Fused tricyclic compound and use thereof

Abstract

Provided are a fused tricyclic compound as shown in formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing same, and the use thereof as a WRN inhibitor in the prevention or treatment of related diseases.

Description

Tricyclic compounds and their applications

[0001] Cross-references to related applications

[0002] This disclosure claims Chinese Patent Application No. 202410486263.1, filed April 22, 2024; Chinese Patent Application No. 202410540628.4, filed April 30, 2024; Chinese Patent Application No. 202410572238.5, filed May 10, 2024; Chinese Patent Application No. 202410702225.5, filed June 1, 2024; and Chinese Patent Application No. 202410754608.7, filed June 12, 2024. The priority and interests of Chinese patent applications No. 202410957616.1 filed on July 17, 2024, Chinese patent application No. 202411297630.X filed on September 18, 2024, Chinese patent application No. 202411493772.3 filed on October 24, 2024, and Chinese patent application No. 202411782492.4 filed on December 5, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This disclosure relates to tricyclic compounds or their stereoisomers or pharmaceutically acceptable salts thereof, methods of their preparation, pharmaceutical compositions containing them, and their use as WRN inhibitors in the prevention or treatment of related diseases. Background Technology

[0004] Helicases are a class of enzymes that untangle double-stranded nucleotides. They utilize the energy released during the hydrolysis of nucleoside triphosphates to move along the nucleic acid chain, thereby untangling and separating the nucleic acid double strands. As representatives of DNA helicases, the RecQ helicase family plays a significant role in maintaining chromosomal genome and telomere stability. During DNA replication, they catalyze the unwinding of double-stranded DNA, completing DNA repair and thus maintaining genome integrity.

[0005] RecQ family helicases are highly conserved throughout evolution, and their functions are involved in a variety of DNA metabolic processes, playing a crucial role in maintaining genome stability. The loss of function of BLM, WRN, and RecQ4, three of the five RecQ family helicases in humans, leads to BLM syndrome, WRN syndrome, and Rothmund-Thomson syndrome, respectively. These diseases all exhibit high genomic instability, chromosomal abnormalities, and susceptibility to DNA damage at the molecular level.

[0006] WRN and RecQ1 are two of the most representative RecQ helicases. WRN includes not only its unwinding domain but also an exonuclease domain. Werner syndrome (WS) is an autosomal recessive genetic disorder characterized by accelerated clinical aging, resulting in a lifespan of less than 50 years. WS helicase nuclease (WRN) is involved in many important pathways, including DNA replication, recombination, and repair. WRN can unwind non-canonical secondary DNA structures encountered during replication and recombination; mutations in WRN can cause chromosomal instability disorders such as Werner syndrome. Under normal circumstances, WRN depletion leads to DNA double-strand breaks in MSI cells, resulting in cell cycle arrest and / or apoptosis. Studies have shown a strong synthetic lethal relationship between WRN helicase and microsatellite instability-high (MSI-H) cancers. WRN forms a synthetic lethal relationship with the DNA mismatch repair protein MutL homolog 1 (MLH1), and MLH1 loss is associated with microsatellite instability (MSI).

[0007] Microsatellites are simple repetitive sequences of less than 10 nucleotides in the genome. Defects in DNA mismatch repair (MMR) caused by gene mutations or promoter hypermethylation can lead to hypermutations in nucleotide repeat regions (microsatellites), a condition known as microsatellite instability (MSI). MSI can promote the development of various cancers, including colon cancer (15%), gastric cancer (22%), endometrial cancer (20%-30%), and ovarian cancer (12%), with 45%-60% of these not responding to immune checkpoint blockade. Therefore, MSI tumors require novel treatment approaches. In two genome-wide gene inactivation studies using CRISPR or RNA interference, wRN was identified as the most significant dependency in MSI-H cells, and this dependency was associated with unwinding activity but not with the nuclease function of wRN. These findings, coupled with the good tolerance of microsatellite stable (MSS) cancer cells to wRN silencing, demonstrate that wRN is a potential specific target for the treatment of MSI tumors.

[0008] Given the huge unmet clinical needs, the development of small molecule inhibitors targeting WRN has broad application prospects. Summary of the Invention

[0009] This disclosure relates to compounds of formula (I) or their stereoisomers or pharmaceutically acceptable salts.

[0010] Among them, X 2 and X 4 One is N, and the other is C;

[0011] R 1Selected from 4-12 membered heterocyclic groups, 5-10 membered heteroaryl groups, or C6-C 14 arylene, the 4-12 membered heterocyclic arylene, 5-10 membered heterocyclic arylene, or C6-C 14 Alpha-aryl optional R 6’ replace;

[0012] R 6’ Selected from halogens, hydroxyl groups, C1-C6 alkyl groups, amino groups, oxo groups, cyano groups, C3-C6 cycloalkyl groups, or C1-C6 alkoxy groups;

[0013] R 6 Selected from hydrogen, hydroxyl, amino, halogen, oxo, cyano, C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl, the C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl optional R 6a replace;

[0014] R 6a Selected from oxo, halogen, cyano, amino, OH, C1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), NH(C3-C6 cycloalkyl), N(C1-C6 alkyl)(C3-C6 cycloalkyl), C1-C6 alkoxy, 4-12 membered heterocyclic groups, C3-C 10 Cycloalkyl or COOH, wherein the C1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), NH(C3-C6 cycloalkyl), N(C1-C6 alkyl)(C3-C6 cycloalkyl), C1-C6 alkoxy, 4-12 membered heterocyclic or C3-C 10 cycloalkyl optional R 6b replace;

[0015] R 6b Selected from deuterium, halogen, oxo, hydroxyl, amino, cyano, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl or C1-C6 alkoxy is optionally R 6c replace;

[0016] R 6c Selected from halogen, hydroxyl, amino, or cyano groups;

[0017] R 2 Selected from phenyl or 5-10 heteroaryl groups, wherein the phenyl or 5-10 heteroaryl group is optionally R 2a replace;

[0018] R2a Selected from halogens, C1-C6 alkyl groups, SF5 or C(O)H, wherein the C1-C6 alkyl group is optionally substituted with a halogen;

[0019] R 3 Selected from halogens or C1-C4 alkyl groups, wherein the C1-C4 alkyl group is optionally R 3a Substitution; or two R atoms attached to the same ring atom 3 Together with the atoms they are attached to, they form a C3-C6 cycloalkane ring;

[0020] R 3a Selected from halogens or hydroxyl groups;

[0021] x is selected from 0, 1, or 2;

[0022] 'e' can be a single or double bond.

[0023] When e is a single bond, Y is N or CH;

[0024] When e is a double bond, Y is C;

[0025] y is selected from 0, 1, or 2;

[0026] R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a C3-C6 cycloalkane ring;

[0027] R 4 Selected from C1-C4 alkyl, 5-10 heteroaryl, or 4-12 heterocyclic groups, wherein the C1-C4 alkyl, 5-10 heteroaryl, or 4-12 heterocyclic group is optionally R 4a replace;

[0028] R 4a Selected from halogens, hydroxyl groups, oxo groups, C1-C4 alkyl groups, or C1-C4 alkoxy groups, wherein the C1-C4 alkyl groups or C1-C4 alkoxy groups are optionally substituted with halogens.

[0029] In some implementation schemes, X 2 Let C, X 4 Let N be the number of elements in the array.

[0030] In some implementation schemes, X 2 Let N, X 4 The answer is C.

[0031] In some embodiments, Not for

[0032] In some implementation schemes, R 1 Selected from 4-10 heterocyclic groups, 5-10 heteroaryl groups, or C6-C10 arylene, the 4-10 member heterocyclic arylene, 5-10 member heterocyclic arylene or C6-C 10 Alpha-aryl optional R 6’ replace.

[0033] In some implementation schemes, R 1 Selected from 4-6-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 4-6-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally coated with R. 6’ replace.

[0034] In some implementation schemes, R 1 Selected from 6-8-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 6-8-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally coated with R. 6’ replace.

[0035] In some implementation schemes, R 1 Selected from 6-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 6-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally R-coated. 6’ replace.

[0036] In some implementation schemes, R 1 Selected from 6-membered heterocyclic group, 6-membered heteroaryl group, or phenylene group, wherein the 6-membered heterocyclic group, 6-membered heteroaryl group, or phenylene group is optionally R-coated. 6’ replace.

[0037] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the tetrahydropyridyl group, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Optional R 6’ replace.

[0038] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Where # represents X 2 Connect, and with R 6 The linking site can be any location where hydrogen is present, the tetrahydropyridyl group, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Optional R6’ replace.

[0039] In some implementation schemes, R 1 Selected from phenylene Where # represents X 2 Connect, and with R 6 The connection site can be any location where hydrogen is present, the phenylene, Optional R 6’ replace.

[0040] In some implementation schemes, R 1 Selected from Where # represents X 2 Connect, and with R 6 The connection site can be any location where hydrogen is present. Optional R 6’ replace.

[0041] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Pyridyl, phenylene, or thiazolyl, where * represents R 6 The # symbol represents a connection to X. 2 Connection, the tetrahydropyridyl group, Pyridyl, phenylene, or thiazolyl are optionally replaced by R 6’ replace.

[0042] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Pyridyl or phenylene, where * represents the group with R 6 The # symbol represents a connection to X. 2 Connection, the tetrahydropyridyl group, Pyridyl or phenylene optionally R 6’ replace.

[0043] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Pyridyl or phenylene, where * represents the group with R 6 The # symbol represents a connection to X. 2 Connection, the tetrahydropyridyl group, Pyridyl or phenylene optionally R 6’ replace.

[0044] In some implementation schemes, R 1Selected from dihydropyridyl, pyridylene, or phenylene, wherein the dihydropyridyl, pyridylene, or phenylene is optionally R 6’ replace.

[0045] In some implementation schemes, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the Optional R 6’ replace.

[0046] In some implementation schemes, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the Optional R 6’ replace.

[0047] In some implementation schemes, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 connect.

[0048] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Or 5-10 methyl aryl groups, where * represents R 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or 5-10 heteroaryl group is optionally R 6’ replace.

[0049] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Or a 6-membered heteroaryl group, where * represents a group with R. 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or 6-membered heteroaryl group is optionally R 6’ replace.

[0050] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Or pyridyl group, where * represents the group with R 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or pyridyl group is optionally R 6’ replace.

[0051] In some implementation schemes, R1 Selected from dihydropyridyl or 5-10-membered heteroaryl, wherein the dihydropyridyl or 5-10-membered heteroaryl is optionally R 6’ replace.

[0052] In some implementation schemes, R 1 Selected from dihydropyridyl or 6-membered heteroaryl, wherein the dihydropyridyl or 6-membered heteroaryl is optionally R 6’ replace.

[0053] In some implementation schemes, R 1 Selected from dihydropyridyl or pyridylene, wherein the dihydropyridyl or pyridylene is optionally R 6’ replace.

[0054] In some implementation schemes, R 6’ Selected from halogens or hydroxyl groups.

[0055] In some implementation schemes, R 6’ Selected from halogens or C1-C3 alkyl groups.

[0056] In some implementation schemes, R 6’ Selected from fluorine, chlorine, or methyl.

[0057] In some implementation schemes, R 6’ Selected from halogen.

[0058] In some implementation schemes, R 6’ Selected from fluorine.

[0059] In some implementation schemes, R 6 Selected from hydroxyl, amino, halogen, cyano, C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl, the C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl optional R 6a replace.

[0060] In some implementation schemes, R 6Selected from C(O)C3-C8 cycloalkyl, C(O)C1-C8 alkyl, C(O)4-10 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C8 cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C8 alkyl, C1-C8 alkoxy, -O-C3-C8 cycloalkyl, -S-C3-C8 cycloalkyl, -O-4-10 membered heterocyclic, -S-4-10 membered heterocyclic, -S-C1-C8 alkyl, C3-C8 cycloalkyl, -S(O)-C3-C8 cycloalkyl, -S(O)-C1-C8 alkyl, -S(O)2-C3-C8 cycloalkyl, -S(O)2-C1-C8 alkyl or 4-12 membered heterocyclic, The following C(O)C3-C8 cycloalkyl, C(O)C1-C8 alkyl, C(O)4-10 heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C8 cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C8 alkyl, C1-C8 alkoxy, -O-C3-C8 cycloalkyl, -S-C3-C8 cycloalkyl, -O-4-10 heterocyclic, -S-4-10 heterocyclic, -S-C1-C8 alkyl, C3-C8 cycloalkyl, -S(O)-C3-C8 cycloalkyl, -S(O)-C1-C8 alkyl, -S(O)2-C3-C8 cycloalkyl, -S(O)2-C1-C8 alkyl or 4-12 heterocyclic groups are optionally R 6a replace.

[0061] In some implementation schemes, R 6 Selected from C3-C6 cycloalkyl, 4-6 membered heterocyclic, C1-C6 alkyl, or C1-C6 alkoxy groups, wherein the C3-C6 cycloalkyl, 4-6 membered heterocyclic, C1-C6 alkyl, or C1-C6 alkoxy group is optionally R 6a replace.

[0062] In some implementation schemes, R 6 Selected from C3-C6 cycloalkyl, 4-6 membered heterocyclic or C1-C6 alkyl, wherein the C3-C6 cycloalkyl, 4-6 membered heterocyclic or C1-C6 alkyl is optionally R 6a replace.

[0063] In some implementation schemes, R 6 Selected from C3-C4 cycloalkyl, 4-membered heterocyclic, C1-C3 alkyl, or C1-C3 alkoxy groups, wherein the C3-C4 cycloalkyl, 4-membered heterocyclic, C1-C3 alkyl, or C1-C3 alkoxy group is optionally R 6a replace.

[0064] In some implementation schemes, R 6Selected from cyclopropyl, cyclobutyl, oxetyl, azirone, methyl, or methoxy, wherein the cyclopropyl, cyclobutyl, oxetyl, azirone, methyl, or methoxy group is optionally R-coated. 6a replace.

[0065] In some implementation schemes, R 6 Selected from C3-C4 cycloalkyl, 4-membered heterocyclic, or C1-C3 alkyl, wherein the C3-C4 cycloalkyl, 4-membered heterocyclic, or C1-C3 alkyl is optionally R 6a replace.

[0066] In some implementation schemes, R 6a Selected from halogens, cyano groups, amino groups, OH groups, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), 4-10 membered heterocyclic groups, or C3-C8 cycloalkyl groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl), 4-10 membered heterocyclic groups, or C3-C8 cycloalkyl groups are optionally modified by R. 6b replace.

[0067] In some implementation schemes, R 6a Selected from halogens, N(C1-C6 alkyl)2, 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, 4-10 membered heterocyclic groups are optionally R 6b replace.

[0068] In some implementation schemes, R 6a Selected from halogens, N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b replace.

[0069] In some implementation schemes, R 6a Selected from halogens, N(C1-C3 alkyl) 2, 4-7 membered heterocyclic groups, wherein the 4-7 membered heterocyclic group is optionally R 6b replace.

[0070] In some implementation schemes, R 6a Selected from fluorine, N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, The N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, Optional R 6b replace.

[0071] In some implementation schemes, R 6a Selected from halogen.

[0072] In some implementation schemes, R 6b Selected from deuterium, halogen, cyano, C1-C6 alkoxy, or C1-C6 alkyl, wherein the C1-C6 alkoxy or C1-C6 alkyl is optionally R6c replace.

[0073] In some implementation schemes, R 6b Selected from deuterium, halogens, C1-C6 alkoxy or C1-C6 alkyl, wherein the C1-C6 alkoxy or C1-C6 alkyl is optionally R 6c replace.

[0074] In some implementation schemes, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c replace.

[0075] In some implementation schemes, R 6b Selected from deuterium, halogen, C1-C3 alkoxy or C1-C3 alkyl, wherein the C1-C3 alkyl is optionally R 6c replace.

[0076] In some implementation schemes, R 6b Selected from deuterium, fluorine, cyano, methoxy, or methyl, wherein the methoxy or methyl group is optionally R 6c replace.

[0077] In some implementation schemes, R 6c Selected from halogen.

[0078] In some implementation schemes, R 6 Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl group is R 6a Replace, R 6b Selected from N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic group is optionally R 6b The substitution is made, and the atoms connected to the 4-10 membered heterocyclic group and the C1-C6 alkyl group are heteroatoms.

[0079] In some implementation schemes, R 6 Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl group is R 6a Replace, R 6b Selected from N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic group is optionally R 6b The substitution is made such that the atom connected to the 4-10 membered heterocyclic group and the C1-C6 alkyl group is a nitrogen atom.

[0080] In some implementation schemes, R 6 Selected from hydrogen, methyl, methoxy, cyclopropyl,

[0081] In some implementation schemes, R 6 Selected from methyl, methoxy, cyclopropyl,

[0082] In some embodiments, Selected from

[0083] In some embodiments, Selected from

[0084] In some implementation schemes, R 1 Selected from: 6-membered heterocyclic group, R 6 Selected from C3-C4 cycloalkyl or 4-membered heterocyclic groups, wherein the C3-C4 cycloalkyl or 4-membered heterocyclic group is optionally R 6a Replace; or,

[0085] R 1 Selected from 8-membered heterocyclic groups, R 6 Selected from hydrogen; or,

[0086] R 1 Selected from phenyl, wherein the phenyl is optionally R 6’ Replace, R 6 Selected from C1-C3 alkyl groups, wherein the C1-C3 alkyl group is substituted with N(C1-C3 alkyl)2 or 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic groups are optionally replaced with R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl may optionally be substituted with a halogen; or,

[0087] R 1 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group is optionally replaced by R 6’ Replace, R 6 Selected from C1-C3 alkyl, C1-C3 alkoxy, or C3-C4 cycloalkyl, wherein the C1-C3 alkyl group is substituted with a halogen or a 4-7 membered heterocyclic group, wherein the 4-7 membered heterocyclic group is optionally replaced with R. 6b Replace, R 6b It is selected from deuterium, halogen, cyano, C1-C3 alkoxy or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally substituted with halogen.

[0088] In some implementation schemes, R 6Selected from C3-C8 cycloalkyl, 4-10 membered heterocyclic groups or -T-NR 7 R 8 The C3-C8 cycloalkyl or 4-10 heterocyclic group is optionally R 6a Replacement, wherein T is selected from (CH2). n O, S, or C (=O), the (CH2) n Optional R T Replace, two R T Together with their respective connected atoms, they form a C3-C6 cycloalkane ring, where n is selected from 0, 1, or 2, and R... 7 R 8 Each is independently selected from H or arbitrarily selected by R. 8a Substituted C1-C6 alkyl groups, or R 7 R 8 Together with the N atom it is attached to, they form a 4-12 membered heterocyclic group, which is optionally converted by R. 7a Replace; R 8a Selected from deuterium, halogen, hydroxyl, amino, or cyano groups; R 7a Selected from halogen, oxo, hydroxyl, amino, cyano, C1-C6 alkoxy, or C1-C6 alkyl, wherein the C1-C6 alkoxy or C1-C6 alkyl is optionally R 7b Replace; R 7b Selected from halogen, hydroxyl, amino or cyano groups.

[0089] In some implementation schemes, R 6 Selected from C3-C4 cycloalkyl, 4-membered heterocyclic or -T-NR 7 R 8 The C3-C4 cycloalkyl group or the 4-membered heterocyclic group is optionally replaced by R. 6a replace.

[0090] In some implementations, T is selected from (CH2). n The (CH2) n Optional R T Replace, two R T Together with their respective connected atoms, they form a C3-C6 cycloalkane ring.

[0091] In some implementations, T is selected from (CH2). n .

[0092] In some implementations, n is selected from 1.

[0093] In some implementation schemes, R 7 R 8 Each is independently selected from C1-C6 alkyl groups, or R 7 R 8Together with the N atom it is attached to, they form a 4-10 membered heterocyclic group, which is optionally converted by R. 7a replace.

[0094] In some implementation schemes, R 7 R 8 Each is independently selected from C1-C3 alkyl groups, or R 7 R 8 Together with the N atom it is attached to, they form a 4-7 membered heterocyclic group, which is optionally converted by R. 7a replace.

[0095] In some implementation schemes, R 8a Selected from deuterium.

[0096] In some implementation schemes, R 7a Selected from halogens, C1-C6 alkoxy groups, or C1-C6 alkyl groups, wherein the C1-C6 alkoxy groups or C1-C6 alkyl groups are optionally R 7b replace.

