane (10 mL), and trifluoroacetic acid (5 mL, 65.30 mmol) were added to a reaction flask. The mixture was stirred at room temperature for 2 hours, then evaporated to dryness. 4 mL of methanol was added, and the pH was adjusted to 7 with saturated sodium bicarbonate solution. A large amount of dichloromethane was added, and the mixture was dried over anhydrous sodium sulfate. The mixture was filtered and evaporated to dryness to obtain compound 31-i (673 mg, 100%), which was used directly in the next reaction without purification. LC-MS (ESI): m / z 207.0 (M+H) + .

[0408] Synthesis of compound 31-h

[0409] Compound 31-i (670 mg, 3.25 mmol), dichloromethane (30 mL), tert-butyldimethylsiloxane acetaldehyde (1258 mg, 6.50 mmol), and sodium triacetoxyborohydride (2056 mg, 9.75 mmol) were added to a reaction flask and stirred overnight at room temperature under nitrogen. The solution was evaporated to dryness and purified by column chromatography (ethyl acetate: petroleum ether = 0–30%) to give compound 31-h (485 mg, 41%). LC-MS (ESI): m / z 365.6 (M+H) + .

[0410] Synthesis of compound 31-e

[0411] Compound 31-h (485 mg, 1.33 mmol), tetrahydrofuran (20 mL), and 10% Pd / C (113 mg, 0.11 mmol) were added to reaction flask B. The mixture was purged with hydrogen three times and stirred at room temperature for 1.5 hours. The mixture was then filtered to obtain compound 31-g, which was quickly used in the next step.

[0412] Compound 29-i (200 mg, 0.53 mmol), dried acetonitrile (20 mL), N,N-diisopropylethylamine (0.66 mL, 4.00 mmol), and phosphorus oxychloride (0.75 mL, 8.00 mmol) were added to reaction flask A. The mixture was stirred at 80 °C for 2 hours under nitrogen, and then evaporated to dryness to give compound 31-f. Dried tetrahydrofuran (20 mL), N,N-diisopropylethylamine (0.66 mL, 4.00 mmol), and 31-g were added to 31-f. The mixture was stirred overnight at room temperature under nitrogen. The next day, the mixture was evaporated to dryness and purified by column chromatography (ethyl acetate: petroleum ether = 0–13%) to give compound 31-e (254 mg, 81%). LC-MS (ESI): m / z 587.1 (M+H) + .

[0413] Synthesis of compound 31-d

[0414] Compound 31-e (254 mg, 0.43 mmol) was added to a reaction flask, along with 15 mL of dried N,N-dimethylformamide and cesium fluoride (1970 mg, 12.97 mmol). The mixture was stirred at 55 °C for 2 hours under nitrogen atmosphere. Water was added under ice-water bath cooling, and the mixture was extracted twice with ethyl acetate, washed three times with saturated brine, evaporated to dryness, and purified by column chromatography (ethyl acetate: petroleum ether = 0–30%) to give compound 31-d (163 mg, 83%). LC-MS (ESI): m / z 452.9 (M+H) + .

[0415] Synthesis of compound 31-c

[0416] Compound 31-d (163 mg, 0.36 mmol), ethyl acetate (30 mL), and m-chloroperoxybenzoic acid (233 mg, 1.08 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 2 hours. A saturated sodium sulfite solution was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution, then with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness to give compound 31-c (174 mg, 100%). LC-MS (ESI): m / z 484.9 (M+H) + .

[0417] Synthesis of compound 31-b

[0418] Compound 31-c (174 mg, 0.36 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (114 mg, 0.72 mmol), and dry dichloromethane (20 mL) were added to a reaction flask. Sodium tert-butoxide (103 mg, 1.08 mmol) was added under ice-water bath conditions, and the mixture was stirred for 1 hour at this temperature. The reaction solution was immediately purified by column chromatography (methanol:dichloromethane = 0–6%) to give compound 31-b (150 mg, 74%). LC-MS (ESI): m / z 564.1 (M+H) + .

[0419] Synthesis of compound 31-a

[0420] Compound 31-b (150 mg, 0.27 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (269 mg, 0.66 mmol), cesium carbonate (433 mg, 1.33 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (20 mg, 0.029 mmol), and toluene (18 mL) were added to a microwave tube. The tube was sealed, purged with nitrogen three times, and stirred at 100 °C for 3 hours. The solution was evaporated to dryness and purified by column chromatography (methanol:dichloromethane = 0–10%) to give compound 31-a (182 mg, 45%). LC-MS (ESI): m / z 776.2 (M+H) + .

[0421] Synthesis of Compound 31

[0422] Compound 31-a (182 mg, 0.12 mmol), dichloromethane (8 mL), and trifluoroacetic acid (8 mL) were added to a reaction flask and stirred at room temperature for 1 hour. The mixture was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 31 (27 mg, 34%). LC-MS (ESI): m / z 676.2 (M+H) + ; 1 H NMR(400M,DMSO_d6)δ8.04(2H,s),7.15-7.02(2H,m),5.28(1H,d,J=53.6Hz),4.54-4.27(2H,m),4.17-3.96 (2H,m),3.87(2H,t,J=7.2Hz),3.23-2.97(7H,m),2.91-2.76(1H,m),2.31-1.94(5H,m),1.91-1.69(3H,m).

[0423] Synthesis of Compound 32

[0424] Following the synthetic route of compound 25, starting with 24-g, and replacing [1,4,5]oxadiazazepoxide with hexahydropyridazine dihydrochloride, compound 32 was synthesized. LC-MS (ESI): m / z 642.2 (M+H) + ; 1 H NMR(400M,DMSO_d6)δ8.09(2H,s),7.26-7.19(1H,m),7.18-7.10(1H,m),5.42-5.18(3H,m),4 .43-3.93(4H,m),3.27-3.00(5H,m),2.96-2.80(1H,m),2.27-1.97(3H,m),1.95-1.55(7H,m).

[0425] Synthetic route of compound 33

[0426] Synthesis of compound 33-f

[0427] Compound 24-i (980 mg, 2.87 mmol), acetonitrile (30 mL), N,N-diisopropylethylamine (3.56 mL, 21.52 mmol), and phosphorus oxychloride (4.01 mL, 43.04 mmol) were added to a reaction flask, and the mixture was stirred at 80 °C for 2 hours under nitrogen. The solution was evaporated to dryness to obtain the crude product. Tetrahydrofuran (30 mL) was added to the crude product, followed by N,N-diisopropylethylamine (3.56 mL, 21.52 mmol) and hexahydropyridazine dihydrochloride (548 mg, 3.44 mmol) under an ice-water bath. After the addition was complete, the mixture was allowed to warm to room temperature and stirred overnight. The next day, the reaction solution was evaporated to dryness and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–20%) to give compound 33-f (450 mg, 38%). LC-MS (ESI): m / z 408.6 (M+H) + .

[0428] Synthesis of compound 33-e

[0429] Compound 33-f (450 mg, 1.10 mmol), dichloromethane (30 mL), tert-butyldimethylsiloxane acetaldehyde (425 mg, 2.20 mmol), and sodium triacetoxyborohydride (695 mg, 3.30 mmol) were added to a reaction flask and stirred overnight at room temperature under nitrogen. The next day, the reaction mixture was evaporated to dryness and purified by column chromatography (ethyl acetate: petroleum ether = 0–5%, UV 254 nm) to give compound 33-e (600 mg, 96%). LC-MS (ESI): m / z 567.0 (M+H) + .

[0430] Synthesis of compound 33-d

[0431] Compound 33-e (600 mg, 1.06 mmol), N,N-dimethylformamide (30 mL), and cesium fluoride (1604 mg, 10.56 mmol) were added to a reaction flask, and the mixture was stirred at 60 °C for 24 hours under nitrogen atmosphere. Ice water was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate. The organic phase was washed three times with saturated brine, evaporated to dryness, and purified by column chromatography (ethyl acetate: petroleum ether = 0–30%) to give compound 33-d (137 mg, 30%). LC-MS (ESI): m / z 432.8 (M+H) + .

[0432] Synthesis of compound 33-c

[0433] Compound 33-d (137 mg, 0.32 mmol), ethyl acetate (15 mL), and m-chloroperoxybenzoic acid (192 mg, 0.95 mmol) were added to a reaction flask, and the mixture was stirred at room temperature under nitrogen for 1 hour. A saturated sodium sulfite solution was added, and the mixture was extracted twice with ethyl acetate. The organic phase was washed with saturated sodium bicarbonate solution, then with saturated brine, dried over anhydrous sodium sulfate, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–50%) to give compound 33-c (125 mg, 85%). LC-MS (ESI): m / z 464.9 (M+H) + .

[0434] Synthesis of compound 33-b

[0435] Compound 33-c (125 mg, 0.27 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (128 mg, 0.81 mmol), dry dichloromethane (15 mL), and sodium tert-butoxide (90 mg, 0.94 mmol) were added to a reaction flask under ice-water bath conditions. The mixture was stirred at this temperature for 1 hour. The reaction solution was then directly purified by column chromatography (mobile phase: methanol:dichloromethane = 0–10%) to give compound 33-b (97 mg, 66%). LC-MS (ESI): m / z 543.9 (M+H) + .

[0436] Synthesis of compound 33-a

[0437] Compound 33-b (97 mg, 0.18 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (144 mg, 0.36 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (13 mg, 0.018 mmol), cesium carbonate (290 mg, 0.89 mmol), and toluene (15 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 33-a (135 mg, 100%). LC-MS (ESI): m / z 756 (M+H) + .

[0438] Synthesis of Compound 33

[0439] Trifluoroacetic acid (2 mL) was added to a solution of compound 33-a (135 mg, 0.18 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 33 (63 mg, 54%). LC-MS (ESI): m / z 656.2 (M+H) + . 1 H NMR(400M,DMSO-d6)δ8.05(2H,d,J=8.8Hz),7.29-7.20(1H,m),7.18-7.07(1H,m),5.30(1H,d,J=54.0Hz),4.81-4.65(1H,m),4.4 3-3.94(4H,m),3.77-3.61(1H,m),3.25-2.70(8H,m),2.26-1.97(3H,m),1.95-1.71(5H,m),1.70-1.54(1H,m),1.46-1.31(1H,m).

[0440] Synthesis of compounds 33-1 and 33-2

[0441] Compound 33 (60 mg, 0.091 mmol) was chirally resolved to give two isomers, compound 33-1 (26 mg, 43%) and compound 33-2 (23 mg, 38%).

[0442] Synthesis of Compound 34

[0443] Following the synthetic route of compound 24, starting with 3-d and replacing 24-g with 8-g, compound 34 was synthesized. LC-MS (ESI): m / z 642.2 (M+H) + ; 1 H NMR(400M,DMSO-d6)δ8.14(2H,s),7.42-7.31(1H,m),7.21-7.11(1H,m),5.64-5.12(3H,m),4.89-4.57 (1H,m),4.18-3.46(8H,m),3.19-2.96(4H,m),2.90-2.77(1H,m),2.21-1.95(3H,m),1.92-1.68(3H,m).

