Five-membered heteroaromatic compound and use thereof

Abstract

The present disclosure provides a five-membered heteroaromatic compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, a pharmaceutical composition containing same, and a use thereof as a WRN inhibitor in the prevention or treatment of related diseases.

Description

Five-membered heteroaromatic ring compounds and applications thereof

[0001] Cross-reference to Related Applications

[0002] This application claims priority to and the benefit of the following Chinese patent applications: Chinese Patent Application No. 202410812440.0, filed on June 21, 2024; Chinese Patent Application No. 202411083296.8, filed on August 8, 2024; Chinese Patent Application No. 202411385008.4, filed on September 30, 2024; Chinese Patent Application No. 202510050857.2, filed on January 13, 2025. The entire contents of the above-identified patent applications are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0003] The present disclosure relates to five-membered heteroaromatic ring compounds or pharmaceutically acceptable salts thereof or stereoisomers thereof, methods for preparing the same, pharmaceutical compositions containing the same, and uses thereof as WRN inhibitors in the prevention or treatment of related diseases. BACKGROUND

[0004] Helicases are a class of enzymes that can unwind nucleotide double strands, which can move along the nucleic acid chain using the energy released by nucleotide triphosphate hydrolysis, thereby completing the unwinding and separation of nucleic acid double strands. As a representative of DNA helicases, the RecQ helicase family is a class of protein factors that can play a huge role in maintaining the stability of chromosomal genomes and telomeres. In the process of DNA replication, it plays a catalytic role in unwinding double-stranded DNA, completes DNA repair, and thus maintains the integrity of the genome.

[0005] RecQ family helicases are highly conserved throughout evolution, and their functions are involved in various DNA metabolic processes, which are crucial for maintaining genome stability. The loss of functions of BLM, WRN and RecQ4, which are 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 DNA damage sensitivity at the molecular level.

[0006] WRN and RecQ1 are two most representative RecQ helicases, WRN includes an exonuclease domain in addition to the helicase domain. Werner syndrome (WS) is an autosomal recessive disorder that presents with symptoms of accelerated clinical aging, leading to a mean life expectancy of less than 50 years. WRN helicase / nuclease (WRN) is involved in many important pathways, including DNA replication, recombination and repair. WRN can unwind non-canonical secondary DNA structures that can be encountered during replication and recombination processes, and its mutation can cause chromosomal instability diseases such as Werner syndrome. Normally, WRN depletion leads to DNA double-strand breaks in MSL cells, resulting in cell cycle arrest and / or apoptosis. Studies have shown that there is a strong synthetic lethal relationship between WRN helicase and microsatellite instability-high (MSI-H) cancer. WRN forms a synthetic lethal relationship with MutL homolog 1 (MLH1), and MLH1 loss is associated with microsatellite instability (MSI).

[0007] Microsatellites are simple repeat sequences of less than 10 nucleotides in the genome, and DNA mismatch repair (MMR) defects caused by gene mutations or promoter hypermethylation can cause hypermutation of nucleotide repeat regions (microsatellites), i.e. microsatellite instability (MSI). MSI can promote the occurrence of various cancers, including colon cancer (15%), gastric cancer (22%), endometrial cancer (20-30%) and ovarian cancer (12%), of which 45-60% are not responsive to immune checkpoint blockade. Therefore, MSI-type tumors need new treatment methods. In two whole-genome gene inactivation studies using CRISPR or RNA interference, WRN was identified as the most essential dependency of MSI-H cells, and this dependency is related to the helicase activity, but not to the nuclease function of WRN. These findings, combined with the good tolerance of microsatellite stable (MSS) cancer cells to WRN silencing, demonstrate that WRN is a potential specific target for treating MSI tumors.

[0008] In view of the huge unmet clinical needs, the development of small molecule inhibitors targeting WRN has broad application prospects. SUMMARY

[0009] The present disclosure relates to a compound of Formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof,

[0010] wherein Y 1 , Y 2 , Y 3 are each independently selected from N, NH, CH, C, S, Se or O, Y 4Y 5 each independently selected from N or C;

[0011] n is selected from 0, 1, 2 or 3;

[0012] R 2 selected from halogen, cyano, NHC(=0)Ci-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 10 aryl, Ci-C6alkyl, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, NH(C3-C8cycloalkyl), N(Ci-C6alkyl)(C3-C8cycloalkyl), C2-C6alkenyl, -S-Ci-C6alkyl, Ci-C6alkoxy, -O-C3-C8cycloalkyl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl, said NHC(=0)Ci-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 10 aryl, Ci-C6alkyl, amino, NH(Ci-C6alkyl), N(Ci-C6alkyl)2, NH(C3-C8cycloalkyl), N(Ci-C6alkyl)(C3-C8cycloalkyl), C2-C6alkenyl, -S-Ci-C6alkyl, Ci-C6alkoxy, -O-C3-C8cycloalkyl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl, said NHC(=0)Ci-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 2a substituted; or two R 2 and the atom to which they are attached together form a C5-C8cycloalkenyl, C6-C 10 aryl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl, said C5-C8cycloalkenyl, C6-C 10 aryl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl, said C5-C8cycloalkenyl, C6-C 2A substituted;

[0013] R 2a selected from hydroxyl, oxo, halogen, Ci-C6alkyl, phenyl, Ci-C6alkoxy, C3-C6cycloalkyl, cyano, amino, carboxyl, -S-Ci-C6alkyl, -S-C3-C8cycloalkyl, or -O-C3-C8cycloalkyl, said Ci-C6alkyl, phenyl, Ci-C6alkoxy, C3-C6cycloalkyl, amino, -S-Ci-C6alkyl, -S-C3-C8cycloalkyl, or -O-C3-C8cycloalkyl optionally substituted with R 2b substituted;

[0014] R 2b selected from halogen or Ci-C6alkyl;

[0015] R 2Ahalogen, cyano, NHC(=O)C1-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 10 aryl, C1-C6alkyl, amino, NH(C1-C6alkyl), N(C1-C6alkyl)2, NH(C3-C8cycloalkyl), N(C1-C6alkyl)(C3-C8cycloalkyl), C2-C6alkenyl, -S-C1-C6alkyl, C1-C6alkoxy, -O-C3-C8cycloalkyl, 4- to 12-membered heterocyclyl, or 5- to 10-membered heteroaryl, said NHC(=O)C1-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 10 aryl, C1-C6alkyl, amino, NH(C1-C6alkyl), N(C1-C6alkyl)2, NH(C3-C8cycloalkyl), N(C1-C6alkyl)(C3-C8cycloalkyl), C2-C6alkenyl, -S-C1-C6alkyl, C1-C6alkoxy, -O-C3-C8cycloalkyl, 4- to 12-membered heterocyclyl, or 5- to 10-membered heteroaryl, said NHC(=O)C1-C6alkyl, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C6-C 2B substituted;

[0016] R 2B selected from the group consisting of hydroxy, oxo, halogen, C1-C6alkyl, phenyl, C1-C6alkoxy, C3-C6cycloalkyl, cyano, amino, carboxyl, -S-C1-C6alkyl, -S-C3-C8cycloalkyl, or -O-C3-C8cycloalkyl, said C1-C6alkyl, phenyl, C1-C6alkoxy, C3-C6cycloalkyl, amino, -S-C1-C6alkyl, -S-C3-C8cycloalkyl, or -O-C3-C8cycloalkyl optionally substituted with R 2C substituted;

[0017] R 2C selected from the group consisting of halogen or C1-C6alkyl;

[0018] L 1 selected from the group consisting of a bond, O, S, NH, CH2, or OCH2;

[0019] R 1 selected from the group consisting of C3-C8cycloalkyl, C1-C6alkyl, C6-C 10 aryl, 5- to 10-membered heteroaryl, or 4- to 12-membered heterocyclyl, said C3-C8cycloalkyl, C1-C6alkyl, C6-C 10 aryl, 5- to 10-membered heteroaryl, or 4- to 12-membered heterocyclyl optionally substituted with R 1a substituted;

[0020] R 1aselected from halogen, deuterium, cyano, hydroxy, amino, C1-C6alkyl, C3-C6cycloalkyl, C1-C6alkoxy, oxo, -S-C1-C6alkyl, -O-C3-C8cycloalkyl, -S-C3-C8cycloalkyl, or carboxy;

[0021] L 2 selected from -C(=O)NH-, -C(=S)NH-, -S(=O)2NH-, 1,2,3-triazidylidene, oxadiazidylidene, imidazidylidene, tetrazolylidene, pyrazolylidene, -NHC(=O)NH-, -NHC(=O)O-, -CH(CF3)NH-, or

[0022] R 3 selected from hydrogen, C1-C6alkyl, C3-C8cycloalkyl, 4-12 membered heterocyclyl, C6-C10aryl, or 5-10 membered heteroaryl, said C1-C6alkyl, C3-C8cycloalkyl, 4-12 membered heterocyclyl, C6-C10aryl, or 5-10 membered heteroaryl optionally substituted with R 10 ; 10 ; 3a ;

[0023] R 3a selected from halogen, cyano, hydroxy, amino, carboxy, oxo, -S-C3-C8cycloalkyl, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, -S-C1-C6alkyl, -O-C3-C6cycloalkyl, C(=O)NH2, C(=O)NH(C1-C6alkyl), C(=O)N(C1-C6alkyl)2, or phenoxy, said amino, -S-C3-C8cycloalkyl, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, -S-C1-C6alkyl, -O-C3-C6cycloalkyl, C(=O)NH2, C(=O)NH(C1-C6alkyl), C(=O)N(C1-C6alkyl)2, or phenoxy optionally substituted with R 3b ;

[0024] R 3b selected from halogen or C1-C6alkyl;

[0025] Q is selected from Q 1 , Q 2 , Q 3 , or Q 4 ;

[0026] Q 1 is wherein, represents is in the (Z) configuration or in the (E) configuration;

[0027] X 3is selected from O, NH or NR 5 ;

[0028] R 4 is selected from C1-C6alkyl, C3-C6cycloalkyl, 4-6 membered heterocyclyl or NR 8 R 9 , said C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl being optionally substituted with R 4a ;

[0029] R 4a is selected from halogen, cyano, hydroxy, amino, C1-C6alkyl or C1-C6alkoxy;

[0030] R 11 is selected from hydrogen, or R 11 , R 4 and the atom to which they are attached together form a 4-6 membered heterocyclyl, said 4-6 membered heterocyclyl being optionally substituted with R 11a ;

[0031] R 11a is selected from halogen, cyano, hydroxy, amino, C1-C6alkyl, C1-C6alkoxy or C3-C6cycloalkyl;

[0032] R 8 , R 9 are each independently selected from hydrogen, C1-C6alkyl, C3-C6cycloalkyl, 4-6 membered heterocyclyl or C(O)C1-C6alkyl, said C1-C6alkyl, C3-C6cycloalkyl, 4-6 membered heterocyclyl or C(O)C1-C6alkyl being optionally substituted with R 8a ; or R 8 , R 9 and the nitrogen atom to which they are attached together form a 4-12 membered heterocyclyl, said 4-12 membered heterocyclyl being optionally substituted with R 9a ;

[0033] R 8a , R 9a are each independently selected from halogen, hydroxy, amino, cyano, C1-C6alkoxy or C1-C6alkyl;

[0034] R 5 is selected from cyano, C(=O)C1-C6alkyl, C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl, said C(=O)C1-C6alkyl, C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl being optionally substituted with R 5a ; or, said R 5 , R 4 and the atom to which they are attached together form a 4-10 membered heterocyclyl, said 4-10 membered heterocyclyl being optionally substituted with R 5a ;

[0035] R 5a is selected from halogen, cyano, hydroxy, amino, C1-C6alkyl or C1-C6alkoxy;

[0036] Q 2 is wherein, represents is in the (Z) configuration or in the (E) configuration;

[0037] Q 3 is

[0038] Q 4 is wherein, R a is C1-C6alkyl, which is optionally substituted.

[0039] In some embodiments, Y 1 , Y 2 , Y 3 are each independently selected from N, NH, CH, C, S or O, Y 4 , Y 5 are each independently selected from N or C;

[0040] Q 1 is wherein, represents is in the (Z) configuration or in the (E) configuration;

[0041] R 4 is selected from C1-C6alkyl, C3-C6cycloalkyl, 4-6 membered heterocyclyl or NR 8 R 9 , which C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl is optionally substituted with R 4a ;

[0042] R 4a is selected from halogen, cyano, hydroxy, amino, C1-C6alkyl or C1-C6alkoxy;

[0043] R 5 is selected from cyano, C(=O)C1-C6alkyl, C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl, which C(=O)C1-C6alkyl, C1-C6alkyl, C3-C6cycloalkyl or 4-6 membered heterocyclyl is optionally substituted with R 5a .

[0044] In some embodiments, Y 1 , Y 2 , Y 3are each independently selected from N, NH, CH, C, S, or O, Y 4 , Y 5 are each independently selected from N or C.

[0045] In some embodiments, Y 1 , Y 2 , Y 3 are each independently selected from N, NH, CH, C, or S, Y 4 , Y 5 are each independently selected from N or C.

[0046] In some embodiments, Y 1 , Y 2 , Y 3 are each independently selected from N, C, CH, S, Se, or NH, Y 4 , Y 5 are each selected from C.

[0047] In some embodiments, Y 1 , Y 2 , Y 3 are each independently selected from N, CH, C, S, or Se, Y 4 , Y 5 are each selected from C.

[0048] In some embodiments, Y 1 , Y 2 , Y 3 are each independently selected from N, CH, C, or S, Y 4 , Y 5 are each selected from C.

[0049] In some embodiments, Y 1 is selected from N, C, or CH, Y 2 is selected from N or C, Y 3 is selected from CH, C, S, N, Se, or NH, Y 4 and Y 5 are each selected from C.

[0050] In some embodiments, Y 1 is selected from N or C, Y 2 is selected from N or C, Y 3 is selected from CH, C, S, N, or Se, Y 4 and Y 5 are each selected from C.

[0051] In some embodiments, Y 1 is selected from N or C, Y 2 is selected from N or C, Y 3 is selected from CH, C, or S, Y 4 and Y 5each is selected from C.

[0052] In some embodiments, Y is selected from 1 In some embodiments, Y is selected from 2 In some embodiments, Y is selected from 3 In some embodiments, Y is selected from 4 In some embodiments, Y is selected from 5 In some embodiments, at least one of Y is a heteroatom selected from a nitrogen atom, a sulfur atom, or a selenium atom.

[0053] In some embodiments, Y is selected from 1 In some embodiments, Y is selected from 2 In some embodiments, Y is selected from 3 In some embodiments, Y is selected from 4 In some embodiments, Y is selected from 5 In some embodiments, at least one of Y is a nitrogen atom or a sulfur atom.

[0054] In some embodiments, Y is selected from 1 In some embodiments, Y is selected from 2 In some embodiments, Y is selected from 3 In some embodiments, Y is selected from 4 In some embodiments, Y is selected from 5 In some embodiments, at least one of Y is a nitrogen atom.

[0055] In some embodiments, Y is selected from 1 In some embodiments, Y is selected from 2 In some embodiments, Y is selected from 3 In some embodiments, at least one of Y is N.

[0056] In some embodiments, n is selected from 1 or 2.

[0057] In some embodiments, n is selected from 1.

[0058] In some embodiments, n is selected from 2.

[0059] In some embodiments, is selected from

[0060] In some embodiments, is selected from

[0061] In some embodiments, is selected from

[0062] In some embodiments, is selected from wherein * represents the atom to which L 1 is attached.