[0097] In some implementation schemes, R 7a Selected from halogens, C1-C3 alkoxy groups, or C1-C3 alkyl groups, wherein the C1-C3 alkoxy groups or C1-C3 alkyl groups are optionally R 7b replace.

[0098] In some implementation schemes, R 7a Selected from halogens, C1-C3 alkoxy groups, or C1-C3 alkyl groups, wherein the C1-C3 alkyl group is optionally R-coated. 7b Replacement. In some implementations, R 7b Selected from halogens.

[0099] In some implementation schemes, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the Optional R 6’ replace.

[0100] In some embodiments, Selected from

[0101] In some implementation schemes, R 2 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 2a replace.

[0102] In some implementation schemes, R 2 Selected from phenyl or 9-membered heteroaryl, wherein the phenyl or 9-membered heteroaryl group is optionally R 2areplace.

[0103] In some implementation schemes, R 2 Selected from phenyl, benzofuranyl, or benzothiophene, wherein the phenyl, benzofuranyl, or benzothiophene group is optionally R 2a replace.

[0104] In some implementation schemes, R 2 Selected from phenyl, wherein the phenyl is optionally R 2a replace.

[0105] In some implementation schemes, R 2a Selected from halogens or C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with halogens.

[0106] In some implementation schemes, R 2a Selected from halogens or C1-C3 alkyl groups, wherein the C1-C3 alkyl groups are optionally substituted with halogens.

[0107] In some implementation schemes, R 2a Selected from chlorine or methyl, wherein the methyl group is optionally substituted with a halogen.

[0108] In some implementation schemes, R 2 Selected from

[0109] In some implementation schemes, R 3 Selected from halogens or C1-C4 alkyl groups, wherein the C1-C4 alkyl group is optionally R 3a replace.

[0110] In some implementation schemes, R 3 Selected from C1-C4 alkyl groups.

[0111] In some implementation schemes, R 3 Selected from methyl.

[0112] In some implementations, x is selected from 1 or 2.

[0113] In some implementations, x is selected from 1.

[0114] In some implementations, 'e' is a single bond or a double bond.

[0115] When e is a single bond, Y is N or CH;

[0116] When e is a double bond, Y is C;

[0117] y is selected from 0, 1, or 2;

[0118] R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5Together with the atoms and bonds they are connected to, they form a C3-C4 cycloalkane ring.

[0119] In some implementations, e is a single bond and Y is N or CH;

[0120] y is selected from 0, 1, or 2;

[0121] R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5 Together with the atoms and bonds they are attached to, they form a C3-C4 cycloalkane ring. In some embodiments, R 5 Selected from methyl groups, or two R atoms attached to adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a C3-C4 cycloalkane ring.

[0122] In some embodiments, Selected from

[0123] In some embodiments, Selected from

[0124] In some embodiments, Selected from

[0125] In some embodiments, e is a single bond or a double bond,

[0126] When e is a single bond, Y is CH, and y is selected from 2, where R is attached to two adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a cyclopropane ring;

[0127] When e is a double bond, Y is C.

[0128] In some implementations, e is a single bond, Y is CH, y is selected from 2, and two R atoms are attached to adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a cyclopropane ring.

[0129] In some embodiments, Selected from

[0130] In some embodiments, Selected from

[0131] In some implementation schemes, R 4 Selected from 5-10-membered heteroaryl or 4-10-membered heterocyclic groups, wherein the 5-10-membered heteroaryl or 4-10-membered heterocyclic group is optionally R4a replace.

[0132] In some implementation schemes, R 4 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group is optionally replaced by R 4a replace.

[0133] In some implementation schemes, R 4 Selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group is optionally replaced by R 4a replace.

[0134] In some implementation schemes, R 4 Selected from pyrimidinyl groups, wherein the pyrimidinyl group is optionally R 4a replace.

[0135] In some implementation schemes, R 4a Selected from halogens, hydroxyl groups, or C1-C4 alkyl groups, wherein the C1-C4 alkyl groups are optionally substituted with halogens.

[0136] In some implementation schemes, R 4a Selected from hydroxyl or C1-C4 alkyl.

[0137] In some implementation schemes, R 4a Selected from hydroxyl or methyl.

[0138] In some implementation schemes, R 4 Selected from 5-10-membered heteroaryl groups, wherein at least one ortho-position of the 5-10-membered heteroaryl group is substituted with a hydroxyl group, and the 5-10-membered heteroaryl group substituted with at least one ortho-position hydroxyl group is further optionally R 4a replace.

[0139] In some implementation schemes, R 4 Selected from 5-10-membered heteroaryl groups, wherein one ortho-position of the 5-10-membered heteroaryl group is substituted with a hydroxyl group, and the 5-10-membered heteroaryl group substituted with a hydroxyl group at the ortho-position is further optionally R 4a replace.

[0140] In some implementation schemes, R 4 Selected from

[0141] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (I'):

[0142] Where e, x, y, X 2 X 4 R 1 R 2 R 3 R 4 R 5 R6 Y is as defined by compound (I).

[0143] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (Ia):

[0144] Among them, e, y, X 2 X 4 R 1 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0145] In some embodiments, the compounds of formula (I) of the present disclosure are selected from compounds of formula (II):

[0146] Where e, x, y, R 1 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0147] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (II'):

[0148] Where e, x, y, R 1 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0149] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (IIa):

[0150] Among them, e, y, R 1 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0151] In some embodiments, the compounds of formula (I) of the present disclosure are selected from compounds of formula (III):

[0152] Where x and X 2 X 4 R 1 R 2 R 3 R 4 and R 6 As defined by compound (I).

[0153] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (III'):

[0154] Where x and X 2 X 4 R 1 R 2 R 3 R 4 and R 6 As defined by compound (I).

[0155] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (IIIa):

[0156] Among them, X 2 X 4 R 1 R 2 R 3 R 4 and R 6 As defined by compound (I).

[0157] In some embodiments, the compounds of formula (I) of the present disclosure are selected from compounds of formula (IV):

[0158] Where e, x, y, X 2 X 4 R 1 R 2 R 3 R 4 R 5 R 6a Y is as defined in compound (I), wherein q1 is selected from 1, 2, 3, 4, 5, 6, 7 or 8, and q2 is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0159] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (IV'):

[0160] Where e, x, y, X 2 X 4 R 1 R2 R 3 R 4 R 5 R 6a Y is as defined in compound (I), wherein q1 is selected from 1, 2, 3, 4, 5, 6, 7 or 8, and q2 is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0161] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (IVa):

[0162] Among them, e, y, X 2 X 4 R 1 R 2 R 3 R 4 R 5 R 6a Y is as defined in compound (I), wherein q1 is selected from 1, 2, 3, 4, 5, 6, 7 or 8, and q2 is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0163] In some implementation schemes, R 1 Selected from 6-8 member subheterocyclic groups, wherein the 6-8 member subheterocyclic group is optionally R 6’ replace.

[0164] In some implementation schemes, R 1 Selected from tetrahydropyridyl, Tetrahydropyranyl or Where # represents X 2 Connection, the tetrahydropyridyl group, Tetrahydropyranyl or Optional R 6’ replace.

[0165] In some implementation schemes, R 1 Selected from tetrahydropyridyl, wherein the tetrahydropyridyl group is optionally R 6’ replace.

[0166] In some implementation schemes, R 1 Selected from tetrahydropyridyl group.

[0167] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (IVa-1):

[0168] Among them, e, y, X 2 X 4 R 2 R 3 R4 R 5 R 6a Y is as defined in compound (I), wherein q1 is selected from 1, 2, 3, 4, 5, 6, 7 or 8, and q2 is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8.

[0169] In some implementations, q1 is selected from 1 or 2.

[0170] In some implementations, q2 is selected from 0 or 2.

[0171] In some implementation schemes, R 6a It is selected from halogen, cyano, amino, OH or C1-C3 alkyl.

[0172] In some implementation schemes, R 6a Selected from halogens.

[0173] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (V):

[0174] Where e, x, y, X 2 X 4 R 2 R 3 R 4 R 5 R 6 R 6’ And Y is as defined in compound of formula (I), and z1 is selected from 0, 1, 2, 3 or 4.

[0175] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (V'):

[0176] Where e, x, y, X 2 X 4 R 2 R 3 R 4 R 5 R 6 R 6’ And Y is as defined in compound of formula (I), and z1 is selected from 0, 1, 2, 3 or 4.

[0177] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (Va):

[0178] Among them, e, y, X 2 X 4 R 2 R 3 R4 R 5 R 6 R 6’ And Y is as defined in compound of formula (I), and z1 is selected from 0, 1, 2, 3 or 4.

[0179] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (VI) or formula (VI'):

[0180] Where e, x, y, X 2 X 4 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0181] In some embodiments, the compounds of formula (I) disclosed herein are selected from compounds of formula (VIa):

[0182] Among them, e, y, X 2 X 4 R 2 R 3 R 4 R 5 R 6 Y is as defined by compound (I).

[0183] In some implementations, z1 is selected from 0, 1, or 2.

[0184] In some implementation schemes, R 6 Selected from C1-C3 alkyl groups, wherein the C1-C3 alkyl group is R 6a Replace, R 6a Selected from N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c Replace, R 6c Selected from halogen.

[0185] In some implementation schemes, R 6a The atom attached to the C1-C3 alkyl group is an N atom.

[0186] In some implementation schemes, R 6 Selected from methyl, said methyl is R 6a Replace, R6a Selected from N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c Replace, R 6c Selected from halogen.

[0187] In some implementation schemes, R 6a The atom bonded to the methyl group is an N atom.

[0188] In some implementation schemes, R 6 Selected from methyl, said methyl is R 6a Replace, R 6a Selected from N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, The N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, Optional R 6b Replace, R 6b Selected from deuterium, fluorine, cyano, methoxy, or methyl, wherein the methoxy or methyl group is optionally R 6c Replace, R 6c Selected from halogen.

[0189] In some embodiments, the compounds of formula (I) of the present disclosure are selected from the following compounds:

[0190] On the other hand, this disclosure provides pharmaceutical compositions comprising a compound of formula (I) of this disclosure or a stereoisomer thereof or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0191] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for inhibiting WRN.

[0192] In some implementations, the intended use is for treating cancer.

[0193] On the other hand, this disclosure provides a method for inhibiting WRN, which includes the step of contacting a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, with WRN.

[0194] On the other hand, this disclosure provides a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, which is used as a WRN inhibitor.

[0195] On the other hand, this disclosure provides a method for treating diseases mediated by WRN helicase in mammals, comprising administering to a mammal in need of treatment, preferably a human, a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0196] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for the prevention or treatment of WRN helicase-mediated diseases.

[0197] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the prevention or treatment of WRN helicase-mediated diseases.

[0198] On the other hand, this disclosure provides a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the prevention or treatment of WRN helicase-mediated diseases.

[0199] In some embodiments, the WRN helicase-mediated disease is selected from cancer or Werner syndrome.

[0200] In some embodiments, the cancer is colon cancer.

[0201] On the other hand, this disclosure provides that the compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt can be used as a medicine, particularly for treating conditions or diseases that can be treated by WRN inhibition.

[0202] On the other hand, this disclosure provides that compounds of formula (I) or their stereoisomers or pharmaceutically acceptable salts, or pharmaceutical compositions thereof, are used to treat cancer.

[0203] On the other hand, this disclosure provides a method for modulating WRN activity in a patient, wherein the method comprises administering to the patient a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0204] On the other hand, this disclosure provides a method for inhibiting WRN in a patient, wherein the method comprises administering to the patient a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0205] On the other hand, this disclosure provides a method for treating a condition or disease that can be treated in a patient by WRN inhibition, comprising administering to the patient a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0206] On the other hand, this disclosure provides a method of treating a patient with cancer, comprising administering to the patient a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0207] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or its pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament.

[0208] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for treating cancer.

[0209] On the other hand, this disclosure provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for treating diseases that can be treated by WRN inhibition.

[0210] Definitions and Explanations of Terms

[0211] Unless otherwise indicated, the terms used in this disclosure have the following meanings. The definitions of groups and terms described in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions described in tables, and definitions of specific compounds in the examples, may be combined and coupled with each other in any manner. A particular term should not be considered as undefined or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

[0212] In this article Indicates the connection site.

[0213] In this disclosure, "*" or "#" indicates that the atom it identifies is a linking site, such as a linking group. This indicates that the N atom in the linking group is the linking site.

[0214] In the ring This indicates that the corresponding ring is an aromatic ring. Structural unit. It can be understood as an aromatic ring with a cyclic conjugated system.

[0215] In this text, the keys are depicted by solid and dashed lines. Indicates a single bond or a double bond. For example, structural unit. Include

[0216] Unless otherwise specified, use wedge keys and virtual wedge keys. The absolute configuration of a solid center is represented by black solid bonds and imaginary bonds. It indicates the relative configuration of a stereocenter (such as the cis-trans configuration of alicyclic compounds).

[0217] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds disclosed herein can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer typically yield a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form is dominant; while in phenols, the enol form is dominant. This disclosure encompasses all tautomeric forms of the compounds.

[0218] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0219] The compounds disclosed herein may have asymmetric atoms such as carbon, sulfur, nitrogen, and phosphorus atoms, or asymmetric double bonds, and therefore may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E- and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof or other mixtures, such as mixtures enriched with enantiomers or diastereomers. All such isomers and mixtures thereof are within the scope of the definition of the compounds disclosed herein. Alkyl groups or other substituents may contain additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms. All such isomers involved in all substituents, and mixtures thereof, are also included within the scope of the definition of the compounds disclosed herein. The compounds containing asymmetric atoms disclosed herein can be isolated in optically active pure form or in racemic form. The optically active pure form can be separated from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0220] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (ie, =O), it means that two hydrogen atoms are replaced.

[0221] The terms “optional” or “optionally” mean that the event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, “optionally” substituted with a halogen means that the ethyl group can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible and / or cannot be synthesized is introduced.

[0222] When any variable (e.g., R) a R b When a group appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is surrounded by two R... b Replaced, then each R b Each has its own independent options.

[0223] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a bond.

[0224] When a substituent is cross-bonded to two atoms on a ring, it can bond to any atom on that ring. For example, structural units. R represents 5 Substitution can occur at any position on the benzene ring.

[0225] C in this article m -C n It refers to having an integer number of carbon atoms in the range mn. For example, "C1-C 10 "" means that the group can have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0226] The term "alkyl" refers to a compound with the general formula C1. n H 2n+1 The alkyl group can be straight-chain or branched. The term "C1-C" refers to a hydrocarbon group. 10"Alkyl" can be understood as representing a straight-chain or branched saturated hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, and 1,1-dimethylbutyl. 2,3-Dimethylbutyl, 1,3-Dimethylbutyl, or 1,2-Dimethylbutyl, etc.; the term "C1-C8 alkyl" can be understood as referring to an alkyl group having 1 to 8 carbon atoms. The term "C1-C6 alkyl" can be understood as referring to an alkyl group having 1 to 6 carbon atoms, specific examples including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" can be understood as referring to a straight-chain or branched saturated alkyl group having 1 to 3 carbon atoms. The "C1-C" 10 "alkyl" can include the range of "C1-C8 alkyl", "C1-C6 alkyl" or "C1-C3 alkyl", and the "C1-C6 alkyl" can further include "C1-C3 alkyl".

[0227] The term "alkoxy" refers to a group formed by the loss of a hydrogen atom from a hydroxyl group in straight-chain or branched alcohols; it can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C"... 10 "Alkoxy" can be understood as "C1-C" 10 "alkyloxy" or "C1-C" 10 Alkyl-O-”; the term “C1-C8 alkoxy” can be understood as “C1-C8 alkyloxy” or “C1-C8 alkyl-O-”; the term “C1-C6 alkoxy” can be understood as “C1-C6 alkyloxy” or “C1-C6 alkyl-O-”. The “C1-C”... 10 "Alkoxy" can include the range of "C1-C8 alkoxy", "C1-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" can further include "C1-C3 alkoxy".

[0228] The term "cycloalkyl" refers to a fully saturated carbon ring existing in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 10-membered ring. The term "C3-C" is also used. 10"Cycloalkyl" can be understood as referring to a saturated monocyclic, fused, spirocyclic, or bridged ring having 3 to 10 carbon atoms. Specific examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, etc. The term "C3-C" 10 "Cycloalkyl" can include "C3-C8 cycloalkyl", "C3-C6 cycloalkyl", "C3-C4 cycloalkyl", etc. The term "C3-C6 cycloalkyl" can be understood as indicating a saturated monocyclic or bicyclic hydrocarbon ring with 3 to 6 carbon atoms. Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0229] The term "cycloalkylene" refers to a divalent group derived from a "cycloalkyl" group as defined herein.

[0230] The term "heterocyclic group" refers to a fully saturated or partially saturated (not aromatic as a whole) monocyclic, fused, spirocyclic, or bridged ring group containing 1 to 5 heteroatoms or heteroatomic groups (i.e., groups containing heteroatoms). The "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=O)2-, -S(=O)-, -P(=O)2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH-, or -NHC(=O)NH-, etc. The term "4-18 membered heterocyclic group" refers to a heterocyclic group with 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, and whose ring atoms contain 1 to 5 independently selected heteroatoms or heterogroups as described above. The term "4-12 membered heterocyclic group" refers to a heterocyclic group with 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, and whose ring atoms contain 1 to 5 independently selected heteroatoms or heterogroups as described above. "4-10 membered heterocyclic groups" includes "4-7 membered heterocyclic groups", wherein specific examples of 4 membered heterocyclic groups include, but are not limited to, azacyclic butyl or oxacyclic butyl; specific examples of 5 membered heterocyclic groups include, but are not limited to, tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoyl, pyrazolyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6 membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazine, trithiaalkyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; and specific examples of 7 membered heterocyclic groups include, but are not limited to, diazacyclic heptyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include but are not limited to hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include but are not limited to hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group may be a benzo-fused ring group or a heteroaromatic ring-fused ring group of the above-mentioned 4-7-membered heterocyclic group, specific examples of which include but are not limited to dihydroisoquinolinyl. The heterocyclic group may also be a tricyclic group."4-10 membered heterocyclyl" may include "5-10 membered heterocyclyl", "4-7 membered heterocyclyl", "5-6 membered heterocyclyl", "6-8 membered heterocyclyl", "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered heterocycloalkyl", etc., and "4-7 membered heterocyclyl" may further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", etc. Although some bicyclic heterocyclyl groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclyl group as a whole is still non-aromatic.

[0231] The term "subheterocyclic group" refers to a divalent group derived from a "heterocyclic group" as defined herein.

[0232] The term "aryl" refers to an aromatic ring group consisting of an all-carbon monocyclic or fused polycyclic aromatic cyclic group with a conjugated π-electron system. Aryl groups can have 6-20, 6-14, or 6-12 carbon atoms. The term "C6-C"... 20 "Aryl" can be understood as an aryl group having 6 to 20 carbon atoms. Specifically, it refers to a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring with 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl; or a ring with 10 carbon atoms ("C9 aryl"). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl; or rings having 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups; or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. The term "C6-C" is used. 10 "Aryl" can be understood as an aryl group having 6 to 10 carbon atoms. Specifically, it refers to a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring with 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl; or a ring with 10 carbon atoms ("C9 aryl"). 10 Aryl), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0233] The term "aryl" refers to a divalent group derived from the "aryl" group.

[0234] The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, or S, with the remaining ring atoms being C. The term "5-10-membered heteroaryl" can be understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, or 10 ring atoms, particularly 5, 6, 9, or 10 ring atoms, and containing 1-5, preferably 1-3, heteroatoms independently selected from N, O, and S. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzothiazolyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl or isindolyl; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and their benzo[derivatives], such as quinolinyl, quinazolinyl or isoquinolinyl; or acrylinyl, inazinyl, purinyl and their benzo[derivatives]; or cyclolinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphthidyl, pteridinyl, carbazolyl, acrylinyl, phenazinyl, phenothiazinyl or phenothiazinyl. "5-10-membered heteroaryl" includes "5-6-membered heteroaryl". The term "5-6-membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and it contains 1 to 3, preferably 1 to 2, heteroatoms independently selected from N, O and S.

[0235] The term "hybrid aryl" refers to a divalent group derived from the "heteroaryl" group.

