Nitrogen-containing heteroaryl compound, preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a nitrogen-containing heteroaryl compound, a preparation method therefor and the use thereof. The present invention specifically relates to a compound represented by formula (I) for treating cancer, and an isotope-labelled substance, enantiomer, diastereomer, atropisomer, solvate, prodrug or pharmaceutically acceptable salt thereof. The compound of the present invention has good inhibitory activity against WRN, and has good application prospects.

Description

A class of nitrogen-containing heteroaryl compounds, their preparation methods and uses

[0001] This application claims priority to Chinese Patent Application No. 2024119258432, filed on December 25, 2024, and Chinese Patent Application No. 2025102703601, filed on March 7, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention relates to a WRN inhibitor, its preparation method and application, and more specifically to a class of nitrogen-containing heteroaryl compounds, their compositions and their use in the treatment and prevention of cancer. Background Technology

[0003] WRN, the only member of the RecQ helicase family to possess both helicase and exonuclease activities, plays a crucial role in DNA repair, replication, transcription, and telomere maintenance. The protein structure of WRN includes an N-terminal 3'-5' exonuclease domain, an ATPase domain, an RQC domain (RecQ C-terminal), a HRDC domain (Helicase-and-ribonuclease DC-terminal), and a C-terminal nuclear localization signal sequence (Kitano K. Front Genet. 2014, 5:366). WRN can recognize specific DNA secondary structures such as DNA bubbles, replication forks, Holliday junctions, G4-quadruaplexes, and triple helices, and promote their unwinding, thereby participating in DNA replication and repair processes (Constantinou A, et al. EMBO Rep. 2000, 1(1):80-84). WRN has a synthetic lethal relationship with tumors that are microsatellite highly unstable (MSI-high, MSI-H) or mismatch repair deficient (dMMR) (Chan EM, et al. Nature 2019, 568(7753):551-556). Studies have shown that WRN gene knockdown or knockout, WRN helicase loss mutations, and small molecule WRN helicase inhibitors can significantly inhibit the cell growth of MSI-H tumors and the formation of subcutaneous xenografts in mice (Ding Z, et al. Cancer Res. 2020, 80(16_Supplement): 1377-1377; Picco G, et al. Cancer Discov. 2021, 11(8): 1923-1937; Lieb S, et al. Elife. 2019, 8: e43333; Aggarwal M, et al. Proc. Natl. Acad. Sci. 2011, 108(4): 1525-1530). Therefore, highly active and selective WRN inhibitors can be used to treat MSI-H or dMMR cancers.

[0004] Covalent inhibitors are a class of small molecule inhibitors that exert their biological functions by irreversibly binding to target protein residues via covalent bonds. Covalent inhibitors often exhibit high biochemical efficiency, potent and sustained effects, requiring reduced dosage and frequency of administration, maintaining efficacy even after drug clearance, and targeting specific non-conserved residues of proteins, achieving high selectivity. Therefore, the development of covalent inhibitors targeting WRN holds great promise for anti-tumor applications. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, this invention provides a class of compounds, their preparation methods, and applications. The compounds of this invention exhibit good inhibitory activity against WRN.

[0006] This invention provides a compound of formula (I), its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof:

[0007] in,

[0008] X 1 For N or CR 0 ,

[0009] R 0 Selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, C1-C6 alkyl, halo-C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, and deuterated C1-C6 alkoxy.

[0010] X 2 For N or CR 3 ,

[0011] R 3 Selected from hydrogen, halogen, cyano, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, unsubstituted or substituted C3-C6 cycloalkyl, unsubstituted or substituted 3-8 membered heterocyclic groups, -NR 3a R 3b ;R 3 The substitution mentioned herein refers to substitution by one or more substituents selected from group A; group A substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR A1 R A2 -CONR A1 R A2 -NR A1 C(O)R A2 ;

[0012] Y represents O, CR 9 R 10 or

[0013] R 9 R 10 R 11 R 12 R 13R 14 Each group is independently selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, C1-C6 alkyl, halo-C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, and deuterated C1-C6 alkoxy.