[0063] In some embodiments, is selected from

[0064] In some embodiments, is selected from

[0065] In some embodiments, is selected from In some embodiments, is selected from wherein * represents and L 1 the atom to which they are attached.

[0066] In some embodiments, each R 2 is independently selected from halogen, cyano, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C1-C6alkyl, amino, NH(C1-C6alkyl), -S-C1-C6alkyl, C1-C6alkoxy, or -O-C3-C8cycloalkyl, which C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C1-C6alkyl, amino, NH(C1-C6alkyl), -S-C1-C6alkyl, C1-C6alkoxy, or -O-C3-C8cycloalkyl is optionally substituted with R 2a and the atom to which they are attached; or two R 2 and the atom to which they are attached form, together with the carbon atom to which they are attached, a C6-C 10 aryl or 5-10 membered heteroaryl, which C6-C 10 aryl or 5-10 membered heteroaryl is optionally substituted with R 2A .

[0067] In some embodiments, each R 2 is independently selected from C1-C6alkyl, N(C1-C6alkyl)2, C3-C6cycloalkyl, N(C1-C6alkyl)(C3-C6cycloalkyl), or C(O)C3-C6cycloalkyl, which C1-C6alkyl, N(C1-C6alkyl)2, C3-C6cycloalkyl, N(C1-C6alkyl)(C3-C6cycloalkyl), or C(O)C3-C6cycloalkyl is optionally substituted with R 2a and the atom to which they are attached; or two R 2 and the atom to which they are attached form, together with the carbon atom to which they are attached, a 5-10 membered heteroaryl or 4-10 membered heterocyclyl, which 5-10 membered heteroaryl or 4-10 membered heterocyclyl is optionally substituted with R 2A .

[0068] In some embodiments, each R 2independently selected from C1-C6alkyl, N(C1-C6alkyl)2, or C(O)C3-C6cycloalkyl, said C1-C6alkyl, N(C1-C6alkyl)2, or C(O)C3-C6cycloalkyl optionally substituted with R 2a substituted; or two R 2 atoms to which they are attached collectively form a 5-10 membered heteroaryl or 4-10 membered heterocyclyl, said 5-10 membered heteroaryl or 4-10 membered heterocyclyl optionally substituted with R 2A substituted.

[0069] In some embodiments, each R 2 independently selected from C1-C6alkyl, C3-C6cycloalkyl, or C(O)C3-C6cycloalkyl, said C1-C6alkyl, C3-C6cycloalkyl, or C(O)C3-C6cycloalkyl optionally substituted with R 2a substituted; or two R 2 atoms to which they are attached collectively form a 5-10 membered heteroaryl, said 5-10 membered heteroaryl optionally substituted with R 2A substituted.

[0070] In some embodiments, each R 2 independently selected from C1-C4alkyl, N(C1-C3alkyl)2, C3-C6cycloalkyl, N(C1-C3alkyl)(C3-C4cycloalkyl), or -C(O)C3-C4cycloalkyl, said C1-C4alkyl, N(C1-C3alkyl)2, C3-C6cycloalkyl, N(C1-C3alkyl)(C3-C4cycloalkyl), or -C(O)C3-C4cycloalkyl optionally substituted with R 2a substituted; two R 2 atoms to which they are attached collectively form a 6 membered heteroaryl or 5 membered heterocyclyl, said 6 membered heteroaryl or 5 membered heterocyclyl optionally substituted with R 2A substituted.

[0071] In some embodiments, each R 2 independently selected from C1-C4alkyl, N(C1-C3alkyl)2, or -C(O)C3-C4cycloalkyl, said C1-C4alkyl, N(C1-C3alkyl)2, or -C(O)C3-C4cycloalkyl optionally substituted with R 2a substituted; two R 2 atoms to which they are attached collectively form a 6 membered heteroaryl or 5 membered heterocyclyl, said 6 membered heteroaryl or 5 membered heterocyclyl optionally substituted with R 2A substituted.

[0072] In some embodiments, each R 2independently selected from C1-C4alkyl, cyclopropyl, or -C(O)cyclopropyl, said C1-C4alkyl optionally substituted with R 2a two R 2 groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R 2A groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0073] In some embodiments, each R 2 is independently selected from C1-C6alkyl, C3-C6cycloalkyl, N(C1-C6alkyl)2, N(C1-C6alkyl)(C3-C6cycloalkyl), or -C(O)C3-C6cycloalkyl, said C1-C6alkyl, C3-C6cycloalkyl, N(C1-C6alkyl)2, N(C1-C6alkyl)(C3-C6cycloalkyl), or -C(O)C3-C6cycloalkyl optionally substituted with R 2a groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0074] In some embodiments, each R 2 is independently selected from C1-C4alkyl, C3-C6cycloalkyl, N(C1-C3alkyl)2, N(C1-C3alkyl)(C3-C4cycloalkyl), or -C(O)C3-C4cycloalkyl, said C1-C4alkyl, C3-C6cycloalkyl, N(C1-C3alkyl)2, N(C1-C3alkyl)(C3-C4cycloalkyl), or -C(O)C3-C4cycloalkyl optionally substituted with R 2a groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0075] In some embodiments, each R 2 is independently selected from methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or -C(O)cyclopropyl, said methyl, ethyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, or -C(O)cyclopropyl optionally substituted with R 2a groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0076] In some embodiments, R 2a is selected from hydroxyl, halogen, cyano, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or amino, said C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or amino optionally substituted with R 2b groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0077] In some embodiments, R 2a is selected from halogen or C3-C6cycloalkyl, said C3-C6cycloalkyl optionally substituted with R 2b groups, together with the atom to which they are attached, form a 6-membered heteroaryl, said 6-membered heteroaryl optionally substituted with R

[0078] In some implementation schemes, R 2a Selected from C3-C6 cycloalkyl groups, wherein the C3-C6 cycloalkyl group is optionally R 2b replace.

[0079] In some implementation schemes, R 2a Selected from halogens or C3-C4 cycloalkyl groups.

[0080] In some implementation schemes, R 2a Selected from C3-C4 cycloalkyl groups.

[0081] In some implementation schemes, R 2a Selected from halogens, C3-C6 cycloalkyl groups, or C1-C6 alkyl groups, wherein the C3-C6 cycloalkyl groups or C1-C6 alkyl groups are optionally R 2b replace.

[0082] In some implementation schemes, R 2a Selected from halogens, C3-C4 cycloalkyl groups, or C1-C3 alkyl groups, wherein the C3-C4 cycloalkyl group or C1-C3 alkyl group is optionally R 2b replace.

[0083] In some implementation schemes, R 2a Selected from fluorine, cyclopropyl, or methyl, wherein the cyclopropyl or methyl group is optionally R 2b replace.

[0084] In some implementation schemes, R 2b Selected from halogens.

[0085] In some implementation schemes, R 2b Selected from fluorine.

[0086] In some implementations, two R atoms connected to adjacent ring atoms 2 Together with the atoms they are attached to, they form a 5-10 membered heteroaryl group or a 4-10 membered heterocyclic group, wherein the 5-10 membered heteroaryl group or the 4-10 membered heterocyclic group is optionally R 2A replace.

[0087] In some implementations, two R atoms connected to adjacent ring atoms 2 Together with the atoms they are attached to, they form a 6-membered heteroaryl or a 5-membered heterocyclic group, wherein the 6-membered heteroaryl or 5-membered heterocyclic group is optionally converted by R 2A replace.

[0088] In some implementations, two R atoms connected to adjacent ring atoms 2 Together with the atoms they are attached to, they form a pyridyl group or The pyridyl or Optional R 2A replace.

[0089] In some embodiments, selected from wherein * represents and L 1 the atom to which it is attached.

[0090] In some embodiments, R 2A selected from halogen, cyano, C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C1-C6alkyl, amino, NH(C1-C6alkyl), -S-C1-C6alkyl, C1-C6alkoxy, or -O-C3-C8cycloalkyl, said C3-C8cycloalkyl, C(O)C3-C8cycloalkyl, C5-C8cycloalkenyl, C1-C6alkyl, amino, NH(C1-C6alkyl), -S-C1-C6alkyl, C1-C6alkoxy, or -O-C3-C8cycloalkyl optionally substituted with R 2B substituents.

[0091] In some embodiments, R 2A selected from C3-C6cycloalkyl or C1-C6alkyl, said C3-C6cycloalkyl or C1-C6alkyl optionally substituted with R 2B substituents.

[0092] In some embodiments, R 2A selected from C3-C4cycloalkyl or C1-C4alkyl.

[0093] In some embodiments, R 2A selected from cyclopropyl or methyl. In some embodiments, L 1 is selected from O, S, NH, CH2, or OCH2.

[0094] In some embodiments, L 1 is O, S, or NH.

[0095] In some embodiments, L 1 is O.

[0096] In some embodiments, R 1 selected from C3-C8cycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl, or 4-12 membered heterocyclyl, said C3-C8cycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl, or 4-12 membered heterocyclyl optionally substituted with R 1a substituents.

[0097] In some embodiments, R 1 selected from C6-C 10 aryl, 5-6 membered heteroaryl, or 5-6 membered heterocyclyl, said C6-C 10aryl, 5-6 membered heteroaryl, or 5-6 membered heterocyclyl, optionally substituted with R 1a substituted.

[0098] In some embodiments, R 1 is selected from C6-C 10 aryl, said C6-C 10 aryl is optionally substituted with R 1a substituted.

[0099] In some embodiments, R 1 is selected from phenyl.

[0100] In some embodiments, L 2 is selected from -C(=O)NH-, -C(=S)NH-, or -S(=O)2NH-.

[0101] In some embodiments, L 2 is selected from -C(=O)NH-.

[0102] In some embodiments, R 3 is selected from C3-C8cycloalkyl or 4-12 membered heterocyclyl, said C3-C8cycloalkyl or 4-12 membered heterocyclyl optionally substituted with R 3a substituted.

[0103] In some embodiments, R 3 is selected from C3-C6cycloalkyl, said C3-C6cycloalkyl optionally substituted with R 3a substituted.

[0104] In some embodiments, R 3 is selected from C3-C6cycloalkyl or 4-10 membered heterocyclyl, said C3-C6cycloalkyl or 4-10 membered heterocyclyl optionally substituted with R 3a substituted.

[0105] In some embodiments, R 3 is selected from C3-C4cycloalkyl or 4-6 membered heterocyclyl.

[0106] In some embodiments, R 3 is selected from C3-C4cycloalkyl.

[0107] In some embodiments, R 3 is selected from C3-C4cycloalkyl or 4 membered heterocyclyl.

[0108] In some embodiments, R 3 is selected from cyclopropyl or oxetanyl.

[0109] In some embodiments, R 3 is selected from cyclopropyl or

[0110] In some embodiments, R 3 is selected from cyclopropyl or azetidinyl.

[0111] In some embodiments, R 3 is selected from methyl, cyclopropyl or ethyl, optionally substituted with R 5a .

[0112] In some embodiments, Q 1 is selected from Q 1 .

[0113] In some embodiments, Q 1 is selected from Q 3 .

[0114] In some embodiments, X 3 is selected from O.

[0115] In some embodiments, X 3 is selected from NH.

[0116] In some embodiments, X 3 is selected from NR 5 .

[0117] In some embodiments, R 5 is selected from C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclyl, optionally substituted with R 5a .

[0118] In some embodiments, R 5 is selected from C1-C6 alkyl or C3-C6 cycloalkyl, optionally substituted with R 5a .

[0119] In some embodiments, R 5 is selected from C1-C3 alkyl or C3-C4 cycloalkyl, optionally substituted with R 5a .

[0120] In some embodiments, R 5 is selected from methyl, cyclopropyl or ethyl, optionally substituted with R 5a . In some embodiments, R 5 is selected from methyl, cyclopropyl or ethyl, optionally substituted with R 5a .

[0121] In some embodiments, R 5a is selected from halogen, C1-C6 alkyl or C1-C6 alkoxy.

[0122] In some embodiments, R 5a is selected from C1-C3alkoxy.

[0123] In some embodiments, R 5a is selected from methoxy.

[0124] In some embodiments, R 5 is selected from methyl, cyclopropyl or

[0125] In some embodiments, R 4 is selected from C1-C6alkyl or 4-6 membered heterocyclyl, said C1-C6alkyl or 4-6 membered heterocyclyl optionally substituted with R 4a .

[0126] In some embodiments, R 4 is selected from C1-C6alkyl or NR 8 R 9 , said C1-C6alkyl optionally substituted with R 4a .

[0127] In some embodiments, R 4 is selected from C1-C6alkyl, said C1-C6alkyl optionally substituted with R 4a .

[0128] In some embodiments, R 4 is selected from C1-C3alkyl or NR 8 R 9 , said C1-C3alkyl optionally substituted with R 4a .

[0129] In some embodiments, R 4 is selected from C1-C3alkyl.

[0130] In some embodiments, R 4 is selected from methyl.

[0131] In some embodiments, R 8 , R 9 are each independently selected from hydrogen, C1-C6alkyl or C3-C6cycloalkyl, said C1-C6alkyl or C3-C6cycloalkyl optionally substituted with R 8a .

[0132] In some embodiments, R 8 , R 9 are each independently selected from hydrogen.

[0133] In some embodiments, R 4a is selected from halogen, cyano, hydroxyl or amino.

[0134] In some embodiments, R 4a is selected from halogen.

[0135] In some embodiments, R 4a is selected from fluorine.

[0136] In some embodiments, X 3 is selected from O, while R 4 is selected from C1-C6 alkyl or NR 8 R 9 , said C1-C6 alkyl being optionally substituted with R 4a , said R 8 , R 9 are each independently selected from hydrogen, C1-C6 alkyl or C3-C6 cycloalkyl, said C1-C6 alkyl or C3-C6 cycloalkyl being optionally substituted with R 8a .

[0137] In some embodiments, X 3 is selected from O, while R 4 is selected from C1-C3 alkyl or NR 8 R 9 , said R 8 , R 9 are each independently selected from hydrogen.

[0138] In some embodiments, X 3 is selected from O, while R 4 is selected from methyl or NH2.

[0139] In some embodiments, X 3 is selected from NH, while R 4 is selected from C1-C6 alkyl, said C1-C6 alkyl being optionally substituted with R 4a , R 4a is selected from halogen, cyano, hydroxyl or amino.

[0140] In some embodiments, X 3 is selected from NH, while R 4 is selected from C1-C3 alkyl, said C1-C3 alkyl being optionally substituted with R 4a , R 4a is selected from halogen.

[0141] In some embodiments, X 3 is selected from NH, while R 4 is selected from methyl, said methyl being optionally substituted with halogen.

[0142] In some embodiments, X 3 is selected from NR 5 , R 5selected from C1-C6alkyl or C3-C6cycloalkyl, said C1-C6alkyl or C3-C6cycloalkyl being optionally substituted with R 5a substituted with R 5a selected from C1-C6alkoxy, while R 4 is selected from C1-C6alkyl, said C1-C6alkyl being optionally substituted with R 4a substituted.

[0143] In some embodiments, X 3 is selected from NR 5 , R 5 is selected from C1-C3alkyl or C3-C6cycloalkyl, said C1-C3alkyl or C3-C6cycloalkyl being optionally substituted with R 5a substituted with R 5a selected from C1-C3alkoxy, while R 4 is selected from C1-C3alkyl.