[0236] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., nucleophilic substitution). For example, representative leaving groups include, but are not limited to, trifluoromethanesulfonates, chlorine, bromine, iodine, sulfonate groups (e.g., methanesulfonates, toluenesulfonates, p-bromobenzenesulfonates, p-toluenesulfonates), or acyloxy groups (e.g., acetoxy, trifluoroacetoxy).

[0237] The term "halogen" or "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0238] The term "hydroxyl group" refers to the -OH group.

[0239] The term "amino" refers to the -NH2 group.

[0240] The term "cyano" refers to the -CN group.

[0241] The term "C(O)" refers to a -C(=O) group.

[0242] The term "WRN inhibitor" or "WRN helicase inhibitor" refers to a compound that inhibits Werner Syndrome RecQ DNA helicase (WRN). The term "WRN" refers to the protein of Werner Syndrome RecQ DNA helicase. The term "WRN" includes mutants, fragments, variants, isoforms, and homologs of full-length wild-type WRN. "Diseases or conditions mediated by WRN" include diseases or conditions that are treated by WRN inhibition, such as cancer.

[0243] The term "therapeutic effective amount" means (i) the amount of the disclosed compound used to treat a particular disease, condition, or disorder, or (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder. The amount of the disclosed compound constituting a "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by someone skilled in the art based on their own knowledge and the content of this disclosure.

[0244] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0245] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of an acid or base, including salts formed by a compound with an inorganic or organic acid, and salts formed by a compound with an inorganic or organic base.

[0246] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or their salts and a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present disclosure to an organism.

[0247] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0248] The word “comprise” or “include” and its English variants such as comprises or comprising can be understood as having an open, non-exclusive meaning, that is, “including but not limited to”.

[0249] This disclosure also includes compounds of this disclosure that are identical to those described herein, but in which one or more atoms are labeled with isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, etc.

[0250] Certain isotope-labeled compounds of this disclosure (e.g., using...) 3 H and 14 C-labeling can be used in the analysis of compound and / or substrate tissue distribution. Tritiumization (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. The isotopically labeled compounds of this disclosure can typically be prepared by replacing the unlabeled reagent with an isotopically labeled reagent using a procedure similar to those disclosed in the schemes and / or examples below.

[0251] The pharmaceutical compositions of the present disclosure can be prepared by combining the compounds of the present disclosure with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

[0252] Typical routes of administration of the disclosed compounds, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0253] The pharmaceutical compositions disclosed herein can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, emulsification, freeze drying, etc.

[0254] In some embodiments, the pharmaceutical composition is in an oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of this disclosure to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0255] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, flow aids, or flavoring agents.

[0256] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0257] In all methods of administration of the compound of general formula I described herein, the daily dose may be from 0.01 mg / kg to 100 mg / kg body weight, in the form of single or separate doses.

[0258] The compounds disclosed herein can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments disclosed herein.

[0259] The chemical reactions in the specific embodiments of this disclosure are carried out in a suitable solvent, which must be suitable for the chemical changes of this disclosure and the reagents and materials required therefor. In order to obtain the compounds of this disclosure, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0260] In some embodiments, compounds of general formula (I) of this disclosure can be prepared by those skilled in the art of organic synthesis via route 1:

[0261] <General Route 1>

[0262] In some embodiments, compounds of general formula (II) of this disclosure can be prepared by those skilled in the art of organic synthesis via route 2:

[0263] <General Route 2> Detailed Implementation

[0264] The invention is described in detail below with reference to embodiments, but this does not imply any adverse limitation of the present disclosure. The present disclosure has been described in detail, including specific embodiments thereof. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present disclosure without departing from the spirit and scope thereof. All reagents used in this disclosure are commercially available and can be used without further purification.

[0265] Unless otherwise stated, the proportions of mixed solvents are volume-based.

[0266] Compounds are processed manually or Software naming conventions are used; commercially available compounds use supplier catalog names.

[0267] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are measured in units of 10⁻⁶. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., with tetramethylsilane (TMS) as the internal standard; "IC 50 "Half-inhibitory concentration" refers to the concentration at which half of the maximum inhibitory effect is achieved.

[0268] The eluent described below may be a mixture of two or more solvents, with the ratio being the volume ratio of each solvent.

[0269] The stereostructure of the compound disclosed herein can be confirmed by single-crystal X-ray diffraction analysis. Single-crystal preparation method: Weigh 20 mg of the compound into a 3 ml glass bottle, add methanol (2.5 ml), and sonicate appropriately to promote solid dissolution. Filter the solution, transfer the filtrate to a 4 ml single-crystal growth flask, seal the flask with a single-crystal stopper, and make a small hole in the stopper. Place the single-crystal flask in a fume hood to allow the solvent to evaporate slowly. As the solvent decreases, the crystal grows slowly within the single-crystal flask. Observe the crystals under a microscope, and select those of suitable size and with well-formed crystals for single-crystal XRD analysis. The parameters of the single-crystal X-ray diffractometer used in this disclosure are as follows:

[0270] Synthesis of intermediate Int1-9: 4-(6-bromo-5-ethyl-7-oxo-4,7-dihydro-2H-[1,2,3]triazolo[4,5-b]pyridin-2-yl)-2-fluorobenzaldehyde

[0271] Step 1: Synthesis of compound Int1-1

[0272] SM1 (5 g) and methoxymethylamine (5.77 g) were dispersed in tetrahydrofuran (80 mL) under nitrogen protection. Isopropyl magnesium chloride and lithium chloride (1.3 M, 136.43 mL) were slowly added dropwise at -20 °C. The reaction was continued under low temperature with stirring for 4 hours. The reaction was quenched by adding saturated ammonium chloride solution. The mixture was extracted with dichloromethane (200 mL), separated, concentrated, and subjected to column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound Int1-1 (5 g).

[0273] Step 2: Synthesis of compound Int1-2

[0274] Compound Int1-1 (1.42 g) was dissolved in 15 mL of 6 M hydrochloric acid aqueous solution. A solution of 545 mg sodium nitrite dissolved in 3 mL of water was added dropwise at 0 °C, and the reaction was carried out at 0 °C for 30 minutes. Then, 89 mg urea was added to consume excess sodium nitrite. The above solution was then added to a solution of 2,6-diamino-4-methoxypyridine (2 g) dissolved in 15 mL of water, and the reaction was carried out at room temperature for 1.5 hours. Next, 100 mL of 10.78 g sodium acetate aqueous solution was added dropwise, and the mixture was stirred at room temperature for 12 hours. The mixture was extracted with dichloromethane (200 mL x 2), separated, concentrated, and subjected to column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound Int1-2 (2.5 g).

[0275] Step 3: Synthesis of compound Int1-3

[0276] Compound Int1-2 (2.5 g) and lead tetraacetate (4.77 g) were dispersed in dichloromethane (50 mL) under nitrogen protection and stirred at 50 °C for 2 hours. The pH was adjusted to about 9 by adding saturated sodium bicarbonate solution, and the mixture was extracted with dichloromethane. The liquid-liquid phase was separated, concentrated, and subjected to column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound Int1-3 (2.4 g).

[0277] Step 4: Synthesis of compound Int1-4

[0278] Compound Int1-3 (2 g) was dispersed in acetonitrile (40 mL) under nitrogen protection. Tert-butyl nitrite (1.49 g) and cuprous bromide (4.14 g) were added in an ice bath. The mixture was heated to 80 °C and stirred for 1 hour. The mixture was filtered, and the filtrate was concentrated under reduced pressure and subjected to column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound Int1-4 (900 mg).

[0279] Step 5: Synthesis of compound Int1-5

[0280] Compound Int1-4 (900 mg), pinacol vinylborate (676 mg), potassium phosphate (1.86 g), and Pd(dppf)Cl2 (240 mg) were dispersed in dioxane (40 mL) / water (10 mL) under nitrogen protection and stirred at 90 °C for 6 hours. The mixture was then diluted with water (60 mL), extracted with ethyl acetate, separated, concentrated in organic phase, and subjected to column chromatography (dichloromethane / methanol = 20 / 1) to obtain compound Int1-5 (500 mg).

[0281] Step 6: Synthesis of compound Int1-6

[0282] Compound Int1-5 (400 mg) was dispersed in tetrahydrofuran (4 mL) / methanol (10 mL), then palladium / carbon (129 mg) was added, hydrogen was purged three times, the reaction was stirred for 1 hour under hydrogen balloon pressure, filtered, the filtrate was dried with anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound Int1-6 (400 mg).

[0283] Step 7: Synthesis of compound Int1-7

[0284] Compound Int1-6 (250 mg) was dissolved in acetonitrile (20 mL), and trimethylchlorosilane (1.07 g) and potassium iodide (1.15 g) were added. The mixture was stirred at 80 °C for 1 hour. Saturated sodium bicarbonate was added to adjust the pH to weakly alkaline. Iodine was quenched with saturated anhydrous sodium sulfite solution. The mixture was extracted with dichloromethane, separated, and the organic phase was dried over anhydrous sodium sulfate and concentrated to obtain compound Int1-7 (230 mg).

[0285] Step 8: Synthesis of compound Int1-8

[0286] Compound Int1-7 (240 mg) was dissolved in tetrahydrofuran (5 mL) under nitrogen protection. Diisobutylaluminum hydride (1 M, 4.86 mL) was slowly added dropwise at -20 °C. The reaction was quenched by slowly adding saturated ammonium chloride solution (4 mL) and water (10 mL) at -20 °C. The reaction was then stirred at room temperature for 1 hour. The mixture was filtered, and the filter cake was redissolved in dichloromethane / methanol = 10 / 1. The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound Int1-8 (190 mg).

[0287] Step 9: Synthesis of compound Int1-9

[0288] Compound Int1-8 (200 mg) was dissolved in acetonitrile (10 mL), and then N-bromosuccinimide (124 mg) was added. The mixture was stirred at 25 °C for 1 hour. LCMS showed that the substrate was completely converted. The mixture was diluted with water (40 mL), filtered, and the filter cake was redissolved in dichloromethane / methanol (10 / 1) solution. The solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound Int1-9 (250 mg).

[0289] LC-MS (ESI): m / z = 365.1 [M+H] +

[0290] Synthesis of intermediate Int4-3: 1-(3,3-difluorocyclobutyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoborhexacyclopentan-2-yl)-1,2,3,6-tetrahydropyridine

[0291] Step 1: Synthesis of compound Int4-2

[0292] Weigh 335 mg of N,N-diisopropylethylamine and dissolve it in 5 mL of dichloromethane. Under argon protection, stir at -78°C. Slowly add 630 mg of trifluoromethanesulfonic anhydride and stir for 10 minutes. Then slowly add 200 mg of the starting material Int4-1. Gradually raise the temperature from -78°C to 25°C and stir for 3 hours until the reaction is complete. TLC (petroleum ether / ethyl acetate = 1:1) confirms that the starting material has reacted completely and can be used directly in the next reaction without further treatment.

[0293] Step 2: Synthesis of compound Int4-3

[0294] Weigh 222 mg of N,N-diisopropylethylamine and dissolve it in the above reaction solution. Slowly add 120 mg of compound Int4-4 at 0°C, stir for 30 minutes, then heat to 25°C and stir for 12 hours until the reaction is complete. Add 20 mL of dichloromethane and 10 mL of water to the reaction solution for extraction. Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate to obtain compound Int4-3 (150 mg).

[0295] LC-MS (ESI): m / z = 300.2 [M+H] +

[0296] Synthesis of intermediate 1-8-R

[0297] Compounds 1-6 (2 g) were dissolved in dichloromethane (10 mL), loaded onto the solution using a wet chromatography method, and subjected to column chromatography (120 g silica gel column, 10 mL feed solution injected, flow rate 80 mL / min, ethyl acetate / petroleum ether = 0-85% gradient elution for 50 min). The first peak fraction 1 (900 mg) eluted first, with a LCMS retention time of 1.725 min, and was collected and concentrated. Then the second peak fraction 2 (320 mg) eluted, was collected and concentrated, with a LCMS retention time of 1.752 min. The LCMS analytical conditions were as follows: (Symmetry C18 column, 50*4.6mm, 3.5µm; mobile phase A: water (0.1% formic acid); mobile phase B: acetonitrile; gradient: 0-0.50 min 10% B-75% B, 0.50-1.00 min 75% B-95% B, 1.00-2.20 min 95% B, 2.20-2.21 min 95% B-10% B, 2.21-3.00 min 10% B).

[0298] Component 1 is a mixture of the following isomers:

[0299] LC-MS(ESI):m / z=574.1 / 576.1[M+H-100] +

[0300] 1 H NMR (400MHz, DMSO-d6) δ10.45(s,1H),8.03–7.91(m,2H),7.74(dd,J=8.7,2.1Hz,1H),5.61(dd,J=8.7,5.3Hz,1H),3. 83–3.62(m,2H),3.51–3.35(m,3H),3.20–2.75(m,4H),2.61(m,1H),2.40(m,1H),1.48(d,J=7.0Hz,3H),1.43(s,9H).

[0301] Component 2 is a mixture of the following isomers:

[0302] LC-MS(ESI):m / z=574.1 / 576.1[M+H-100] +

[0303] 1H NMR (400MHz, DMSO-d6) δ10.49(s,1H),8.02–7.90(m,2H),7.78–7.66(m,1H),5.53(dd,J=10.5,2.2Hz,1H),3. 72–3.67(m,2H),3.48–3.29(m,3H),3.20–2.72(m,5H),2.15–2.11(m,1H),1.43(s,9H),1.39(d,J=7.2Hz,3H).

[0304] Component 2 (9.0 g) was separated by chiral SFC (DAICEL CHIRALCEL OD-3 column, 0.46 cm * 5 cm; CO2: methanol = 60:40 as mobile phase) and two peaks were obtained. The first peak was intermediate 1-8-S (4.79 g, RT = 2.241 min, ee%: 100.0) and the second peak was intermediate 1-8-R (4.22 g, RT = 3.362 min, ee%: 99.9).

[0305] Synthesis of control molecules

[0306] Synthesis of ref-A

[0307] Referring to the preparation method 2 in Example 4A, ref-A was prepared using 1-8-R as the raw material (ref-A is the “control molecule” used in the following biological test examples).

[0308] LC-MS (ESI): m / z = 714.1 [M+H] +

[0309] 1H NMR (400MHz, DMSO-d6)

[0310] δ10.52(s,1H),8.46(s,1H),8.04–7.85(m,2H),7.74(dd,J=8.7,2.1Hz,1H),6.7 9(d,J=2.6Hz,1H),5.53(dd,J=10.3,2.3Hz,1H),4.57–4.35(m,2H),4.31–4.20( m,2H),3.79(t,J=6.1Hz,2H),3.75–3.64(m,2H),3.44–3.39(m,3H),3.25–3.08( m,4H),3.05–2.94(m,2H),2.41(s,3H),2.15–2.07(m,1H),1.41(d,J=7.2Hz,3H).

[0311] Synthesis of ref-B

[0312] Referring to Example 4B, ref-B was prepared from intermediate 1-8-S (see the preparation of 1-8-R, which is the first peak obtained by further chiral separation of component 2 after chiral separation of compound 1-6 to obtain component 2, RT = 2.241 min).

[0313] LC-MS (ESI): m / z = 714.1 [M+H] +

[0314] 1H NMR (400MHz, DMSO-d6)

[0315] δ10.52(s,1H),8.46(s,1H),8.04–7.85(m,2H),7.74(dd,J=8.7,2.1Hz,1H),6.7 9(d,J=2.6Hz,1H),5.53(dd,J=10.3,2.3Hz,1H),4.57–4.35(m,2H),4.31–4.20( m,2H),3.79(t,J=6.1Hz,2H),3.75–3.64(m,2H),3.44–3.39(m,3H),3.25–3.08( m,4H),3.05–2.94(m,2H),2.41(s,3H),2.15–2.07(m,1H),1.41(d,J=7.2Hz,3H).

[0316] Synthesis of ref-CD

[0317] Referring to Example 4CD, ref-CD was prepared using component 1 (see preparation of 1-8-R, LCMS retention time of 1.725 min) initially separated from compounds 1-6 as the starting material.

[0318] LC-MS (ESI): m / z = 714.1 [M+H] +

[0319] 1H NMR (400MHz, DMSO-d6)

[0320] δ10.47(s,1H),8.57(s,1H),8.03–7.90(m,2H),7.74(dd,J=8.6,2.1Hz,1H), 6.77(s,1H),5.61(dd,J=8.6,5.3Hz,1H),4.60–3.35(m,1H),4.27–4.20(m,2 H),3.84–3.76(m,2H),3.75–3.67(m,1H),3.47–3.32(m,4H),3.28–2.98(m,4 H),2.72–2.54(m,2H),2.44(s,3H),2.39–2.31(m,1H),1.50(d,J=6.9Hz,3H).

[0321] According to Example 14 and the activity data described in WO2024079623A, the stereoconfiguration of the most active isomer in Example 14 is (7R, 9R). From the WRN ATP hydrolase inhibitor activity data in Table 1 of Test Example 1 of this disclosure, ref-A is the most active isomer. Therefore, the absolute configuration of ref-A can be determined to be (7R, 9R), and the absolute configuration of compound 1-8-R used to prepare ref-A can also be obtained as (7R, 9R).

[0322] Since compounds 1-6 were separated into component 1 and component 2 by a normal column (i.e. silica gel column), and the absolute configuration of 1-8-R in component 2 is (7R,9R), the absolute configuration of compound 1-8-S can be obtained as (7S,9S). Component 1, which was initially separated from compounds 1-6, is a racemic mixture composed of isomers with absolute configurations of (7R,9S) and (7S,9R).

[0323] Example 1: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin)-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide (Compound 1)

[0324] Step 1: Synthesis of Compound 1-1. Starting material SM (15.0 g) was dissolved in dichloromethane (150 mL). Potassium iodide (5.83 g), 2-(chloromethoxy)ethyltrimethylsilane (7.02 g), and N,N-diisopropylethylamine (15.12 g) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 50 °C for 2 hours. The reaction was monitored by LCMS until completion. The reaction solution was cooled to room temperature, filtered, concentrated, and subjected to column chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain compound 1-1 (7.0 g).

[0325] Step 2: Synthesis of Compounds 1-2

[0326] Compound 1-1 (4.42 g) was dissolved in tetrahydrofuran (80 mL), and nitrogen gas was introduced. Sodium bis(trimethylsilyl)amino (9.51 mL, 1 mol / L) was added at -78 °C. After the addition was complete, the reaction mixture was incubated at -78 °C for 2 hours, followed by the addition of allyl iodine (4.42 g). The reaction was monitored by LCMS until completion. The reaction was quenched by adding saturated ammonium chloride aqueous solution. Extraction was performed at 0 °C with ethyl acetate (200 mL) and water (100 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (ethyl acetate / petroleum ether = 0-30%) to give compound 1-2 (3.8 g).

[0327] Step 3: Synthesis of Compounds 1-3

[0328] Compounds 1-2 (3.8 g) were dissolved in dichloromethane (50 mL), and peracetic acid (24.18 g, 40%) solution was added dropwise at 0 °C. The reaction was carried out at 0 °C for 10 minutes, and then allowed to return to room temperature for 15 hours. The reaction was monitored by LCMS until completion. The reaction solution was then slowly added dropwise to a mixed solution of saturated sodium bicarbonate aqueous solution (100 mL), saturated sodium thiosulfate aqueous solution (120 mL), and dichloromethane (100 mL) at 0 °C and stirred for 10 minutes for extraction. The organic layer was separated by adding saturated sodium thiosulfate (100 mL) and stirring for 10 minutes. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain compounds 1-3 (2.8 g).

[0329] Step 4: Synthesis of Compounds 1-4

[0330] Compounds 1-3 (2.8 g) were dissolved in a mixed solution of water (1.0 mL) and N,N-dimethylformamide (50.0 mL), and potassium fluoride (1.33 g) was added. The reaction was then carried out at 60 °C for 18 h. The reaction solution was diluted with water (50.0 mL) and extracted with ethyl acetate (80.0 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (ethyl acetate / petroleum ether = 0-75%) to give compounds 1-4 (1.0 g).