[0014] R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted C3-C8 cycloalkyl groups, unsubstituted or substituted C6-C10 aryl groups, unsubstituted or substituted 3-8 membered heterocyclic groups, unsubstituted or substituted 5-10 membered heteroaryl groups, and unsubstituted or substituted C3-C8 cycloalkenyl groups; R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;

[0015] R 2 For -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 -OR 17 ;R 16 and R 17 Selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl, unsubstituted or substituted C2-C6 ynyl, unsubstituted or substituted C3-C8 cycloalkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 3-8 membered heterocyclic, and unsubstituted or substituted 5-10 membered heteroaryl; R 16 and R17 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR D1 R D2 -CONR D1 R D2 -NR D1 C(O)R D2 ;

[0016] R 4 It is hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C3-C8 cycloalkyl, or unsubstituted or substituted 5-6 membered heterocyclic group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR E1 R E2 -CONR E1 R E2 -NR E1 C(O)R E2 ;

[0017] R 5 It is hydrogen or deuterium;

[0018] R 6 It is hydrogen or C1-C6 alkyl;

[0019] R 7 It is hydrogen, C1-C6 alkyl, or halogen;

[0020] R 8 It is hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C3-C8 cycloalkyl, or unsubstituted or substituted 4-6 membered heterocyclic group; R 8The substitution mentioned herein refers to substitution by one or more substituents selected from group F; group F substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR F1 R F2 -CONR F1 R F2 -NR F1 C(O)R F2 ;

[0021] R 2a R 2b R 3a R 3b R A1 R A2 R B1 R B2 R C1 R C2 R D1 R D2 R E1 R E2 R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, halo-C1-C6 alkyl, and C3-C6 cycloalkyl;

[0022] With S atoms and R 7 Connected separately express It can be a geometric isomer of (Z) or (E); This indicates the connection point of the group with other locations;

[0023] The *-marked C atom can be a chiral carbon atom or a non-chiral carbon atom; when the *-marked C atom is a chiral carbon atom, the configuration of the chiral carbon atom is S configuration or R configuration;

[0024] The heteroatoms in the 3-8 membered heterocyclic group, 5-6 membered heterocyclic group, 4-6 membered heterocyclic group, and 5-10 membered heteroaryl group are independently selected from 1, 2, 3, or 4 of N, O, and S.

[0025] In one implementation method

[0026] X 1 For N or CR 0 ,

[0027] R 0Selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, C1-C6 alkyl, halo-C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, and deuterated C1-C6 alkoxy.

[0028] X 2 For N or CR 3 ,

[0029] R 3 Selected from hydrogen, halogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, -NR 3a R 3b ;R 3 The substitution mentioned herein refers to substitution by one or more substituents selected from group A; group A substituents include: deuterium, halogen, C1-C6 alkoxy, halo-C1-C6 alkoxy, -NR A1 R A2 -CONR A1 R A2 -NR A1 C(O)R A2 ;R 3a R 3b R A1 R A2 Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl.

[0030] In one implementation, X 1 For N or CH; X 2 For N or CR 3 R 3 It is selected from hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, amino, -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl).

[0031] In one implementation, X 1 For N or CH; X 2 For N or CR 3 R 3 Selected from hydrogen, amino, -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl).

[0032] In one implementation, X 1 For N or CH; X 2 For N or CR 3 R 3 Selected from hydrogen, amino, -NHCH3, -NH-cyclopropyl.

[0033] In one implementation, X 1 For N or CH; X2 It can be N or CH.

[0034] In one implementation, X 1 It can be N or CH; preferably N.

[0035] In one implementation, X 2 It can be N or CH; preferably CH.

[0036] In one implementation, Y is CR 9 R 10 or

[0037] In one implementation, R 9 R 10 R 11 R 12 R 13 R 14 Each is independently selected from hydrogen.

[0038] In one embodiment, Y is -CH2- or -(CH2)2-; preferably -CH2-.

[0039] In one implementation, R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted cyclobutyl, unsubstituted or substituted cyclopentyl, unsubstituted or substituted cyclohexyl, unsubstituted or substituted phenyl, unsubstituted or substituted... No substitution or substitution No substitution or substitution No substitution or substitution R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 RC2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halogenated C1-C6 alkyl, and C3-C6 cycloalkyl; R 1s It is a C1-C6 alkyl or a halogenated C1-C6 alkyl.

[0040] In one implementation, R 1 The phenyl group is either unsubstituted or substituted; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, and C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halogenated C1-C6 alkyl, and C3-C6 cycloalkyl.

[0041] In one implementation, R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3.

[0042] In one implementation, R 1 For R15 ;R 15 It is an unsubstituted or substituted C6-C10 aryl group; R 15 The substitution mentioned herein refers to being replaced by one or more substituents selected from group B; the substituents in group B are deuterium.

[0043] In one implementation, R 1 In this context, the substituent in group B is deuterium.