[0444] Synthesis of Compound 35

[0445] Following the synthetic route of compound 25, starting with 25-e, and replacing paraformaldehyde with an aqueous acetaldehyde solution (49%), compound 35 was synthesized. LC-MS (ESI): m / z 672.2 (M+H) + ; 1 H NMR(400M, DMSO-d6)δ8.08(2H,s),7.37-7.20(1H,m),7.20-7.09(1H,m),5.75-5.50(1H,m),5.29(1H,d,J=54.0Hz),4.89-4.58(1H,m),4.20-3. 84(4H,m),3.83-3.61(1H,m),3.60-3.43(2H,m),3.21-2.95(4H,m),2.9 3-2.69(2H,m),2.22-1.95(3H,m),1.93-1.71(3H,m),1.66-1.33(3H,m).

[0446] Synthesis of Compound 36

[0447] Following the synthetic route of compound 6, starting with 6-c, compound 36 was synthesized by replacing ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol with (1-(morpholinomethyl)cyclopropyl)methanol. LC-MS (ESI): m / z 676.2 (M+H) + ; 1H NMR(400M, DMSO-d6)δ9.84(1H,s),8.07(2H,s),7.27-7.20(1H,m),7.17-7.09(1H,m),6.16(1H,t,J=6.0Hz),4.34-4.20(3H,m),4.19-4.05( 1H,m),4.01-3.91(2H,m),3.79-3.69(2H,m),3.59-3.48(4H,m),3.16- 3.00(2H,m),2.46-2.20(6H,m),0.70-0.56(2H,m),0.47-0.34(2H,m).

[0448] Synthetic route of compound 37

[0449] Synthesis of compound 37-b

[0450] Compound 7-c (250 mg, 0.47 mmol), (2-(difluoromethylene)tetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (357 mg, 1.89 mmol), acetonitrile (15 mL), and N,N-diisopropylethylamine (390 μL, 2.36 mmol) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and stirred at 90 °C for 2 days. The reaction mixture was evaporated to dryness and purified by column chromatography (ethyl acetate: petroleum ether = 0–100%) to give compound 37-b (210 mg, 65%). LC-MS (ESI): m / z 682.3 (M+H) + .

[0451] Synthesis of compound 37-a

[0452] Compound 37-b (100 mg, 0.15 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoborhexane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (178 mg, 0.44 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (10 mg, 0.015 mmol), and cesium carbonate (239 mg, 0.73 mmol) were added to a microwave tube. 1,4-dioxane (15 mL) was dried, the tube was sealed, purged three times with nitrogen, and heated to 120 °C for 3 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (ethyl acetate: petroleum ether = 0–100%) to give compound 37-a (101 mg, 77%). LC-MS (ESI): m / z 894.8 (M+H) + .

[0453] Synthesis of Compound 37

[0454] Trifluoroacetic acid (2 mL) was added to a 4 mL solution of compound 37-a (100 mg, 0.11 mmol) in dichloromethane at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 37 (13 mg, 16%). LC-MS (ESI): m / z 694.2 (M+H) + ; 1 H NMR(400M,DMSO-d6)δ9.84(1H,s),8.07(2H,s),7.29-7.20(1H,m),7.18-7 .08(1H,m),6.17(1H,t,J=6.4Hz),4.36-4.20(1H,m),4.19-4.03(3H,m),4. 01-3.91(2H,m),3.80-3.70(2H,m),3.69-3.59(1H,m),3.35-3.26(1H,m),3 .17-2.95(3H,m),2.69-2.53(2H,m),2.46-2.34(1H,m),2.04-1.70(4H,m).

[0455] Synthetic route of compound 38

[0456] Synthesis of compound 38-e

[0457] At room temperature, tert-butyl 1,4,5-oxadiaza-4-carboxylic acid (202 mg, 1.00 mmol), methanol (10 mL), sodium cyanoborohydride (251 mg, 4.00 mmol), acetic acid (60 mg, 1.00 mmol), and paraformaldehyde (300 mg, 10.00 mmol) were added to a reaction flask. The mixture was purged with nitrogen three times and stirred at room temperature for 12 hours. The reaction solution was directly evaporated to dryness and purified by column chromatography (mobile phase: dichloromethane / methanol = 100 / 0; 10 / 1) to give compound 38-e (163 mg, 75%). LC-MS (ESI): m / z = 217.4 (M+H) + .

[0458] Synthesis of compound 38-d

[0459] 38-e (163 mg, 0.75 mmol), dichloromethane (6 mL), and trifluoroacetic acid (2 mL) were added to the reaction flask, and the mixture was stirred at room temperature for 3 hours. The solution was then evaporated to dryness to give compound 38-d (88 mg, 100%). LC-MS (ESI): m / z = 117.2 (M+H) + .

[0460] Synthesis of compound 38-c

[0461] Dry tetrahydrofuran (20 mL), N,N-diisopropylethylamine (294 mg, 2.27 mmol), and 38-d (88 mg, 0.76 mmol) were added to the reaction flask under an ice-water bath, and the mixture was stirred for 10 minutes. 7-Bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (303 mg, 0.83 mmol) was added to the reaction mixture under nitrogen protection. The reaction mixture was slowly brought to room temperature and stirred for 4 hours. The reaction mixture was evaporated to dryness and purified by column chromatography (petroleum ether / ethyl acetate = 100 / 0; 1 / 3) to give compound 38-c (300 mg, 89%). LC-MS (ESI): m / z = 443.1 (M+H) + .

[0462] Synthesis of compound 38-b

[0463] 38-c (310 mg, 0.70 mmol), N,N-diisopropylethylamine (271 mg, 2.10 mmol), dry acetonitrile (30 mL), and ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (445 mg, 2.80 mmol) were added to a reaction flask. The mixture was heated to 90 °C under nitrogen and stirred overnight. The solution was evaporated to dryness and purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1; 1 / 3) to give compound 38-b (149 mg, 38%).

[0464] Synthesis of compound 38-a

[0465] 38-b (50 mg, 0.09 mmol), tert-butyl(3-cyano-4-(5,5-dimethyl-1,3,2-dioxaboronic-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate (71 mg, 0.18 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (13 mg, 0.02 mmol), cesium carbonate (144 mg, 0.44 mmol), and 1,4-dioxane (16 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 95 °C for 4 hours. The reaction mixture was evaporated to dryness and purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 38-a (38 mg, 55%). LC-MS (ESI): m / z = 778.0 (M+H) + .

[0466] Synthesis of Compound 38

[0467] Trifluoroacetic acid (3 mL) was added to a solution of 38-a (39 mg, 0.05 mmol) in dichloromethane (9 mL) at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction solution was evaporated to dryness and purified by HPLC to give compound 38 (8.6 mg, 25%). LC-MS (ESI): m / z = 678.4 (M+H) + .

[0468] Synthesis of compounds 38-1 and 38-2

[0469] Trifluoroacetic acid (3 mL) was added to a dichloromethane (9 mL) solution of compound 38-a (210 mg, 0.27 mmol) at room temperature, and the mixture was stirred at room temperature for 3 hours. The mixture was evaporated to dryness, and saturated sodium bicarbonate solution (20 mL) was added. The mixture was extracted with ethyl acetate (20 mL x 3), concentrated, and purified by HPLC. Chiral resolution yielded compound 38-1 (27.2 mg, 15%) and compound 38-2 (24.6 mg, 13%).

[0470] Compound 38-1: LC-MS (ESI): m / z = 678.4 (M+H) + ; 1 H NMR (400MHz, DMSO_d6): δ9.53(1H,s),8.07(2H,s),7.23(1H,t,J=5.2Hz),7 .12(1H,t,J=8.4Hz),5.27(1H,d,J=52.4Hz),4.62(1H,d,J=11.6Hz),3.90- 4.15(5H,m),3.64-3.76(2H,m),3.46-3.60(1H,m),2.95-3.17(4H,m),2.78 -2.88(1H,m),2.73(3H,d,J=10.4Hz),1.93-2.07(3H,m),1.73-1.88(3H,m).

[0471] Compound 38-2: LC-MS (ESI): m / z = 678.4 (M+H) + ; 1H NMR (400MHz, DMSO_d6): δ9.53(1H,s),8.07(2H,s),7.24(1H,t,J=6.8Hz),7 .12(1H,t,J=7.6Hz),5.27(1H,d,J=52.4Hz),4.62(1H,d,J=12.0Hz),3.90- 4.15(5H,m),3.64-3.76(2H,m),3.46-3.60(1H,m),2.97-3.15(4H,m),2.78 -2.88(1H,m),2.73(3H,d,J=11.6Hz),1.93-2.11(3H,m),1.73-1.88(3H,m).

[0472] Synthesis of Compound 39

[0473] Following the synthetic route of compound 38, starting with 38-c, compound 39 was synthesized by replacing ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a(5H)-yl)methanol with (2-(difluoromethylene)tetrahydro-1H-pyrrolazin-7a-yl)methanol. LC-MS (ESI): m / z 708.2 (M+H) + ; 1 H NMR(400M,DMSO-d6)δ9.54(1H,s),8.08(2H,s),7.29-7.20(1H,m),7.13(1H,t,J=9.6Hz),4.68-4.56(1H,m),4.18-3.86(4H,m),3.77-3.50(4H ,m),3.48-3.40(1H,m),3.34-3.25(1H,m),3.09-2.93(2H,m),2.73(3H, d,J=10.4Hz),2.68-2.53(2H,m),2.45-2.31(1H,m),2.04-1.70(4H,m).

[0474] Synthetic route of compound 40

[0475] Synthesis of compound 40-e

[0476] 5,7-Dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(1H)-one (1000 mg, 3.57 mmol), dry acetonitrile (20 mL), N,N-diisopropylethylamine (4.43 mL, 26.78 mmol), and phosphorus oxychloride (4.99 mL, 53.55 mmol) were added to a reaction flask, and the mixture was stirred at 80 °C for 2 hours under nitrogen. The reaction solution was evaporated to dryness, and dry dichloromethane (20 mL) was added. Under dry ice and acetone bath cooling, N,N-diisopropylethylamine (4.43 mL, 26.78 mmol) and hexahydropyridazine dihydrochloride (568 mg, 3.57 mmol) were added. After the addition was complete, the mixture was naturally heated and stirred for 1 hour. The reaction solution was directly purified by column chromatography (ethyl acetate: petroleum ether = 0–30%) to give compound 40-e (730 mg, 59%). LC-MS(ESI): m / z 348.0(M+H) + .

[0477] Synthesis of compound 40-d

[0478] Compound 40-e (730 mg, 2.10 mmol), dichloromethane (40 mL), tert-butyldimethylsiloxane acetaldehyde (812 mg, 4.19 mmol), and sodium borohydride acetate (1327 mg, 6.29 mmol) were added to a reaction flask and stirred overnight at room temperature under nitrogen. A saturated sodium bicarbonate solution was added to the reaction mixture, and the mixture was extracted with dichloromethane, evaporated to dryness, and purified by column chromatography (ethyl acetate: petroleum ether = 0–20%) to give compound 40-d (600 mg, 57%). LC-MS (ESI): m / z 506.2 (M+H) + .