[0331] Step 5: Synthesis of Compounds 1-5

[0332] Compounds 1-4 (500 mg) were dispersed in dichloromethane (15 mL). Sodium bicarbonate aqueous solution (16 mL, 5%), potassium bromide (12.31 mg), methyltrioctylammonium chloride (20.09 mg), and 2,2,6,6-tetramethylpiperidine oxide (1.64 mg) were added at 0 °C. Sodium hypochlorite aqueous solution (3.85 g) was slowly added dropwise. The reaction mixture was incubated at 0 °C for 0.5 hours. The reaction was monitored by LCMS until completion. Dichloromethane (20 mL) was added to the reaction mixture, and 5 N hydrochloric acid aqueous solution was slowly added dropwise until the pH reached 3. The mixture was stirred for 5 minutes to separate the phases. The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain compounds 1-5 (530 mg, crude product).

[0333] Step 6: Synthesis of Compounds 1-6

[0334] Compounds 1-5 (500 mg) were dissolved in pyridine (8 mL), and 3-chloro-4-aminotrifluorotoluene (393.23 mg) was added. After the addition was complete, under nitrogen protection, phosphorus oxychloride (402.45 mg) reaction solution was added at 0 °C and the reaction was carried out at 0 °C for 1 hour. The reaction was monitored by LCMS until completion. The mixture was extracted with saturated sodium bicarbonate aqueous solution (10 mL) and ethyl acetate (20 mL) at 0 °C. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated, and subjected to column chromatography (ethyl acetate / petroleum ether = 0-75%) to give compounds 1-6 (380 mg).

[0335] Step 7: Synthesis of Compounds 1-7

[0336] Compounds 1-6 (300 mg) were dissolved in dichloromethane (7.5 mL), and trifluoroacetic acid (1.28 g) was added at 0 °C. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The residue was extracted with saturated sodium carbonate aqueous solution (10 mL) at 0 °C. The organic phase was dried over anhydrous sodium sulfate and concentrated to give compounds 1-7 (320 mg).

[0337] Step 8: Synthesis of Compounds 1-8

[0338] Compounds 1-7 (320 mg) were dissolved in N,N-dimethylformamide (8 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (106.18 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (237.52 mg), and 1-hydroxybenzotriazole (77.58 mg) were added. The reaction mixture was reacted at 25 °C for 2 hours. After the reaction was monitored by LCMS, water (10 mL) was added, followed by extraction with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated, and then subjected to column chromatography (methanol / dichloromethane = 0-10%) to give compounds 1-8 (300 mg).

[0339] Step Nine: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin)-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 1) Compounds 1-8 (150 mg) were dissolved in 1 In a mixed solution of 4-dioxane (5 mL) and water (0.8 mL), 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoborhexacyclopentan-2-yl)-1,2,3,6-tetrahydropyridine (63.09 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (25.85 mg), and potassium phosphate (134.37 mg) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 90 °C for 4 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-7%) to obtain the crude product. Further preparation was performed (column: YMC C). 18 Mobile phase: A was 0.7% NH4HCO3 aqueous solution; B%: 55%, B was acetonitrile, 25 mL / min) Purification yielded 60 mg of the title compound.

[0340] LC-MS (ESI): m / z = 753.2 [M+H] +

[0341] 1H NMR(400MHz,DMSO-d6)δ10.49(s,1H),8.51(s,1H),8.04–7.89(m,2H),7.76–7 .72(m,1H),6.75–6.71(m,1H),5.62–5.51(m,1H),4.44(s,1H),3.73–3.66(m,3 H),3.25(m,6H),3.03–2.95(m,2H),2.82–2.74(m,3H),2.63–2.56(m,1H),2.42 –2.33(s,4H),1.73(m,1H),1.50–1.40(m,3H),0.44(m,2H),0.36–0.33(m,2H).

[0342] Step 10: Synthesis of Compound 1A

[0343] Compound 1 (56 mg) was chirally resolved (DAICEL CHIRALCEL OD-3 column, 100*4.6 mm ID, 3 μm; mobile phase: 40% ethanol (0.05% DEA) in CO2) to give compound 1B (peak 3, 12.10 mg, RT = 1.626 min, ee% = 97%) and compound 1A (peak 4, 6.50 mg, RT = 4.782 min, ee% = 99.9%). Peak 1 (RT = 0.961 min) and peak 2 (RT = 1.037 min) were collected together.

[0344] Data for compound 1A:

[0345] LC-MS (ESI): m / z = 753.2 [M+H] +

[0346] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.49(s,1H),8.05–7.85(m,2H),7.74(dd,J= 8.7,2.1Hz,1H),6.82–6.67(m,1H),5.52(dd,J=10.4,2.2Hz,1H),4.61–4.29(m,1H) ,3.71(m,2H),3.41–3.36(m,5H),3.27–3.22(m,3H),2.99(m,2H),2.78(m,3H),2.42 (s,3H),2.11(m,1H),1.73(m,1H),1.40(d,J=7.2Hz,3H),0.44(m,2H),0.35(m,2H).

[0347] Data for compound 1B:

[0348] LC-MS (ESI): m / z = 753.2 [M+H] +

[0349] The mixture of peak1 and peak2 was subjected to a second chiral resolution (Chiralpak AD-3 column, 100*4.6mm ID, 3µm; mobile phase: 40% isopropanol (0.05% DEA) in CO2) to obtain compound 1D (peak1, 3.30 mg, RT = 1.423 min, ee% = 98.2%) and compound 1C (peak2, 8.10 mg, RT = 1.811 min, ee% = 96.7%).

[0350] Data for compound 1C:

[0351] LC-MS (ESI): m / z = 753.2 [M+H] +

[0352] Data for compound 1D:

[0353] LC-MS (ESI): m / z = 753.2 [M+H] +

[0354] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.49(s,1H),8.05–7.85(m,2H),7.74(dd,J= 8.7,2.1Hz,1H),6.82–6.67(m,1H),5.52(dd,J=10.4,2.2Hz,1H),4.61–4.29(m,1H) ,3.71(m,2H),3.41–3.36(m,5H),3.27–3.22(m,3H),2.99(m,2H),2.78(m,3H),2.42 (s,3H),2.11(m,1H),1.73(m,1H),1.40(d,J=7.2Hz,3H),0.44(m,2H),0.35(m,2H).

[0355] Example 1a: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0356] Step 1, Synthesis of 1a-1

[0357] 1-8-R (500 mg, 740.85 μmol) was dissolved in a mixture of dioxane (50 mL) and water (10 mL). At room temperature, 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1,2,3,6-tetrahydropyridine (276.88 mg, 1.11 mmol), tetraphenylphosphine palladium (128.46 mg, 111.13 μmol), and cesium fluoride (1.69 g, 11.11 mmol) were added. After addition, the mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. After cooling to room temperature, ethyl acetate (50 mL) and saturated brine (20 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (methanol / dichloromethane = 0.7%) to give 1a-1 (430 mg).

[0358] Step 2, Synthesis of 1a-2

[0359] 1a-1 (400 mg, 558.44 μmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (3.34 g, 29.28 mmol) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a 10 / 1 dichloromethane / methanol mixture (40 mL). The mixture was extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The aqueous layer was then extracted again with a 10 / 1 dichloromethane / methanol mixture (20 mL). The combined organic phases were dried and concentrated to obtain compound 1a-2 (334 mg).

[0360] Step 3, Synthesis of 1a

[0361] 1a-2 (320 mg, 518.69 μmol) was dissolved in dichloromethane (10 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (103.90 mg, 674.16 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (214.54 mg, 1.09 mmol), and 1-hydroxybenzotriazole (70.14 mg, 519.06 μmol) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 in H2O; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 1a (200 mg).

[0362] LC-MS (ESI): m / z = 753.2 [M+H]+

[0363] 1 H NMR (400MHz, DMSO-d6) δ10.51(s,1H),8.49(s,1H),8.05–7.85(m,2H),7.74(dd,J= 8.7,2.1Hz,1H),6.82–6.67(m,1H),5.52(dd,J=10.4,2.2Hz,1H),4.61–4.29(m,1H) ,3.71(m,2H),3.41–3.36(m,5H),3.27–3.22(m,3H),2.99(m,2H),2.78(m,3H),2.42 (s,3H),2.11(m,1H),1.73(m,1H),1.40(d,J=7.2Hz,3H),0.44(m,2H),0.35(m,2H).

[0364] Compound 1a, prepared from intermediate 1-8-R, and compound 1A, obtained by chiral resolution of compound 1 in Example 1, were identified as the same compound by liquid chromatography and activity testing, with an absolute configuration of (7R, 9R). See below for details:

[0365] Compounds 1a and 1A were mixed and separated by liquid chromatography. The liquid chromatography conditions were: YMC-Triart-C18 column, 100*4.6mm, phase A 0.1% H3PO4 in H2O, phase B acetonitrile, flow rate 1.0mL / min, phase B elution 10-90% for 15min. The results showed that 1a and 1A had the same retention time, retention time: RT = 6.959min.

[0366] Compounds 1a, 1A, and HRO761 were subjected to WRN ATP hydrolase inhibition experiments in the same batch using the method described in Test Example 1. The results are shown below:

[0367] Test results show that compounds 1a and 1A have comparable activity and are the same compound with the same absolute configuration.

[0368] Example 2: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-3-fluorophenyl)-5-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide (compound 2)

[0369] Step 1: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-6-bromo-5-ethyl-2,4-dihydro-7H-[1,2,3]triazolo[4,5-b]pyridin-7-one

[0370] The starting intermediate Int1-9 (960 mg, 2.63 mmol) was dissolved in toluene (5 mL), and ethylene glycol (327 mg, 5.26 mmol) and p-toluenesulfonic acid (91 mg, 525.80 μmol) were added. The reaction was carried out at 100 °C for 5 hours, and the reaction was monitored by LCMS until completion. A solid precipitated out, and 10 mL of petroleum ether was added to slurry the mixture. The mixture was filtered, and the filter cake was evaporated to dryness to obtain compound 2-1 (1 g).

[0371] Step 2: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-6-bromo-5-ethyl-4-((2-(trimethylsilyl)ethoxy)methyl)-2,4-dihydro-7H-[1,2,3]triazolo[4,5-b]pyridin-7-one

[0372] Compound 2-1 (1.08 g, 2.64 mmol) was dissolved in dichloromethane (10 mL), and N,N-diisopropylethylamine (1.02 g, 7.92 mmol, 1.38 mL), potassium iodide (438 mg, 2.64 mmol), and 2-(trimethylsilyl)ethoxymethyl chloride (660 mg, 3.96 mmol) were added. The mixture was heated to 50 °C and reacted for 10 hours, with the reaction monitored by LC-MS until completion. The reaction mixture was evaporated to dryness with silica gel and purified by column chromatography using petroleum ether:ethyl acetate (5:1). Compound 2-2 (820 mg) was obtained.

[0373] Step 3: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-6-bromo-5-(pent-4-en-2-yl)-4-((2-(trimethylsilyl)ethoxy)methyl)-2,4-dihydro-7H-[1,2,3]triazolo[4,5-b]pyridin-7-one

[0374] Compound 2-2 (820 mg, 1.52 mmol) was dissolved in tetrahydrofuran (10 mL) under argon protection and cooled to -78 °C. Lithium bis(trimethylsilyl)amino (1 M, 7.60 mL, 20% purity) was added dropwise, and the mixture was stirred for 2 hours. Allyl iodine (1.28 g, 7.60 mmol) was then added dropwise. The mixture was then heated to -20 °C and reacted for 10 hours. LCMS monitoring showed that the starting material was not completely reacted, so the mixture was heated to 0 °C and reacted for another 10 hours. LMCS monitoring showed that the reaction was complete. Partial deprotection occurred. The system was quenched with saturated ammonium chloride aqueous solution, and 20 mL of water was added. The mixture was extracted with 40 mL of ethyl acetate, separated, and the organic phase was washed once with saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure. The solution was then subjected to column chromatography with petroleum ether:ethyl acetate (5:1 ratio) to obtain compound 2-3 (440 mg, 50% yield).

[0375] Step 4: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-6-bromo-5-(1-(epoxyethylene-2-yl)propan-2-yl)-4-((2-(trimethylsilyl)ethoxy)methyl)-2,4-dihydro-7H-[1,2,3]triazolo[4,5-b]pyridin-7-one

[0376] Compound 2-3 (440 mg, 759.23 μmol) was dissolved in 10 mL of dichloromethane, and peracetic acid (3.85 g, 15.18 mmol, 30% purity) was added at 0 °C. The mixture was allowed to rise naturally to room temperature for 2 days, and the reaction was monitored by LC-MS until completion. 20 mL of dichloromethane was added to the mixture, and the mixture was separated. The organic phase was adjusted to neutral pH with saturated sodium bicarbonate solution, quenched with saturated sodium thiosulfate solution, and separated again. The aqueous phase was extracted with 20 mL of dichloromethane, separated again, and the organic phase was washed once with saturated sodium chloride solution, separated again, and dried over anhydrous sodium sulfate. The mixture was filtered and evaporated to dryness. Compound 2-4 (370 mg) was obtained.

[0377] Step 5: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-5-bromo-8-(hydroxymethyl)-6-methyl-7,8-dihydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-4(6H)-one

[0378] Compound 2-4 (371 mg, 622.97 μmol) was dissolved in N,N-dimethylformamide (20 mL), and a solution of potassium fluoride (361.95 mg, 6.23 mmol) in water (4 mL) was added. The mixture was heated to 60 °C and reacted for 3 days. LC-MS monitoring showed that approximately 10% of the starting material remained unreacted. The system was cooled, and 40 mL of water was added. The mixture was extracted with 200 mL of methyl tert-butyl ether, separated, and the organic phase was washed once with saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain compound 2-5 (164 mg).

[0379] Step Six: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-5-bromo-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxylic acid

[0380] Compound 2-5 (164 mg, 352.48 μmol) was dissolved in dichloromethane (5 mL). At 0 °C, an aqueous solution of sodium bicarbonate (5.92 g, 3.52 mmol, 5% purity), potassium bromide (4.2 mg, 35.25 μmol), methyltrioctylammonium chloride (72 mg, 176.24 μmol), and 2,2,6,6-tetramethylpiperidine N-oxide (5.5 mg, 35.25 μmol) were added dropwise. Sodium hypochlorite (660.90 mg, 881.20 μmol, 10% purity) was added slowly, and the reaction was carried out at 0 °C for 30 min. The reaction was monitored by LC-MS until completion. The pH of the system was adjusted to 3 with dilute hydrochloric acid, and the mixture was extracted with 20 mL of dichloromethane. The liquid was separated, and the organic phase was washed once with saturated sodium chloride aqueous solution. The liquid was then separated again, dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. Compound 2-6 (210 mg, crude product) was obtained.

[0381] Step 7: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-5-bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide

[0382] Compound 2-6 (180 mg, 375.58 μmol) was dissolved in acetonitrile (5 mL), and N,N,N',N'-tetramethylchloromethanemidane hexafluorophosphate (317 mg, 1.13 mmol) and 2,4,6-trimethylpyridine (227 mg, 1.88 mmol) were added. The mixture was stirred for 5 min, and then 2-chloro-4-(trifluoromethyl)aniline (111 mg, 563.37 μmol) was added. The reaction was brought to 60 °C and carried out for 3 h, with the reaction monitored by LC-MS until completion. 20 mL of water was added to the system, and the mixture was extracted with 40 mL of ethyl acetate. The liquid phase was separated, washed once with saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate. The mixture was filtered, and the solvent was removed under reduced pressure. The solution was then subjected to column chromatography with dichloromethane:methanol = 50:1 to give compound 2-7 (80 mg, crude product).

[0383] Step 8: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-N-(2-chloro-4-(trifluoromethyl)phenyl)-6-methyl-4-oxo-5-(piperazin-1-yl)-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide

[0384] Compound 2-7 (70 mg, 106.58 μmol) was dissolved in N-methylpyrrolidone (4 mL), and piperazine (184 mg, 2.13 mmol) was added. The reaction was carried out under argon protection and microwaved at 140 °C for 45 min. The reaction was monitored by LC-MS until completion. 20 mL of water was added to the system, and the mixture was extracted with 40 mL of ethyl acetate. The liquid phase was separated, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. Compound 2-8 (50 mg, crude product) was obtained.

[0385] LC-MS (ESI): m / z = 662.2 [M+H] +

[0386] Step Nine: Synthesis of 2-(4-(1,3-dioxacyclopentan-2-yl)-3-fluorophenyl)-N-(2-chloro-4-(trifluoromethyl)phenyl)-5-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide. Compound 2-8 (50 mg, 75.52 μmol) was dissolved in N,N-dimethylformamide (2 mL), and 5-hydroxy-6-methyl-pyrimidin-4-carboxylic acid (12 mg, 75.52 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (29 mg, 151.05 μmol), and 1-hydroxybenzotriazole (2 mg, 15.10 μmol) were added. The reaction was carried out at room temperature for 1 hour, and the reaction was monitored by LCMS until completion. 20 mL of water was added to the system, and the mixture was extracted with 40 mL of ethyl acetate. The liquid phase was separated, washed once with saturated sodium chloride aqueous solution, and dried over anhydrous sodium sulfate. The mixture was filtered, and the solvent was removed under reduced pressure. The solution was then subjected to column chromatography with dichloromethane:methanol (50:1). Compound 2-9 (20 mg, crude product) was obtained.

[0387] Step 10: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(3-fluoro-4-formylphenyl)-5-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide

[0388] Compound 2-9 (10 mg, 12.53 μmol) was dissolved in tetrahydrofuran (1 mL), and hydrochloric acid (3 M, 1 mL) was added. The reaction was heated to 55 °C and carried out for 15 hours, with the reaction monitored by LC-MS until completion. 3 mL of dichloromethane was added to the system, and the pH was adjusted to neutral with 3N sodium bicarbonate aqueous solution. 5 mL of water was added to the system, and the mixture was extracted with 20 mL of dichloromethane. The liquid-liquid phase was separated, washed once with saturated sodium chloride aqueous solution, separated again, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated under reduced pressure to obtain compound 2-10 (6 mg, crude product).

[0389] Step 11: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-3-fluorophenyl)-5-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-6-methyl-4-oxo-4,6,7,8-tetrahydro-2H-[1,2,3]triazolo[4,5-e]indoleazine-8-carboxamide

[0390] Dimethylamine hydrochloride (5 mg, 61.3 μmol) was dissolved in tetrahydrofuran (2 mL), and triethylamine (7 mg, 66.31 μmol, 9.25 μL) was added. The mixture was stirred for 15 minutes, followed by the addition of acetic acid (6 mg, 99.9 μmol) and compound 2-10 (6 mg, 7.96 μmol). The mixture was stirred for 5 minutes. Sodium cyanoborohydride (4 mg, 66.31 μmol) was then added. The reaction was allowed to proceed at room temperature for 1 hour, and the reaction was monitored by LC-MS until completion. 20 mL of water was added to the system, and the mixture was extracted with 40 mL of dichloromethane. The liquid phase was separated, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The solution was then purified by column chromatography using dichloromethane:methanol at a ratio of 50:1 to 10:1. Compound 2 (5 mg) was obtained.

[0391] LC-MS (ESI): m / z = 783.2 [M+H] +

[0392] 1 H NMR(400MHz, CDCl3)δ8.52(s,1H),7.90–7.86(m,3H),7.47–7.45(m,2H),7.42–7.38(m,1H),5.66(m,1H),3.81–3.80(m,1H),3.73–3.72 (m,1H),3.42(m,3H),2.95–2.94(m,2H),2.73–2.70(m,4H),2.44–2.41(m,3H),2.40–2.38(m,2H),2.18–2.13(m,6H),1.61–1.59(m,3H).

[0393] Example 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-3-fluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 3)

[0394] Compounds 1-8 (150 mg) were dissolved in a mixture of 1,4-dioxane (5 mL) and water (0.8 mL). At room temperature, (4-((dimethylamino)methyl)-3-fluoro-phenyl)boronic acid (49.88 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (25.85 mg), and potassium phosphate (134.37 mg) were added. After addition, the mixture was reacted under nitrogen protection at 90 °C for 4 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparative chromatography (column: YMC C) was performed. 18 Mobile phase: A was 0.7% NH4HCO3 aqueous solution; B%: 55%, B was acetonitrile, 25 mL / min) to obtain 50 mg of the title compound after purification.

[0395] LC-MS (ESI): m / z = 783.2 [M+H] +

[0396] 1 H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.52(s,1H),8.03–7.86(m,3H),7.79–7.67(m,2H),7.57–7.52(m,1H),5.70–5.59(m,1H),4.43(s,1H),3. 79–3.74(m,2H),3.49(m,5H),3.07–2.99(m,3H),2.68–2.59(m,1H),2.5 6–2.52(m,1H),2.47–2.39(m,4H),2.27–2.08(m,6H),1.53–1.43(m,3H).