[0144] In some embodiments, X 3 is selected from NR 5 , R 5 is selected from methyl, cyclopropyl or ethyl, said methyl, cyclopropyl or ethyl being optionally substituted with methoxy, while R 4 is selected from methyl.

[0145] In some embodiments, Q 1 is

[0146] In some embodiments, Q 1 is

[0147] In some embodiments, the compound of formula (I) is selected from a compound of formula (II):

[0148] wherein R 2 , n, L 1 , R 1 , L 2 , R 3 and Q are as defined above.

[0149] In some embodiments, the compound of formula (I) is selected from a compound of formula (III):

[0150] wherein Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 2 , n, L 1 , R1 , L 2 , R 3 and Q are as defined above.

[0151] In some embodiments, the compound of Formula (I) is selected from a compound of Formula (IV):

[0152] wherein Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , R 2 , n, L 1 , R 1 , L 2 , R 3 and R 4 are as defined above.

[0153] In some embodiments, the compound of Formula (I) of the present disclosure is selected from the following compounds:

[0154] In some embodiments, the compound of Formula (I) of the present disclosure is selected from the following compounds:

[0155] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of Formula (I) of the present disclosure, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable excipient.

[0156] In another aspect, the present disclosure provides a method of inhibiting WRN, comprising the step of contacting a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition thereof, with WRN.

[0157] In another aspect, the present disclosure provides a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition thereof, for use as a WRN inhibitor.

[0158] In another aspect, the present disclosure provides a method of treating a disease mediated by WRN helicase in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition thereof.

[0159] In another aspect, the present disclosure provides the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition thereof, in the manufacture of a medicament for preventing or treating a disease mediated by WRN helicase.

[0160] In another aspect, the present disclosure provides a use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, or a pharmaceutical composition thereof, in the prevention or treatment of a WRN helicase-mediated disease.

[0161] In another aspect, the present disclosure provides a compound of Formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, or a pharmaceutical composition thereof, for use in the prevention or treatment of a WRN helicase-mediated disease.

[0162] In some embodiments, the WRN helicase-mediated disease is a disease in which WRN helicase is inhibited.

[0163] In some embodiments, the WRN helicase-mediated disease is selected from cancer or Werner syndrome.

[0164] In some embodiments, the WRN helicase-mediated disease is selected from cancer.

[0165] In some embodiments, the cancer is colorectal cancer.

[0166] Definitions and explanations of terms

[0167] Unless otherwise indicated, the terms used in the present disclosure have the following meanings. The definitions of groups and terms recited in the present disclosure, including the definitions thereof as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in examples, etc., can be combined and incorporated with each other arbitrarily. A particular term should not be considered indefinite or unclear without a specific definition, but should be understood according to the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or the active ingredient thereof.

[0168] Herein represents a point of attachment.

[0169] Herein represents that the corresponding ring is an aromatic ring.

[0170] Unless otherwise indicated, a wedge-shaped bond and a dotted wedge-shaped bond represents the absolute configuration of a stereocenter.

[0171] Herein represents the (Z) configuration or the (E) configuration, for example represents the (Z) configuration or the (E) configuration

[0172] The term "tautomer" refers to isomers of a functional group that result from the movement of a certain atom in a molecule to two positions. The compounds of the present disclosure can exhibit tautomerism. Compounds that tautomerize can exist in two or more interconvertible forms. Tautomers generally exist in equilibrium and attempts to isolate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates; in phenols, the enol form predominates. The present disclosure includes all tautomeric forms of the compounds.

[0173] The term "stereoisomer" refers to isomers that have the same molecular formula but different physical properties, such as different melting points, different retention times on a chiral column, and / or different optical properties. The term "stereoisomers" includes enantiomers, diastereomers, and geometric isomers.

[0174] The compounds of the present disclosure can have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms, or asymmetric double bonds, and therefore the compounds of the present disclosure can exist in particular geometric or stereoisomeric forms. The particular geometric or stereoisomeric form can be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as the racemic mixtures, or other mixtures thereof, such as those that are substantially free of ingredients of natural isomeric abundance, and mixtures thereof. Additional asymmetric carbon, sulfur, nitrogen, or phosphorus atoms or asymmetric double bonds can be present in a substituent such as an alkyl group, and the present disclosure includes all such isomers, as well as mixtures thereof. The compounds of the present disclosure containing an asymmetric atom can be isolated in optically active form by separation of composition of matter, or by absolute stereochemical synthesis from a racemic mixture of the compounds, or as achieved by using chiral reagents in a synthesis.

[0175] The term "substituted" means that any one or more hydrogen atoms on the particular atom is replaced with a substituent group, provided that the valency of the particular atom is not exceeded and that the substituted compound is stable. When the substituent is oxo (i.e., =0), it means that two hydrogen atoms are replaced.

[0176] 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.

[0177] 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.

[0178] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a bond.

[0179] When one of the variables is selected as a chemical bond or does not exist, it means that the two groups it is connected to are directly connected. For example, when L in ALZ represents a bond, it means that the structure is actually AZ.

[0180] If the linking group mentioned in this article does not specify its linking direction, then its linking direction is arbitrary. For example, when the structural unit... L in 1 When selected from "C1-C3 alkylene-O", L 1 Both loops Q and R can be connected in a left-to-right direction. 1 Composed of "cyclo-Q-C1-C3 alkylene-OR" 1 Alternatively, rings Q and R can be connected from right to left. 1 Composed of "cyclo-QO-C1-C3 alkylene-R" 1 ".

[0181] 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.

[0182] C in this article m -C nIt 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.

[0183] The term "alkyl" refers to a compound with the general formula C10. 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, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, etc.; the term "C1-C6 alkyl" can be understood as referring to alkyl groups 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 straight-chain or branched saturated alkyl groups having 1 to 3 carbon atoms. The "C1-C6"... 10 "Alkyl" can encompass the range of "C1-C8 alkyl", "C1-C6 alkyl", or "C1-C3 alkyl", and the "C1-C6 alkyl" can further include "C1-C3 alkyl". The term "alkoxy" refers to a group formed by the loss of a hydrogen atom from a hydroxyl group in a straight-chain or branched alcohol, and 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-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-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" can further include "C1-C3 alkoxy".

[0184] The term "alkenyl" refers to an unsaturated aliphatic hydrocarbon group consisting of a straight or branched chain of carbon and hydrogen atoms and having at least one double bond. The term "C2-C"... 10 "Alkenyl" can be understood as representing a straight-chain or branched unsaturated hydrocarbon group containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, "C2-C". 10 The term "alkenyl" may include "C2-C8 alkenyl," preferably "C2-C6 alkenyl," further preferably "C2-C4 alkenyl," and even more preferably C2 or C3 alkenyl. It is understood that when the alkenyl group contains more than one double bond, the double bonds may be separable or conjugated. Specific examples of the alkenyl group include, but are not limited to, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl, 2-methylprop-1-enyl, (E)-1-methylprop-1-enyl, or (Z)-1-methylprop-1-enyl, etc.

[0185] 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" and "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" can be understood as indicating a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms. Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0186] The term "cycloalkenyl" refers to a non-aromatic carbon ring that is not fully saturated and exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 5- to 8-membered ring. Specific examples of cycloalkenyl include, but are not limited to, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, cycloheptenyl, or cycloheptadienyl.

[0187] The term "heterocyclyl" refers to a monocyclic, bicyclic, spiro, or bridged ring radical which is completely saturated or partially saturated, having from 1 to 5 heteroatoms or heteroatom groups (i.e., an atom group containing a heteroatom) in the ring atoms, including, but not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=0)2-, -S(=0)-, -P(=0)2-, -P(=0)-, -NH-, -S(=0)(=NH)-, -C(=0)NH-, or -NHC(=0)NH-, and the like, in the ring atoms. The term "4-18 membered heterocyclyl" refers to a heterocyclyl having 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, and having from 1 to 5 ring atoms independently selected from the heteroatoms or heteroatom groups described above. The term "4-12 membered heterocyclyl" refers to a heterocyclyl having 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, and having from 1 to 5 ring atoms independently selected from the heteroatoms or heteroatom groups described above. "4-10 membered heterocyclyl" includes "4-7 membered heterocyclyl", wherein specific examples of 4-membered heterocyclyl include, but are not limited to, azetidinyl or oxetanyl; specific examples of 5-membered heterocyclyl include, but are not limited to, tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-lH-pyrrolyl; specific examples of 6-membered heterocyclyl include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl, or 4H-[l,3,4]thiadiazinyl; and specific examples of 7-membered heterocyclyl include, but are not limited to, diazepanyl. The heterocyclyl can also be a bicyclic radical, wherein specific examples of 5,5 membered bicyclic radical include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(lH)-yl; specific examples of 5,6 membered bicyclic radical include, but are not limited to, hexahydropyrrolo[l,2-a]pyrazin-2(lH)-yl, 5,6,7,8-tetrahydro-[l,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[l,5-a]pyrazinyl. Optionally, the heterocyclyl can be a benzo-fused or heteroaromatic annulated radical of the aforementioned 4-7 membered heterocyclyl, including, but not limited to, dihydroisoquinolinyl, and the like. The heterocyclyl can also be a tricyclic radical. "4-10 membered heterocyclyl" can 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", and the like. "4-7 membered heterocyclyl" can further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", and the like.In the present disclosure, although some bicyclic heterocyclyl moieties contain a benzene ring or a heteroaromatic ring partially, the heterocyclyl is still overall non-aromatic.

[0188] The term "aryl" refers to all-carbon monocyclic or fused polycyclic ring systems having a conjugated pi-electron system. Aryl groups can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 20 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 13 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 14 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 The term "aryl" can be understood as aryl having 6 to 20 carbon atoms. In particular a ring having 6 carbon atoms ("C6-aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9-aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C

[0189] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromaticity, which contains at least one ring atom selected from N, O, S, the remaining ring atoms being C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and which contain 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. The term "5-9 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8 or 9 ring atoms, in particular 5 or 6 or 9 ring atoms, and which contain 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and the like and their benzo derivatives, such as, for example, benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and the like and their benzo derivatives, such as, for example, quinolinyl, quinazolinyl or isoquinolinyl and the like; or azocinyl, indolizinyl, purinyl and the like and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenoxazinyl, phenothiazinyl or phenoxazinyl and the like. "5-10 membered heteroaryl" includes "5-9 membered heteroaryl", "5-6 membered heteroaryl". The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and which contains 1-3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0190] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromaticity, which contains at least one ring atom selected from N, O, S, the remaining ring atoms being C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and which contain 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. The term "5-9 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8 or 9 ring atoms, in particular 5 or 6 or 9 ring atoms, and which contain 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and the like and their benzo derivatives, such as, for example, benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and the like and their benzo derivatives, such as, for example, quinolinyl, quinazolinyl or isoquinolinyl and the like; or azocinyl, indolizinyl, purinyl and the like and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenoxazinyl, phenothiazinyl or phenoxazinyl and the like. "5-10 membered heteroaryl" includes "5-9 membered heteroaryl", "5-6 membered heteroaryl". The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and which contains 1-3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0191] The term "leaving group" refers to a functional group or atom which can be displaced by another functional group or atom through a substitution reaction, such as a nucleophilic displacement reaction. For example, representative leaving groups include, but are not limited to, triflate, chloro, bromo, iodo, sulfonate (e.g., methanesulfonate, tosylate, brosylate, p-tosylate) or acyloxy (e.g., acetoxy, trifluoroacetoxy) and the like.

[0192] The term "halo" or "halogen" refers to fluoro, chloro, bromo or iodo.

[0193] The term "hydroxy" refers to an -OH group.

[0194] The term "amino" refers to an -NH2 group.

[0195] The term "C(O)" means a -C(=0) group.

[0196] The term "WRN inhibitor" or "WRN helicase inhibitor" means a compound that inhibits Werner Syndrome RecQ DNA helicase (WRN). The term "WRN" means the protein Werner Syndrome RecQ DNA helicase. The term "WRN" includes mutants, fragments, variants, isoforms, and homologs of full-length wild-type WRN. A "disease or disorder mediated by WRN" includes diseases or disorders that are treated by WRN inhibition, for example, cancer.

[0197] The term "therapeutically effective amount" means an amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, or (ii) attenuates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder. The amount of a compound of the present disclosure that will constitute a "therapeutically effective amount" will vary depending on the compound, the disease state and its severity, the manner of administration, and the age of the mammal to be treated, but can be determined routinely by the skilled practitioner as an initial consideration.

[0198] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0199] The term "pharmaceutically acceptable salt" refers to salts of a compound that are pharmaceutically acceptable, including salts of inorganic acids or organic acids, and salts of inorganic bases or organic bases.

[0200] The term "pharmaceutical composition" means a mixture of one or more compounds of the present disclosure or salts thereof with a pharmaceutically acceptable excipient. The objective of a pharmaceutical composition is to facilitate administration of a compound of the present disclosure to an organism.

[0201] The term "pharmaceutically acceptable excipient" means an excipient that is not biologically or otherwise undesirable, i.e., the excipient can be administered to an organism without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained. The suitable excipient is well known to a person skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable, polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0202] The words "comprise" or "comprising" and variations thereof such as "comprises" or "comprising", when used in this document, can be understood to encompass the terms "consisting of" or "consisting of" and variations thereof, such as "consists" or "consisting of".

[0203] The present disclosure also includes isotopically-labeled compounds of the present disclosure which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be 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.

[0204] Certain isotopically-labeled compounds of the present disclosure (for example, with 3 H and 14 C) can be used in compound and / or substrate tissue distribution analysis. Tritiated (i.e., with 3 H), and carbon-14 (i.e., with 14 C) isotopes are particularly preferred for 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. Isotopically-labeled compounds of the present disclosure can generally be prepared by substituting a readily available isotopically-labeled reagent for a non-isotopically labeled reagent in a procedure of synthesis of a compound disclosed herein according to the procedures disclosed below and / or in the examples.

[0205] The pharmaceutical compositions of the present disclosure can be prepared by combining a compound of the present disclosure with suitable pharmaceutically acceptable excipients, for example, can be formulated into solid, semi-solid, liquid, or gaseous dosage forms, such as tablets, pills, capsules, powders, granules, creams, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres, aerosols, and the like.

[0206] Typical routes of administering a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition thereof, include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0207] The pharmaceutical compositions of the present disclosure can be manufactured in a manner that is generally known, e.g., by means of conventional mixing, dissolving, granulating, emulsifying, lyophilizing, or drilling processes.

[0208] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by combining the active compounds with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present disclosure to be formulated into tablets, pills, dragees, sugar-coated tablets, capsules, liquids, gels, slurries, suspensions, etc., for oral administration to a patient.

[0209] Solid oral compositions can be prepared by conventional mixing or compaction methods. For example, the active compounds can be mixed with a solid excipient, optionally ground, and then filled into a capsule, if desired with the addition of additional excipients. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, or flavoring agents.

[0210] The pharmaceutical compositions can also be adapted for parenteral administration, such as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

[0211] In all methods of administration of the compounds of Formula I described herein, the dosage administered daily can range from 0.01 mg / kg to 100 mg / kg of body weight in single or divided doses.

[0212] The compounds of the present disclosure can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the other embodiments and equivalents thereof well known to those skilled in the art, and preferred embodiments including, but not limited to, the examples of the present disclosure.