[0479] Synthesis of compound 40-c

[0480] Compound 40-d (600 mg, 1.19 mmol), N,N-dimethylformamide (15 mL), and cesium fluoride (540 mg, 3.55 mmol) were added to a reaction flask, and the mixture was stirred at 55 °C for 3 hours under nitrogen atmosphere. The reaction solution was cooled to room temperature, water was added, and the mixture was extracted twice with ethyl acetate. The organic phase was washed three times with saturated brine, evaporated to dryness, and purified by column chromatography (ethyl acetate: petroleum ether = 0–40%), followed by column chromatography (methanol: dichloromethane = 0–2%) to give compound 40-c (110 mg, 26%).

[0481] Synthesis of compound 40-b

[0482] Compound 40-c (110 mg, 0.31 mmol) and ethyl acetate (30 mL) were added to a reaction flask. Then, m-chloroperoxybenzoic acid (200 mg, 0.93 mmol) was added under ice-water bath conditions. The mixture was stirred at room temperature for 2 hours, resulting in the appearance of a large amount of white solid. Saturated sodium sulfite and sodium bicarbonate solutions were added. The mixture was extracted with ethyl acetate (100 mL x 2), followed by extraction with dichloromethane (100 mL x 2). The mixed organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to obtain compound 40-b (149 mg, 124%), which was used directly in the next reaction without purification.

[0483] Synthesis of compound 40-a

[0484] Compound 40-b (149 mg, 0.38 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (73 mg, 0.46 mmol), and dichloromethane (30 mL) were added to a reaction flask. Sodium tert-butoxide (92 mg, 0.96 mmol) was added under ice-water bath conditions, and the mixture was stirred for 1 hour at this temperature. The mixture was then purified directly by column chromatography (methanol:dichloromethane = 0–10%) to give compound 40-a (150 mg, 84%). LC-MS (ESI): m / z 467.2 (M+H) + .

[0485] Synthesis of Compound 40

[0486] Compound 40-a (50 mg, 0.11 mmol), 3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxobenzofuran-2-yl)-4-(trifluoromethyl)aniline (69 mg, 0.21 mmol), potassium phosphate (68 mg, 0.32 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.021 mmol), tetrahydrofuran (10 mL), and water (2 mL) were added to a microwave tube. The tube was sealed, purged with nitrogen three times, and stirred overnight at 65 °C. The next day, the mixture was evaporated to dryness, purified by column chromatography (methanol:dichloromethane = 0–10%), and then purified by HPLC (ammonium bicarbonate). The purified compound was lyophilized to give compound 40 (7.5 mg, 11%). LC-MS(ESI): m / z 626.2(M+H) + ; 1H NMR(400M,DMSO-d6)δ6.84(1H,s),6.66-6.15(3H,m),5.28(1H,d,J=54.0 Hz),4.92-4.76(1H,m),4.43-4.27(1H,m),4.18-4.03(2H,m),3.97(1H,t ,J=10.8Hz),3.75-3.62(1H,m),3.18-2.90(6H,m),2.89-2.76(1H,m),2. 36-1.93(4H,m),1.92-1.68(5H,m),1.67-1.49(1H,m),1.44-1.32(1H,m).

[0487] Synthetic route of compound 41

[0488] Synthesis of compound 41-a

[0489] Compound 40-a (100 mg, 0.21 mmol), 6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-5-((triisopropylsilyl)ethynyl)naphth-2-amine (150 mg, 0.32 mmol), potassium phosphate (136 mg, 0.64 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (31 mg, 0.043 mmol), water (2 mL), and tetrahydrofuran (10 mL) were added to a microwave tube. The tube was sealed, purged with nitrogen three times, and stirred overnight at 65 °C. The reaction mixture was evaporated to dryness and purified by column chromatography (methanol:dichloromethane = 0–10%) to give compound 41-a (90 mg, 54%). LC-MS (ESI): m / z 772.4 (M+H) + .

[0490] Synthesis of Compound 41

[0491] Compound 41-a (90 mg, 0.12 mmol), N,N-dimethylformamide (10 mL), and cesium fluoride (531 mg, 3.50 mmol) were added to a reaction flask and stirred at room temperature for 1 hour. The mixture was purified by HPLC (ammonium bicarbonate) and lyophilized to give compound 41 (30 mg, 42%). LC-MS (ESI): m / z 616.3 (M+H) + .

[0492] Synthetic route of compound 42

[0493] Synthesis of Compound 42

[0494] Compound 41 (50 mg, 0.081 mmol), ethanol (20 mL), tetrahydrofuran (20 mL), and 10% Pd / C (86 mg, 0.081 mmol) were added to the reaction flask. The mixture was purged with hydrogen three times and stirred at room temperature for 2 hours. The mixture was filtered, washed twice with methanol, and then three times with N,N-dimethylformamide. The filtrates were combined, evaporated to dryness, and purified by HPLC (ammonium bicarbonate). The purified compound was lyophilized to give compound 42 (1.2 mg, 2%). LC-MS (ESI): m / z 620.3 (M+H) + .

[0495] Synthetic route of compound 43

[0496] Synthesis of compound 43-g

[0497] At room temperature, 5-g (1000 mg, 3.08 mmol), acetonitrile (20 mL), phosphorus oxychloride (7074 mg, 46.14 mmol), and DIPEA (7951 mg, 61.52 mmol) were added sequentially to a reaction flask. Under nitrogen protection, the mixture was heated to 80°C and reacted for 3 hours. The reaction solution was concentrated, stirred with silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 8 / 2) to give 43-g (900 mg, 85%) of the compound.

[0498] Synthesis of compound 43-f

[0499] 43-g (900 mg, 2.62 mmol), pyrazolidine dihydrochloride (570 mg, 3.93 mmol), and DCM (20 mL) were added sequentially to a reaction flask at room temperature. Under nitrogen purging protection, DIPEA (1693 mg, 13.10 mmol) was added in an ice bath. The reaction mixture was allowed to react overnight at room temperature. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 43-f (600 mg, 60%). LC-MS (ESI): m / z 378.8 (M+H) + .

[0500] Synthesis of compound 43-e

[0501] 43-f (600 mg, 1.58 mmol), tert-butyldimethylsiloxane acetaldehyde (552 mg, 3.17 mmol), and DCM (30 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement, and sodium borohydride acetate (1001 mg, 4.75 mmol) was added. The reaction mixture was allowed to react overnight at room temperature. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 9 / 1) to give compound 43-e (700 mg, 82%). LC-MS (ESI): m / z = 537.0 (M+H) + .

[0502] Synthesis of compound 43-d

[0503] 43-e (700 mg, 1.30 mmol), cesium fluoride (1978 mg, 13.02 mmol), and DMF (30 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by N2 displacement and heated to 60 °C overnight. The reaction solution was quenched with ice water and extracted with ethyl acetate. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 43-d (300 mg, 57%). LC-MS (ESI): m / z = 402.5 (M+H) + .

[0504] Synthesis of compound 43-c

[0505] 43-d (300 mg, 0.74 mmol) and DCM (20 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement, and m-chloroperoxybenzoic acid (300 mg, 1.48 mmol) was added under ice bath conditions. The reaction mixture was reacted for half an hour. The reaction solution was alkalized with saturated sodium bicarbonate solution, extracted with DCM, and the organic phases were combined, washed with water, washed with saturated NaCl, dried over anhydrous Na2SO4, and concentrated to give crude compound 43-c (320 mg). LC-MS (ESI): m / z = 419.0 (M+H) + .

[0506] Synthesis of compound 43-b

[0507] At room temperature, 43-c (320 mg, 0.74 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (355 mg, 2.23 mmol), and DCM (20 mL) were added sequentially to a reaction flask. The mixture was protected by N2 displacement, and sodium tert-butoxide (286 mg, 2.98 mmol) was added under ice bath conditions. The reaction mixture was reacted for 1 hour. The reaction solution was stirred into silica gel and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 43-b (200 mg, 52%). LC-MS (ESI): m / z 514.0 (M+H) + .

[0508] Synthesis of compound 43-a

[0509] At room temperature, 43-b (100 mg, 0.19 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (236 mg, 0.58 mmol), toluene (8 mL), bis(diphenylphosphine ether)palladium dichloride (28 mg, 0.04 mmol), and cesium carbonate (253 mg, 0.78 mmol) were added sequentially to a microwave-safe mixture. The mixture was protected by N2 displacement and heated to 100 °C overnight. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 43-a (130 mg, 92%). LC-MS (ESI): m / z 726.8 (M+H) + .

[0510] Synthesis of Compound 43

[0511] 43-a (130 mg, 0.18 mmol), DCM (10 mL), and trifluoroacetic acid (5 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to obtain compound 43 (50 mg, 45%). LC-MS (ESI): m / z 626.2 (M+H) + ; 1 HNMR(CDCl3,400MHz)δ7.30-7.17(1H,m),7.12-6.90(1H,t,J=8.8Hz),6.27-6.20(2H,m) ,5.48-5.14(1H,d,J=53.2Hz),4.60-3.80(6H,m),3.48-2.90(8H,m),2.45-1.80(8H,m).

[0512] Synthesis of compounds 43-1 and 43-2

[0513] A new batch of compound 43 was synthesized. Compound 43 (270 mg) was chirally resolved to give two isomers, compound 43-1 (120 mg) and compound 43-2 (80 mg).

[0514] Compound 43-1: LC-MS (ESI): m / z 626.2 (M+H) + ; 1 HNMR(CDCl3,400MHz)δ7.40-7.17(1H,m),7.12-6.90(1H,t,J=8.8Hz),6.08(2H,s),5 .48-5.14(1H,d,J=53.2Hz),4.60-3.80(6H,m),3.56-2.80(8H,m),2.49-1.80(8H,m).

[0515] Compound 43-2: LC-MS (ESI): m / z 626.2 (M+H) + ; 1 HNMR(CDCl3,400MHz)δ7.38-7.17(1H,m),7.12-6.90(1H,t,J=8.8Hz),5.89(2H,s),5 .48-5.14(1H,d,J=52.8Hz),4.60-3.80(6H,m),3.48-2.80(8H,m),2.45-1.80(8H,m).

[0516] Synthetic route of compound 44

[0517] Synthesis of compound 44-f

[0518] 24-i (1000 mg, 2.93 mmol), acetonitrile (20 mL), phosphorus oxychloride (7074 mg, 46.14 mmol), and DIPEA (7951 mg, 61.52 mmol) were added sequentially to a reaction flask at room temperature. The reaction mixture was heated to 80 °C for 3 hours under nitrogen protection. The reaction solution was concentrated, and pyrazolidine dihydrochloride (435 mg, 3.00 mmol) and DCM (20 mL) were added. Under nitrogen purging protection, DIPEA (2586 mg, 20.00 mmol) was added in an ice bath. The reaction mixture was allowed to react overnight at room temperature. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 44-f (600 mg, 52%). LC-MS (ESI): m / z 394.8 (M+H) + .