[0397] Example 3A: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-3-fluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 3A)

[0398] The title compound was prepared from intermediate 1-8-R and (4-((dimethylamino)methyl)-3-fluoro-phenyl)boronic acid according to the method of Example 19. Since the absolute configuration of intermediate 1-8-R is (7R,9R), the absolute configuration of compound 3A is (7R,9R).

[0399] LC-MS (ESI): m / z = 783.2 [M+H]+

[0400] 1 H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.53(s,1H),8.03–7.86(m,3H),7.79–7.67(m,2H),7.57–7.52(m,1H),5.70–5.59(m,1H),3.79–3.74(m ,2H),3.49(m,4H),3.80-3.75(m,2H),3.07–2.99(m,3H),2.68–2.59(m ,1H),2.56–2.52(m,1H),2.46(s,3H),2.24(s,6H),1.50–1.48(m,3H).

[0401] Example 4: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazole[1,5-a]pyrimidin-9-carboxamide (compound 4)

[0402] Compounds 1-8 (100 mg) were dissolved in a mixture of dioxane (4 mL) and water (0.5 mL). At room temperature, 1-(3,3-difluorocyclobutyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1,2,3,6-tetrahydropyridine Int4-3 (168.33 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (17.23 mg), and potassium phosphate (89.58 mg) were added. After addition, the reaction mixture was kept under nitrogen protection and reacted at 90 °C for 4 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparative chromatography purification (column: YMC C) was performed. 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 64%, B is acetonitrile, 25 mL / min) to obtain 40 mg of the title compound.

[0403] LC-MS (ESI): m / z = 803.3 [M+H] +

[0404] 1H NMR (400MHz, DMSO-d6) δ10.50(s,1H),8.52(s,1H),8.04–7.88(m,2H),7.73(dd,J=8.7,2.2Hz,1H),6.75–6.67(m,1H),5.61–5.50(m,1H),4.65–4 .30(m,1H),3.73–3.66(m,2H),3.20–3.14(m,1H),3.11–2.95(m,5H),2.8 1–2.67(m,5H),2.61–2.52(m,5H),2.47–2.32(m,5H),1.50–1.39(m,3H).

[0405] Example 4A: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazol[1,5-a]pyrimidin-9-carboxamide (compound 4A)

[0406] Preparation method 1:

[0407] Step 1: Synthesis of 4A-1

[0408] 1-8-R (700 mg) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (593 mg) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a 10 / 1 dichloromethane / methanol mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The aqueous layer was extracted again with a 10 / 1 dichloromethane / methanol mixture (20 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give compound 4A-1 (584 mg).

[0409] Step 2, Synthesis of 4A-2

[0410] 4A-1 (584 mg) was dissolved in dichloromethane (20 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (204 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (391 mg), and 1-hydroxybenzotriazole (138 mg) were added. The reaction solution was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-7%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 in H2O; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 4A-2 (405 mg).

[0411] Step 3: Synthesis of 4A

[0412] Compound 4A-2 (400 mg) was dissolved in a mixed solution of dioxane (16 mL) and water (2 mL). At room temperature, 1-(3,3-difluorocyclobutyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1,2,3,6-tetrahydropyridine Int4-3 (673.32 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (68.92 mg), and potassium phosphate (358.32 mg) were added. After addition, the reaction mixture was kept under nitrogen protection and reacted at 90 °C for 4 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparative chromatography purification (column: YMC C) was performed. 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 64%, B is acetonitrile, 25 mL / min) to obtain 161 mg of the title compound.

[0413] LC-MS (ESI): m / z = 803.3 [M+H] +

[0414] 1 H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.50(s,1H),8.03–7.93(m,2H),7.75-7.73( d,J=8.0,1H),6.75–6.73(m,1H),5.54–5.51(m,1H),4.43(m,1H),3.75–3.67(m,2H) ,3.55–3.49(m,3H),3.25(s,1H),3.07–3.04(m,3H),3.03-2.95(m,2H),2.68–2.59( m,4H),2.66-2.52(m,4H),2.47–2.39(m,4H),2.13-2.10(m,1H),1.51–1.47(m,3H).

[0415] Preparation method two:

[0416] Step 1: Synthesis of 4A-3

[0417] 1-8-R (700.0 mg, 1.04 mmol) was dissolved in a mixture of dioxane (50 mL) and water (10 mL). At room temperature, 1-(3,3-difluorocyclobutyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1,2,3,6-tetrahydropyridine Int4-3 (930.87 mg, 3.12 mmol), tetraphenylphosphine palladium (179.85 mg, 155.58 μmol), and cesium fluoride (2.36 g, 15.56 mmol) were added. After the addition was complete, the reaction mixture was incubated under nitrogen protection at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. Cool to room temperature, add ethyl acetate (100 mL) and saturated brine (50 mL) for extraction, concentrate the organic layer, and perform column chromatography (methanol / dichloromethane = 0-5%) to obtain 4A-3 (700 mg).

[0418] LC-MS (ESI): m / z = 767.2 [M+H] +

[0419] Step 2: Synthesis of 4A-4

[0420] 4A-3 (700 mg, 912.42 μmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (520 mg, 4.56 mmol) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a 10 / 1 dichloromethane / methanol mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The aqueous layer was extracted again with a 10 / 1 dichloromethane / methanol mixture (20 mL). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to give compound 4A-4 (601 mg).

[0421] LC-MS (ESI): m / z = 667.2 [M+H] +

[0422] Step 3: Synthesis of 4A

[0423] 4A-4 (597 mg, 896.74 μmol) was dissolved in dichloromethane (20 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (180.2 mg, 1.17 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (343.1 mg, 1.79 mmol), and 1-hydroxybenzotriazole (121.2 mg, 896.74 μmol) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-7%) to obtain a yellow solid. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 in H2O; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 4A (403 mg).

[0424] LC-MS (ESI): m / z = 803.2 [M+H] +

[0425] 1 H NMR(400MHz,DMSOd6)δ10.51(s,1H),8.50(s,1H),8.03–7.93(m,2H),7.75–7.73(d, J=8.0Hz,1H),6.75–6.73(m,1H),5.54–5.51(m,1H),4.43(m,1H),3.75–3.67(m,2H) ,3.55-3.49(m,3H),3.25(s,1H),3.07–3.04(m,3H),3.03–2.95(m,2H),2.68–2.59( m,4H),2.66–2.52(m,4H),2.47–2.39(m,4H),2.13–2.10(m,1H),1.51–1.47(m,3H).

[0426] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of the compound 4A prepared from it is also (7R, 9R).

[0427] Example 4B: Synthesis of (7S,9S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0428] Following the synthesis method of preparation method 2 of 4A and the above route, 4B was prepared using 1-8-S (see the preparation of 1-8-R, which is the first peak obtained by further chiral separation of component 2 after chiral resolution of compound 1-6, RT = 2.241 min) as raw material.

[0429] LC-MS (ESI): m / z = 803.2 [M+H] +

[0430] 1H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.50(s,1H),8.03–7.93(m,2H),7.75–7.73(d ,J=8.0Hz,1H),6.75–6.73(m,1H),5.54–5.51(m,1H),4.43(m,1H),3.75–3.67(m,2H ),3.55-3.49(m,3H),3.25(s,1H),3.07–3.04(m,3H),3.03–2.95(m,2H),2.68–2.59 (m,4H),2.66–2.52(m,4H),2.47–2.39(m,4H),2.13–2.10(m,1H),1.51–1.47(m,3H).

[0431] Since the absolute configuration of intermediate 1-8-S is (7S, 9S), the absolute configuration of 4B obtained from it is (7S, 9S).

[0432] Example 4CD: (7S,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxyl A mixture of amine and (7R,9S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0433] Following the synthesis method of preparation method 2 of 4A and the above route, 4CD was prepared using component 1 initially separated from compounds 1-6 (see preparation of intermediate 1-8-R, LCMS retention time is 1.725 min) as raw material.

[0434] LC-MS (ESI): m / z = 803.2 [M+H] +

[0435] 1H NMR (400MHz, DMSO-d6) δ10.47(s,1H),8.57(s,1H),7.99–7.89(m,2H),7.73(dd,J=8. 8,2.1Hz,1H),6.79–6.67(m,1H),5.60(dd,J=8.6,5.3Hz,1H),4.63–4.18(m,1H),3.90 –2.58(m,2H),3.50–3.34(m,3H),3.24–3.18(m,1H),3.14–2.98(m,3H),2.87–2.65(m ,4H),2.63–2.58(m,1H),2.57–2.52(m,5H),2.47–2.34(m,5H),1.50(d,J=7.0Hz,3H).

[0436] Since component 1, which was initially isolated from compounds 1-6, is a racemic mixture composed of isomers with absolute configurations of (7R,9S) and (7S,9R), the 4CD obtained from it is a racemic mixture composed of isomers with absolute configurations of (7R,9S) and (7S,9R).

[0437] Example 5: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-6-oxo-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 5)

[0438] Step 1: Synthesis of Compound 5-2

[0439] Compound 5-1 (100 mg) and 4-dimethylaminopyridine (15.95 mg) were dissolved in dichloromethane (5 mL), and triethylamine (198.18 mg) was added. Under argon protection, trifluoromethanesulfonic anhydride (276.29 mg) was slowly added dropwise at 0 °C, and the mixture was stirred for 10 minutes. The temperature was raised to 25 °C and stirred for 3 hours. The reaction solution was subjected to column chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain compound 5-2 (115 mg).

[0440] Step 2: Synthesis of Compound 5-3

[0441] Compound 5-2 (110 mg) and pinacol diborate (117.51 ​​mg) were dissolved in dioxane (5 mL). Potassium acetate (113.38 mg) and 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (15.75 mg) were added at room temperature. After the addition was complete, the reaction mixture was reacted at 90 °C for 12 hours under nitrogen protection. After cooling to room temperature, the mixture was filtered and used directly in the next step.

[0442] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-6-oxo-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0443] Compounds 1-8 (100 mg) were dissolved in a mixed solution of dioxane (5 mL) and water (0.5 mL). Compounds 5-3 (74.03 mg) in a dioxane solution (2 mL), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (17.23 mg), and potassium phosphate (89.58 mg) were added at room temperature. Under nitrogen protection, the reaction mixture was incubated at 90 °C for 4 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was performed (column: YMC C). 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 65%, B is acetonitrile, 25 mL / min) to obtain 20 mg of the title compound.

[0444] LC-MS (ESI): m / z = 767.2 [M+H] +

[0445] 1 H NMR (400MHz, DMSO-d6) δ10.49(s,1H),8.55(s,1H),7.99(d,J=2.0Hz,1H),7.91(d,J=8.5 Hz,1H),7.74(dd,J=8.8,2.0Hz,1H),6.51(d,J=1.7Hz,1H),5.56(dd,J=10.4,2.3Hz,1H) ,4.22(m,1H),3.74(m,1H),3.46(m,3H),3.23-3.18(m,1H),3.00(m,3H),2.81(t,J=6.8H z,2H),2.70(m,2H),2.43(s,3H),2.13(m,1H),1.43(d,J=7.2Hz,3H),0.83–0.53(m,4H).

[0446] Example 6: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((3-fluoroazacyclobutan-1-yl)methyl)phenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 6)

[0447] Step 1: Synthesis of Compound 6-2

[0448] Compound 6-1 (100 mg) and 3-fluorozacriane hydrochloride (115 mg) were dispersed in acetonitrile (5 mL), and potassium carbonate (464.45 mg) was added. The mixture was heated to 70 °C and stirred for 2 hours until the reaction was complete. The mixture was filtered, the filtrate was concentrated, and column chromatography (ethyl acetate / petroleum ether = 0-50%) was performed to give compound 6-2 (80 mg).

[0449] Step 2: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((3-fluoroazacyclobutan-1-yl)methyl)phenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0450] Compounds 1-8 (45 mg) were dissolved in a mixed solution of dioxane (5 mL) and water (0.8 mL). Compounds 6-2 (36.86 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (7.75 mg), and potassium phosphate (40.31 mg) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was performed (column: YMC C). 18 Mobile phase: A was 0.7% NH4HCO3 aqueous solution; B%: 50%, B was acetonitrile, 25 mL / min) to obtain 20 mg of the title compound.

[0451] LC-MS (ESI): m / z = 795.2 [M+H] +

[0452] 1H NMR(400MHz,DMSO-d6)δ10.57(s,1H),8.46(s,1H),8.06–7.90(m,4H),7.74 (d,J=9.3Hz,1H),7.42–7.39(m,2H),5.65–5.49(m,1H),5.28–5.08(m,1H),3 .77–3.71(m,1H),3.68(s,2H),3.60–3.52(m,5H),3.22–3.16(m,2H),3.14–3 .11(m,2H),3.07–2.98(m,2H),2.41(s,3H),2.15(m,1H),1.55–1.43(m,3H).

[0453] Example 6A: (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((3-fluoroazacyclobutane-1-yl)methyl)phenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0454] The title compound was prepared from intermediate 1-8-R and compound 6-2 according to the method of Example 15. Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 6A is (7R, 9R).

[0455] LC-MS (ESI): m / z = 795.2 [M+H] +

[0456] 1H NMR (400MHz, DMSO-d6) δ10.57(s,1H),8.48(s,1H),8.06–7.90(m,4H),7.74(d,J=9.3Hz,1H),7.42–7.38(m,2H),5.65–5.49(m ,1H),5.28–5.08(m,1H),4.54-4.39(m,1H),3.77–3.71(m,1H),3.68(s,2H),3.60–3.50(m,4H),3.28–3.19(m,3H),3.12–3.04 (m,2H), 3.01–2.98(m,3H), 2.41(s,3H), 2.18(m,1H), 1.50–1.46(m,3H). Example 7: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(6-((3-fluoroazacyclobutan-1-yl)methyl)pyridin-3-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 7).

[0457] Step 1: Synthesis of Compound 7-2. Compound 7-1 (200 mg) and 3-fluorozacriane hydrochloride (330.85 mg) were dispersed in acetonitrile (5 mL), and potassium carbonate (802.06 mg) was added. The mixture was heated to 70 °C and stirred for 2 hours until the reaction was complete. The mixture was filtered, the filtrate was concentrated, and column chromatography (ethyl acetate / petroleum ether = 0-50%) was performed to obtain compound 7-2 (220 mg).

[0458] Step 2: Synthesis of Compound 7-3

[0459] Compound 7-2 (90 mg) and pinacol diboronate (186.50 mg) were dissolved in dioxane (5 mL). Potassium acetate (107.96 mg) and tetraphenylphosphine palladium (63.67 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and filtered. The filtrate was used directly in the next step.

[0460] LC-MS(ESI): m / z = 211.1[M+H] + ;

[0461] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(6-(((3-fluoroazacyclobutan-1-yl)methyl)pyridin-3-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide. Compounds 1-8 (40 mg) were dissolved in a mixture of dioxane (4 mL) and water (0.8 mL). At room temperature, a dioxane solution (47.27 mg) of compound 7-3 obtained in Step 2, tetraphenylphosphine palladium (13.79 mg), and potassium phosphate (95.55 mg) were added. After addition, the reaction was carried out under nitrogen protection at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was then performed (column: YMC C). 18 Mobile phase: A was 0.7% NH4HCO3 aqueous solution; B%: 50%, B was acetonitrile, 25 mL / min) to obtain 15 mg of the title compound.

[0462] LC-MS (ESI): m / z = 796.2 [M+H] +

[0463] 1 H NMR(400MHz,DMSO-d6)δ10.58(s,1H),9.13(m,1H),8.46(s,1H),8.39–8.34(m,1H),8 .02–7.90(m,2H),7.75–7.72(m,1H),7.51(d,J=8.2Hz,1H),5.61(m,1H),5.21(m,1H), 3.82(s,2H),3.76(m,1H),3.69–3.53(m,4H),3.30–3.26(m,3H),3.25–3.21(m,2H),3 .20–3.19(m,2H),2.69–2.59(m,1H),2.41(s,3H),2.16(m,1H),1.44(d,J=7.2Hz,3H).

[0464] Example 8: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-2-oxo-1,2-dihydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 8)

[0465] Step 1: Synthesis of Compound 8-1

[0466] 4-Bromopyridin-2-ol (1.8 g), cyclopropylboronic acid (2.0 g), copper acetate (2.0 g), 2,2'-bipyridine (1.7 g), and sodium carbonate (2.5 g) were dissolved in 1,2-dichloroethane (75 mL) and reacted at room temperature in air for 18 h. After the reaction was complete, water (100 mL) and dichloromethane (100 mL) were added to the reaction solution, and the mixture was stirred. The mixture was filtered through diatomaceous earth, the filter cake was washed with dichloromethane, the filtrate was separated into layers, the organic phase was dried over anhydrous sodium sulfate, the solvent was removed by concentration under reduced pressure, and the residue was purified by Flash column chromatography (ethyl acetate: petroleum ether 0-50%) to give compound 8-1 (900 mg).

[0467] Step 2: Synthesis of Compound 8-2

[0468] Compound 8-1 (780 mg), pinacol diboronate (1.4 g), tris(dibenzylacetone)palladium (166.8 mg), tricyclohexylphosphine (102.2 mg), and potassium acetate (715.2 mg) were dissolved in anhydrous dioxane (8 mL). The mixture was heated at 80 °C for 6 h under nitrogen protection. After the reaction was completed, the reaction solution was concentrated to dryness under reduced pressure to obtain crude product 8-2, which was used directly in the next reaction without further purification.

[0469] LC-MS (ESI): m / z = 180.1 [M+H] + .

[0470] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-cyclopropyl-2-oxo-1,2-dihydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0471] Compound 1-8 (40 mg) was dissolved in a mixed solution of dioxane (4 mL) and water (0.8 mL). Compound 8-2 (30.21 mg), tetraphenylphosphine palladium (13.79 mg), and potassium phosphate (95.55 mg) were added at room temperature. After addition, the mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was performed (column: YMC C). 18 Mobile phase: A was 0.7% NH4HCO3 aqueous solution; B%: 60%, B was acetonitrile, 25 mL / min) to obtain 12 mg of the title compound.

[0472] LC-MS (ESI): m / z = 765.1 [M+H] +

[0473] 1 H NMR(400MHz,DMSO-d6)δ10.60(s,1H),8.25(s,1H),8.06–7.86(m,2H),7.79–7.6 3(m,2H),6.95(m,1H),6.77(d,J=7.1Hz,1H),5.58(m,1H),4.60–4.35(m,1H),3.7 6(m,2H),3.65–3.45(m,3H),3.21–3.14(m,1H),3.01(m,2H),2.63(m,1H),2.35( s,3H),2.22–2.09(m,1H),1.44(d,J=7.2Hz,3H),1.07–0.95(m,2H),0.87(m,2H).

[0474] Example 9: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2-fluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 9)

[0475] Step 1: Synthesis of Compound 9-2

[0476] Compound 9-1 (200 mg) and dimethylamine hydrochloride (186 mg) were dispersed in acetonitrile (5 mL), and potassium carbonate (619 mg) was added. The mixture was heated to 70 °C and stirred for 4 hours until the reaction was complete. The mixture was filtered, the filtrate was concentrated, and column chromatography (ethyl acetate / petroleum ether = 0-50%) was performed to give compound 9-2 (80 mg).

[0477] Step 2: Synthesis of Compound 9-3

[0478] Compound 9-2 (40 mg) and pinacol diboronate (87.53 mg) were dissolved in dioxane (5 mL). Potassium acetate (50.67 mg) and tetraphenylphosphine palladium (29.88 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and used directly in the next step.

[0479] LC-MS (ESI): m / z = 198.1 [M+H] + ;

[0480] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2-fluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide. Compounds 1-8 (40 mg) were dissolved in a mixture of dioxane (4 mL) and water (0.8 mL). Compound 9-3 (15 mg) in the same dioxane (1 mL) solution, along with tetraphenylphosphine palladium (17.60 mg) and potassium carbonate (52.53 mg), were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. Cool to room temperature, adjust pH to approximately 5 with 2N hydrochloric acid in an ice bath, extract with ethyl acetate, dry the organic layer with anhydrous sodium sulfate, and perform column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product, which is then further processed (column: YMC C). 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 55%, B is acetonitrile, 25 mL / min) to obtain 10 mg of the title compound.