[0213] The chemical reactions of the specific embodiments of the present disclosure are performed in solvents appropriate to the reagents and materials employed and to the desired product, in a manner known to those skilled in the art. For obtaining the compounds of the present disclosure, it can be necessary to modify the synthetic sequence or the reaction conditions employed in the embodiments described above, as is well known to those skilled in the art.

[0214] An important consideration in the planning of synthetic routes in the art is the selection of an appropriate protecting group for a reactive functional group, for example, see Greene's Protective Groups in Organic Synthesis (4th Ed). Hoboken, New Jersey: John Wiley & Sons, Inc. All references cited in the present disclosure are incorporated herein in their entirety.

[0215] The following abbreviations are used in the present disclosure: DETAILED DESCRIPTION

[0216] The present disclosure is described in detail by the following examples, but it does not mean any unfavorable limitation to the present disclosure. The present disclosure has been described in detail, and the specific embodiment manner thereof has also been disclosed, and it will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiment of the present disclosure without departing from the spirit and scope of the present disclosure. All reagents used in the present disclosure are commercially available, and can be used without further purification.

[0217] Unless otherwise specified, the ratio indicated by the mixed solvent is the volume mixing ratio.

[0218] The compounds are named by hand or by software, and the commercially available compounds are named by the supplier's catalog name.

[0219] The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 (ppm). The solvent for NMR determination is deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS); "IC 50 " refers to the half maximal inhibitory concentration, which refers to the concentration at which the maximum inhibitory effect is halved.

[0220] The eluent hereinafter can be formed into a mixed eluent by two or more solvents, and the ratio is the volume ratio of each solvent.

[0221] The "reverse column chromatography" hereinafter refers to a C18 column unless otherwise specified.

[0222] Example 1, Synthesis of (S,E)-1-(tert-butyl)-N-(1-cyclopropyl-(3-methanesulfonyl)allyl)-3-phenoxy-1H-pyrazole-4-carboxamide (Compound 1)

[0223] Step 1: Synthesis of Compound 1b

[0224] ​Compound 1a (21.5 g, 99.89 mmol) was dissolved in tetrahydrofuran (100 mL) under argon protection, and stirred at 0-5 °C in an ice bath. Red-Al (70% solution in toluene, 100 mL, 349.6 mmol) was added slowly dropwise to the above system. The reaction was allowed to proceed at 0-5 °C with stirring. TLC was used to monitor the reaction. Methanol was added to quench the reaction, and potassium sodium tartrate (40 mL, 1 g / mL) was added. The aqueous phase was extracted twice with ethyl acetate, and the organic phase was washed once with saturated sodium bicarbonate solution and once with saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 1b (20.1 g).

[0225] Step two: synthesis of compound 1c

[0226] Compound 1b (5.0 g, 24.89 mmol) was dissolved in DCM (150 mL), and sodium bicarbonate (12.55 g, 149.36 mmol) was added. The system was protected by argon, and stirred at 0-5 °C. Then DMP (21.12 g, 49.79 mmol) was added in batches. The reaction was allowed to proceed at 0-5 °C with stirring. TLC was used to monitor the reaction. The system was diluted with DCM, and washed twice with saturated sodium thiosulfate solution and twice with saturated sodium bicarbonate solution. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel with gradient elution (0-35% EA) to give compound 1c (1.67 g).

[0227] Step three: synthesis of compound 1e

[0228] Compound 1c (1.04 g, 5.22 mmol) was dissolved in THF (15 mL) under argon protection, and stirred at 0 °C. Sodium hydride (225.32 mg, 5.63 mmol, 60% dispersion in mineral oil) was added to the reaction system, which was allowed to react at 0 °C for 0.5 h. Then compound 1d (748.3 mg, 3.25 mmol) in THF (10 mL) was added, and the reaction was allowed to proceed at room temperature with stirring. TLC was used to monitor the reaction. The reaction system was quenched by adding saturated ammonium chloride solution, and the organic phase was extracted with dichloromethane. The organic phase was washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography with gradient elution (0-35% EA) to give compound 1e (644.1 mg).

[0229] Step four: synthesis of compound 1f

[0230] Compound 1e (293 mg, 1.06 mmol) was dissolved in acetonitrile (5 mL), p-toluenesulfonic acid (201.55 mg, 1.17 mmol) was added, and the reaction was stirred at room temperature until completion. The reaction was directly concentrated to dryness under reduced pressure to obtain compound 1f (crude, p-toluenesulfonic acid salt, 369.7 mg), which was directly used in the next step.

[0231] Step five: synthesis of compound 1i

[0232] Compound 1g (300 mg, 1.13 mmol) was added to 1h (1 mL), and concentrated hydrochloric acid (0.2 mL) was slowly added dropwise. The mixture was reacted at 80°C until completion. Water (3 mL) was added to the reaction, and ethyl acetate (2 mL*3) was used for extraction. The combined organic phase was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and rotary evaporated to obtain compound 1i (326 mg).

[0233] MS m / z (ESI): 323.1 [M+H] + .

[0234] Step six: synthesis of compound 1k

[0235] Compound 1i (150 mg, 466 μmol), 1j (65.7 mg, 699 μmol), and potassium carbonate (128 mg, 932 μmol) were added to acetonitrile (3 mL), and the reaction was carried out at 60°C until completion. The reaction was purified by flash column chromatography (PE:EA = 5:1) to obtain compound 1k (83.2 mg).

[0236] MS m / z (ESI): 289.2 [M+H] + .

[0237] Step seven: synthesis of compound 1l

[0238] Compound 1k (83.2 mg, 288 μmol) and lithium hydroxide monohydrate (36.4 mg, 866 μmol) were added to methanol (0.4 mL) and water (0.1 mL). The mixture was reacted at room temperature until completion. The reaction was purified by reverse phase column chromatography (acetonitrile:water) to obtain compound 1l (68.0 mg).

[0239] MS m / z (ESI): 261.2 [M+H] + .

[0240] Step eight: synthesis of compound 1

[0241] Compound 1 1 (20.0 mg, 76.9 μmol), compound 1 f (p-toluenesulfonate, 32.0 mg, 92.3 μmol), 2,4,6-trimethylpyridine (46.5 mg, 385 μmol) and TCFH (64.6 mg, 231 μmol) were added into acetonitrile (0.1 mL). The reaction was carried out at 50 °C thoroughly. The reaction solution was purified by reverse column chromatography (C18 column, acetonitrile: water) to give compound 1 (11.0 mg).

[0242] MS m / z (ESI): 418.3 [M+H] + .

[0243] 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.39 - 7.28 (m, 2H), 7.1 1 - 7.00 (m, 3H), 6.78 (dd, J = 15.3, 4.6 Hz, 1H), 6.71 (dd, J = 15.3, 1.2 Hz, 1H), 4.00 (td, J = 8.5, 4.4 Hz, 1H), 2.99 (s, 3H), 1.51 (s, 9H), 1.04 (ddq, J = 13.0, 8.6, 4.7 Hz, 1H), 0.60 - 0.13 (m, 4H).

[0244] Example 2, Synthesis of (S,E)-2-(tert-butyl)-N-(1-cyclopropyl-3- (methylsulfonyl)allyl)-4-phenoxythiazole-5-carboxamide (Compound 2)

[0245] Step one: Synthesis of compound 2c

[0246] Compound 2a (2.0 g, 8.36 mmol) and compound 2b (3.05 g, 26.1 mmol) were dissolved in anhydrous ethanol (10 mL), stirred, and pyridine (2.7 g, 34.15 mmol) was added. The system was stirred at 80 °C thoroughly. Then the volatile was removed by concentration under reduced pressure. The residue was diluted with ethyl acetate (30 mL), washed with water (30 mL) and saturated brine (30 mL) respectively, dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure. Purification by column chromatography (petroleum ether: ethyl acetate = 1 : 1) gave compound 2c (1.5 g).

[0247] Step two: Synthesis of compound 2d

[0248] Compound 2c (1.19 g, 5.19 mmol) was dissolved in dichloromethane (10 mL) under argon, and the system was cooled to 0-5°C and stirred. Trifluoromethanesulfonic anhydride (1.90 g, 6.75 mmol) was added dropwise, followed by the addition of DIEA (1.34 g, 10.3 mmol), and the system was stirred at room temperature until the reaction was complete. The system was diluted with dichloromethane (20 mL) and washed once with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. Purification by column chromatography (petroleum ether: ethyl acetate = 1:1) gave compound 2d (1.6 g).

[0249] Step three: synthesis of compound 2f

[0250] Compound 2d (1.4 g, 3.87 mmol) was dissolved in NMP (15 mL), and compound 2e (1.59 g, 11.6 mmol) and DIEA (2.50 g, 19.3 mmol) were added to the system, which was stirred at 90°C until the reaction was complete. The reaction solution was diluted with ethyl acetate (10 mL) and washed once with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. Purification by flash column chromatography (petroleum ether: ethyl acetate = 0-45%) gave compound 2f (950 mg).

[0251] Step four: synthesis of compound 2g

[0252] Compound 2f (0.90 g, 2.69 mmol) was dissolved in trifluoroacetic acid (6 mL), and the system was heated and stirred at 80°C until the reaction was complete. The system was directly concentrated under reduced pressure to remove the trifluoroacetic acid, and the residue was mixed with silica gel and purified by column chromatography (petroleum ether: ethyl acetate = 0-65%) to give compound 2g (500 mg).

[0253] Step five: synthesis of compound 2h

[0254] Tert-butyl nitrite (481 mg, 4.67 mmol) and cuprous bromide (267 mg, 1.87 mmol) were added to a 10 mL reaction bottle, and acetonitrile (10 mL) was added. The system was heated at 55°C for 1 hour, and then cooled to room temperature. To the above system was added a solution of compound 2g (200 mg, 877 μmol) in acetonitrile (3 mL), and the system was stirred at room temperature until the reaction was complete. The system was diluted with ethyl acetate (30 mL) and washed once with saturated sodium bicarbonate solution (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent. Purification by flash column chromatography (petroleum ether: ethyl acetate = 0-40%) gave compound 2h (220 mg).

[0255] Step six: synthesis of compound 2i

[0256] Compound 2h (220 mg, 752 μmol), phenol 1j (106 mg, 1.13 mmol), N, N- dimethylglycine (46.58 mg, 451 μmol), cuprous iodide (28.68 mg, 150 μmol) and potassium carbonate (208 mg, 1.51 mmol) were dissolved in DMF (5 mL), the system was protected by argon, and stirred at 105 °C for sufficient reaction. After the reaction was completed, the reaction solution was diluted with ethyl acetate (20 mL), and washed with water (20 mL) and saturated brine (20 mL) once respectively, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent, and purified by column chromatography (petroleum ether: ethyl acetate = 0-30%) to obtain compound 2i (71 mg).

[0257] Step seven: synthesis of compound 2j

[0258] Compound 2i (71 mg, 232 μmol) was dissolved in tetrahydrofuran (3 mL) and stirred to dissolve, then lithium hydroxide monohydrate (29.3 mg, 697.47 μmol) aqueous solution (3 mL) was added to the system, and the system was stirred at room temperature for sufficient reaction. After the reaction was completed, the system was diluted with water (10 mL), adjusted to pH 3-4 with 2N hydrochloric acid, and then extracted with ethyl acetate (10 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, and concentrated to dryness under reduced pressure to obtain compound 2j (63 mg). The crude product was directly used in the next step reaction.

[0259] Step eight: synthesis of compound 2

[0260] HATU (61.2 mg, 162 μmol) and DIEA (41.9 mg, 324 μmol) were added to a DMF (2 mL) solution of compound 2j (30.0 mg, 108 μmol) and compound 1f (p-toluenesulfonate, 59 mg, 171 μmol), and the mixture was stirred at room temperature for sufficient reaction. The system was diluted with ethyl acetate (20 mL), and washed with water (20 mL) and saturated brine (20 mL) once respectively, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent, and the residue was purified by reversed-phase column chromatography (C18 column, acetonitrile: water = 10-70%) to obtain compound 2 (8 mg, yield 17%).

[0261] LC-MS: m / z (ESI): 435.1 [M+H] + .

[0262] 1H NMR (400 MHz, DMSO-d6) δ 8.01 (d, J = 8.3 Hz, 1H), 7.46 - 7.38 (m, 2H), 7.23 - 7.15 (m, 3H), 6.85 (dd, J = 15.3, 5.1 Hz, 1H), 6.76 (dd, J = 15.2, 1.2 Hz, 1H), 4.06 - 4.00 (m, 1H), 2.99 (s, 3H), 1.32 (s, 9H), 1.21 - 1.14 (m, 1H), 0.57 - 0.51 (m, 1H), 0.47 - 0.35 (m, 2H), 0.31 - 0.24 (m, 1H).

[0263] Example 3, Synthesis of (S,E)-N-(l-cyclopropyl-3-(methylsulfonyl)allyl)-2- (cyclopropyldifluoromethyl)-4-phenoxythiazole-5-carboxamide (Compound 3)

[0264] Step one: Synthesis of compound 3b

[0265] Compound 3a (2.0 g, 11.0 mmol) was put into a 100 mL reaction flask, toluene (20 mL) was added, stirred, then TsOH (189.2 mg, 1.1 mmol) and ethylene glycol (2.0 g, 32.9 mmol) were added, the system was fully reacted at 120 °C. The reaction liquid was directly poured into saturated sodium bicarbonate solution (40 mL) to quench, the aqueous phase was extracted twice with ethyl acetate (15 mL x 2), dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, the residue was silica gel mixed sample, column chromatography purification, gradient elution (0-30% EA / PE), the product was collected to give compound 3b (2.2 g).

[0266] Step two: Synthesis of compound 3d

[0267] Compound 3b (1.8 g, 8.0 mmol) was put into a 50 mL reaction flask, protected by argon, and THF (30 mL) was added, stirred, the system was cooled to -70 °C and stirred, then n-BuLi (3.8 mL, 9.5 mmol, 2.5 M) was added, after addition, the system was incubated at -70 °C for 30 min; compound 3c (1.2 g, 9.5 mmol) was added to the above system, after addition, the system was fully reacted at -70 °C. The system was quenched with saturated ammonium chloride solution (20 mL), stirred for 10 min, separated into layers, the aqueous phase was extracted twice with ethyl acetate (30 mL x 2), the combined organic phases were dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, the residue was silica gel mixed sample, column chromatography purification, gradient elution (0-40% EA / PE), to give compound 3d (2.0 g).

[0268] Step three: synthesis of compound 3e

[0269] Compound 3d (1.0 g, 3.85 mmol) was put into a 25 mL reaction bottle, stirred with THF (8 mL), then 2N aqueous HC1 solution (2.5 mL) was added, and the system was fully reacted at 50°C. The system was directly poured into saturated sodium bicarbonate solution (30 mL) to quench, and the system was extracted with ethyl acetate (15 mL x 3) three times, the organic phase was combined, washed with saturated brine (10 mL) once, dried with anhydrous sodium sulfate, and concentrated to dryness under reduced pressure to obtain compound 3e (800 mg). The crude product was directly used in the next step reaction.