[0519] Synthesis of compound 44-e

[0520] 44-f (600 mg, 1.52 mmol), tert-butyldimethylsiloxane acetaldehyde (529 mg, 3.03 mmol), and DCM (30 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected by nitrogen displacement, and sodium borohydride acetate (960 mg, 4.55 mmol) was added. The reaction mixture was allowed to react overnight at room temperature. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 9 / 1) to give compound 44-e (600 mg, 71%). LC-MS (ESI): m / z = 553.1 (M+H) + .

[0521] Synthesis of compound 44-d

[0522] 44-e (600 mg, 1.08 mmol), cesium fluoride (1645 mg, 10.83 mmol), and DMF (30 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement and heated to 60 °C overnight. The reaction solution was extracted with ethyl acetate after adding ice water. The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 7 / 3) to give compound 44-d (280 mg, 62%). LC-MS (ESI): m / z = 418.9 (M+H) + .

[0523] Synthesis of compound 44-c

[0524] 44-d (280 mg, 0.67 mmol) and DCM (20 mL) were added sequentially to a reaction flask at room temperature. The mixture was protected with N2 displacement, and m-chloroperoxybenzoic acid (300 mg, 1.48 mmol) was added under ice bath conditions. The reaction mixture was reacted for half an hour. The reaction solution was alkalized with saturated sodium bicarbonate solution, extracted with DCM, and the organic phases were combined, washed with water, washed with saturated NaCl, dried over anhydrous Na2SO4, and concentrated to give crude compound 44-c (300 mg). LC-MS (ESI): m / z = 434.9 (M+H) + .

[0525] Synthesis of compound 44-b

[0526] At room temperature, 44-c (300 mg, 0.67 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (319 mg, 2.00 mmol), and DCM (20 mL) were added sequentially to a reaction flask. The mixture was protected by N2 displacement, and sodium tert-butoxide (256 mg, 2.67 mmol) was added under ice bath conditions. The reaction mixture was reacted for 1 hour. The reaction solution was stirred into silica gel and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 44-b (200 mg, 56%). LC-MS (ESI): m / z 530.0 (M+H) + .

[0527] Synthesis of compound 44-a

[0528] At room temperature, 44-b (200 mg, 0.38 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (457 mg, 1.13 mmol), toluene (8 mL), bis(diphenylphosphine ether)palladium dichloride (54 mg, 0.08 mmol), and cesium carbonate (491 mg, 1.51 mmol) were added sequentially to a microwave-safe mixture. The mixture was protected by N2 displacement and heated to 100 °C overnight. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 44-a (120 mg, 43%). LC-MS (ESI): m / z 742.3 (M+H) + .

[0529] Synthesis of Compound 44

[0530] 44-a (211 mg, 0.16 mmol), DCM (10 mL), and trifluoroacetic acid (5 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to give compound 44 (29 mg, 28%). LC-MS (ESI): m / z 642.2 (M+H) + ; 1 H NMR(CDCl3,400MHz)δ7.30-7.17(1H,m),7.05-6.90(1H,m),5.97-5.65(2H,m),5.4 8-5.14(1H,d,J=52.8Hz),4.60-3.80(6H,m),3.70-2.90(8H,m),2.54-1.90(8H,m).

[0531] Synthesis of compounds 44-1 and 44-2

[0532] A new batch of compound 44 was synthesized. Compound 44 (270 mg) was chirally resolved to give two isomers, compound 44-1 (77.1 mg) and compound 44-2 (76.6 mg).

[0533] Compound 44-1: LC-MS (ESI): m / z 642.2 (M+H) + ; 1 H NMR (DMSO-d6, 400MHz) δ8.06 (2H, s), 7.23-7.17 (1H, m), 7.12 (1H, t, J = 9.2Hz), 5.27 (1H, d, J = 53.6Hz), 4.38 (2H, d, J = 16.4Hz), 4.08 (1H, d, J = 1 0.0Hz), 3.98 (1H, d, J = 10.4Hz), 3.86 (2H,, s), 3.03-3.15 (6H, m), 2.76 -2.88(1H,m), 2.08-2.27(3H,m), 1.96-2.06(3H,m), 1.73-1.89(3H,m).

[0534] Compound 44-2: LC-MS (ESI): m / z 642.2 (M+H) + ; 1 H NMR (DMSO-d6, 400MHz) δ8.06 (2H, s), 7.23-7.17 (1H, m), 7.12 (1H, t, J = 9.2Hz), 5.28 (1H, d, J = 53.6Hz), 4.38 (2H, d, J = 15.6Hz), 4.10 (1H, d, J = 1 0.4Hz), 3.99 (1H, d, J = 10.8Hz), 3.87 (2H,, s), 3.03-3.15 (6H, m), 2.78 -2.88(1H,m), 2.08-2.26(3H,m), 1.98-2.06(3H,m), 1.74-1.88(3H,m).

[0535] Synthetic route of compound 45

[0536] Synthesis of compound 45-h

[0537] 10 g (37.55 mmol) of compound 2-(tert-butyl)hydrazine-1,2-dicarboxylic acid-1-benzyl ester was dissolved in 100 mL of DMF at room temperature. 1,4-dibromobutane (4.49 mL, 37.55 mmol) and cesium carbonate (36.71 g, 112.66 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (100 mL x 2). The organic phase was washed with saturated brine (200 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in 30 mL of petroleum ether and stirred for 1 hour, resulting in the precipitation of a large amount of solid. The solid was filtered, washed with petroleum ether (10 mL), collected, and dried under vacuum to give compound 45-h (9.8 g, 82%). LC-MS (ESI): m / z = 321.1 [M+H] + .

[0538] Synthesis of compound 45-g

[0539] Compound 45-h (800 mg, 2.50 mmol) was dissolved in 15 mL of DCM at room temperature. TFA (5 mL) was added under ice-water bath conditions, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure. 2 mL of saturated sodium bicarbonate solution was added to the residue, followed by solid sodium bicarbonate until no more bubbles were produced. A mixture of dichloromethane and methanol (10:1, 100 mL) was added to the mixture. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in 15 mL of dichloromethane. tert-butyldimethylsiloxane acetaldehyde (1.09 g, 6.24 mmol) and sodium triacetoxyborohydride (1.58 g, 7.49 mmol) were added, and the mixture was stirred overnight at room temperature. 50 mL of saturated sodium bicarbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane (100 mL). The organic phase was washed with saturated sodium bicarbonate solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid separation column chromatography (EA / PE = 0–30%) to give 45 g (800 mg, 85%) of the compound. LC-MS (ESI): m / z = 379.2 [M+H] + .

[0540] Synthesis of compound 45-f

[0541] Compound 45-g (400 mg, 1.06 mmol) was dissolved in 30 mL of ethanol at room temperature, and palladium on carbon (112 mg, 10%, 50% wet) was added. The mixture was stirred at room temperature under hydrogen atmosphere for 2 hours. The reaction solution was filtered, the filtrate was concentrated under reduced pressure, and dried under vacuum to give compound 45-f (258 mg). The crude product was used directly in the next reaction without purification. LC-MS (ESI): m / z = 245.3 [M+H] + .

[0542] Synthesis of compound 45-e

[0543] Compound 29-i (100 mg, 0.27 mmol) was dissolved in 10 mL of anhydrous acetonitrile at room temperature. DIPEA (0.33 mL, 2.0 mmol) and phosphorus oxychloride (0.37 mL, 4.0 mmol) were added, and the mixture was stirred at 80 °C under nitrogen for 2 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and the residue was dried under vacuum for 20 minutes. The resulting residue was dissolved in anhydrous THF (20 mL), and DIPEA (0.33 mL, 2.0 mmol) and compound 45-f (261 mg, 0.53 mmol, 50%) were added under ice-water bath conditions. The mixture was heated to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (EA / PE = 0–15%) to give compound 45-e (120 mg, 74%). LC-MS (ESI): m / z = 601.0 [M+H] + .

[0544] Synthesis of compound 45-d

[0545] Compound 45-e (120 mg, 0.20 mmol) was dissolved in 10 mL of anhydrous DMF at room temperature, and cesium fluoride (909 mg, 5.99 mmol) was added. The reaction mixture was stirred at 55 °C under nitrogen for 3 hours. The reaction solution was cooled to room temperature, and an ice-water mixture (50 mL) was added. Extraction was performed with ethyl acetate (100 mL). The organic phase was washed with saturated brine (100 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid separation column chromatography (EA / PE = 0–30%) to give compound 45-d (50 mg, 54%). LC-MS (ESI): m / z = 467.0 [M+H] + .

[0546] Synthesis of compound 45-c

[0547] Compound 45-d (50 mg, 0.11 mmol) was dissolved in 10 mL of ethyl acetate at room temperature. Under an ice-water bath, m-chloroperoxybenzoic acid (58 mg, 0.27 mmol, 80%) was added, and the reaction mixture was heated to room temperature and stirred for 3 hours. The reaction solution was cooled under an ice-water bath, and saturated sodium thiosulfate solution (50 mL) was added. Extraction was performed with ethyl acetate (100 mL). The organic phase was washed successively with saturated sodium thiosulfate solution (100 mL) and saturated sodium bicarbonate solution (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and dried under vacuum for 1 hour to give compound 45-c (60 mg). The crude product was used directly in the next reaction without further purification. LC-MS (ESI): m / z = 499.0 [M+H] + .

[0548] Synthesis of compound 45-b

[0549] Compound 45-c (60 mg, 0.12 mmol) and compound ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (38 mg, 0.24 mmol) were dissolved in 10 mL of anhydrous dichloromethane at room temperature. Sodium tert-butoxide (35 mg, 0.36 mmol) was added under ice-water bath conditions, and the reaction mixture was stirred for 1 hour. A saturated ammonium chloride solution (50 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid column chromatography (MeOH / DCM = 0–10%) to give compound 45-b (36 mg, 52%). LC-MS (ESI): m / z = 578.0 [M+H] + .

[0550] Synthesis of compound 45-a

[0551] Compound 45-b (36 mg, 0.062 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (75 mg, 0.19 mmol), cesium carbonate (122 mg, 0.37 mmol), dichloro[bis(diphenylphosphine)ether]palladium(II) (22 mg, 0.031 mmol), and anhydrous toluene (8 mL) were added to a microwave-safe tube at room temperature. The mixture was purged with nitrogen three times, and then stirred at 100 °C for 16 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by rapid separation column chromatography (MeOH / DCM = 0–10%) to give compound 45-a (49 mg). LC-MS (ESI): m / z = 790.3 [M+H] + .

[0552] Synthesis of Compound 45

[0553] Compound 45-a (49 mg, 0.062 mmol) was dissolved in 3 mL of dichloromethane at room temperature. Trifluoroacetic acid (3 mL) was added under ice-water bath conditions, and the reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by Prep-HPLC (alkaline method, ammonium bicarbonate system). The residue was then lyophilized to give compound 45 (3.7 mg, 8.6%). LC-MS (ESI): m / z = 690.2 [M+H] + .