[0481] LC-MS (ESI): m / z = 783.2 [M+H] +

[0482] 1 H NMR(400MHz,DMSO-d6)δ10.56(s,1H),8.58(s,1H),8.04–7.89(m,3H),7. 78–7.69(m,1H),7.32(s,2H),5.60(m,1H),3.79–3.69(m,1H),3.58(s,2H ),3.42–3.35(m,3H),3.20–3.15(m,1H),3.08–2.99(m,3H),2.96–2.86(m ,1H),2.70–2.62(m,1H),2.45(s,3H),2.27(s,6H),1.44(d,J=7.2Hz,3H).

[0483] Example 10: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2,5-difluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 10)

[0484] Step 1: Synthesis of Compound 10-2

[0485] Compound 10-1 (120 mg) and dimethylamine hydrochloride (162.10 mg) were dispersed in acetonitrile (5 mL), and potassium carbonate (422.11 mg) was added. The mixture was heated to 70 °C and stirred for 4 hours until the reaction was complete. The mixture was filtered, the filtrate was concentrated, and column chromatography (ethyl acetate / petroleum ether = 0-50%) was performed to give compound 10-2 (100 mg). LC-MS (ESI): m / z = 250.0 / 252.0 [M+H] + ;

[0486] Step 2: Synthesis of compound 10-3

[0487] Compound 10-2 (50 mg) and pinacol diboronate (101.54 mg) were dissolved in dioxane (5 mL). Potassium acetate (58.78 mg) and tetraphenylphosphine palladium (34.67 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and used directly in the next step.

[0488] LC-MS (ESI): m / z = 216.1 [M+H] + ;

[0489] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2,5-difluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide. Compounds 1-8 (40 mg) were dissolved in a mixture of dioxane (4 mL) and water (0.8 mL). Compound 10-3 (16.37 mg) in the same dioxane (1 mL) solution, along with tetraphenylphosphine palladium (17.60 mg) and potassium carbonate (52.53 mg), were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. Cool to room temperature, adjust pH to approximately 5 with 2N hydrochloric acid in an ice bath, extract with ethyl acetate, dry the organic layer with anhydrous sodium sulfate, and perform column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product, which is then further processed (column: YMC C). 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 60%, B is acetonitrile, 25 mL / min) to obtain 8 mg of the title compound.

[0490] LC-MS (ESI): m / z = 801.2 [M+H] +

[0491] 1H NMR(400MHz,DMSO-d6)δ10.58(s,1H),10.24(s,1H),8.58(s,1H),8.03–7 .92(m,2H),7.87–7.62(m,3H),5.60(m,1H),4.15(s,1H),3.86–3.66(m,2H ),3.53–3.38(m,3H),3.27–3.21(m,1H),3.20–3.13(m,1H),3.04(m,2H), 2.80–2.54(m,7H),2.45(s,3H),2.20–2.12(m,1H),1.45(d,J=7.2Hz,2H).

[0492] Example 11: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2,3-difluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 11)

[0493] Step 1: Synthesis of Compound 11-2

[0494] Compound 11-1 (200 mg) was dissolved in THF (5 mL), and a tetrahydrofuran solution of dimethylamine (81.60 mg, 2 M) was added. Acetic acid (81.82 mg) was added, and the mixture was stirred at 25 °C for 0.5 hours. Sodium triacetoxyborohydride (575.41 mg) was added, and the mixture was stirred at 25 °C for 2 hours until the reaction was complete. The mixture was filtered, the filtrate was concentrated, and column chromatography (ethyl acetate / petroleum ether = 0-50%) was performed to give compound 11-2 (150 mg).

[0495] LC-MS(ESI):m / z=250.0 / 252.0[M+H] + ;

[0496] Step 2: Synthesis of Compound 11-3

[0497] Compound 11-2 (50 mg) and pinacol diboronate (101.54 mg) were dissolved in dioxane (5 mL). Potassium acetate (58.78 mg) and tetraphenylphosphine palladium (34.67 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and used directly in the next step.

[0498] LC-MS (ESI): m / z = 216.1 [M+H] + ;

[0499] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(4-((dimethylamino)methyl)-2,3-difluorophenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide. Compounds 1-8 (40 mg) were dissolved in a mixture of dioxane (4 mL) and water (0.8 mL). At room temperature, 11-3 (16.37 mg) of the above dioxane solution (1 mL), tetra(triphenylphosphine)palladium (17.60 mg), and potassium carbonate (52.53 mg) were added. After addition, the reaction was carried out under nitrogen protection at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. Cool to room temperature, adjust pH to approximately 5 with 2N hydrochloric acid in an ice bath, extract with ethyl acetate, dry the organic layer with anhydrous sodium sulfate, and perform column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product, which is then further processed (column: YMC C). 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 60%, B is acetonitrile, 25 mL / min) to obtain 20 mg of the title compound.

[0500] LC-MS (ESI): m / z = 801.2 [M+H] +

[0501] 1 H NMR(400MHz,DMSO-d6)δ8.46(s,1H),7.99–7.93(m,2H),7.80(m,1H),7.73(m,1H),7.35(m,1H),5.60(m,1H),3.82–3.72(m,1H),3.54 (s,2H),3.45–3.36(m,3H),3.18–3.12(m,1H),3.08–2.99(m,3H),2.70–2.59(m,3H),2.41(s,3H),2.18(s,6H),1.44(d,J=7.2Hz,3H).

[0502] Example 12: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-2-oxo-1,2-dihydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 12)

[0503] Step 1: Synthesis of Compound 12-2. Compound 12-1 (200 mg) and 3,3-difluorocyclobutanol (124.24 mg) were dissolved in toluene (5 mL). Triphenylphosphine (452.24 mg) and diisopropyl azodicarboxylate (348.64 mg) were added. The mixture was stirred at 100 °C for 15 hours until the reaction was complete. After cooling to room temperature, the reaction solution was concentrated and subjected to column chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain compound 12-2 (15 mg).

[0504] Step 2: Synthesis of Compound 12-3

[0505] Compound 12-2 (15 mg) and pinacol diboronate (28.85 mg) were dissolved in dioxane (5 mL). Potassium acetate (16.70 mg), tricyclohexylphosphine (1.59 mg), and tris(dibenzylacetone)palladium (2.60 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and used directly in the next step.

[0506] Step 3: Synthesis of N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-(3,3-difluorocyclobutyl)-2-oxo-1,2-dihydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide. Compounds 1-8 (40 mg) were dissolved in a mixed solution of dioxane (4 mL) and water (0.8 mL). 12-3 (16.37 mg) of the above dioxane (1 mL) solution, tetratriphenylphosphine palladium (17.60 mg), and cesium fluoride (132.67 mg) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. After cooling to room temperature, the mixture was extracted with ethyl acetate (10 mL) and saturated brine (5 mL). The organic layer was dried over anhydrous sodium sulfate and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was then performed (column: YMC C). 18 Mobile phase: 0.7% NH4HCO3 aqueous solution; B%: 55%, B is acetonitrile, 25 mL / min) to obtain 10 mg of the title compound.

[0507] LC-MS (ESI): m / z = 815.2 [M+H] +

[0508] 1H NMR (400MHz, DMSO-d6) δ10.56 (s, 1H), 8.49 (s, 1H), 7.99 (d, J = 2.1Hz, 1H), 7.90 (m, 2H), 7.7 4(dd,J=8.7,2.1Hz,1H),6.99(d,J=1.9Hz,1H),6.89(dd,J=7.2,1.9Hz,1H),5.59(m,1H),4 .87–4.77(m,1H),3.80–3.73(m,1H),3.50–3.41(m,3H),3.24–3.16(m,1H),3.15–3.05(m,5 H),3.04–2.96(m,2H),2.75–2.53(m,1H),2.42(s,3H),2.16(m,1H),1.44(d,J=7.2Hz,3H).

[0509] Example 13: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-fluoro-2-methoxypyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 13)

[0510] Step 1: Synthesis of Compound 13-2

[0511] Intermediate 1-8-R (100.0 mg) was dissolved in a mixture of dioxane (5 mL) and water (1 mL). At room temperature, (5-fluoro-2-methoxypyridin-4-yl)boronic acid (32.93 mg), tetrakis(triphenylphosphine)palladium (25.69 mg), and cesium fluoride (450.15 mg) were added. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. After cooling to room temperature, ethyl acetate (10 mL) and saturated brine (5 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (ethyl acetate / petroleum ether = 0-100%) to give compound 13-2 (90 mg).

[0512] LC-MS (ESI): m / z = 721.2 [M+H] +

[0513] Step 2: Synthesis of Compound 13-3

[0514] Compound 13-2 (90.0 mg) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (711.54 mg) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The organic phase was dried and concentrated to obtain compound 13-3 (75 mg).

[0515] LC-MS (ESI): m / z = 621.2 [M+H] +

[0516] Step 3: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-fluoro-2-methoxypyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0517] Compound 13-3 (75.0 mg) was dissolved in dichloromethane (8 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (24.20 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (49.96 mg), and 1-hydroxybenzotriazole (16.33 mg) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain a solid. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 25 mg of the title compound.

[0518] LC-MS (ESI): m / z = 757.2 [M+H] +

[0519] 1H NMR (400MHz, DMSO-d6) δ10.56 (s, 1H), δ8.49 (s, 1H), 8.36 (d, J = 2.3Hz, 1H), 8.03–7.88(m,2H),7.74(d,J=8.5Hz,1H),7.32(d,J=4.8Hz,1H),5.63–5.56( m,1H),4.60–4.28(m,1H),3.88(s,3H),3.78(m,2H),3.54–3.51(m,3H)3.30 –3.27(m,1H),3.03(m,2H),2.42(s,3H),2.16(m,1H),1.44(d,J=7.2Hz,3H).

[0520] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 13 is (7R, 9R).

[0521] Example 14: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(3-fluoro-2-methoxypyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 14)

[0522] Step 1: Synthesis of Compound 14-2

[0523] Intermediate 1-8-R (100.0 mg) was dissolved in a mixture of dioxane (5 mL) and water (1 mL). (3-fluoro-2-methoxypyridin-4-yl)boronic acid (32.93 mg), tetrakis(triphenylphosphine)palladium (25.69 mg), and cesium fluoride (450.15 mg) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LC-MS until completion. After cooling to room temperature, ethyl acetate (10 mL) and saturated brine (5 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (ethyl acetate / petroleum ether = 0-100%) to give compound 14-2 (90 mg).

[0524] LC-MS (ESI): m / z = 721.2 [M+H] +

[0525] Step 2: Synthesis of Compound 14-3

[0526] Compound 14-2 (90.0 mg) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (711.54 mg) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The organic phase was dried and concentrated to obtain compound 14-3 (75 mg).

[0527] LC-MS (ESI): m / z = 621.2 [M+H] +

[0528] Step 3: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(3-fluoro-2-methoxypyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide. Compound 14-3 (75.0 mg) was dissolved in dichloromethane (8 mL), and 5-hydroxy-6-methylpyrimidin-4-carboxylic acid (24.20 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (49.96 mg), and 1-hydroxybenzotriazole (16.33 mg) were added. The reaction solution was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction solution was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Preparative chromatography (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 20 mg of the title compound.

[0529] LC-MS (ESI): m / z = 757.2 [M+H] +

[0530] 1 H NMR (400MHz, DMSO-d6) δ10.57(s,1H),8.46(s,1H),8.09(d,J=5.3Hz,1H),8.02–7.88(m,2H),7.74(m,1H),7.54(m,1H),5.61(m,1H),4.60– 4.37(m,1H),4.00(s,3H),3.77(m,3H),3.50–3.45(m,3H),3.23–3.20 (m,1H), 3.03(m,2H),2.42(s,3H),2.16(m,1H),1.44(d,J=7.2Hz,3H).

[0531] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 14 is (7R, 9R).

[0532] Example 15: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(6-cyclopropylpyridin-3-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 15)

[0533] Step 1: Synthesis of Compound 15-2

[0534] Intermediate 1-8-R (100.0 mg) was dissolved in a mixture of dioxane (5 mL) and water (1 mL). 2-Cyclopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)pyridine (47.22 mg), tetraphenylphosphine palladium (25.69 mg), and cesium fluoride (450.15 mg) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LCMS until completion. After cooling to room temperature, ethyl acetate (10 mL) and saturated brine (5 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (ethyl acetate / petroleum ether = 0-100%) to give compound 15-2 (90 mg).

[0535] LC-MS (ESI): m / z = 713.2 [M+H] +

[0536] Step 2: Synthesis of Compound 15-3

[0537] Compound 15-2 (90.0 mg) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (711.54 mg) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The organic phase was dried and concentrated to give compound 15-3 (75 mg).

[0538] LC-MS (ESI): m / z = 613.2 [M+H] +

[0539] Step 3: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(6-cyclopropylpyridin-3-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0540] Compound 15-3 (80.0 mg) was dissolved in dichloromethane (10 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (24.20 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (53.99 mg), and 1-hydroxybenzotriazole (17.65 mg) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 35 mg of the title compound.

[0541] LC-MS (ESI): m / z = 749.2 [M+H] +

[0542] 1 H NMR (400MHz, DMSO-d6) δ10.53(s,1H),8.97(d,J=2.2Hz,1H),8.46(s,1H),8.17(m,1H),8.00–7.82(m,2H),7.67(m,1H),7.37(d,J=8.2Hz,1H),5.5 4(m,1H),4.53–4.22(m,1H),3.68(m,2H),3.50–3.45(m,2H),3.20–3.17( m,2H),2.96(m,2H),2.37(s,3H),2.10(m,2H),1.37(m,3H),0.94(m,4H).

[0543] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 15 is (7R, 9R).

[0544] Example 16: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-((1r,3R)-3-fluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide (compound 16)

[0545] Step 1: Synthesis of Compound 16-1

[0546] Weigh 100 mg of N,N-diisopropylethylamine and dissolve it in 2.5 mL of dichloromethane. Under nitrogen protection, stir at -78 °C. Slowly add 170 mg of trifluoromethanesulfonic anhydride and stir for 10 minutes. Then slowly add 50 mg of (1s,3s)-3-fluorocyclobutane-1-ol. Gradually raise the temperature from -78 °C to 25 °C and stir for 12 hours until the reaction is complete. TLC (petroleum ether / ethyl acetate = 1:1) confirms that the starting material has reacted completely and can be used directly in the next reaction without further treatment.

[0547] Step 2: Synthesis of compound 16-3

[0548] Weigh 286 mg of N,N-diisopropylethylamine and dissolve it in the above reaction solution. Slowly add compound 16-2 (123 mg) at 0 °C, stir for 30 minutes, then heat to 25 °C and stir for 12 hours until the reaction is complete. Add 20 mL of dichloromethane and 10 mL of water to the reaction solution for extraction. Dry the organic layer with anhydrous sodium sulfate and concentrate to obtain compound 16-3 (120 mg).

[0549] LC-MS(ESI): m / z = 282.2[M+H] + ;

[0550] Step 3: Synthesis of Compound 16-4

[0551] Intermediate 1-8-R (100 mg) was dissolved in a mixture of dioxane (5 mL) and water (1 mL). Compound 16-3 (104 mg), tetraphenylphosphine palladium (34 mg), and N,N-diisopropylethylamine (957 mg) were added at room temperature. After addition, the reaction mixture was kept under nitrogen protection and reacted at 85 °C for 6 hours. The reaction was monitored by LCMS until completion. The mixture was cooled to room temperature, extracted with ethyl acetate (10 mL) and saturated brine (5 mL), and the organic layer was concentrated and subjected to column chromatography (methanol / dichloromethane = 0.7%) to give compound 16-4 (100 mg).

[0552] LC-MS (ESI): m / z = 749.3 [M+H] +

[0553] Step 4: Synthesis of Compound 16-5

[0554] Compound 16-4 (90 mg) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (791 mg, 6.93 mmol) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The organic phase was dried and concentrated to give compound 16-5 (80 mg).

[0555] LC-MS (ESI): m / z = 649.3 [M+H] +

[0556] Step 5: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-((1r,3R)-3-fluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0557] Compound 16-5 (80 mg) was dissolved in dichloromethane (10 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (26 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (54 mg), and 1-hydroxybenzotriazole (17 mg) were added. The reaction mixture was reacted at 25 °C for 2 hours. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation was performed (column: YMC TA; mobile phase: 7 mmol / L).

[0558] NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) to give 21 mg of the title compound.

[0559] LC-MS (ESI): m / z = 785.3 [M+H] +

[0560] 1H NMR(400MHz,DMSO-d6)δ10.53(s,1H),8.45(s,1H),8.03–7.90(m,2H),7.74( m,1H),6.74(d,J=3.7Hz,1H),5.53(m,1H),5.24–5.02(m,1H),4.48(s,1H),3. 70(m,2H),3.61–3.45(m,3H),3.33–3.18(m,5H),3.12–2.89(m,5H),2.48(s,1 H),2.41(s,3H),2.35–2.17(m,4H),2.15–2.06(m,1H),1.40(d,J=7.2Hz,3H).

[0561] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 16 is (7R, 9R).

[0562] Example 17: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-((1s,3S)-3-fluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidine-9-carboxamide (compound 17)

[0563] Step 1: Synthesis of Compound 17-1

[0564] Weigh 100.34 mg of N,N-diisopropylethylamine and dissolve it in 2.5 mL of dichloromethane. Under nitrogen protection, stir at -78°C. Slowly add 170.23 mg of trifluoromethanesulfonic anhydride and stir for 10 minutes. Then slowly add 50 mg of (1r,3r)-3-fluorocyclobutane-1-ol. Gradually raise the temperature from -78°C to 25°C and stir for 12 hours until the reaction is complete. TLC (petroleum ether / ethyl acetate = 1:1) confirms complete reaction of the starting material. The reactants were used directly in the next reaction without further treatment.

[0565] Step 2: Synthesis of Compound 17-3

[0566] Weigh out N,N-diisopropylethylamine (286.23 mg), dissolve it in the above reaction solution, and slowly add compound 17-2 (123 mg) at 0 °C. Stir for 30 minutes, then heat to 25 °C and stir for 12 hours until the reaction is complete. Add 20 mL of dichloromethane and 10 mL of water to the reaction solution for extraction. Dry the organic layer with anhydrous sodium sulfate and concentrate to obtain compound 17-3 (120 mg).

[0567] LC-MS(ESI): m / z = 282.2[M+H] + ;

[0568] Step 3: Synthesis of Compound 17-4

[0569] Intermediate 1-8-R (100 mg) was dissolved in a mixture of dioxane (5 mL) and water (1 mL). Compound 17-3 (104.15 mg), tetraphenylphosphine palladium (34.26 mg), and N,N-diisopropylethylamine (957.48 mg) were added at room temperature. After addition, the mixture was kept under nitrogen protection and reacted at 85 °C for 6 hours. The reaction was monitored by LCMS until completion. The mixture was cooled to room temperature, extracted with ethyl acetate (10 mL) and saturated brine (5 mL), and the organic layer was concentrated and subjected to column chromatography (methanol / dichloromethane = 0.7%) to give compound 17-4 (100 mg).

[0570] LC-MS (ESI): m / z = 749.3 [M+H] +

[0571] Step 4: Synthesis of Compound 17-5

[0572] Compound 17-4 (92.65 mg) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (790.60 mg) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (20 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (10 mL). The organic phase was dried and concentrated to give compound 17-5 (75 mg).

[0573] LC-MS (ESI): m / z = 649.3 [M+H] +

[0574] Step 5: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(1-((1s,3S)-3-fluorocyclobutyl)-1,2,3,6-tetrahydropyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0575] Compound 17-5 (85 mg) was dissolved in dichloromethane (10 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (26.24 mg), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (54.18 mg), and 1-hydroxybenzotriazole (17.71 mg) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 30 mg of the title compound.

[0576] LC-MS (ESI): m / z = 785.3 [M+H] +

[0577] 1 H NMR (400MHz, DMSO-d6) δ10.52(s,1H),8.51(s,1H),8.02–7.88(m,2H),7.74(dd,J=8.8, 2.1Hz,1H),6.74(d,J=3.7Hz,1H),5.53(m,1H),4.86(m,1H),4.56–4.43(m,1H),3.71(m ,2H),3.55–3.43(m,3H),3.29–3.24(m,3H),3.19–3.15(m,1H),3.08–2.91(m,4H),2.62 –2.51(m,4H),2.46–2.31(m,4H),2.11(m,1H),2.06–1.88(m,2H),1.41(d,J=7.2Hz,3H).