[0270] Step four: synthesis of compound 3f

[0271] Compound 3e (1.1 g, 5.1 mmol) was put into a 100 mL reaction bottle, stirred with THF (18 mL) and water (12 mL), then 4-aminobenzenesulfonamide (1.1 g, 6.6 mmol) and NaC102 (6.6 mmol) aqueous solution (10 mL) were added, and the system was fully reacted at room temperature. The system was diluted with ethyl acetate (80 mL), and washed with water (20 mL) once, saturated brine (20 mL) once, and the organic phase was concentrated to dryness under reduced pressure, the crude product was dissolved with ethyl acetate (60 mL), then washed with saturated aqueous sodium bicarbonate solution (40 mL x 2) twice, and the organic phase was discarded; the aqueous phase was adjusted to pH 2-3 with 2N HC1, then extracted with ethyl acetate (20 mL x 3) three times, the organic phase was combined, dried with anhydrous sodium sulfate, and concentrated to dryness under reduced pressure to obtain compound 3f (726 mg), which was directly used in the next step reaction.

[0272] Step five: synthesis of compound 3g

[0273] Compound 3f (720 mg, 3.1 mmol) was put into a 50 mL reaction bottle, sodium bicarbonate (1.0 g, 12.4 mmol) was added, and DMF (10 mL) was added, stirred, then iodoethane (967 mg, 6.2 mmol) was added, and the system was fully reacted at room temperature. The system was diluted with ethyl acetate (60 mL), washed with water (20 mL x 2) twice, saturated brine (20 mL) once, and the organic phase was concentrated to remove the solvent under reduced pressure, the residue was silica gel mixed sample, and column chromatography was used for purification, gradient elution (0-40% EA / PE), and the product was collected to obtain compound 3g (680 mg).

[0274] Step six: synthesis of compound 3h

[0275] Compound 3g (400 mg, 1.5 mmol) and compound 1j (169 mg, 1.8 mmol) were charged into a 20 mL microwave tube, followed by the addition of sodium carbonate (488 mg, 4.6 mmol) and NMP (12 mL), and the mixture was subjected to microwave reaction at 140 °C for 2 h. The mixture was diluted with ethyl acetate (50 mL), washed with water (15 mL x 2) twice, and saturated brine (15 mL) once. The organic phase was concentrated under reduced pressure to remove the solvent, and the residue was mixed with silica gel and purified by column chromatography with gradient elution (0-30% EA / PE). Compound 3h (115 mg) was collected.

[0276] Step Seven: Synthesis of compound 3i

[0277] Compound 3h (100 mg, 315 μmol) was charged into a 10 mL reaction bottle, followed by the addition of THF (5 mL) and methanol (2 mL), and stirring. Then, NaBH4 (35.8 mg, 945 μmol) was added, and the mixture was stirred at room temperature until the reaction was completed. The reaction was quenched with 2N aqueous HCl solution (0.4 mL) and diluted with ethyl acetate (40 mL). The mixture was washed with water (15 mL) once and saturated brine (15 mL) once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to dryness. Compound 3i (90.0 mg) was obtained.

[0278] Step Eight: Synthesis of compound 3j

[0279] Compound 3i (90.0 mg, 282 μmol) was charged into a 8 mL reaction bottle, followed by the addition of dichloromethane (5 mL) and stirring. Then, DAST (136 mg, 845 μmol) was added, and the mixture was stirred at room temperature until the reaction was completed. The mixture was diluted with dichloromethane (50 mL) and washed with saturated aqueous sodium bicarbonate solution (15 mL x 2) twice and saturated brine (15 mL) once. The mixture was dried over anhydrous sodium sulfate and concentrated under reduced pressure to remove the solvent. The residue was mixed with silica gel and purified by column chromatography with gradient elution (0-55% EA / PE). Compound 3j (45.0 mg) was obtained.

[0280] Step Nine: Synthesis of compound 3k

[0281] Compound 3j (40.0 mg, 125 μmol) was put into a 25 mL reaction vial, followed by THF (5 mL), argon protection, the system was cooled to -78 °C and stirred, then LDA (0.15 mL, 2M) was added dropwise, after addition, the system was incubated at -78 °C for 1 h; then the temperature was raised to 20-25 °C and reacted for 15 min; the system was cooled to -78 °C again and stirred, then a solution of N-fluorobenzenesulfonimide (58.9 mg, 187 μmol) in THF (1 mL) was added, and the system was naturally warmed to room temperature for sufficient reaction. The system was quenched with saturated aqueous sodium bicarbonate solution (4 mL), diluted with ethyl acetate (40 mL), the organic phase was washed with water (15 mL) once, saturated brine (15 mL) once, dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, the residue was mixed with silica gel and purified by column chromatography, gradient elution (0-30% EA / PE), the product was collected and concentrated under reduced pressure to dryness to give compound 3k (25.0 mg).

[0282] Step Ten: Synthesis of compound 3l

[0283] Compound 3k (25.0 mg, 73.7 μmol) was put into a 10 mL reaction vial, THF (2 mL) and water (2 mL) were added and stirred, then lithium hydroxide (1.8 mg, 73.7 μmol) was added, the system was stirred at room temperature for sufficient reaction. The system was diluted with water (10 mL), then the pH was adjusted to 2-3 with 2N HCl, then extracted with ethyl acetate (10 mL x 3) three times, the organic phase was combined, washed with saturated brine (10 mL) once, dried over anhydrous sodium sulfate, concentrated under reduced pressure to dryness to give compound 3l (30.0 mg). The crude product was directly used in the next step reaction.

[0284] Step Eleven: Synthesis of compound 3

[0285] HATU (58.3 mg, 153 μmol) and DIEA (40.0 mg, 307 μmol) were added to a solution of compound 3l (30.0 mg, 96.4 μmol) and compound If (p-toluenesulfonate, 32.2 mg, 92.7 μmol) in DMF (4 mL), and the mixture was stirred at room temperature for sufficient reaction. The system was diluted with ethyl acetate (30 mL) and washed with water (10 mL x 2) twice, saturated brine (10 mL) once, dried over anhydrous sodium sulfate, concentrated under reduced pressure to remove the solvent, and the residue was purified by reverse phase chromatography (C18 column, acetonitrile / water = 30%-50%) to give compound 3 (3 mg).

[0286] LC-MS: m / z (ESI): 469.1 [M+H] + .

[0287] LC-MS: m / z (ESI): 469.1 [M+H]1 H NMR (400 MHz, CDC13) δ 7.47-7.40 (m, 2H), 7.30-7.24 (m, 2H), 7.22-7.17 (m, 2H), 6.98 (dd, J = 15.2, 4.7 Hz, 1H), 6.52 (dd, J = 15.2, 1.7 Hz, 1H), 4.16 (m, 1H), 2.93 (s, 3H), 1.75-1.63 (m, 1H), 1.04-1.01 (m, 1H), 0.83-0.77 (m, 2H), 0.74-0.66 (m, 3H), 0.64-0.61 (m, 1H), 0.50-0.39 (m, 2H).

[0288] Example 4, Synthesis of (S,E)-N-(l-cyclopropyl-3-(methylsulfonyl)allyl)-6- methyl-3-phenoxythieno[2,3-b]pyridine-2-carboxamide (Compound 4)

[0289] Step one: Synthesis of compound 4c

[0290] Compound 4a (500 mg, 3.28 mmol), 4b (348 mg, 3.28 mmol) and sodium methoxide (355 mg, 6.58 mmol) were added into DMF (5 mL) and reacted thoroughly at room temperature. The reaction solution was purified by reverse column chromatography (acetonitrile: water) to obtain compound 4c (436 mg).

[0291] MS m / z (ESI): 223.4 [M+H] + .

[0292] Step two: Synthesis of compound 4e

[0293] Compound 4c (436 mg, 1.96 mmol) and cuprous bromide (410 mg, 2.86 mmol) were added into acetonitrile (5 mL), and tert-butyl nitrite 4d (236 mg, 2.29 mmol) was added at 80 °C, and then reacted thoroughly at 80 °C. The reaction solution was filtered and purified by flash column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain compound 4e (332 mg).

[0294] MS m / z (ESI): 286.3 / 288.3 [M+H] + .

[0295] Step three: Synthesis of compound 4f

[0296] Compound 4e (332 mg, 1.16 mmol), 1j (164 mg, 1.74 mmol), potassium phosphate (492 mg, 2.32 mmol), cuprous iodide (221 mg, 1.16 μmol) and N1,N2-bis([1,1'-biphenyl]-2-yl)oxalamide (455 mg, 1.16 μmol) were added into DMF (5 mL), and the mixture was reacted thoroughly at 100 °C. The reaction solution was purified by reverse phase column chromatography (acetonitrile / water = 50%-95%) to obtain compound 4f (90.2 mg).

[0297] MS m / z (ESI): 300.4 [M+H] + .

[0298] Step four: synthesis of compound 4g

[0299] Compound 4f (90.2 mg, 0.30 mmol) and lithium hydroxide (36 mg, 1.50 mmol) were added into methanol (1 mL) and water (0.5 mL), and the mixture was reacted thoroughly at room temperature. The reaction solution was purified by reverse phase column chromatography (acetonitrile / water = 30%-80%) to obtain compound 4g (62.4 mg).

[0300] MS m / z (ESI): 286.3 [M+H] + .

[0301] Step five: synthesis of compound 4

[0302] By the method consistent with Example 1, eighth step, using compound 4g (20.0 mg, 70.2 μmol) and compound 1f (p-toluenesulfonate, 24.3 mg, 70.2 μmol) as raw materials, compound 4 (6.2 mg) was synthesized and purified.

[0303] MS m / z (ESI): 443.5 [M+H] + .

[0304] 1H NMR (400 MHz, DMSO-d6) δ 8.29 (d, J = 8.2 Hz, 1H), 7.67 (d, J = 8.3 Hz, 1H), 7.43 - 7.35 (m, 2H), 7.32 (d, J = 8.4 Hz, 1H), 7.15 (t, J = 7.4 Hz, 1H), 7.11 - 7.04 (m, 2H), 6.80 (dd, J = 15.3, 5.3 Hz, 1H), 6.66 (dd, J = 15.3, 1.3 Hz, 1H), 4.01 (q, J = 8.1 Hz, 1H), 2.94 (s, 3H), 2.61 (s, 3H), 1.10 (ddd, J = 12.3, 10.1, 5.9 Hz, 1H), 0.54 - 0.10 (m, 4H).

[0305] Example 5, Synthesis of (S,E)-N-(l-cyclopropyl-3-(methylsulfonyl)allyl)-2- ethyl(methyl)amino)-4-phenoxythiazole-5-carboxamide (Compound 5)

[0306] Step one: Synthesis of compound 5c

[0307] Compound 5a (500 mg, 1.74 mmol) and compound 5b (544 mg, 3.49 mmol) were dissolved in DMF (5 mL), then sodium bicarbonate (586 mg, 6.97 mmol) was added. The resulting reaction solution was stirred at room temperature for sufficient time to react. Water was added to quench, and ethyl acetate (20 mL) was added to extract. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (petroleum ether / ethyl acetate = 15%) to obtain compound 5c (427 mg).

[0308] Step two: Synthesis of compound 5e

[0309] Compound 5c (400.0 mg, 1.3 mmol), compound 5d (112 mg, 1.9 mmol), and triethylamine (384 mg, 3.8 mmol) were dissolved in DMF (4 mL) and stirred at 60 °C for sufficient time to react. Water was added to quench, and ethyl acetate (20 mL) was added to extract. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (silica gel, petroleum ether / ethyl acetate = 30%) to obtain compound 5e (358 mg).

[0310] Step three: Synthesis of compound 5f

[0311] Compound 5e (358 mg, 1.22 mmol), compound 1j (172 mg, 1.83 mmol), N, N- dimethylglycine (101 mg, 976 μmol), cuprous iodide (93 mg, 488 μmol) and cesium carbonate (795 mg, 2.44 mmol) were dissolved in dry DMF (2 mL) and stirred at 115 °C until the reaction was complete. The reaction mixture was diluted with water and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with saturated brine once, dried over anhydrous sodium sulfate, concentrated and purified by column chromatography (petroleum ether / ethyl acetate = 20%) to give compound 5f (360 mg).

[0312] Step four: synthesis of compound 5g

[0313] Compound 5f (74 mg, 242 μmol) was dissolved in tetrahydrofuran (1 mL) and methanol (1 mL), then lithium hydroxide (7 mg, 290 μmol) in water (2 mL) was added. The reaction mixture was stirred at 60 °C until the reaction was complete. The reaction mixture was concentrated under reduced pressure and then lyophilized to give compound 5g (lithium salt, 80 mg). The crude product was used directly in the next step.

[0314] Step five: synthesis of compound 5

[0315] Compound 5 (20 mg) was synthesized and purified by the method consistent with Example 2, step eight, using compound 5g (lithium salt, 45 mg, 158 μmol) and compound 1f (p-toluenesulfonate, 54.6 mg, 158 μmol) as starting materials.

[0316] LC-MS: m / z (ESI): 436.44 [M+H] + .

[0317] 1 H NMR (400 MHz, CDCl3) δ 7.38 (t, J = 7.8 Hz, 2H), 7.23 - 7.13 (m, 3H), 6.98 (dd, J = 15.2, 4.4 Hz, 1H), 6.88 (d, J = 7.6 Hz, 1H), 6.47 (d, J = 15.1 Hz, 1H), 4.22 - 4.08 (m, 1H), 3.42 (q, J = 7.2 Hz, 2H), 2.99 (s, 3H), 2.90 (s, 3H), 1.17 (t, J = 7.1 Hz, 3H), 1.01 - 0.87 (d, J = 8.1 Hz, 1H), 0.71 - 0.49 (m, 2H), 0.46-0.32 (m, 2H).

[0318] Example 6, Synthesis of (E)-2-(cyclopropyldifluoromethyl)-N-(3-(methylsulfonyl)- 1 -(oxetan-3-yl)allyl)-4-phenoxythiazole-5-carboxamide (Compound 6)

[0319] Step one: Synthesis of Compound 6c

[0320] Compound 6a (1.0 g, 11.6 mmol) was dissolved in dry DCM (10 mL). Then compound 6b (1.7 g, 13.9 mmol) and titanium tetraisopropoxide (4.6 g, 16.3 mmol) were added. The resulting reaction solution was stirred at room temperature overnight. Concentration, the crude product was purified by silica gel column chromatography (EA / PE = 0% - 50%) to give compound 6c (1.6 g).

[0321] Step two: Synthesis of Compound 6e

[0322] Compound 6c (1.6 g, 8.45 mmol) was dissolved in dry DCM (30 mL). The temperature was lowered to -60 °C, then compound 6d (1M THF solution, 25 mL) was added dropwise. After the addition was completed, the resulting reaction solution was allowed to react at room temperature. Water (30 mL) was added to quench. Celite was added to filter, the filter cake was washed with DCM. The organic layer was separated, the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. Concentration, the crude product was purified by silica gel column chromatography (EA / PE = 45%) to give compound 6e (1.6 g).

[0323] Step three: Synthesis of Compound 6f

[0324] Compound 6e (1.5 g, 6.9 mmol) was dissolved in tetrahydrofuran (15 mL), then 5 mL of water, sodium carbonate (732 mg, 6.9 mmol), and iodine (876 mg, 3.45 mmol) were added. The resulting reaction solution was stirred at 50 °C until the reaction was complete. The reaction solution was cooled to room temperature, saturated sodium thiosulfate solution (10 mL) and ethyl acetate (10 mL) were added. The organic layer was separated, the organic layer was washed with saturated sodium bicarbonate aqueous solution (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give compound 6f (0.8 g, crude). The resulting crude product was used directly in the subsequent reaction.