[0554] Synthesis of Compound 47

[0555] Following the synthetic route of compound 40, compound 47 was synthesized using 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidin-4(1H)-one as the starting material, and pyrazolidine dihydrochloride was used instead of hexahydropyridazine dihydrochloride. LC-MS (ESI): m / z = 612.2 (M+H) + .

[0556] Synthetic route of compound 48

[0557] Synthesis of compound 47-a

[0558] Following the synthetic route of compound 40-a, compound 47-a was synthesized using 5,7-dichloro-8-fluoro-2-(methylthio)pyrido[4,3-d]pyrimidine-4(1H)-one as the starting reactant and replacing hexahydropyridazine dihydrochloride with pyrazolidine dihydrochloride.

[0559] Synthesis of compound 48-a

[0560] At room temperature, 47-a (34 mg, 0.08 mmol), tetrahydrofuran (1 mL), potassium phosphate (48 mg, 0.23 mmol), water (0.2 mL), cataCXium A Pd G3 (11 mg, 0.02 mmol), and ((6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5-((triisopropylsilyl)ethynyl)naphthyl-2-amine) (53 mg, 0.11 mmol) were added to a reaction flask. The mixture was purged with nitrogen three times and stirred in an oil bath preheated to 65 °C for 12 hours. The reaction solution was concentrated and purified by column chromatography (mobile phase: methanol / dichloromethane = 0 / 100; 1 / 10) to give compound 48-a (32 mg, 56%). LC-MS (ESI): m / z = 758.4 (M+H) + .

[0561] Synthesis of Compound 48

[0562] 48-a (32 mg, 0.04 mmol), DMF (2 mL), and cesium fluoride (500 mg, 3.29 mmol) were added to a reaction flask at room temperature, and the mixture was stirred at room temperature for 3 hours. The reaction solution was filtered, and the mixture was purified by preparative HPLC to give compound 48 (8.6 mg, 34%). LC-MS (ESI): m / z = 602.2 (M+H) + .

[0563] Synthetic route of compound 49

[0564] Synthesis of compound 49-b

[0565] At room temperature, 1-ethoxycarbonylcyclobutane-1-carboxylic acid (5000 mg, 29.04 mmol), (R)-3-fluoropyrrolidine hydrochloride (3647 mg, 29.04 mmol), DCM (80 mL), EDCI (6680 mg, 34.85 mmol), HOBt (4709 mg, 34.85 mmol), and DIPEA (11260 mg, 87.12 mmol) were added sequentially to a reaction flask. The reaction mixture was purged with nitrogen and reacted overnight at room temperature. Ice water was added to the reaction mixture, and the mixture was extracted with DCM. The organic phases were combined, washed with water and brine, dried, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 49-b (5000 mg, 71%).

[0566] Synthesis of compound 49-a

[0567] 49-b (5000 mg, 20.55 mmol) and THF (60 mL) were added sequentially to a reaction flask at room temperature. Under nitrogen purging protection, a lithium aluminum hydride THF solution (1 M, 35 mL) was added dropwise under dry ice and acetone bath. After the addition was complete, the reaction proceeded at room temperature for 2 hours. A 10% sodium hydroxide aqueous solution (8 mL) was added to the reaction mixture, and the mixture was filtered through diatomaceous earth. The filtrate was concentrated and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 49-a (2900 mg, 75%). LC-MS (ESI): m / z 188.2 (M+H) + .

[0568] Synthesis of Compound 49

[0569] Following the synthetic route of compound 38, starting with 38-c, compound 49 was synthesized by replacing ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol with 49-a. LC-MS (ESI): m / z 706.7 (M+H)+ .

[0570] Synthesis of Compound 50

[0571] Following the synthetic route of compound 37, starting with 7-c, and replacing (2-(difluoromethylene)tetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol with 49-a, compound 50 was synthesized.

[0572] Synthesis of compounds 50-1 and 50-2

[0573] Compound 50 was resolved by chirality to give compound 50-1 (24.8 mg), LC-MS (ESI): m / z = 692.7 (M+H). + Compound 50-2 (20.8 mg), LC-MS (ESI): m / z = 692.7 (M+H) + .

[0574] Synthesis of Compound 51

[0575] Following the synthetic route of compound 43, starting with 5-g, compound 51 was synthesized by replacing pyrazolidine dihydrochloride with hexahydropyridazine dihydrochloride. LC-MS (ESI): m / z = 640.2 (M+H) + .

[0576] Synthetic route of compound 52

[0577] Synthesis of compound 52-b

[0578] At room temperature, 7-c (60 mg, 0.14 mmol), (1S)-1-[(2S)-1-methylpyrrolidone-2-yl]ethanol (54 mg, 0.42 mmol), acetonitrile (10 mL), and DIPEA (90 mg, 0.70 mmol) were added sequentially to a reaction flask. Under nitrogen purging protection, the mixture was heated to 90 °C and reacted overnight. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 52-b (50 mg, 69%). LC-MS (ESI): m / z 622.2 (M+H) + .

[0579] Synthesis of compound 52-a

[0580] At room temperature, 52-b (50 mg, 0.10 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoboron-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (77 mg, 0.19 mmol), bis(diphenylphosphine ether)palladium dichloride (14 mg, 0.02 mmol), cesium carbonate (156 mg, 0.48 mmol), and toluene (3 mL) were added sequentially to a microwave-safe tube. The mixture was purged with nitrogen and heated to 100 °C for 3 hours. The reaction solution was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 52-a (60 mg, 85%). LC-MS (ESI): m / z 834.8 (M+H) + .

[0581] Synthesis of Compound 52

[0582] 52-a (60 mg, 0.08 mmol), DCM (6 mL), and trifluoroacetic acid (3 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to obtain compound 52 (13 mg, 25%). LC-MS (ESI): m / z 634.8 (M+H) + .

[0583] Synthesis of Compound 53

[0584] Following the synthetic route of compound 8, using tert-butyl 1,2-diazazo-1-carboxylate as the starting reactant and substituting compound 29-g for 8-g, compound 53 was synthesized. LC-MS (ESI): m / z 690.6 (M+H) + .

[0585] Synthetic route of compound 54

[0586] Synthesis of compound 54-d

[0587] 7-Bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline (1627 mg, 3.86 mmol), 1,4,5-oxadiazacycloheptan-4-carboxylic acid tert-butyl ester (600 mg, 2.97 mmol), DCM (30 mL), and DIPEA (1150 mg, 8.90 mmol) were added sequentially to a reaction flask at room temperature under nitrogen purging protection. The reaction mixture was extracted with water and ethyl acetate. The organic phases were combined, washed with water and brine, dried, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 1 / 1) to give compound 54-d (1100 mg, 63%). LC-MS (ESI): m / z 586.9 (M+H) + .

[0588] Synthesis of compound 54-c

[0589] At room temperature, 54-d (1100 mg, 1.87 mmol), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (596 mg, 3.74 mmol), THF (20 mL), DMF (20 mL), triethylenediamine (840 mg, 7.49 mmol), and cesium carbonate (1830 mg, 5.62 mmol) were added sequentially to a reaction flask. The mixture was protected by N2 displacement and reacted overnight at room temperature. The reaction solution was quenched with water (100 mL) and extracted with ethyl acetate (100 mL * 2). The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 54-c (1000 mg, 75%). LC-MS(ESI): m / z = 710.1(M+H) + .

[0590] Synthesis of compound 54-b

[0591] At room temperature, 54-c (710 mg, 1.00 mmol), pinacol vinylborate (616 mg, 4.00 mmol), Pd(dppf)Cl2.CH2Cl2 (163 mg, 0.20 mmol), potassium carbonate (553 mg, 4.00 mmol), dioxane (10 mL), and water (2 mL) were added sequentially to a microwave-safe tube. The mixture was protected by N2 displacement and heated to 60 °C overnight. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: PE / EA 10 / 0 to 0 / 10) to give compound 54-b (530 mg, 87%). LC-MS (ESI): m / z 610.2 (M+H) + .

[0592] Synthesis of compound 54-a

[0593] At room temperature, 54-b (220 mg, 0.36 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxoborhexane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl (291 mg, 0.72 mmol), 1,4-dioxane (4 mL), bis(diphenylphosphine ether)palladium dichloride (52 mg, 0.07 mmol), and cesium carbonate (470 mg, 1.44 mmol) were added sequentially to a microwave-safe tube. The mixture was purged with N2 and heated to 100 °C overnight. The reaction mixture was concentrated, stirred into silica gel, and purified by column chromatography (mobile phase: DCM / MeOH 10 / 0 to 10 / 1) to give compound 54-a (120 mg, 41%). LC-MS (ESI): m / z 822.7 (M+H) + .

[0594] Synthesis of Compound 54

[0595] 54-a (70 mg, 0.09 mmol), DCM (6 mL), and trifluoroacetic acid (3 mL) were added to a reaction flask at room temperature. The mixture was reacted at room temperature for 3 hours. The reaction solution was concentrated at room temperature and purified by preparative HPLC to give compound 54 (15 mg, 28%). LC-MS (ESI): m / z 622.8 (M+H) + ; 1 HNMR(DMSO_d6,400MHz)δ9.60(1H,s),8.03(2H,s),7.31-7.03(2H,m),6.34-6.15(1H,m),6. 12-5.95(1H,m),5.72-5.03(3H,m),4.40-3.64(8H,m),3.25-2.70(6H,m),2.23-1.63(6H,m).

[0596] Synthesis of Compound 55

[0597] Following the synthetic routes of compounds 13 and 14, compound 55 was synthesized using 7-bromo-2,4-dichloro-8-fluoro-6-iodoquinazoline as the starting material and replacing tert-butyl 1,2-diazacycloheptan-1-carboxylate with [1,4,5]oxodiazazoline. LC-MS (ESI): m / z = 610.2 [M+H] + .

[0598] Synthesis of compounds 55-1 and 55-2

[0599] Compound 55 (110 mg) was chirally resolved to give compound 55-1 (41 mg) and compound 55-2 (28 mg).

[0600] Compound 55-1: LC-MS (ESI): m / z = 610.0 [M+H] + ee% = 92.82%; 1 HNMR(CDCl3, 400MHz) δ8.96(1H,s),7.13-7.06(1H,m),6.95(1H,t,J=8.8Hz),6.00(2H,brs),5.29(1H,d,J=52.4Hz),4.39-4.13(4H,m) ,4.04(2H,t,J=4.8Hz),3.84(2H,t,J=4.8Hz),3.50-3.15(5H,m),3.11-2.96(1H,m),2.42-2.12(3H,m),2.08(3H,s),2.04-1.80(4H,m).

[0601] Compound 55-2: LC-MS (ESI): m / z = 610.0 [M+H] + ee% = 97.94%; 1 HNMR (CDCl3, 400MHz) δ8.99 (1H, s), 7.13-7.06 (1H, m), 6.95 (1H, t, J = 8.8Hz), 6.01 (2H, brs), 5.37 (1H, d, J = 53.6Hz), 4.50-4.16 (4H ,m),4.11-3.99(2H,m),3.91-3.78(2H,m),3.62-3.15(5H,m),3.15-3.01(1H,m),2.54-2.22(3H,m),2.05(3H,s),2.03-1.79(4H,m).