[0578] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 17 is (7R, 9R).

[0579] Example 18: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-cyclopropylpyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 18)

[0580] Step 1: Synthesis of Compound 18-2

[0581] Compound 18-1 (40 mg, 201.96 μmol) and pinacol diboronate (102.57 mg, 403.92 μmol) were dissolved in dioxane (1.5 mL). Potassium acetate (59.47 mg, 605.88 μmol) and tetraphenylphosphine palladium (35.02 mg, 30.29 μmol) were added at room temperature. After the addition was complete, the reaction was carried out under nitrogen protection at 110 °C for 6 hours. The mixture was then cooled to room temperature and used directly in the next step.

[0582] Step 2: Synthesis of Compound 18-3

[0583] Intermediate 1-8-R (24.85 mg, 36.81 μmol) was dissolved in a mixture of dioxane (2 mL) and water (0.4 mL). The above solution of compound 18-2, tetraphenylphosphine palladium (8.51 mg, 7.36 μmol), and cesium fluoride (111.84 mg, 736.28 μmol) were added at room temperature. After addition, the mixture was kept under nitrogen protection and reacted at 80 °C for 12 hours. The reaction was monitored by LCMS until completion. After cooling to room temperature, ethyl acetate (10 mL) and saturated brine (5 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (methanol / dichloromethane = 0.5%) to give compound 18-3 (20 mg).

[0584] Step 3: Synthesis of Compound 18-4

[0585] Compound 18-3 (20 mg, 28.04 μmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (239.82 mg, 2.10 mmol) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (10 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (5 mL). The organic phase was dried and concentrated to give compound 18-4 (17 mg).

[0586] Step 4: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-cyclopropylpyridin-4-yl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0587] Compound 18-4 (20 mg, 32.62 μmol) was dissolved in dichloromethane (5 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (6.54 mg, 42.41 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (13.13 mg, 68.51 mmol), and 1-hydroxybenzotriazole (4.41 mg, 32.65 μmol) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 6.86 mg of the title compound.

[0588] LC-MS (ESI): m / z = 749.2 [M+H] +、

[0589] 1 H NMR(400MHz,DMSO-d6)δ8.54(d,J=5.1Hz,1H),8.36(s,1H),8.02–7.89(m,3H ),7.78–7.68(m,2H),5.61(m,1H),4.56–4.35(m,1H),3.76(m,2H),3.45–3.4 3(m,1H),3.20–3.15(m,1H),3.03(m,3H),2.67(m,1H),2.38(s,3H),2.33(m, 1H), 2.26 (m, 1H), 2.17 (d, J = 13.5Hz, 1H), 1.44 (d, J = 7.2Hz, 3H), 0.97 (m, 4H).

[0590] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 18 is (7R, 9R).

[0591] Example 19: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-2-(2-methoxypyridin-4-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide (compound 19)

[0592] Step 1: Synthesis of Compound 19-1

[0593] Intermediate 1-8-R (22.97 mg, 34.03 μmol) was dissolved in a mixture of dioxane (2 mL) and water (0.4 mL). At room temperature, (2-methoxypyridin-4-yl)boronic acid (12 mg, 78.46 μmol), tetraphenylphosphine palladium (7.87 mg, 6.81 μmol), and cesium fluoride (77.54 mg, 510.45 mmol) were added. After addition, the reaction mixture was kept under nitrogen protection and reacted at 80 °C for 10 hours. The reaction was monitored by LC-MS until completion. After cooling to room temperature, ethyl acetate (10 mL) and saturated brine (5 mL) were added for extraction. The organic layer was concentrated and subjected to column chromatography (ethyl acetate / petroleum ether = 0-100%) to give compound 19-1 (20 mg).

[0594] Step 2: Synthesis of Compound 19-2

[0595] Compound 19-1 (20 mg, 28.45 μmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (162.17 mg, 1.42 mmol) was added. After the addition was complete, the reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated, and the residue was redissolved in a dichloromethane / methanol (20 / 1) mixture (10 mL). The mixture was then extracted at 0 °C with a saturated sodium carbonate aqueous solution (5 mL). The organic phase was dried and concentrated to obtain compound 19-2 (17 mg).

[0596] Step 3: Synthesis of (7R,9R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-6-(4-(5-hydroxy-6-methylpyrimidin-4-carbonyl)piperazin-1-yl)-2-(2-methoxypyridin-4-yl)-7-methyl-5-oxo-5,7,8,9-tetrahydropyrrolo[1,2-c][1,2,4]triazolo[1,5-a]pyrimidin-9-carboxamide

[0597] Compound 19-2 (17 mg, 28.19 μmol) was dissolved in dichloromethane (4 mL), and 5-hydroxy-6-methylpyrimidine-4-carboxylic acid (5.65 mg, 36.65 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (11.66 mg, 60.82 mmol), and 1-hydroxybenzotriazole (3.81 mg, 28.22 μmol) were added. The reaction mixture was reacted at 25 °C for 1 hour. The reaction was monitored by LCMS until completion. The reaction mixture was concentrated and subjected to column chromatography (methanol / dichloromethane = 0-10%) to obtain the crude product. Further preparation (column: YMC TA; mobile phase: 7 mmol / L NH4HCO3 aqueous solution; B%: 30%–70%, B is acetonitrile, 25 mL / min) yielded 2.5 mg of the title compound.

[0598] LC-MS (ESI): m / z = 739.2 [M+H]+

[0599] 1 H NMR(400MHz,DMSO-d6)δ8.32(d,J=5.3Hz,1H),8.20(s,1H),8.01–7.90(m,2H ),7.73(d,J=8.7Hz,1H),7.60(d,J=5.4Hz,1H),7.34(s,1H),5.59(d,J=10.1 Hz,1H),3.90(s,3H),3.81–3.70(m,3H),3.31–3.25(m,1H),3.17(m,1H),3.0 5–2.94(m,3H),2.67(m,2H),2.33(s,3H),2.16(m,1H),1.44(d,J=7.3Hz,3H).

[0600] Since the absolute configuration of intermediate 1-8-R is (7R, 9R), the absolute configuration of compound 19 is (7R, 9R).

[0601] Biological activity and related property test examples

[0602] The compounds in the following test examples were all prepared using the methods described in the embodiments of this disclosure.

[0603] Test Example 1: WRN ATP hydrolase inhibitory activity assay

[0604] Experimental Principle

[0605] WRN unwinding is driven by ATP hydrolysis by the ATP hydrolase domain of the WRN protein. ATP hydrolysis generates ADP, which releases energy to promote WRN unwinding. The ADP generated during the reaction is detected using the ADP-Glo ​​assay kit (Promega). The signal value after adding the assay reagent is positively correlated with the amount of ADP generated in the reaction system, and changes in the signal value reflect changes in the ATP hydrolase activity of the WRN protein.

[0606] Experimental instruments

[0607] Experimental materials

[0608] Experimental methods

[0609] Test compounds were dissolved in DMSO at a stock concentration of 10 mM. Serial dilutions of the compound stock solution were performed using the dose-response program on the Echo instrument. The dilution program consisted of 4 μL of total volume, starting at 10 μM, followed by 3-fold dilutions over 10 concentration points. The total volume of compound and DMSO was 160 nL, with a final DMSO concentration of 0.4%. 2 μL of 2X WRN-ATP mixture (buffer: 25 mM Tris-HCl (pH 8.0), 50 mM NaCl, 2 mM MgCl2, 1 mM DTT, 0.01% Tween-20, 0.00025% BSA; WRN: 20 nM; ATP: 600 μM) was added to the experimental plate and incubated at room temperature for 3 hours. Then, 2 μL of 2X Hec1 (0.4 nM) was added and incubated at room temperature for 0.5 hours. Next, 4 μL of ADP-Glo ​​reagent (ADP-Glo ​​detection kit) was added and incubated for another 1 hour. Finally, 8 μL of Detection reagent (ADP-Glo ​​detection kit) was added and incubated at room temperature for 1 hour. The chemiluminescence signal values ​​were read on an Envision microscope. The signal value of the WRN reaction well was defined as the 0% inhibition control, and the signal value of the well without WRN was defined as the 100% inhibition control. The inhibition rate of the compound-treated wells was calculated. The IC50 was calculated using a four-parameter fitting method based on the inhibition rate. 50 .

[0610] The experimental results are shown in Table 1 below:

[0611] Table 1

[0612] The compounds disclosed in this embodiment exhibit good WRN ATP hydrolase inhibitory activity.

[0613] Test Example 2: HCT-116, Caco-2, DLD-1WRN-KO Cell Proliferation Experiment

[0614] Experimental Principle

[0615] The cells used in the experiment were HCT-116 (MSI, microsatellite unstable) cell line, Caco-2 (MSS, microsatellite stable) cell line, and DLD-1WRN-KO (DLD-1 cells stabilized by CRSIPR editing to knock out the WRN gene). Proliferation experiments used kits (…). The 2.0 Luminescent Cell Viability Assay quantifies ATP in living cells and tests the inhibitory effect of compounds on cell proliferation. The kit generates a luminescent signal that is directly proportional to the amount of ATP, which in turn is directly proportional to the number of cells.

[0616] Experimental instruments

[0617] Experimental materials

[0618] Experimental methods

[0619] HCT-116 cells were cultured in McCoy's 5A medium with 10% FBS, Caco-2 cells in MEM medium with 20% FBS, and DLD-1WRN-KO cells in RPMI-1640 medium with 10% FBS. After trypsin digestion, cell concentrations were adjusted to 300 / 40 μL / well for HCT-116, 600 / 40 μL / well for Caco-2, and 500 / 40 μL / well for DLD-1WRN-KO. Cells were seeded in 384-well plates and cultured overnight at 37°C in a 5% CO2 incubator. The test compounds were dissolved in DMSO, with a stock solution concentration of 10 mM. The stock solutions were serially diluted using the Echo instrument's dose-response program with DMSO as the dilution solvent. The program conditions were: total experimental volume 40 μL, starting at 10 μM, 3-fold dilution, 10 concentration points, and a total volume of 200 nL for both the compound and DMSO. After incubating the cell culture plates for another 4 days, the plates were equilibrated to room temperature. 20 μL of CCL reagent was added to each well, and the plates were shaken in the dark for 60 minutes. Luminescence signals were then read using Envision. Cell control wells were defined as 0% inhibition controls, and culture medium wells as 100% inhibition controls. The inhibition rate of each well was calculated. The IC50 was calculated using a four-parameter fitting method based on the inhibition rate. 50 (XLfit, Formula 205).

[0620] The experimental results are shown in Tables 2 and 3 below:

[0621] Table 2

[0622] Table 3

[0623] The compounds disclosed herein exhibit good anti-cell proliferation activity against microsatellite unstable cells and high selectivity for microsatellite stable cells.

[0624] Test Example 3: Canine Pharmacokinetics Study

[0625] 1. Test Materials

[0626] The beagle was purchased from Jiangsu Mas Biotechnology Co., Ltd.

[0627] HP-β-CD (hydroxypropyl-β-cyclodextrin) and verapamil were purchased from Sigma, while acetonitrile and formic acid were purchased from Merck (USA). K2EDTA anticoagulant blood collection tubes were purchased from Jiangsu Xinkang Medical Device Co., Ltd.

[0628] II. Test Methods

[0629] 1. Animal testing

[0630] For each test compound, six male beagle dogs (10-12 kg, age ≥6 months) were selected and randomly divided into two groups of three. Group 1 received the compound via intravenous injection in the hind limbs, with 10% HP-β-CD as the solvent (prepared drug concentration 0.5 mg / ml). Group 2 received the corresponding dose of the compound orally, with 10% HP-β-CD as the solvent (prepared drug concentration 0.4 mg / ml). Before the experiment, the animals were fasted overnight but given water normally. During the experiment, blood was collected from the forelimb veins of each group of dogs before administration and at 0.083 h (Group 1 only), 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h, and 48 h after administration. The collected whole blood samples were placed in K2EDTA anticoagulant tubes, centrifuged for 5 min (4000 rpm, 4℃), and plasma was collected for testing. Four hours after administration, the animals were given normal food and water.

[0631] 2. Sample processing and bioanalysis

[0632] Take 10 μL of canine plasma sample, add 150 μL of acetonitrile solvent (containing internal standard, 10 ng / mL verapamil) to precipitate proteins, vortex for 5 min, centrifuge for 5 min (14000 rpm, 4℃), and take 70 μL of supernatant and mix thoroughly with 70 μL of water containing 0.1% formic acid (v / v). Quantitative detection is performed using an LC-MS / MS system (AB Sciex Triple Quad 6500+). A standard curve of male beagle plasma and quality control samples are used concurrently with the sample concentration determination. For 20× diluted samples, take 2 μL of the plasma sample to be tested and add it to 38 μL of beagle blank plasma. Vortex for 1 min, add 600 μL of acetonitrile solvent (containing internal standard, 10 ng / mL verapamil) to precipitate proteins, and the remaining processing steps are the same as above.

[0633] Canine tissues (heart, liver, spleen, lung, kidney, brain, stomach, duodenum, jejunum, ileum, cecum, etc.) were homogenized with PBS buffer at a weight-to-volume ratio of 9 (w / v). 40 μL of the homogenized tissue sample was added to 600 μL of an acetonitrile:methanol mixture (1:1, v / v) containing 10 ng / mL internal standard (verapamil). The mixture was vortexed for 5 minutes and then centrifuged at 14,000 rpm for 5 minutes. 70 μL of the supernatant was thoroughly mixed with 70 μL of water containing 0.1% formic acid (v / v) and quantitatively analyzed using an LC-MS / MS system (AB Sciex Triple Quad 6500+).

[0634] 3. Data Processing

[0635] The pharmacokinetic parameters were calculated using the non-compartmental statistical moment method using Phoenix WinNonlin 8.0 software (Certara, USA).

[0636] 4. Test Results

[0637] The results of the canine pharmacokinetic studies are shown in Tables 4 and 5 below.

[0638] Table 4: Canine pharmacokinetic (PK) of the compounds (intravenous administration)

[0639] Table 5: Canine pharmacokinetics of the compounds (oral administration) Note: (1) " / " indicates that it was not detected;

[0640] (2) The structural formula of HRO761 is:

[0641] The compounds disclosed herein exhibit high drug exposure and oral bioavailability, and good pharmacokinetic properties. They show good distribution in various tissues, particularly the brain, suggesting potential for the treatment of brain cancer.

[0642] Test Example 4: Caco-2 Permeability Test

[0643] The apparent permeability (P) of the drug was determined using liquid chromatography-tandem mass spectrometry (LC-MS / MS) in a Caco-2 cell model. app ).

[0644] In this test case, Caco-2 cells were purchased from the American Type Culture Collection (ATCC), 4-hydroxyethylpiperazine ethanesulfonic acid (HEPES) was purchased from Beijing Solarbio Science & Technology Co., Ltd., Hank's balanced salt solution (HBSS) and non-essential amino acids (NEAA) were purchased from Thermo Fisher Scientific, penicillin, streptomycin, and trypsin / EDTA were purchased from Solarbio, fetal bovine serum (FBS) and DMEM medium were purchased from Corning, HTS-96-well Transwell plates and other sterile consumables were purchased from Corning, and the Millicell electrical resistance measurement system was purchased from Millipore. Purchased from Nexcelom Bioscience; Infinite 200PRO microplate reader purchased from Tecan; MTS2 / 4orbital shaker purchased from IKA Labortechnik.

[0645] Step 1: Cell Culture and Seeding

[0646] Caco-2 cells were cultured in cell culture flasks. The incubator was set to 37°C, 5% CO2, and 95% relative humidity. Cells were ready for Transwell inoculation when confluence reached 70-90%. Before inoculation, 50 μL of cell culture medium was added to each well in the upper chamber of the Transwell, and 25 mL of cell culture medium was added to the lower culture plate. The culture plate was incubated at 37°C, 5% CO2 for 1 hour before inoculation. After cell digestion, the cell suspension was transferred to a round-bottom centrifuge tube and centrifuged at 120 g for 5 minutes. The cells were resuspended in culture medium to a final concentration of 6.86 × 10⁻⁶. 5 cells / mL. Add 50 μL of cell suspension to each well of a 96-well Transwell plate, resulting in a final seeding density of 2.4 × 10⁶ cells / mL. 5 cells / cm2. Change the medium 24 hours after inoculation and culture for 14-18 days, changing the medium every other day. The medium change procedure is as follows: Separate the Transwell chamber from the receiving plate. First discard the medium in the receiving plate, then discard the medium in the Transwell chamber. Finally, add 75 μL of fresh medium to each chamber and 25 mL of fresh medium to the receiving plate.

[0647] The second step is the evaluation of the integrity of the cell monolayer.

[0648] After approximately 14 days of culture, Caco-2 cells reach convergence and complete differentiation. At this point, they can be used for breakthrough experiments. The monolayer membrane resistance is measured using a resistance meter (Millipore, USA), and the resistance of each well is recorded. After the measurement, the Transwell plate is returned to the incubator. The resistance value is calculated as: measured resistance (ohms) × membrane area (cm²). 2 ) = TEER value (ohm·cm) 2 If the TEER value is <230 ohms·cm 2 If so, the hole cannot be used for a penetration test.

[0649] Step 3: Solution Preparation

[0650] Weigh out 2.38g HEPES and 0.35g sodium bicarbonate, add 900mL of pure water to dissolve them, then add 100mL of 10×HBSS and stir well. Adjust the pH to 7.4, and finally filter to obtain 1L of transfer buffer (HBSS, 10mM HEPES, pH 7.4).

[0651] A 1 mM DMSO stock solution of the test compound was diluted with transfer buffer to obtain a 5 μM test solution. The control compound digoxin or minoxidil was diluted to 2 mM with DMSO and then to 10 μM with the same transfer buffer to obtain the control compound test solution. Additionally, DMSO was also diluted with the same transfer buffer to a receiving end solution containing 0.5% DMSO.

[0652] Step 4 Drug Penetration Test

[0653] Remove the Transwell culture plate from the incubator. Rinse the cell monolayer twice with transport buffer (10 mM HEPES, pH 7.4) and incubate at 37°C for 30 minutes.

[0654] The transport rate of the compound from the tip to the substrate was determined. 125 μL of test solution was added to each well in the upper chamber (tip), and immediately 50 μL of the solution was transferred from the tip to 200 μL of acetonitrile containing the internal standard (0.1 μM tolbutamide) as the initial sample from the tip to the substrate. 235 μL of receiver solution was added to each well in the lower chamber (substrate).

[0655] The transport rate of the compound from the base to the top was determined. 285 μL of the receiving end solution was added to each well in the upper chamber (top), and immediately 50 μL of the solution was transferred from the top to 200 μL of acetonitrile containing the internal standard (0.1 μM tolbutamide) as the initial sample from the base to the top. 75 μL of the test solution was added to each well in the lower chamber (base).

[0656] After merging the upper and lower transfer devices, incubate at 37°C for 2 hours.

[0657] After incubation, 50 μL of sample was taken from each well of the upper and lower chambers of the Transwell plate and added to a new sample tube. 200 μL of acetonitrile containing the internal standard (0.1 μM tolbutamide) was added to the sample tube, vortexed for 10 minutes, and then centrifuged at 3220 g for 40 minutes. 150 μL of the supernatant was collected, diluted with 150 μL of water, and analyzed by LC-MS / MS. All samples were prepared in triplicate.

[0658] The integrity of the cell monolayer was assessed after 2 hours of incubation using fluorescein leakage. Fluorescein stock solution was diluted to a final concentration of 100 μM using transport buffer (10 mM HEPES, pH 7.4). 100 μL of fluorescein solution was added to each well of the upper Transwell plate, and 300 μL of transport buffer (10 mM HEPES, pH 7.4) was added to each well of the lower receiving plate. After incubation at 37°C for 30 minutes, 80 μL of solution was aspirated from both the upper and lower layers of each well into a new 96-well plate. Fluorescence was measured using a microplate reader at an excitation wavelength of 485 nm and an emission wavelength of 530 nm.

[0659] Step 5: Data Analysis

[0660] All calculations were performed using Microsoft Excel. Peak areas were determined using the extracted ion chromatograms.