[0325] Step four: Synthesis of Compound 6g

[0326] Into a 25-mL flask, was placed compound 3l (78.0 mg, 251 μmol), compound 6f (56.7 mg, 501 μmol), and HATU (105 mg, 276 μmol), followed by DMF (6 mL). The mixture was stirred under argon. DIEA (97 mg, 753 μmol) was then added. The resulting reaction mixture was stirred at room temperature for sufficient time. The reaction mixture was diluted with ethyl acetate (50 mL), washed with water (15 mL) once, saturated brine (15 mL) once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluting with ethyl acetate / petroleum ether = 0-85%) to give compound 6g (65 mg).

[0327] Step five: synthesis of compound 6h

[0328] Into a 25-mL flask, was placed compound 6g (65.0 mg, 160 μmol), followed by THF (4 mL) and water (2 mL). The mixture was stirred, and N-methylmorpholine N-oxide (56.2 mg, 480 μmol) and potassium osmate dihydrate (5.9 mg, 16 μmol) were added. The resulting reaction mixture was stirred at room temperature for sufficient time. The reaction mixture was diluted with ethyl acetate (40 mL), washed with water (15 mL x 2) twice, saturated brine (15 mL x 1) once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to dryness to give compound 6h (70 mg). The crude product was used directly in the next step.

[0329] Step six: synthesis of compound 6i

[0330] Into a 25-mL flask, was placed compound 6h (70.0 mg, 159 μmol), followed by THF (3 mL) and water (1 mL). The mixture was stirred, and NaIO4(85.7 mg, 397 μmol) was added. The resulting reaction mixture was stirred at room temperature for sufficient time. The reaction mixture was diluted with ethyl acetate (50 mL) and water (15 mL), and the organic phase was washed with saturated sodium thiosulfate solution (15 mL x 1) and saturated brine (15 mL x 1) once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to dryness to give compound 6i (64 mg). The crude product was used directly in the next step.

[0331] Step six: synthesis of compound 6

[0332] Compound 6i (64 mg, 157 μmol) and compound Id (47.4 mg, 206 μmol) were put into a 25 mL reaction vial, THF (6 mL) was added, stirred, argon protection, then anhydrous lithium chloride (6.66 mg, 157 μmol) and DIEA (66.3 mg, 514 μmol) were added. The resulting reaction solution was stirred at room temperature for sufficient reaction. The system was diluted with ethyl acetate (40 mL), then washed with water (15 mL x 1) and saturated brine (15 mL x 1) once respectively, dried over anhydrous sodium sulfate, and the solvent was removed by concentration under reduced pressure. The residue was purified by reverse phase chromatography (C18 column, acetonitrile / water = 5%-70%) to give compound 6 (24 mg).

[0333] LC-MS: m / z (ESI): 484.8 [M+H] + .

[0334] 1 H-NMR (400MHz, CDC13) δ 8.32 (d, J = 8.4 Hz, 1H), 7.48-7.42 (m, 2H), 7.27-7.18 (m, 3H), 6.88 (dd, J = 15.2, 1.6 Hz, 1H), 6.71 (dd, J = 15.2, 5.6 Hz, 1H), 5.14-5.04 (m, 1H), 4.65-4.60 (m, 1H), 4.55-4.48 (m, 1H), 4.39-4.34 (m, 1H), 4.29-4.21 (m, 1H), 4.13-4.04 (m, 1H), 2.98 (s, 3H), 1.87-1.75 (m, 1H), 0.77-0.67 (m, 4H).

[0335] Example 7, Synthesis of N-((lS,E)-l-cyclopropyl-3-(S-methylsulfonylimino)allyl)-2- (cyclopropyldifluoromethyl)-4-phenoxythiazole-5-carboxamide (Compound 7)

[0336] Step one: Synthesis of compound 7b

[0337] Compound 7a (1.0 g, 10.7 mmol) was dissolved in anhydrous THF (50 mL), sodium hydride (644 mg, 16.1 mmol, 60% dispersion in mineral oil) was added portionwise at 0 °C, stirred at 0 °C for 0.5 h, di-tert-butyl dicarbonate (3.1 g, 14.0 mmol) was added dropwise at 0 °C, and the reaction was stirred at room temperature for sufficient reaction. The reaction solution was poured into water, extracted with ethyl acetate (20 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was directly purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give compound 7b (274 mg).

[0338] Step two: synthesis of compound 7c

[0339] Compound 7b (100 mg, 517 μmol) was dissolved in anhydrous THF (2 mL) under argon protection, and diisopropylamine lithium (0.5 mL, 2 N, 1.1 mmol) was added dropwise at 0 °C. The reaction was stirred at 0 °C for 0.5 h, and then diphenyl chlorophosphate (139 mg, 517 μmol) was added dropwise. The reaction was stirred at 0 °C until the reaction was completed. The reaction was poured into water (20 mL), and extracted with ethyl acetate (20 mL x 2). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was directly purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 2) to give compound 7c (74 mg).

[0340] Step three: synthesis of compound 7d

[0341] Compound 1c (600 mg, 3.01 mmol) was dissolved in anhydrous acetonitrile (10 mL), and anhydrous lithium chloride (128 mg, 3.01 mmol), compound 7c (2.57 g, 6.04 mmol), and DIEA (1.95 g, 15.1 mmol) were added successively. The reaction was stirred at room temperature until the reaction was completed. The reaction was poured into water, and extracted with ethyl acetate (30 mL x 3). The organic phase was combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase column chromatography (C18 column, acetonitrile / water = 5%-80%), and lyophilized to give compound 7d (650 mg).

[0342] Step four: synthesis of compound 7e

[0343] Compound 7d (500 mg, 1.33 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (2 mL) was added. The reaction was stirred at 40 °C until the reaction was completed. The reaction was concentrated under reduced pressure, and then lyophilized to give compound 7e (trifluoroacetate, 210 mg).

[0344] Step five: synthesis of compound 7

[0345] Compound 3l (90 mg, 289 μmol) was dissolved in anhydrous THF (3 mL), and isopropyl chloroformate (35.4 mg, 289 μmol) in THF (1 mL) and triethylamine (234 mg, 2.31 mmol) were added successively. The reaction was stirred at room temperature for 30 min. Then compound 7e (trifluoroacetate, 86.2 mg, 295 μmol) was added. The reaction was stirred at room temperature until the reaction was completed. The reaction was purified by reverse phase column chromatography (C18 column, acetonitrile / water = 5%-70%), and lyophilized to give compound 7 (45 mg).

[0346] LC-MS: m / z (ESI): 468.2 [M+H] + .

[0347] 1 H NMR (400 MHz, Chloroform-d) δ 7.48 - 7.36 (m, 2H), 7.30 - 7.09 (m, 5H), 6.91 (dt, J = 14.9, 5.3 Hz, 1H), 6.62 (dd, J = 15.0, 7.1 Hz, 1H), 4.24 - 4.10 (m, 1H), 2.99 (d, J = 4.4 Hz, 3H), 1.79 - 1.54 (m, 1H), 1.08 - 0.96 (m, 1H), 0.84 - 0.76 (m, 2H), 0.75 - 0.56 (m, 4H), 0.51 - 0.37 (m, 2H).

[0348] Example 8, Synthesis of (S,E)-2-cyclobutyl-N-(l-cyclopropyl-3- (methylsulfonyl)allyl)-4-phenoxythiazole-5-carboxamide (Compound 8)

[0349] Step one: Synthesis of compound 8b

[0350] Compound 3b (3.0 g, 13.27 mmol) was placed in a 100 mL reaction flask, which was protected by argon, and anhydrous tetrahydrofuran (20 mL) was added, stirred, and the system was cooled to -70°C and stirred, then n-BuLi (1.6 M, 9.95 mL) was added, after addition, the system was incubated at -70°C for 30 min; cyclobutanone (1.02 g, 14.60 mmol) was added to the above system, after addition, the system was stirred at -70°C and reacted sufficiently. The system was quenched with saturated aqueous ammonium chloride solution (15 mL), and ethyl acetate (40 mL) and water (20 mL) were added and stirred, the layers were separated, the aqueous phase was extracted twice with ethyl acetate (20 mL x 2), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by reduced pressure concentration. The residue was purified by silica gel column chromatography, gradient elution (0-45% EA), to give compound 8b (2.23 g).

[0351] Step two: Synthesis of compound 8c

[0352] Compound 8b (2.23 g, 8.51 mmol) was placed in a 100 mL reaction flask, tetrahydrofuran (12 mL) was added, stirred, then HCl (2 M, 15 mL) was added, the system was heated and stirred at 60°C and reacted sufficiently. The system was added with water (20 mL) and ethyl acetate (40 mL) and stirred, the layers were separated, the aqueous phase was extracted with ethyl acetate (20 mL), the combined organic phase was washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, concentrated to dryness by reduced pressure, to give compound 8c (1.8 g).

[0353] Step three: synthesis of compound 8e

[0354] Compound 8c (2.1 g, 9.65 mmol) was put into a 25 mL reaction bottle, stirred with tetrahydrofuran (20 mL) and water (20 mL), then compound 8d (2.16 g, 12.54 mmol) and an aqueous solution of NaClO2 (1.13 g, 12.54 mmol) (5 mL) were added, and the system was stirred at room temperature for sufficient reaction. The system was added with saturated aqueous sodium bicarbonate solution (60 mL), while adding ethyl acetate (40 mL), stirring, separating the layers, discarding the organic layer, and the aqueous phase was adjusted to pH 2-3 with 4N aqueous hydrochloric acid solution, then extracted twice with ethyl acetate (40 mL x 2), the combined organic phase was washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The obtained crude product was treated with dichloromethane (50 mL) to form a slurry, filtered, and the solid was discarded, and the mother liquor was concentrated to dryness under reduced pressure to obtain compound 8e (1.45 g). It was directly used in the next step reaction.

[0355] Step four: synthesis of compound 8f

[0356] Compound 8e (1.45 g, 6.21 mmol) was put into a 100 mL reaction bottle, added with DMF (15 mL) and stirred, then added with sodium bicarbonate (1.56 g, 18.6 mmol) and compound 5b (1.55 g, 9.93 mmol), and the system was stirred at room temperature for sufficient reaction. The system was diluted with ethyl acetate (120 mL), then washed twice with water (20 mL x 2) and once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the solvent was removed by concentration under reduced pressure. The residue was mixed with silica gel and purified by column chromatography on silica gel with gradient elution (0-25% EA), and the product was collected to obtain compound 8f (1.1 g).

[0357] Step five: synthesis of compound 8g

[0358] Compound 8f (200 mg, 764 μmol) was put into a 25 mL reaction bottle, added with dichloromethane (3 mL) and stirred, then added with dichlorosulfoxide (909 mg, 7.64 mmol), and the system was stirred at room temperature for sufficient reaction. The system was directly concentrated to dryness under reduced pressure to obtain compound 8g (210 mg). The crude product was directly used in the next step reaction.

[0359] Step six: synthesis of compound 8h

[0360] Compound 8g (160 mg, 571 μmol) was charged into a 25 mL reaction flask, followed by tetrahydrofuran (3 mL) and methanol (3 mL), stirred, and then 10% Pd / C (16.0 mg, 10% m / m) was added. The system was replaced with hydrogen for three times, and stirred at room temperature for sufficient reaction. The system was filtered with celite, and the filtrate was directly concentrated to dryness under reduced pressure to obtain compound 8h (140.0 mg).

[0361] Step Seven: Synthesis of compound 8i

[0362] Compound 8h (140 mg, 570 μmol), compound 1j (215 mg, 2.28 mmol) and sodium carbonate (242 mg, 2.28 mmol) were charged into a 10 mL microwave tube, followed by N-methylpyrrolidone (2 mL), and the system was subjected to microwave reaction at 140 °C for 2 h. The system was diluted with ethyl acetate (40 mL), washed with water (20 mL) and saturated brine (20 mL) once respectively, and the organic phase was concentrated to remove the solvent under reduced pressure. The residue was mixed with silica gel, and purified by silica gel column chromatography with gradient elution (0-20% EA), and the product was collected to obtain compound 8i (90.0 mg).

[0363] Step Eight: Synthesis of compound 8j

[0364] Compound 8i (90.0 mg, 297 μmol) was charged into a 25 mL reaction flask, followed by tetrahydrofuran (3 mL) and water (3 mL), stirred, and then lithium hydroxide monohydrate (37.4 mg, 890 μmol) was added. After the addition was completed, the system was stirred at room temperature for sufficient reaction. 2N aqueous hydrochloric acid (2 mL) was added to the system to adjust the pH value to 2-3, and then water (10 mL) and ethyl acetate (20 mL) were added, stirred, and the layers were separated. The aqueous phase was extracted with ethyl acetate (10 mL x 2) twice, and the combined organic phase was washed with saturated brine (10 mL) once, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain compound 8j (43.0 mg).

[0365] Step Nine: Synthesis of compound 8

[0366] By the method consistent with Example 2, Step Eight, using compound 8j (42.0 mg, 153 μmol) and compound 1f (p-toluenesulfonate, 63.6 mg, 183 μmol) as raw materials, compound 8 (19 mg) was synthesized and purified.

[0367] LC-MS: m / z (ESI): 432.9 [M+H] + .

[0368] 1H NMR (400 MHz, CDC13) δ 7.42 (dd, J = 8.6, 7.3 Hz, 2H), 7.25 - 7.18 (m, 2H), 7.18 - 7.13 (m, 2H), 6.98 (dd, J = 15.1, 4.6 Hz, 1H), 6.47 (dd, J = 15.1, 1.7 Hz, 1H), 4.19 - 4.11 (m, 1H), 3.77 - 3.67 (m, 1H), 2.91 (s, 3H), 2.51 - 2.36 (m, 2H), 2.32 - 2.19 (m, 2H), 2.12 - 1.99 (m, 1H), 1.97 - 1.88 (m, 1H), 1.0 - 0.95 (m, 1H), 0.71 - 0.63 (m, 1H), 0.61 - 0.55 (m, 1H), 0.47 - 0.39 (m, 2H).

[0369] Example 9, synthesis of compound 9

[0370] Step one: synthesis of compounds 7d-1 and 7d-2

[0371] Compound 7d (2 g) was separated by chiral separation (Daicel ChiralPak IC-H, 5 um, 30 mm diameter, 250 mm length) using a mixture of isopropanol and carbon dioxide kept polar (20%) as eluent to give compound 7d-1 (0.78 g, first peak corresponding to chiral method retention time 2.23 min) and compound 7d-2 (0.4 g, second peak corresponding to chiral method retention time 2.73 min).

[0372] Chiral method: column: IC-3, 4.6 mm diameter, 50 mm length; flow rate: 2.5 mL / min, gradient: 20% mixture of isopropanol and carbon dioxide. First peak retention time 2.23 min, second peak retention time 2.73 min.

[0373] Compound 7d-1: LC-MS: m / z (ESI): 375.4 [M+H] + .

[0374] 1H NMR (400 MHz, Chloroform-d) δ 7.06 (dd, J = 15.0, 3.9 Hz, 1H), 6.58 (dd, J = 15.1, 1.9 Hz, 1H), 4.79 (d, J = 7.7 Hz, 1H), 3.76 (s, 1H), 3.27 (s, 3H), 1.52 (s, 9H), 1.48 (s, 9H), 0.99 - 0.88 (m, 1H), 0.73 - 0.60 (m, 2H), 0.54 - 0.37 (m, 2H).

[0375] Compound 7d-2: LC-MS: m / z (ESI): 375.4 [M+H] + .

[0376] Step two: synthesis of compound 9a

[0377] Compound 7d-1 (300 mg, 801 μmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the reaction was stirred at room temperature until completion. The reaction was rotary evaporated and lyophilized to give compound 9a (trifluoroacetate salt, 250 mg).