[0602] The conditions for chiral analysis and chiral preparation are as follows:

[0603] Compare the synthetic route of compound 1′

[0604] Synthesis of compound 1′-a

[0605] 18-b (100 mg, 0.16 mmol), (3-cyano-4-(5,5-dimethyl-1,3,2-dioxaborane-2-yl)-7-fluorobenzo[b]thiophene-2-yl)carbamate tert-butyl ester (197 mg, 0.49 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (12 mg, 0.016 mmol), cesium carbonate (264 mg, 0.81 mmol), and toluene (15 mL) were added to a microwave tube. The tube was purged three times with nitrogen, sealed, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: dichloromethane:methanol = 10:1 / dichloromethane, 0–60%) to give compound 1′-a (153 mg, 96%). LC-MS (ESI): m / z 828.5 (M+H) + .

[0606] Synthesis of compound 1′

[0607] Trifluoroacetic acid (2 mL) was added to a solution of 1′-a (50 mg, 0.060 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give compound 1′ (9 mg, 24%). LC-MS (ESI): m / z 628.5 (M+H) + ; 1 H NMR(400M,CD3OD)δ9.56(1H,d,J=1.6Hz),7.21-7.15(1H,m),7.02(1H,t,J=9.2Hz),5.40-5.21(1H,m),4.60(3H,bs) ,4.32-4.06(4H,m),3.39-2.96(6H,m),2.42-2.08(3H,m),2.05-1.94(4H,m),1.94-1.84(1H,m),1.82-1.67(4H,m).

[0608] Compare the synthesis of compounds 1′-1 and 1′-2

[0609] A new batch of control compound 1′ was synthesized. Control compound 1′ (130 mg) was chirally resolved to give control compound 1′-1 (45 mg), LC-MS (ESI): m / z 627.9 (M+H). + Comparative compound 1′-2 (41 mg), LC-MS (ESI): m / z 628.6 (M+H) + .

[0610] The conditions for chiral analysis and chiral preparation are as follows:

[0611] Compare the synthetic route of compound 2′

[0612] Synthesis of compound 2′-c

[0613] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-(trifluoromethyl)quinazoline (1 g, 2.75 mmol) and N,N-diisopropylethylamine (1065 mg, 8.24 mmol) in tetrahydrofuran (50 mL), tert-butyl 1,2-diazaphen-1-carboxylic acid (550 mg, 2.75 mmol) was added. The mixture was allowed to warm to room temperature and stirred overnight. The next day, water was added, and the mixture was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with saturated brine (10 mL x 2), dried over anhydrous sodium sulfate, filtered, evaporated to dryness, and purified by column chromatography (mobile phase: ethyl acetate: petroleum ether = 0–10%) to give compound 2′-c (1270 mg, 88%). LC-MS (ESI): m / z 527.1 (M+H) + .

[0614] Synthesis of compound 2′-b

[0615] To a mixed solution of 2′-c (1200 mg, 2.27 mmol) in tetrahydrofuran (25 mL) and N,N-dimethylformamide (25 mL), ((2R,7aS)-2-fluorohexahydro-1H-pyrrolazin-7a-yl)methanol (1086 mg, 6.82 mmol), cesium carbonate (2223 mg, 6.82 mmol), and DABCO (1020 mg, 9.10 mmol) were added, and the mixture was reacted overnight at room temperature. Water was added to the reaction solution, and the mixture was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 2′-b (649 mg, 44%). LC-MS (ESI): m / z 650.3 (M+H) + .

[0616] Synthesis of compound 2′-a

[0617] 2′-b (60 mg, 0.092 mmol), (7-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)benzo[d]thiazolyl)carbamate tert-butyl (57 mg, 0.18 mmol), bis(diphenylphosphine ether)palladium(II) dichloride (7 mg, 0.009 mmol), cesium carbonate (150 mg, 0.46 mmol), and toluene (10 mL) were added to a microwave-safe tube. The tube was sealed, purged three times with nitrogen, and heated to 100 °C for 1.5 hours. The reaction mixture was evaporated to dryness, and the crude product was purified by column chromatography (mobile phase: methanol: dichloromethane = 0–10%) to give compound 2′-a (66 mg, 86%). LC-MS (ESI): m / z 838.7 (M+H) + .

[0618] Synthesis of compound 2′

[0619] Trifluoroacetic acid (2 mL) was added to a solution of 2′-a (66 mg, 0.079 mmol) in dichloromethane (6 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. The reaction solution was evaporated to dryness, purified by HPLC (ammonium bicarbonate), and lyophilized to give the control compound 2′ (21 mg, 42%). LC-MS (ESI): m / z 662.2 (M+H) + ; 1 H NMR (400M, DMSO-d6): δ9.82(1H,s),7.89(2H,s),7.27-7.16(1H,m),7.03(1H,t,J=8.8Hz),5.90-5.77(1H ,m),5.27(1H,d,J=54.0Hz),4.21-3.92(4H,m),3.16-2.93(5H,m),2.89-2.76(1H,m),2.20-1.55(12H,m).

[0620] Compare the synthesis of compounds 2′-1 and 2′-2

[0621] A new batch of comparative compound 2′ was synthesized. Comparative compound 2′ (80 mg) was prepared by chirality, first by rotary evaporation and then by freeze-drying to obtain comparative compound 2′-1 (31 mg) and comparative compound 2′-2 (28 mg).

[0622] Comparative compound 2′-1: LC-MS (ESI): m / z 662.4 (M+H) + ; 1H NMR(400M,DMSO-d6)δ9.83(1H,s),8.06(2H,s),7.29-7.19(1H,m),7.16-7.07(1H,m),5.91-5.82(1H,m ),5.27(1H,d,J=53.2Hz),4.20-3.88(3H,m),3.17-2.92(6H,m),2.88-2.77(1H,m),2.17-1.56(12H,m).

[0623] Comparative compound 2′-2: LC-MS (ESI): m / z 662.7 (M+H) + ; 1 H NMR(400M,DMSO-d6)δ9.84(1H,s),8.07(2H,s),7.28-7.20(1H,m),7.17-7.08(1H,m),5.92-5.84(1H,m ),5.28(1H,d,J=53.2Hz),4.20-3.89(3H,m),3.18-2.93(6H,m),2.88-2.78(1H,m),2.18-1.56(12H,m).

[0624] Example 1: CTG assay to detect the inhibitory effect of the compound on the proliferation of RAS cell lines

[0625] NCI-H358 represents non-small cell lung cancer cells with a KRAS G12C mutation; LS513 represents colon cancer cells with a KRAS G12D mutation; NCI-H441 represents lung cancer cells with a KRAS G12V mutation; GP2D represents colon cancer cells with a KRAS G12D mutation; SW403 represents colon cancer cells with a KRAS G12V mutation; SW480 represents colon cancer cells with a KRAS G12V mutation; and MKN1 represents wild-type gastric cancer cells with KRAS amplification. The inhibitory effects of the compounds on the proliferation of these cell lines were evaluated by detecting their proliferative inhibitory activity.

[0626] The experiments were conducted on 384-well or 96-well plates. The specific procedure is as follows:

[0627] Add the cell suspension to 384 or 96-well plates (excluding the outer wells) (384-well plate: 50 μL; 96-well plate: 100 μL). Incubate the plates overnight in a CO2 incubator. Incubate NCI-H358, GP2D, LS513, MKN1, and NCI-H441 overnight in a 5% CO2 incubator at 37°C, and incubate SW403 and SW480 overnight in a CO2-free incubator at 37°C. Using an HPD300 micro-dosing instrument, add the corresponding concentration of the compound to each well (obtaining 9-10 concentration gradients through 3-fold dilution). Incubate the cell plates in a CO2 incubator for 6 days. Incubate NCI-H358, GP2D, LS513, MKN1, and NCI-H441 in a 5% CO2 incubator at 37°C, and incubate SW403 and SW480 in a CO2-free incubator. On the day of testing, first equilibrate the 384 or 96-well plate to room temperature for 10-30 minutes. Then add CellTiter Glo reagent to the 384 or 96-well plate (25 μL for 384-well plate; 100 μL for 96-well plate), shake in the dark for 10 minutes, and incubate for 10 minutes. Place the culture plate in an ENVIDION or ENVIDION reader, plot the drug efficacy inhibition rate curve using XLFit, and calculate the IC50. 50 Values. The activity results for representative compounds are shown in the table below. Among them, "IC50" is... 50 ">1μM" is represented by "**", and "1μM≥IC" is represented by "**". 50 ">100nM" is represented by "***", "IC" 50 ≤100nM is represented by “****”.

[0628] The results showed that compounds 44 and 44-1 of the present invention had significantly better inhibitory effects on the RAS-related activity of SW480, SW403, H358, and LS513 cells than comparative compounds 1′ and 1′-1. In LS513 cells, compound 44 of the present invention had better activity than comparative compound 2′, while in SW403 and H358 cells, compound 44-1 of the present invention had significantly better activity than comparative compound 2′-2.

[0629] Example 2: Pharmacokinetic Study of Rats After Intravenous and Oral Administration

[0630] Experimental procedure:

[0631] Following intravenous and oral administration, blood samples were collected from male SD rats at nine time points (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h). The collected blood samples were transferred to microcentrifuge tubes containing EDTA-K2 anticoagulant, centrifuged at 4°C and 4000 g for 5 min, and the supernatant was collected and stored at -75°C ± 15°C. The plasma compound concentrations at different time points were detected by LC-MS / MS, and relevant pharmacokinetic parameters were calculated using WinNonlin software. Note: The intravenous injection solvent is 5% DMSO + 10% Solutol HS15 + 85% (20% HP-β-CD in water); the oral administration solvent is 0.1% Tween 80 + 0.5% MC in water. Note: The solvent for intravenous injection is 5% DMSO + 10% Solutol HS15 + 85% (20% HP-β-CD in water); the solvent for oral administration is 0.5% Tween 80 + 0.5% MC in water.

[0632] Example 3: Pharmacokinetic Study of Mice After Intravenous and Oral Administration

[0633] Experimental procedure:

[0634] Following intravenous and oral administration, blood samples were collected from male CD-1 mice at nine time points (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h). The collected blood samples were transferred to microcentrifuge tubes containing EDTA-K2 anticoagulant, centrifuged at 4000 g for 5 min at 4 °C, and the supernatant was collected and stored at -75 °C ± 15 °C. The concentrations of compounds in plasma at different time points were determined by LC-MS / MS, and relevant pharmacokinetic parameters were calculated using WinNonlin software. Note: The solvent for intravenous injection is 5% DMSO + 10% Solutol HS15 + 85% (20% HP-β-CD in water); the solvent for oral administration is 0.5% Tween 80 + 0.5% MC in water.