[0661] Apparent permeability coefficient (P) app Unit: cm / s × 10 -6 The following formula can be used to calculate:

[0662] In the formula: V A The volume of the receiving solution is given (Ap→Bl is 0.3 mL, Bl→Ap is 0.1 mL), and the area (membrane area) is the membrane area of ​​the Transwell-96-well plate (0.143 cm²). 2 ); time (incubation time, in seconds); [drug] receiver ([drug] 接收端 () represents the drug concentration at the receiving end; [drug] initial， donor ([drug] 初始，供体 () represents the initial drug concentration at the dosing end.

[0663] The efflux ratio is calculated using the following formula:

[0664] In the formula: P app(B-A) The apparent permeability coefficient is measured from the base to the tip.

[0665] P app(A-B) The apparent permeability coefficient is calculated from the top to the base.

[0666] The recovery rate ("Percentage recovery" (%)) is calculated using the following formula:

[0667] In the formula: V A V represents the volume of the solution at the receiving end (unit: mL);D The volume of the solution at the donor end (unit: mL).

[0668] The leakage rate (Percentage leakage (%) or LY (%)) is calculated using the following formula:

[0669] In the formula: I receiver (I 接收端 ) refers to the fluorescence density of the receiving aperture (0.3 mL), I donor (I 供体 LY (%) refers to the fluorescence density of the drug-dosing well (0.1 mL). LY < 1.5% indicates an intact monolayer cell membrane. For individual cases where LY > 1.5%, if P app The values ​​are similar to other parallel values, and based on scientific judgment, the final data can be adopted.

[0670] The compounds disclosed in this embodiment have good Caco-2 cell permeability.

Claims

1. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in, X 2 and X 4 One is N, and the other is C; R 1 Selected from 4-12 membered heterocyclic groups, 5-10 membered heteroaryl groups, or C6-C 14 arylene, the 4-12 membered heterocyclic arylene, 5-10 membered heterocyclic arylene, or C6-C 14 Alpha-aryl optional R 6’ replace; R 6’ Selected from halogens, hydroxyl groups, C1-C6 alkyl groups, amino groups, oxo groups, cyano groups, C3-C6 cycloalkyl groups, or C1-C6 alkoxy groups; R 6 Selected from hydrogen, hydroxyl, amino, halogen, oxo, cyano, C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl, the C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl optional R 6a replace; R 6a Selected from oxo, halogen, cyano, amino, OH, C1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), NH(C3-C6 cycloalkyl), N(C1-C6 alkyl)(C3-C6 cycloalkyl), C1-C6 alkoxy, 4-12 membered heterocyclic groups, C3-C 10 Cycloalkyl or COOH, wherein the C1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), NH(C3-C6 cycloalkyl), N(C1-C6 alkyl)(C3-C6 cycloalkyl), C1-C6 alkoxy, 4-12 membered heterocyclic or C3-C 10 cycloalkyl optional R 6b replace; R 6b Selected from deuterium, halogen, oxo, hydroxyl, amino, cyano, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl or C1-C6 alkoxy is optionally R 6c replace; R 6c Selected from halogen, hydroxyl, amino, or cyano groups; R 2 Selected from phenyl or 5-10 heteroaryl groups, wherein the phenyl or 5-10 heteroaryl group is optionally R 2a replace; R 2a Selected from halogens, C1-C6 alkyl groups, SF5 or C(O)H, wherein the C1-C6 alkyl group is optionally substituted with a halogen; R 3 Selected from halogens or C1-C4 alkyl groups, wherein the C1-C4 alkyl group is optionally R 3a Substitution; or two R atoms attached to the same ring atom 3 Together with the atoms they are attached to, they form a C3-C6 cycloalkane ring; R 3a Selected from halogens or hydroxyl groups; x is selected from 0, 1, or 2; 'e' can be a single or double bond. When e is a single bond, Y is N or CH; When e is a double bond, Y is C; y is selected from 0, 1, or 2; R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a C3-C6 cycloalkane ring; R 4 Selected from C1-C4 alkyl, 5-10 heteroaryl, or 4-12 heterocyclic groups, wherein the C1-C4 alkyl, 5-10 heteroaryl, or 4-12 heterocyclic group is optionally R 4a replace; R 4a Selected from halogens, hydroxyl groups, oxo groups, C1-C4 alkyl groups, or C1-C4 alkoxy groups, wherein the C1-C4 alkyl groups or C1-C4 alkoxy groups are optionally substituted with halogens.

2. The compound of formula (I) according to claim 1, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, X 2 Let C, X 4 For N, or Among them, X 2 Let N, X 4 For C, or Where X 2 Let C, X 4 Let N be the number of elements, and... Selected from When x is phenyl, x is selected from 1 or 2; or Where X 2 Let C, X 4 Let N be a subset of x, and R be a subset of x. 3 When selected from methyl, Not for 3. The compound of formula (I) according to claim 1 or 2, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 Selected from 4-10 heterocyclic groups, 5-10 heteroaryl groups, or C6-C 10 arylene, the 4-10 member heterocyclic arylene, 5-10 member heterocyclic arylene or C6-C 10 Alpha-aryl optional R 6’ Replace, or Among them, R 1 Selected from 4-6-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 4-6-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally coated with R. 6’ Replace, or Among them, R 1 Selected from 6-8-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 6-8-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally coated with R. 6’ Replace, or Among them, R 1 Selected from 6-membered heterocyclic groups, 5-6-membered heteroaryl groups, or phenylene groups, wherein the 6-membered heterocyclic group, 5-6-membered heteroaryl group, or phenylene group is optionally R-coated. 6’ Replace, or Among them, R 1 Selected from tetrahydropyridyl, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the tetrahydropyridyl group, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Optional R 6’ Replace, or Among them, R 1 Selected from tetrahydropyridyl, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Where # represents X 2 Connect, and with R 6 The connection site is any location where hydrogen is present, the tetrahydropyridyl group, Pyridyl, phenylene, thiazolyl, tetrahydropyranyl or Optional R 6’ Replace, or Among them, R 1 Selected from phenylene Where # represents X 2 Connect, and with R 6 The connection site is any location where hydrogen is present, the phenylene, Optional R 6’ Replace, or Among them, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 Connection, the Optional R 6’ Replace, or Among them, R 1 Selected from Where # represents X 2 Connect, and with R 6 The connection site is any location where hydrogen is present. Optional R 6’ Replace, or Among them, R 1 Selected from Where * represents R 6 The # symbol represents a connection to X. 2 Connection, or Among them, R 1 Selected from tetrahydropyridyl, Or 5-10 methyl aryl groups, where * represents R 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or 5-10 heteroaryl group is optionally R 6’ Replace, or Among them, R 1 Selected from tetrahydropyridyl, Or a 6-membered heteroaryl group, where * represents a group with R. 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or 6-membered heteroaryl group is optionally R 6’ Replace, or Among them, R 1 Selected from tetrahydropyridyl, Or pyridyl group, where * represents the group with R 6 The # symbol represents a connection to X. 2 The linker, wherein the tetrahydropyridyl or pyridyl group is optionally R 6’ Replace, or Among them, R 1 Selected from dihydropyridyl or 5-10-membered heteroaryl, wherein the dihydropyridyl or 5-10-membered heteroaryl is optionally R 6’ Replace, or Among them, R 1 Selected from dihydropyridyl or 6-membered heteroaryl, wherein the dihydropyridyl or 6-membered heteroaryl is optionally R 6’ Replace, or Among them, R 1 Selected from dihydropyridyl or pyridylene, wherein the dihydropyridyl or pyridylene is optionally R 6’ replace, Among them, R 1 Selected from tetrahydropyridyl, Tetrahydropyranyl or Where # represents X 2 Connection, the tetrahydropyridyl group, Tetrahydropyranyl or Optional R 6’ Replace, or Among them, R 1 Selected from tetrahydropyridyl, wherein the tetrahydropyridyl group is optionally R 6’ replace.

4. The compound of formula (I) according to any one of claims 1-3, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 6’ Selected from halogens or C1-C3 alkyl groups, or Among them, R 6’ Selected from halogens or hydroxyl groups, or Among them, R 6’ Selected from fluorine, chlorine or methyl, or Among them, R 6’ Selected from fluorine.

5. The compound of formula (I) according to any one of claims 1-4, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 6 Selected from hydroxyl, amino, halogen, cyano, C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl, the C(O)C3-C 10 Cycloalkyl, C(O)C1-C 10 Alkyl, 5-10 membered heteroaryl, C(O)4-12 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C 10 Cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C 10 Alkyl, C1-C 10 Alkoxy group, -O-C3-C 10 cycloalkyl, -S-C3-C 10 Cycloalkyl, -O-4-12-membered heterocyclic group, -S-4-12-membered heterocyclic group, -S-Cl-C 10 Alkyl, C3-C 10 Cycloalkyl, -S(O)-C3-C 10 Cycloalkyl, -S(O)-C1-C 10 Alkyl group, -S(O)2-C3-C 10 Cycloalkyl, -S(O)2-C1-C 10 Alkyl, 4-12 membered heterocyclic groups, -N=S(O)(C1-C6 alkyl)2, -S(O)(NH)-C3-C 10 Cycloalkyl, -S(O)(NC1-C6 alkyl)-C3-C 10 Cycloalkyl, -S(O)(NH)-C1-C 10 Alkyl or -S(O)(NC1-C6 alkyl)-C1-C 10 Alkyl optional R 6a Replace, or Among them, R 6 Selected from C(O)C3-C8 cycloalkyl, C(O)C1-C8 alkyl, C(O)4-10 membered heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C8 cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C8 alkyl, C1-C8 alkoxy, -O-C3-C8 cycloalkyl, -S-C3-C8 cycloalkyl, -O-4-10 membered heterocyclic, -S-4-10 membered heterocyclic, -S-C1-C8 alkyl, C3-C8 cycloalkyl, -S(O)-C3-C8 cycloalkyl, -S(O)-C1-C8 alkyl, -S(O)2-C3-C8 cycloalkyl, -S(O)2-C1-C8 alkyl or 4-12 membered heterocyclic, The following C(O)C3-C8 cycloalkyl, C(O)C1-C8 alkyl, C(O)4-10 heterocyclic, C(O)NHC1-C6 alkyl, C(O)OC3-C8 cycloalkyl, C(O)OC1-C6 alkyl, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C8 alkyl, C1-C8 alkoxy, -O-C3-C8 cycloalkyl, -S-C3-C8 cycloalkyl, -O-4-10 heterocyclic, -S-4-10 heterocyclic, -S-C1-C8 alkyl, C3-C8 cycloalkyl, -S(O)-C3-C8 cycloalkyl, -S(O)-C1-C8 alkyl, -S(O)2-C3-C8 cycloalkyl, -S(O)2-C1-C8 alkyl or 4-12 heterocyclic groups are optionally R 6a Replace, or Among them, R 6 Selected from C3-C6 cycloalkyl, 4-6 membered heterocyclic, C1-C6 alkyl, or C1-C6 alkoxy groups, wherein the C3-C6 cycloalkyl, 4-6 membered heterocyclic, C1-C6 alkyl, or C1-C6 alkoxy group is optionally R 6a Replace, or Among them, R 6 Selected from C3-C4 cycloalkyl, 4-membered heterocyclic, C1-C3 alkyl, or C1-C3 alkoxy groups, wherein the C3-C4 cycloalkyl, 4-membered heterocyclic, C1-C3 alkyl, or C1-C3 alkoxy group is optionally R 6a Replace, or Among them, R 6 Selected from cyclopropyl, cyclobutyl, oxetyl, azirone, methyl, or methoxy, wherein the cyclopropyl, cyclobutyl, oxetyl, azirone, methyl, or methoxy group is optionally R-coated. 6a replace.

6. The compound of formula (I) according to any one of claims 1-5, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 6a Selected from halogens, cyano groups, amino groups, OH groups, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), 4-10 membered heterocyclic groups, or C3-C8 cycloalkyl groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl), 4-10 membered heterocyclic groups, or C3-C8 cycloalkyl groups are optionally modified by R. 6b Replace, or Among them, R 6a Selected from halogens, N(C1-C6 alkyl)2, 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, 4-10 membered heterocyclic groups are optionally R 6b Replace, or Among them, R 6a Selected from halogens, N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b Replace, or Among them, R 6a Selected from fluorine, N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, The N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, Optional R 6b replace Among them, R 6b Selected from deuterium, halogen, cyano, C1-C6 alkoxy, or C1-C6 alkyl, wherein the C1-C6 alkoxy or C1-C6 alkyl is optionally R 6c Replace, or Among them, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c Replace, or Among them, R 6b Selected from deuterium, fluorine, cyano, methoxy, or methyl, wherein the methoxy or methyl group is optionally R 6c replace, Among them, R 6c Selected from halogen.

7. The compound of formula (I) according to any one of claims 1-6, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 6 Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl group is R 6a Replace, R 6b Selected from N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic group is optionally R 6b Substitution, wherein the atom connecting the 4-10 membered heterocyclic group to the C1-C6 alkyl group is a heteroatom, or Among them, R 6 Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl group is R 6a Replace, R 6b Selected from N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic groups, wherein the N(C1-C6 alkyl)2, NH(C1-C6 alkyl) or 4-10 membered heterocyclic group is optionally R 6b Substitution, wherein the atom connecting the 4-10 membered heterocyclic group to the C1-C6 alkyl group is a nitrogen atom, or Among them, R 6 Selected from hydrogen, methyl, methoxy, cyclopropyl, or Among them, R 6 Selected from methyl, methoxy, cyclopropyl, or R 6 Selected from C1-C3 alkyl groups, wherein the C1-C3 alkyl group is R 6a Replace, R 6a Selected from N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c Replace, R 6c Selected from halogens, preferably wherein R 6a The atom attached to the C1-C3 alkyl group is a nitrogen atom, or Among them, R 6 Selected from methyl, said methyl group is R 6a Replace, R 6a Selected from N(C1-C3 alkyl)2, 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic group is optionally R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally R 6c Replace, R 6c Selected from halogens, preferably wherein R 6a The atom bonded to the methyl group is a nitrogen atom, or Among them, R 6 Selected from methyl, said methyl is R 6a Replace, R 6a Selected from N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, The N(CH3)2, nitrogen-containing heterocyclic butyl, pyrrolidinyl, Optional R 6b Replace, R 6b Selected from deuterium, fluorine, cyano, methoxy, or methyl, wherein the methoxy or methyl group is optionally R 6c Replace, R 6c Selected from halogen.

8. The compound of formula (I) according to any one of claims 1-7, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Selected from or Selected from or in, Selected from or Among them, R 1 Selected from: 6-membered heterocyclic group, R 6 Selected from C3-C4 cycloalkyl or 4-membered heterocyclic groups, wherein the C3-C4 cycloalkyl or 4-membered heterocyclic group is optionally R 6a Replace, or Among them, R 1 Selected from 8-membered heterocyclic groups, R 6 Selected from hydrogen, or, Among them, R 1 Selected from phenyl, wherein the phenyl is optionally R 6’ Replace, R 6 Selected from C1-C3 alkyl groups, wherein the C1-C3 alkyl group is substituted with N(C1-C3 alkyl)2 or 4-7 membered heterocyclic groups, wherein the N(C1-C3 alkyl)2 or 4-7 membered heterocyclic groups are optionally replaced with R 6b Replace, R 6b Selected from deuterium, halogen, cyano, C1-C3 alkoxy, or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl may optionally be substituted with a halogen, or... Among them, R 1 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group is optionally replaced by R 6’ Replace, R 6 Selected from C1-C3 alkyl, C1-C3 alkoxy, or C3-C4 cycloalkyl, wherein the C1-C3 alkyl group is substituted with a halogen or a 4-7 membered heterocyclic group, wherein the 4-7 membered heterocyclic group is optionally replaced with R. 6b Replace, R 6b It is selected from deuterium, halogen, cyano, C1-C3 alkoxy or C1-C3 alkyl, wherein the C1-C3 alkoxy or C1-C3 alkyl is optionally substituted with halogen.

9. The compound of formula (I) according to any one of claims 1-8, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 2 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 2a Replace, or Among them, R 2 Selected from phenyl or 9-membered heteroaryl, wherein the phenyl or 9-membered heteroaryl group is optionally R 2a Replace, or Among them, R 2 Selected from phenyl, benzofuranyl, or benzothiophene, wherein the phenyl, benzofuranyl, or benzothiophene group is optionally R 2a Replace, or Among them, R 2 Selected from or Among them, R 2a Selected from halogens or C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with halogens, or Among them, R 2a Selected from halogens or C1-C3 alkyl groups, wherein the C1-C3 alkyl groups are optionally substituted with halogens, or Among them, R 2a Selected from chlorine or methyl, wherein the methyl group is optionally substituted with a halogen, or Among them, R 3 Selected from halogens or C1-C4 alkyl groups, wherein the C1-C4 alkyl group is optionally R 3a Replace, or Among them, R 3 Selected from C1-C4 alkyl groups, or Among them, R 3 Selected from methyl, or Where x is selected from 1 or 2, or Where x is selected from 1.

10. The compound of formula (I) according to any one of claims 1-9, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, 'e' can be a single or double bond. When e is a single bond, Y is N or CH; When e is a double bond, Y is C; y is selected from 0, 1, or 2; R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5 The atoms and bonds they are connected to together form a C3-C4 cycloalkane ring, or Where e is a single bond and Y is N or CH; y is selected from 0, 1, or 2; R 5 Selected from C1-C4 alkyl groups, or two R atoms attached to adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a C3-C4 cycloalkane ring, or in, Selected from or Where 'e' is a single or double bond. When e is a single bond, Y is CH, and y is selected from 2, where R is attached to two adjacent ring atoms. 5 Together with the atoms and bonds they are connected to, they form a cyclopropane ring; When e is a double bond, Y is C, or Where e is a single bond, Y is CH, y is selected from 2, and R is two bonds attached to adjacent ring atoms. 5 The atoms and bonds to which it is attached together form a cyclopropane ring, or in, Selected from or in, Selected from 11. The compound of formula (I) according to any one of claims 1-10, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 4 Selected from 5-10-membered heteroaryl or 4-10-membered heterocyclic groups, wherein the 5-10-membered heteroaryl or 4-10-membered heterocyclic group is optionally R 4a Replace, or Among them, R 4 Selected from 5-6-membered heteroaryl groups, wherein the 5-6-membered heteroaryl group is optionally replaced by R 4a Replace, or Among them, R 4 Selected from 6-membered heteroaryl groups, wherein the 6-membered heteroaryl group is optionally replaced by R 4a Replace, or Among them, R 4 Selected from pyrimidinyl groups, wherein the pyrimidinyl group is optionally R 4a Replace, or Among them, R 4a Selected from halogens, hydroxyl groups, or C1-C4 alkyl groups, wherein the C1-C4 alkyl groups are optionally substituted with halogens, or Among them, R 4a Selected from hydroxyl or C1-C4 alkyl, or Among them, R 4a Selected from hydroxy or methyl, or Among them, R 4 Selected from 5-10-membered heteroaryl groups, wherein at least one ortho-position of the 5-10-membered heteroaryl group is substituted with a hydroxyl group, and the 5-10-membered heteroaryl group substituted with at least one ortho-position hydroxyl group is further optionally R 4a Replace, or Among them, R 4 Selected from 5-10-membered heteroaryl groups, wherein one ortho-position of the 5-10-membered heteroaryl group is substituted with a hydroxyl group, and the 5-10-membered heteroaryl group substituted with a hydroxyl group at the ortho-position is further optionally R 4a Replace, or Among them, R 4 Selected from 12. The compound of formula (I) according to any one of claims 1-11, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) is selected from the compounds of formula (IV) or formula (IV'): Where e, x, y, X 2 X 4 R 1 R 2 R 3 R 4 R 5 R 6a And Y as defined in any one of claims 1-11, wherein q1 is selected from 1, 2, 3, 4, 5, 6, 7 or 8, q2 is selected from 0, 1, 2, 3, 4, 5, 6, 7 or 8, or Where q1 is selected from 1 or 2, or Where q2 is selected from 0 or 2, or Wherein, the compound of formula (I) is selected from the compound of formula (VI) or the compound of formula (VI'): Where e, x, y, X 2 X 4 R 2 R 3 R 4 R 5 R 6 Y is as defined in any one of claims 1-11.

13. The compound of formula (I) according to claim 1, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compounds of formula (I) disclosed herein are selected from the following compounds:

14. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

15. A compound of any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof for inhibiting WRN, or a pharmaceutical composition of claim 14.

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