[0378] Step three: synthesis of compound 9b

[0379] Compound 8f (210 mg, 802 μmol) was dissolved in dry dichloromethane (5 mL). The system was cooled to -20 °C, then diethylamine sulfide trifluoride (647 mg, 4.01 mmol) was added. The system was stirred at -20 °C until completion. Ice sodium bicarbonate aqueous solution (15 mL) was added to the system, extracted twice with ethyl acetate (15 mL x 2), the organic phase was combined, washed once with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography, gradient elution (0-15% EA), and the product was collected to give compound 9b (206 mg).

[0380] Step four: synthesis of compound 9c

[0381] Compound 9b (206 mg, 781 μmol) was dissolved in dry DMSO (5 mL). Potassium carbonate (323 mg, 2.34 mmol) and compound 1j (96 mg, 1.02 mmol) were added. The system was stirred at 80 °C until completion. After the reaction was cooled to room temperature, water (15 mL) and ethyl acetate (20 mL) were added, stirred, separated into layers, the aqueous phase was extracted once with ethyl acetate (10 mL), the organic phase was combined, washed once with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography, gradient elution (0-15% EA), and the product was collected to give compound 9c (127 mg).

[0382] Step five: synthesis of compound 9d

[0383] Compound 9d (110 mg) was synthesized and purified by the method consistent with the eighth step of Example 8, using compound 9c (120 mg, 373 μmol) as the starting material.

[0384] Step six: synthesis of compound 9

[0385] Compound 9 (40 mg) was synthesized and purified by the method consistent with the fifth step of Example 7, using compound 9d (45.0 mg, 153 μmol) and compound 9a (trifluoroacetate salt, 44.1 mg, 153 μmol) as the starting materials.

[0386] LC-MS: m / z (ESI): 450.23 [M+H] + .

[0387] 1 H NMR (400 MHz, DMSO-d6) δ 8.12 (d, J = 8.3 Hz, 1H), 7.48 - 7.37 (m, 2H), 7.30 - 7.17 (m, 3H), 6.80 - 6.68 (m, 2H), 4.08 - 3.97 (m, 1H), 2.89 (s, 3H), 2.69 - 2.53 (m, 4H), 2.00 - 1.85 (m, 1H), 1.79 - 1.60 (m, 1H), 1.26 - 1.11 (m, 1H), 0.60 - 0.23 (m, 4H).

[0388] Example 10, synthesis of compound 10

[0389] Step one: synthesis of compound 10b

[0390] Compound 10b (726 mg) was synthesized and purified by the method consistent with the first step of Example 8, using compound 3b (1.0 g, 4.42 mmol) and compound 10a (372 mg, 4.42 mmol) as the starting materials.

[0391] Step two: synthesis of compound 10c

[0392] Compound 10c (524 mg) was synthesized and purified by the method consistent with the second step of Example 8, using compound 10b (726 mg, 2.63 mmol) as the starting material.

[0393] Step three: synthesis of compound 10d

[0394] By the method consistent with the third step of Example 8, using compound 10c (524 mg, 2.26 mmol) and compound 8d (508 mg, 2.95 mmol) as raw materials, compound 10d (426 mg) was synthesized and purified.

[0395] Step four: synthesis of compound 10e

[0396] By the method consistent with the fourth step of Example 8, using compound 10d (426 mg, 1.72 mmol) and compound 5b (429 mg, 2.75 mmol) as raw materials, compound 10e (432 mg) was synthesized and purified.

[0397] Step five: synthesis of compound 10f

[0398] Compound 10e (200 mg, 725 μmol) was put into a 20 mL reaction bottle, dichloromethane (1 mL) was added, stirred, then diethylamine trifluoride DAST (1 mL) was added, the system was stirred at room temperature for sufficient reaction. Saturated aqueous sodium bicarbonate solution was added to the system to adjust the pH value to 8, then water (3 mL) and ethyl acetate (5 mL) were added, stirred, separated, the aqueous phase was extracted twice with ethyl acetate (5 mL x 2), the organic phase was combined, washed once with saturated brine (10 mL), and the organic phase was concentrated under reduced pressure to remove the solvent. The residue was mixed with silica gel and purified by silica gel column chromatography with gradient elution (0-20% EA), and the product was collected to obtain compound 10f (133 mg).

[0399] Step six: synthesis of compound 10g

[0400] By the method consistent with the fourth step of Example 9, using compound 10f (133 mg, 479 μmol) and compound 1j (59 mg, 623 μmol) as raw materials, compound 10g (109 mg) was synthesized and purified.

[0401] Step seven: synthesis of compound 10h

[0402] By the method consistent with the eighth step of Example 8, using compound 10g (109 mg, 325 μmol) as raw material, compound 10h (56 mg) was synthesized and purified.

[0403] Step eight: synthesis of compound 10

[0404] By the method consistent with the fifth step of Example 7, using compound 10h (30.0 mg, 97.6 μmol) and compound 9a (trifluoroacetate, 28.1 mg, 97.6 μmol) as raw materials, compound 10 (9 mg) was synthesized and purified.

[0405] LC-MS: m / z (ESI): 464.3 [M+H] + .

[0406] 1 H NMR (400 MHz, DMSO-d6) δ 8.12 (d, J = 8.4 Hz, 1H), 7.47 - 7.39 (m, 2H), 7.27 - 7.21 (m, 3H), 6.75 - 6.70 (m, 2H), 4.06 - 3.96 (m, 1H), 3.86 (s, 1H), 2.88 (s, 3H), 2.78 - 2.66 (m, 2H), 2.28 - 2.18 (m, 1H), 1.37 - 1.01 (m, 6H), 0.60 - 0.23 (m, 4H).

[0407] Example 11, Synthesis of compound 11

[0408] Step one: Synthesis of compound 11

[0409] Compound 11 (42 mg) was synthesized and purified by the method consistent with Example 7, step five, using compound 3l (42.0 mg, 135 μmol) and compound 9a (trifluoroacetate salt, 38.9 mg, 135 μmol) as starting materials.

[0410] LC-MS: m / z (ESI): 468.2 [M+H] + .

[0411] 1 H NMR (400 MHz, DMSO-d6) δ 8.12 (d, J = 8.4 Hz, 1H), 7.47 - 7.39 (m, 2H), 7.27 - 7.21 (m, 3H), 6.75 - 6.70 (m, 2H), 4.06 - 3.96 (m, 1H), 3.86 (s, 1H), 2.88 (s, 3H), 2.78 - 2.66 (m, 2H), 2.28 - 2.18 (m, 1H), 1.37 - 1.01 (m, 6H), 0.60 - 0.23 (m, 4H).

[0412] Example 12, Synthesis of (compound 12)

[0413] Step one: Synthesis of compound 12a

[0414] Compound 7d-2 (12.5 mg, 33.4 μmol) was dissolved in dichloromethane (0.5 mL), trifluoroacetic acid (0.5 mL) was added and the reaction was stirred at room temperature until completion. The reaction was spin dried and then lyophilized to give compound 12a (trifluoroacetate salt, 11 mg).

[0415] Step two: synthesis of compound 12

[0416] Compound 12 (6.8 mg) was synthesized and purified by the method consistent with Example 7, fifth step, using compound 3l (8.65 mg, 27.8 μmol) and compound 12a (trifluoroacetate salt, 9.6 mg, 33.4 μmol) as starting materials.

[0417] LC-MS: m / z (ESI): 468.4 [M+H] + .

[0418] 1 H NMR (400 MHz, Chloroform-d) δ 7.47-7.33 (m, 3H), 7.22-7.12 (m, 2H), 6.90 (d, J = 14.3 Hz, 1H), 6.64 (d, J = 14.7 Hz, 1H), 4.25-4.10 (m, 1H), 2.99 (s, 3H), 1.07 (m, 2H), 0.90-0.56 (m, 6H), 0.44 (m, 2H).

[0419] Biological activity and related property test examples

[0420] The compounds in the following test examples were prepared according to the methods of the above examples of the present disclosure.

[0421] Test Example 1: WRN ATP hydrolysis enzyme inhibitory activity assay

[0422] Experimental principle

[0423] WRN helicase is driven by ATP hydrolysis in the ATP hydrolysis enzyme domain of the WRN protein. ATP hydrolysis generates ADP, releasing energy to promote WRN helicase. The ADP generated in the reaction is detected by the ADP-Glo Assay Kit (Promega). After adding the detection reagent, the signal value is positively correlated with the amount of ADP generated in the reaction system, and the change in signal value reflects the change in ATP hydrolysis enzyme activity of the WRN protein.

[0424] Experimental instruments

[0425] Experimental materials

[0426] Experimental method

[0427] The analyte was dissolved in DMSO, and the stock solution concentration was 10 mM. The stock solution was serially diluted using the Echo instrument's dose-response program with DMSO as the dilution solvent. The total experimental volume was 4 μL, starting at 10 μM, with 3-fold dilutions and 10 concentration points. The total volume of the compound and DMSO was 160 nL, and the final DMSO concentration was 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 value was 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 .

[0428] The experimental results are shown in the table below:

[0429] The compounds disclosed in this embodiment exhibit good WRN ATP hydrolase inhibitory activity.

[0430] Test Example 2: HCT-116 Cell Proliferation Experiment

[0431] Experimental Principle

[0432] The HCT-116 cell line used in the experiment was an MSI (microsatellite instability) cell line. The proliferation assay used a kit (…). 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.

[0433] Experimental instruments

[0434] Experimental materials

[0435] Experimental methods

[0436] HCT-116 cells were cultured in McCoy's 5A + 10% FBS. After trypsin digestion, the cell concentration was adjusted to 300 / 40 μL / well in 384-well plates and cultured overnight at 37°C in a 5% CO2 incubator. The test compound was dissolved in DMSO, with a stock solution concentration of 10 mM. The stock solution was 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 compound and DMSO. After 4 days of incubation, the cell plates were equilibrated to room temperature, and 20 μL of CCL reagent was added to each well. After shaking in the dark for 60 minutes, the Luminescence signal was read using Envision. Cell control wells were defined as 0% inhibition controls, and culture medium wells as 100% inhibition controls. The inhibition rate of the sample wells was calculated. The IC50 was calculated using a four-parameter fitting method based on the inhibition rate. 50 .

[0437] The experimental results are shown in the table below:

[0438] The compounds disclosed herein exhibit good anti-cell proliferation activity against microsatellite unstable cells.

[0439] Test Example 3: Detection of In Vitro Metabolic Stability of Rat Hepatocytes

[0440] The concentration of compounds in the reaction system was determined by LC / MS / MS to calculate the intrinsic clearance rate of the test compounds and to assess their in vitro metabolic stability in rat hepatocytes.

[0441] 198 μL 0.5 × 10 6 A mixture of rat hepatocytes / mL and 2.0 μL of the test compound or positive control Verapamil was added to the incubation plate to initiate the reaction. Incubation was performed at 37 °C and 900 rpm. At 0, 15, 30, 60, 90, and 120 minutes, 25 μL of the incubation mixture was transferred to a stop plate (each well containing 150 μL of acetonitrile containing 100 nM alprazolam, 200 nM caffeine, and 100 nM tosylate). The mixture was then vortexed for 5 minutes. The stop plate was centrifuged at 3220 g for 45 minutes. 100 μL of the supernatant from each compound was transferred to a 96-well plate, followed by dilution with 100 μL of purified water.

[0442] The obtained samples were quantified by ion chromatogram. The residual rate was calculated based on the peak area of ​​the analyte or positive control. The slope k was determined using Microsoft Excel by linear regression of the natural logarithm of the residual rate against incubation time.

[0443] Intrinsic clearance (in vitro CL int , μL / min / 10 6 cells) was calculated from the slope value k according to the following equation:

[0444] Intrinsic clearance (in vitro CL int = -kV / N

[0445] V = incubation volume (0.25 mL);

[0446] N = number of cells per well (0.125 x 10 6 cells)

[0447] The experimental results are as follows:

[0448] Note: The structure of VVD-133214 (RO7589831) is as follows, which can be prepared according to Example 87 of WO2024010782A1:

[0449] Conclusion: The compound of the present disclosure has a lower in vitro liver cell clearance rate in rats compared with the control molecule.

[0450] Test Example 4: Test of rat in vivo pharmacokinetics of the compound of the present disclosure

[0451] SD rats were used as test animals, and the drug concentration in the plasma of the rats at different times after intravenous and oral administration of the compound of the present disclosure was determined by LC / MS / MS method. The pharmacokinetic behavior of the compound of the present disclosure in rats was studied, and its pharmacokinetic characteristics were evaluated.

[0452] 3 healthy 6-8 week old male SD rats per group.

[0453] Intravenous administration (IV): a certain amount of drug was weighed, 10% volume of N,N-dimethylacetamide, 33% volume of triethylene glycol and 57% volume of physiological saline were added to prepare a colorless, clear and transparent liquid of 1 mg / mL;

[0454] Oral gavage administration (PO): a certain amount of drug was weighed, 0.5% mass of hydroxypropyl methylcellulose, 0.1% volume of Tween 80 and 99.6% volume of physiological saline were added to prepare a white suspension of 1 mg / mL.

[0455] After the SD rats were fasted overnight, intravenous administration or oral gavage administration was performed.

[0456] The compound of the present application is administered to rats by tail vein injection at a dose of 1 mg / kg, and 0.2 mL of blood is collected from the jugular vein at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration, and is placed in a test tube containing EDTA-K2, and is centrifuged at 4°C and 4000 rpm for 5 minutes to separate the plasma, which is stored at -75°C.

[0457] Alternatively, the compound of the present application is administered to rats by gavage at a dose of 5 mg / kg, and 0.2 mL of blood is collected from the jugular vein at 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration, and is placed in a test tube containing EDTA-K2, and is centrifuged at 4°C and 3500 rpm for 10 minutes to separate the plasma, which is stored at -75°C.

[0458] The content of the test compound in the plasma of rats after gavage administration of different concentrations of the drug is determined: 30 μL of rat plasma at each time point after administration is taken, 200 μL of acetonitrile solution of internal standard dexamethasone (50 ng / mL) is added, vortex mixing for 30 seconds, centrifugation at 4°C and 4700 rpm for 15 minutes, the plasma sample is taken and diluted three times with water, and 2.0 μL is taken for LC-MS / MS analysis. The AB SCIEX 1.7.3, run and manage the LC-MS / MS liquid chromatography system, and collect data. The pharmacokinetic parameters are calculated by non-compartment statistical moment method of Phoenix WinNonlin 8.0 software.