[0635] The above results indicate that, compared to comparative compound 2′-2, compounds 43, 44, and 44-1 of the present invention have lower plasma clearance (Cl) and higher plasma exposure (AUC) after intravenous administration, and higher peak plasma concentration (C) after oral administration. max The compound exhibits higher levels of both plasma exposure and AUC, demonstrating that it possesses excellent pharmacokinetic properties.

[0636] Example 4: Study on CYP enzyme inhibitory activity

[0637] The effects of compound 44-1 on five cytochrome P450 enzyme (CYP450) isoforms (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) were evaluated. The test concentration range of the compound group was 0.001 μM to 10.00 μM, with a total of 7 gradient concentrations. A reference inhibitor group and a solvent group were also set up as control experiments.

[0638] Experimental materials:

[0639] Human liver microsomes, purchased from Corning.

[0640] Experimental steps:

[0641] 1. Prepare a 0.1M potassium phosphate buffer (pH 7.4, abbreviated as K-buffer).

[0642] 2. Prepare a series of dilutions (400-fold concentration) for the test compound and the reference inhibitor in a 96-well plate;

[0643] 2.1. For the test compound, take 8 μL of the 10 mM test compound (dissolved in dimethyl sulfoxide) and transfer it to 12 μL of acetonitrile;

[0644] 2.2. For the reference inhibitor solutions of CYP1A2, CYP2C9 and CYP2D6: 12 μL of 1 mM α-naphthoflavone + 10 μL of 40 mM sulfaphenazole + 10 μL of 10 mM quinidine + 8 μL of dimethyl sulfoxide;

[0645] 2.3. For the CYP2C19 reference inhibitor solution: 8 μL of 250 mM omeprazole, add 12 μL of acetonitrile; for the CYP3A4 reference inhibitor solution: 8 μL of 6.25 mM ketoconazole, add 12 μL of acetonitrile.

[0646] 2.4. Dilute each solution from steps 2.1, 2.2, and 2.3 serially with a 1:3 mixture of dimethyl sulfoxide and acetonitrile (v / v: 40:60).

[0647] 3. Prepare an 8 mM NADPH solution: Add 66.7 mg of NADPH to 10 mL of 0.1 M K-buffer.

[0648] 4. Prepare a 0.2 mg / mL human liver microsome solution on ice: Take 10 μL of 20 mg / mL human liver microsomes and add 990 μL of K-buffer.

[0649] 4. Prepare substrates at 4x concentration (2 mL for each subtype), as shown in the table below (if human liver microsomes are to be added, perform the operation on ice);

[0650] 6. Perform on ice: Add 400 μL of human liver microsome solution to the test well, followed by 2 μL of serially diluted test compound solution (from step 2) to the corresponding test well.

[0651] 7. Perform on ice: Add 200 μL of human liver microsomal solution to the test well, followed by 1 μL of serially diluted reference inhibitor solution (from step 2) to the corresponding test well.

[0652] 8. Perform on ice: Add the following solutions to each of the 96-well plates (repeat wells only):

[0653] 8.1. 30 μL of the test compound at twice the concentration, and the reference inhibitor (dissolved in 0.2 mg / mL human liver microsome solution, from steps 6 and 7);

[0654] 8.2. 15 μL of substrate solution at a concentration of 4 (from step 4);

[0655] 9. Pre-incubate the 96-well plate with NADPH solution at 37°C for 10 minutes;

[0656] 10. Add 15 μL of preheated 8 mM NADPH solution (from step 3) to the 96-well plate to start the reaction;

[0657] 11. Incubate the 96-well plate at 37°C: 5 minutes for CYP3A4, 10 minutes for CYP1A2, CYP2C9 and CYP2D6, and 45 minutes for CYP2C19.

[0658] 12. After incubation, at the specified time point, add 120 μL of acetonitrile containing an internal standard (for CYP2C19, the internal standard is 0.1 μM MOH-Mephenytoin-D3; for CYP1A2, the internal standard is 0.1 μM Acetaminophen-D4; for CYP3A4, the internal standard is Osalmid; for CYP2C9, the internal standard is 4'-OH-diclofenac-[ 13C6]; For CYP2D6, the internal standard is 1-OH-bufuralol-maleate-[D9]) to terminate the reaction;

[0659] 13. After terminating the reaction, shake the 96-well plate at 600 rpm for 10 min, then centrifuge at 3220 g for 15 min to precipitate the protein. Take 50 μL of the supernatant into the 96-well plate, dilute with 50 μL of ultrapure water, and then perform LC-MS / MS analysis.

[0660] Experimental results:

[0661] IC50 of CYP enzyme inhibition assay 50 The results are shown in the table below:

[0662] The above IC 50 The results showed that compound 44-1 of the present invention did not exhibit strong inhibition against any of the CYP enzyme subtypes, and the risk of drug interaction was low.

[0663] While specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A hydrazine compound as shown in Formula I, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of a stereoisomer thereof, a prodrug thereof, a deuterated compound thereof, or a PROTAC molecule thereof: Formula I. wherein, R 1 For X 1a is N or CR b1 , R b1 is CN; X 1b and X 1c are independently H, F or CH3; X 2a is N or CH; X 2b is H or F; X 2c is H or F; X 2d is H, F, X 3a and X 3b Independently N or CR b2 R b2 For H, CH3, or F; X 3c For H, F, Cl, CH3, CF3, CN, X 3d CF3, I or X 4a CH3, Cl, CF3, -OCF2Cl or -OCHF2; X 4b is H, F, Cl or CH3; X 4c is H or CN; is a single or double bond; X 5a is CH3, X 6a for X 6b is H or F; X 7a is H or F; X 8a F, X 8b is H or F; Y is O, S, NH, CH2or m1, m2 and n are independently 0, 1, 2, 3 or 4; R 2 is hydrogen, C 1-6 alkyl, C 1-6 alkyl substituted by one or more deuterium, C 1-6 alkyl or -C(=O)R 21 ; R 21 is hydrogen, C 1-6 alkyl, C 1-6 alkyl or C 1-6 alkyl substituted by one or more deuterium; R 4 Independently deuterium, halogen, cyano, hydroxyl, C 1-6 Alkyl, with one or more R 4-1 Replacement C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 cycloalkyl, C 1-6 Alkyl-O-, with one or more R 4-2 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 4-3 Replacement C 1-6 Alkyl-S-, O=, C 3-10 Cycloalkyl-O-,-C(=O)R 4a -NR 4b1 R 4b2 -C(=O)OR 4c -C(=O)NR 4d1 R 4d2 or Alternatively, when n is 2, 3, or 4, any two R values ​​can be chosen. 4 Connected, independently forming 3- to 8-membered carbon rings with the atoms they are connected to, and bounded by one or more R... 4-4 Substituted 3- to 8-membered carbon rings, "4- to 8-membered heterocycles containing 1 to 3 heteroatoms independently selected from O, S, and N", or substituted by one or more R 4-5 The substituted "4- to 8-membered heterocycles containing 1 to 3 heteroatoms, which are independently selected from O, S, and N"; R 5 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 51 Replacement C 1-6 Alkyl, C 3-10 Cycloalkyl, hydroxyl, C 1-6 Alkyl-O-, with one or more R 52 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 53 Replacement C 1-6 Alkyl-S-, amino, C 1-6 Alkyl-NH-, with one or more R 54 Replacement C 1-6 Alkyl-NH-, C 2-6 alkenyl, C 2-6 alkynyl or C 3-10 cycloalkyl-O-; R 6 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 61 Replacement C 1-6 Alkyl, C 3-10 Cycloalkyl, hydroxyl, C 1-6 Alkyl-O-, with one or more R 62 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 63 Replacement C 1-6 Alkyl-S-, C 2-6 alkenyl, C 2-6 alkynyl, amino, C 1-6 Alkyl-NH-, with one or more R 64 Replacement C 1-6 Alkyl-NH- or C 3-10 cycloalkyl-O-; T is N or CR 7 ;R 7 It is hydrogen, deuterium, halogen, cyano, C 1-6 Alkyl, with one or more R 71 Replacement C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 Cycloalkyl, nitro, hydroxyl, C 1-6 Alkyl-O-, with one or more R 72 Replacement C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, with one or more R 73 Replacement C 1-6 Alkyl-S-, C 1-6 Alkyl-NH-, with one or more R 74 Replacement C 1-6 Alkyl-NH- or C 3-10 cycloalkyl-O-; R 51 R 52 R 53 R 54 R 61 R 62 R 63 R 64 R 71 R 72 R 73 and R 74 Independently deuterium, halogen, cyano, C 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C 2-6 Alkenyl, hydroxyl, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, -C(=O)R 51a -NR 51b1 R 51b2 -C(=O)OR 51c or -C(=O)NR 51d1 R 51d2 ;R 51a R 51b1 R 51b2 R 51c R 51d1 and R 51d2 Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl; Or, any R 2 and R 6 Optional R 4 and R 6 Each of these compounds forms a 5-12 membered heterocyclic alkenyl group with the atoms to which it is attached, consisting of 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from O, S, and N, or is bounded by one or more R atoms. a The substituted "contains 1 to 4 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 5 to 12-membered heterocyclic alkenyl groups of O, S and N"; R 4-1 R 4-2 R 4-3 R 4-4 R 4-5 and R a Independent of halogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups, C substituted with one or more deuterium groups 1-6 Alkyl, C 2-6 alkenyl, cyano, hydroxyl, O=, C 1-6 Alkyl-O-, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, -NR 4f1 R 4f2 -C(=O)OR 4g or -C(=O)NR 4h1 R 4h2 ; R 4a R 4b1 R 4b2 R 4c R 4d1 R 4d2 R 4e1 R 4e2 R 4f1 R 4f2 R 4g R 4h1 and R 4h2 Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium groups 1-6 alkyl; L is C 1-6 Alkylene, -OR L-1 -、-SR L-2 -or-NR L-3 (R L-4 )-;R L-1 R L-2 and R L-4 Independently for C 1-6 Alkylene or C substituted with one or more deuterium atoms 1-6 Alkylene; R L-3 For hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium atoms 1-6 alkyl; R 3 C 3-12 cycloalkyl, with one or more R 3-1 Replacement C 3-12 Cycloalkyl, "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S, and N, of a 4 to 12-membered heterocycloalkyl group", and being bound by one or more R 3-2 Substituted "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S, and N in 4 to 12-membered heterocyclic alkyl groups", and being substituted by one or more R 3-3 Replacement C 2-6 alkenyl or -NR 3-41 R 3-42 When there are multiple substituents, they may be the same or different. R 3-41 and R 3-42 Independently hydrogen, C 1-6 Alkyl, C 3-10 cycloalkyl, C 1-6 Alkyl-O- or "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S and N in 4 to 12-membered heterocyclic alkyl groups"; R 3-1 R 3-2 and R 3-3 Independently for C 1-6 Alkyl, with one or more R 3-1-1 Replacement C 1-6 Alkyl, C 1-6 Alkyl-O-, halogen, aryl, with one or more R 3-1-8 Substituted aryl group, "containing 1 to 4 heteroatoms, the heteroatoms being independently selected from O, S, and N, 5 to 12 membered heteroaryl groups", and substituted with one or more R groups. 3-1-9 Substituted "containing 1 to 4 heteroatoms, the heteroatoms being independently selected from O, S, and N 5-12 membered heteroaryl groups" or Or, any two Rs 3-1 Two Rs can be selected at random 3-2 Connected, independently forming 3- to 8-membered carbon rings with the atoms they are connected to, and bounded by one or more R... 3-1-2 Substituted 3- to 8-membered carbon rings, "4- to 8-membered heterocycles containing 1 to 3 heteroatoms independently selected from O, S, and N", or substituted by one or more R 3-1-3 Substituted "4-8 membered heterocycles containing 1-3 heteroatoms, the heteroatoms being independently selected from O, S, and N", C 6-20 aryl, with one or more R 3-1-4 Replacement C 6-20 Aryl, "a 5-12 membered heteroaryl group containing 1-4 heteroatoms independently selected from O, S, and N", and bonded by one or more R 3-1-5 Substituted "5-12-membered heteroaryl groups containing 1-4 heteroatoms, the heteroatoms being independently selected from O, S, and N", C 5-7 Cycloalkenyl, with one or more R 3-1-6 Replacement C 5-7 Cycloalkenyl, "5-7 membered heterocyclic alkenyl groups containing 1-3 heteroatoms independently selected from O, S, and N", or those with one or more R 3-1-7 The substituents are "5-7 membered heterocyclic alkenyl groups containing 1-3 heteroatoms, which are independently selected from O, S, and N"; when there are multiple substituents, they may be the same or different; R 3-1-1 Halogen, -NR 3b1 R 3b2 -OC(=O)NR 3c1 R 3c2 -OR 3d "Contains 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N" or is composed of one or more R 3-1-1-1 The substituted "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N"; R 3d C 6-20 Aryl, "containing 1 to 4 heteroatoms, the heteroatoms being independently selected from O, S, and N, a 5 to 12-membered heteroaryl", and bonded by one or more R 3d1 Replacement C 6-20 aryl, or, by one or more R 3d2 Substituted "containing 1 to 4 heteroatoms, the heteroatoms being independently selected from 5 to 12-membered heteroaryl groups of O, S, and N"; R 3-1-1-1 for halogen or C 1-6 alkyl; R 3a1 , R 3a2 , R 3d1 , and R 3d2 are independently hydrogen, halogen, cyano, C 1-6 alkyl or C 3a11 alkyl substituted by one or more R 1-6 ; R 3a11 is halogen or C 1-6 alkyl-O-; R 3-1-2 R 3-1-3 R 3-1-4 R 3-1-5 R 3-1-6 R 3-1-7 R 3-1-8 and R 3-1-9 Independently for C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups, C substituted with one or more deuterium groups 1-6 Alkyl, halogen, cyano, C 2-6 alkenyl, C 2-6 alkynyl, hydroxyl, -C(=O)R 31a -NR 31b1 R 31b2 -C(=O)OR 31c -C(=O)NR 31d1 R 31d2 -S(O)2NR 31e1 R 31e2 -S(O)2R 31f -OC 1-6 Alkyl groups, -OC groups substituted with one or more halogens 1-6 Alkyl groups, -OC groups substituted with one or more deuterium atoms 1-6 Alkyl, -SC 1-6 Alkyl groups, -SC groups substituted with one or more halogens 1-6 Alkyl groups or -SC substituted with one or more deuterium groups 1-6 Alkyl group; when there are multiple substituents, they may be the same or different; R 3b1 R 3b2 R 3c1 R 3c2 R 31a R 31b1 R 31b2 R 31c R 31d1 R 31d2 R 31e1 R 31e2 and R 31f Independently hydrogen, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups or C groups substituted with one or more deuterium groups 1-6 Alkyl group; when there are multiple substituents, they may be the same or different; Or, R 4b1 and R 4b2 R 4d1 and R 4d2 R 4e1 and R 4e2 R 51b1 and R 51b2 R 51d1 and R 51d2 R 4f1 and R 4f2 R 4h1 and R 4h2 R 3a1 and R 3a2 R 3b1 and R 3b2 R 3c1 and R 3c2 R 31b1 and R 31b2 R 31d1 and R 31d2 R 31e1 and R 31e2 Each of these atoms forms a 4-12 membered heterocyclic alkyl group containing 1 to 3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from O, S, and N, or is formed by one or more R atoms. b The substituted form "contains 1 to 3 heteroatoms, one of which is N, and the other heteroatoms are independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N"; R b For deuterium, halogens, C 1-6 Alkyl groups, C substituted with one or more halogens 1-6 Alkyl groups, C atoms substituted with one or more deuterium atoms 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-10 Cycloalkyl, nitro, hydroxyl, C 1-6 Alkyl-O-, C substituted with one or more halogens 1-6 Alkyl-O-, C substituted with one or more deuterium atoms 1-6 Alkyl-O-, C 1-6 Alkyl-S-, C-substituted with one or more halogens 1-6 Alkyl-S-, C-substituted with one or more deuteriums 1-6 Alkyl-S-, amino, C 1-6 Alkyl-NH-, C substituted with one or more halogens 1-6 Alkyl-NH- or C-substituted with one or more deuterium groups 1-6 Alkyl-NH-; The hydrazine-based compound of the structure of Formula I is not one of the following compounds:

2. The hydrazine-based compound as shown in formula I, stereoisomer thereof, pharmaceutically acceptable salt thereof, pharmaceutically acceptable salt of stereoisomer thereof, prodrug thereof, deuterated compound thereof or PROTAC molecule thereof according to claim 1, wherein, In the structure as shown in Formula I, R 1 is ​ X 1a is N or CR b1 , R b1 is CN; X 1b and X 1c are independently H or F; X 2a is CH; X 2b is H; X 2c is F; X 2d is is a single or double bond, X 5a is CH3or X 8a for X 8b is F; Y is either O or CH2; m1 and m2 are independently 1; n is 0; R 2 is hydrogen or C 1-6 alkyl; R 5 halogen; R 6 is hydrogen, hydroxyl or C 1-6 alkyl-O-; T is N or CR 7 ;R 7 Halogen, cyano, C 1-6 Alkyl, C 3-10 cycloalkyl or with one or more R 71 Replacement C 1-6 alkyl; R 71 independently halogen or cyano; Or, R 2 and R 6 Each of them forms a 5-12 membered heterocyclic alkenyl group with the atoms to which they are attached, consisting of 2 to 4 heteroatoms, two of which are N and the other heteroatoms are independently selected from O, S and N. L is -OR L-1 -; R L-1 is C 1-6 alkylene; R 3 For one or more R 3-1 Replacement C 3-12 cycloalkyl, with one or more R 3-2 The substituted group is "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S, and N"; when there are multiple substituents, they may be the same or different; R 3-1 For one or more R 3-1-1 Replacement C 1-6 alkyl; R 3-1-1 It is defined as "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from O, S, and N in 4- to 12-membered heterocyclic alkyl groups" or being surrounded by one or more R groups. 3- 1-1-1 The substituted "containing 1 to 3 heteroatoms, the heteroatoms being independently selected from 4 to 12-membered heterocyclic alkyl groups of O, S and N"; R 3-1-1-1 is halogen; R 3-2 independently halogen or R 3a1 and R 3a2 is halogen.

3. The hydrazine compound of Formula I as described in claim 1, its stereoisomers, its pharmaceutically acceptable salts, its pharmaceutically acceptable salts, its prodrugs, its deuterated compounds, or its PROTAC molecules, characterized in that, R 1 for and / or, For and / or, For 4. The hydrazine compound of Formula I as described in claim 1, its stereoisomers, its pharmaceutically acceptable salts, its pharmaceutically acceptable salts, its prodrugs, its deuterated compounds, or its PROTAC molecules, characterized in that, For And / or, R 6 For H, hydroxyl, Me, -OMe, -SMe, Br, F, or -NHMe, or R 6 With R 2 form 5. The hydrazine compound of Formula I as described in claim 1, its stereoisomers, its pharmaceutically acceptable salts, its pharmaceutically acceptable salts, its prodrugs, its deuterated compounds, or its PROTAC molecules, characterized in that, The hydrazine compound of the structure as shown in Formula I is any one of the following compounds:

6. The hydrazine-based compound as shown in formula I, a stereoisomer thereof, a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable salt of a stereoisomer thereof, a prodrug thereof, a deuterated compound thereof or a PROTAC molecule thereof according to claim 1, wherein, The hydrazine compound of the structure as shown in Formula I is any one of the following compounds: ​ 7. The hydrazine compound of Formula I as claimed in claim 1, its stereoisomers, its pharmaceutically acceptable salts, its pharmaceutically acceptable salts, its prodrugs, its deuterated compounds, or its PROTAC molecules, characterized in that, The hydrazine-based compound represented by Formula I is any one of the following compounds:

8. A pharmaceutical composition comprising substance A and a pharmaceutical excipient; wherein substance A is a therapeutically effective amount of a hydrazine compound of formula I as described in any one of claims 1-7, its stereoisomer, its pharmaceutically acceptable salt, its stereoisomer pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule.

9. Use of a substance A for the preparation of a RAS inhibitor, characterized in that, The substance A is a hydrazine compound with a structure of Formula I as described in any one of claims 1-7, its stereoisomer, its pharmaceutically acceptable salt, its stereoisomer pharmaceutically acceptable salt, its prodrug, its deuterated compound, or its PROTAC molecule.

10. The application as described in claim 9, characterized in that, The RAS mentioned above includes wild-type RAS and mutant RAS; the wild-type RAS may be wild-type amplified KRAS.

11. The use of a substance A in the preparation of a medicament for the treatment or prevention of RAS-related diseases; wherein the substance A is a hydrazine compound of formula I as described in any one of claims 1-7, its stereoisomers, pharmaceutically acceptable salts thereof, pharmaceutically acceptable salts of its stereoisomers, its prodrugs, its deuterated compounds, or its PROTAC molecules; wherein the RAS-related diseases include cancer; wherein the cancers include one or more of the following: colon cancer, appendix cancer, pancreatic cancer, MYH-related polyposis, blood cancer, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophageal cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia, and melanoma.

12. The use of substance A in the preparation of a medicament for the treatment or prevention of cancer; wherein substance A is a hydrazine compound of formula I as described in any one of claims 1-7, its stereoisomers, pharmaceutically acceptable salts thereof, pharmaceutically acceptable salts of its stereoisomers, its prodrugs, its deuterated compounds, or its PROTAC molecules; wherein the cancers are, for example, colon cancer, appendix cancer, pancreatic cancer, MYH-related polyposis, hematologic malignancies, breast cancer, endometrial cancer, gallbladder cancer, bile duct cancer, prostate cancer, lung cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head or neck cancer, bone cancer, skin cancer, rectal cancer, liver cancer, esophageal cancer, stomach cancer, thyroid cancer, bladder cancer, lymphoma, leukemia, and melanoma, or one or more of these.

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