[0459] The experimental results are as follows:

[0460] Conclusion: Compared with the control molecule, the compound of the present application has a lower apparent clearance and a longer half-life in rats under intravenous injection administration, and has a greater exposure under oral gavage administration.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, in, Y 1 Y 2 Y 3 Each is independently selected from N, NH, CH, C, S, Se or O, Y 4 Y 5 Each is independently selected from N or C; n is selected from 0, 1, 2, or 3; R 2 Selected from halogens, cyano groups, NHC(=O)C1-C6 alkyl groups, C3-C8 cycloalkyl groups, C(O)C3-C8 cycloalkyl groups, C5-C8 cycloalkenyl groups, C6-C 10 Aryl, C1-C6 alkyl, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, NH(C3-C8 cycloalkyl), N(C1-C6 alkyl)(C3-C8 cycloalkyl), C2-C6 alkenyl, -S-C1-C6 alkyl, C1-C6 alkoxy, -O-C3-C8 cycloalkyl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, wherein NHC(=O)C1-C6 alkyl, C3-C8 cycloalkyl, C(O)C3-C8 cycloalkyl, C5-C8 cycloalkenyl, C6-C 10 Aryl, C1-C6 alkyl, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, NH(C3-C8 cycloalkyl), N(C1-C6 alkyl)(C3-C8 cycloalkyl), C2-C6 alkenyl, -S-C1-C6 alkyl, C1-C6 alkoxy, -O-C3-C8 cycloalkyl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, optionally with R 2a Substitution; or two R atoms attached to adjacent ring atoms 2 Together with the atoms they are attached to form C5-C8 cycloalkenyl groups and C6-C... 10 aryl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, wherein the C5-C8 cycloalkenyl, C6-C 10 Aryl, 4-12 heterocyclic or 5-10 heteroaryl groups are optionally replaced by R 2A replace; R 2a Selected from hydroxyl, oxo, halogen, C1-C6 alkyl, phenyl, C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, amino, carboxyl, -S-C1-C6 alkyl, -S-C3-C8 cycloalkyl, or -O-C3-C8 cycloalkyl, wherein the C1-C6 alkyl, phenyl, C1-C6 alkoxy, C3-C6 cycloalkyl, amino, -S-C1-C6 alkyl, -S-C3-C8 cycloalkyl, or -O-C3-C8 cycloalkyl is optionally R 2b replace; R 2b Selected from halogens or C1-C6 alkyl groups; R 2A Selected from halogens, cyano groups, NHC(=O)C1-C6 alkyl groups, C3-C8 cycloalkyl groups, C(O)C3-C8 cycloalkyl groups, C5-C8 cycloalkenyl groups, C6-C 10 Aryl, C1-C6 alkyl, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, NH(C3-C8 cycloalkyl), N(C1-C6 alkyl)(C3-C8 cycloalkyl), C2-C6 alkenyl, -S-C1-C6 alkyl, C1-C6 alkoxy, -O-C3-C8 cycloalkyl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, wherein NHC(=O)C1-C6 alkyl, C3-C8 cycloalkyl, C(O)C3-C8 cycloalkyl, C5-C8 cycloalkenyl, C6-C 10 Aryl, C1-C6 alkyl, amino, NH(C1-C6 alkyl), N(C1-C6 alkyl)2, NH(C3-C8 cycloalkyl), N(C1-C6 alkyl)(C3-C8 cycloalkyl), C2-C6 alkenyl, -S-C1-C6 alkyl, C1-C6 alkoxy, -O-C3-C8 cycloalkyl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, optionally with R 2B replace; R 2B Selected from hydroxyl, oxo, halogen, C1-C6 alkyl, phenyl, C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, amino, carboxyl, -S-C1-C6 alkyl, -S-C3-C8 cycloalkyl, or -O-C3-C8 cycloalkyl, wherein the C1-C6 alkyl, phenyl, C1-C6 alkoxy, C3-C6 cycloalkyl, amino, -S-C1-C6 alkyl, -S-C3-C8 cycloalkyl, or -O-C3-C8 cycloalkyl is optionally R 2C replace; R 2C Selected from halogens or C1-C6 alkyl groups; L 1 Selected from bonds, O, S, NH, CH2, or OCH2; R 1 Selected from C3-C8 cycloalkyl, C1-C6 alkyl, C6-C 10 aryl, 5-10 membered heteroaryl or 4-12 membered heterocyclic, wherein the C3-C8 cycloalkyl, C1-C6 alkyl, C6-C 10 Aryl, 5-10 membered heteroaryl, or 4-12 membered heterocyclic group may be selected as R 1a replace; R 1a Selected from halogen, deuterium, cyano, hydroxyl, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, oxo, -S-C1-C6 alkyl, -O-C3-C8 cycloalkyl, -S-C3-C8 cycloalkyl or carboxyl; L 2 Selected from -C(=O)NH-, -C(=S)NH-, -S(=O)2NH-, 1,2,3-triazolyl, oxadiazolyl, imidazolyl, tetrazolyl, pyrazolyl, -NHC(=O)NH-, -NHC(=O)O-, -CH(CF3)NH- or R 3 Selected from hydrogen, C1-C6 alkyl, C3-C8 cycloalkyl, 4-12 membered heterocyclic groups, C6-C 10 aryl or 5-10 membered heteroaryl, wherein the C1-C6 alkyl, C3-C8 cycloalkyl, 4-12 membered heterocyclic, C6-C 10 Aryl or 5-10 heteroaryl groups are selectively coated with R. 3a replace; R 3a Selected from halogen, cyano, hydroxy, amino, carboxyl, oxo, -S-C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, -S-C1-C6 alkyl, -O-C3-C6 cycloalkyl, C(=O)NH2, C(=O)NH(C1-C6 alkyl), C(=O)N(C1-C6 alkyl)2 or phenoxy, wherein the amino, -S-C3-C8 cycloalkyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, -S-C1-C6 alkyl, -O-C3-C6 cycloalkyl, C(=O)NH2, C(=O)NH(C1-C6 alkyl), C(=O)N(C1-C6 alkyl)2 or phenoxy is optionally R 3b replace; R 3b Selected from halogens or C1-C6 alkyl groups; Q is selected from Q 1 Q 2 Q 3 Or Q 4 ; Q 1 yes in, represent It is either (Z) configuration or (E) configuration; X 3 Selected from O, NH or NR 5 ; R 4 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic groups or NR 8 R 9 The C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclic group is optionally R 4a replace; R 4a Selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy; R 11 Selected from hydrogen, or R 11 R 4 Together with the atoms bonded to it, they form a 4-6 membered heterocyclic group, which is optionally R 11a replace; R 11a Selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl, C1-C6 alkoxy, or C3-C6 cycloalkyl; R 8 R 9 Each is independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic group or C(O)C1-C6 alkyl, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, 4-6 membered heterocyclic group or C(O)C1-C6 alkyl is optionally R 8a Replace; or R 8 R 9 Together with the nitrogen atom attached thereto, they form a 4-12 membered heterocyclic group, wherein the 4-12 membered heterocyclic group is optionally subjected to R 9a replace; R 8a R 9a Each is independently selected from halogen, hydroxyl, amino, cyano, C1-C6 alkoxy or C1-C6 alkyl; R 5 Selected from cyano, C(=O)C1-C6 alkyl, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclic groups, wherein the C(=O)C1-C6 alkyl, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclic group is optionally R 5a Replace; or, the R 5 R 4 Together with the atoms bonded to it, they form 4-10 membered heterocyclic groups, wherein the 4-10 membered heterocyclic groups are optionally subjected to R 5a replace; R 5a Selected from halogen, cyano, hydroxyl, amino, C1-C6 alkyl or C1-C6 alkoxy; Q 2 yes in, represent It is either (Z) configuration or (E) configuration; Q 3 yes Q 4 yes Among them, R a It is a C1-C6 alkyl group, wherein the C1-C6 alkyl group may optionally be substituted.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein, Y 1 Y 2 Y 3 Each is independently selected from N, C, CH, S, Se or NH, Y 4 Y 5 All are selected from C; or Y 1 Selected from N, C or CH, Y 2 Selected from N or C, Y 3 Selected from CH, C, S, N, Se or NH, Y 4 and Y 5 All are selected from C; or Structural unit Selected from Where * represents and L 1 Connected atoms; or Structural unit Selected from Where * represents and L 1 Connected atoms.

3. The compound of formula (I) according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, n is selected from 1 or 2; or, n is selected from 1. Alternatively, n can be selected from 2.

4. The compound of formula (I) according to any one of claims 1-3, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, Each R 2 Independently selected from C1-C6 alkyl, N(C1-C6 alkyl)2, C3-C6 cycloalkyl, N(C1-C6 alkyl)(C3-C6 cycloalkyl) or C(O)C3-C6 cycloalkyl, wherein the C1-C6 alkyl, N(C1-C6 alkyl)2, C3-C6 cycloalkyl, N(C1-C6 alkyl)(C3-C6 cycloalkyl) or C(O)C3-C6 cycloalkyl is optionally R 2a Substitution; or two R atoms attached to adjacent ring atoms 2 Together with the atoms they are attached to, they form a 5-10 membered heteroaryl group or a 4-10 membered heterocyclic group, wherein the 5-10 membered heteroaryl group or the 4-10 membered heterocyclic group is optionally R 2A Replace; or Each R 2 Independently selected from C1-C4 alkyl, N(C1-C3 alkyl)2, C3-C6 cycloalkyl, N(C1-C3 alkyl)(C3-C4 cycloalkyl) or -C(O)C3-C4 cycloalkyl, wherein the C1-C4 alkyl, N(C1-C3 alkyl)2, C3-C6 cycloalkyl, N(C1-C3 alkyl)(C3-C4 cycloalkyl) or -C(O)C3-C4 cycloalkyl is optionally R 2a Substitution; two R atoms attached to adjacent ring atoms 2 Together with the atoms they are attached to, they form a 6-membered heteroaryl or a 5-membered heterocyclic group, wherein the 6-membered heteroaryl or 5-membered heterocyclic group is optionally converted by R 2A Replace; or Each R 2 Independently selected from methyl, ethyl, isopropyl, tert-butyl, Cyclopropyl, cyclobutyl, cyclopentyl, or -C(O)cyclopropyl, wherein the methyl, ethyl, isopropyl, tert-butyl, Cyclopropyl, cyclobutyl, cyclopentyl, or -C(O)cyclopropyl is optionally R 2a Replace; or Two R atoms connected to adjacent ring atoms 2 Together with the atoms they are attached to, they form a pyridyl group or The pyridyl or Optional R 2A replace.

5. The compound of formula (I) according to any one of claims 1-4, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, R 2a Selected from hydroxyl, halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or amino, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or amino group is optionally R 2b Replace; or R 2a Selected from halogens, C3-C6 cycloalkyl groups, or C1-C6 alkyl groups, wherein the C3-C6 cycloalkyl groups or C1-C6 alkyl groups are optionally R 2b replace.

6. The compound of formula (I) according to any one of claims 1-4, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, R 2A Selected from halogens, cyano groups, C3-C8 cycloalkyl groups, C(O)C3-C8 cycloalkyl groups, C5-C8 cycloalkenyl groups, C1-C6 alkyl groups, amino groups, NH (C1-C6 alkyl groups), -S-C1-C6 alkyl groups, C1-C6 alkoxy groups, or -O-C3-C8 cycloalkyl groups, wherein the C3-C8 cycloalkyl, C(O)C3-C8 cycloalkyl, C5-C8 cycloalkenyl, C1-C6 alkyl, amino, NH (C1-C6 alkyl), -S-C1-C6 alkyl, C1-C6 alkoxy groups, or -O-C3-C8 cycloalkyl groups are optionally R 2B Replace; or R 2A Selected from C3-C6 cycloalkyl or C1-C6 alkyl, wherein the C3-C6 cycloalkyl or C1-C6 alkyl is optionally R 2B replace.

7. The compound of formula (I) according to any one of claims 1-6, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, L 1 Selected from O, S, NH, CH2, or OCH2; or, where L 1 It is O, S, or NH; or, where L 1 It is O.

8. The compound of formula (I) according to any one of claims 1-7, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, R 1 Selected from C3-C8 cycloalkyl, C6-C 10 aryl, 5-10 membered heteroaryl or 4-12 membered heterocyclic, wherein the C3-C8 cycloalkyl, C6-C 10 Aryl, 5-10 membered heteroaryl, or 4-12 membered heterocyclic group may be selected as R 1a Replace; or, where R 1 Selected from C6-C 10 Aryl, 5-6 membered heteroaryl or 5-6 membered heterocyclic, wherein the C6-C 10 Aryl, 5-6 membered heteroaryl or 5-6 membered heterocyclic group are optionally replaced by R 1a Replace; or R 1 Selected from C6-C 10 Aryl, the C6-C 10 Aryl optional R 1a Replace; or R 1 Selected from phenyl.

9. The compound of formula (I) according to any one of claims 1-8, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, L 2 Selected from -C(=O)NH-, -C(=S)NH-, Or -S(=O)2NH-; or, L 2 Selected from -C(=O)NH-.

10. The compound of formula (I) according to any one of claims 1-9, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, R 3 Selected from C3-C8 cycloalkyl or 4-12 membered heterocyclic groups, wherein the C3-C8 cycloalkyl or 4-12 membered heterocyclic group is optionally R 3a Replace; or R 3 Selected from C3-C6 cycloalkyl or 4-10 membered heterocyclic groups, wherein the C3-C6 cycloalkyl or 4-10 membered heterocyclic group is optionally R 3a Replace; or R 3 Selected from C3-C4 cycloalkyl or 4-membered heterocyclic groups; or R 3 Selected from cyclopropyl or oxetyl; or R 3 Selected from cyclopropyl or 11. The compound of formula (I) according to any one of claims 1-10, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, Q is selected from Q 1 Q 1 yes in, represent It is either (Z) configuration or (E) configuration; or Q is 12. The compound of formula (I) according to any one of claims 1-11, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, X 3 Selected from O; or X 3 Selected from NH; or X 3 Selected from NR 5 ; and / or R 5 Selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclic group is optionally R 5a Replace; or R 5 Selected from C1-C3 alkyl or C3-C4 cycloalkyl, wherein the C1-C3 alkyl or C3-C4 cycloalkyl is optionally R 5a Replace; or R 5 Selected from methyl, cyclopropyl, or ethyl, wherein the methyl, cyclopropyl, or ethyl group is optionally R 5a replace; and / or R 5a Selected from halogens, C1-C6 alkyl or C1-C6 alkoxy groups; or R 5a Selected from C1-C3 alkoxy groups; or R 5a Selected from methoxy groups.

13. The compound of formula (I) according to any one of claims 1-12, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, R 4 Selected from C1-C6 alkyl or NR 8 R 9 The C1-C6 alkyl group is optionally R 4a Replace; or R 4 Selected from C1-C3 alkyl or NR 8 R 9 The C1-C3 alkyl group is optionally R 4a replace; and / or R 8 R 9 Each is independently selected from hydrogen, C1-C6 alkyl, or C3-C6 cycloalkyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally R 8a Replace; or R 8 R 9 Each is independently selected from hydrogen; and / or R 4a Selected from halogen, cyano, hydroxy, or amino groups; or R 4a Selected from halogens.

14. The compound of formula (I) according to any one of claims 1-13, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, The compound of formula (I) is selected from the compound of formula (II): Among them, R 2 , n, L 1 R 1 L 2 R 3 And Q as defined in any one of claims 1-13.

15. The compound of formula (I) according to any one of claims 1-13, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, The compound of formula (I) is selected from the compound of formula (III): Among them, Y 1 Y 2 Y 3 Y 4 Y 5 R 2 , n, L 1 R 1 L 2 R 3 And Q as defined in any one of claims 1-13.

16. The compound of formula (I) according to any one of claims 1-13, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, wherein, The compound of formula (I) is selected from the compound of formula (IV): Among them, Y 1 Y 2 Y 3 Y 4 Y 5 R 2 , n, L 1 R 1 L 2 R 3 and R 4 As defined in any one of claims 1-13.

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

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, and a pharmaceutically acceptable excipient.

19. Use of the compound of any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof or a stereoisomer thereof, or the pharmaceutical composition of claim 18, in the preparation of a medicament for inhibiting WRN.

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