[0044] In one implementation, R 1 It is a phenyl group.

[0045] In one implementation, R 1 for Preferred

[0046] In one implementation, R 2 Selected from -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl.

[0047] In one implementation, R 2 For -NR 2a R 2b or -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl.

[0048] In one implementation, R 2 -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups and unsubstituted or substituted 3-8 membered heterocyclic groups; R 16 The substitution mentioned herein refers to being replaced by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups or C3-C6 cycloalkyl groups.

[0049] In one implementation, R 2 Selected from methyl, ethyl, tert-butyl, Preferred More preferably For example

[0050] In one implementation, R 4 It is an unsubstituted or substituted C1-C6 alkyl group, an unsubstituted or substituted C3-C6 cycloalkyl group, or an unsubstituted or substituted 4-6 membered heterocyclic group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR E1 R E2 -CONR E1 R E2 -NR E1 C(O)R E2 ;R E1 R E2 Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl.

[0051] In one implementation, R 4 It is an unsubstituted or substituted C1-C6 alkyl group, an unsubstituted or substituted cyclopropyl group, or an unsubstituted or substituted cyclobutyl group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, and halo-C1-C6 alkoxy.

[0052] In one implementation, R 4 It is an unsubstituted C3-C8 cycloalkyl group.

[0053] In one implementation, R 4 for

[0054] In one implementation, R 5 For hydrogen, R 6 For hydrogen, R 7 It is hydrogen.

[0055] In one implementation, R 8 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 8 The substitution mentioned herein refers to being replaced by one or more substituents selected from the group F; the substituents of the group F include: deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, amino, -N(CH3)2.

[0056] In one implementation, R 8 It is an unsubstituted C1-C6 alkyl group.

[0057] In one implementation, R 8 Selected from methyl, ethyl, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2,

[0058] In one implementation, R 8 It is a methyl group.

[0059] In one embodiment, the compound represented by formula (I) has a structure as shown in formula (II):

[0060] in,

[0061] X 1 X 2 Y, R 1 R 2 and R 8 The definition is the same as described above.

[0062] In one embodiment, the compound represented by formula (I) has a structure as shown in formula (III), (IV), (V) or (VI):

[0063] in,

[0064] Y, R 1 R 2 and R 8 The definition is the same as described above.

[0065] Preferably, in formulas (III), (IV), (V), and (VI),

[0066] Y is O, CH2, or -(CH2)2-;

[0067] R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted cyclobutyl, unsubstituted or substituted cyclopentyl, unsubstituted or substituted cyclohexyl, unsubstituted or substituted phenyl, unsubstituted or substituted... No substitution or substitution No substitution or substitution No substitution or substitution R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halogenated C1-C6 alkyl, and C3-C6 cycloalkyl; R 1s It is a C1-C6 alkyl or a halo-C1-C6 alkyl;

[0068] R 2 Selected from -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 ;R 16Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl;

[0069] R 8 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 8 The substitution mentioned herein refers to being replaced by one or more substituents selected from the group F; the substituents of the group F include: deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, amino, -N(CH3)2.

[0070] Preferably, in formulas (III), (IV), (V), and (VI),

[0071] Y is CH2 or -(CH2)2-;

[0072] R 1 For R 15 ;R 15 It is an unsubstituted or substituted C6-C10 aryl group; R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; the substituents in group B are deuterium;

[0073] R 2 -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups and unsubstituted or substituted 3-8 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups, or C3-C6 cycloalkyl groups;

[0074] R 8 It is an unsubstituted C1-C6 alkyl group.

[0075] Preferably, in formulas (III), (IV), (V), and (VI),

[0076] Y is O, CH2, or -(CH2)2;

[0077] R 1It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3;

[0078] R 2 For -NR 2a R 2b or -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl;

[0079] R 8 Selected from methyl, ethyl, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2,

[0080] Preferably, in formulas (III), (IV), (V), and (VI),

[0081] Y is O, CH2, or -(CH2)2;

[0082] R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being replaced by one or more substituents selected from group B; the substituents in group B are: deuterium;

[0083] R 2 For R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups, or C3-C6 cycloalkyl groups;

[0084] R 8 Selected from methyl.

[0085] Preferably, in formulas (III), (IV), (V), and (VI),

[0086] Y is CH2;

[0087] R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3;

[0088] R 2 Selected from methyl, ethyl, tert-butyl,

[0089] R 8 It is a methyl group.

[0090] Preferably, in formulas (III), (IV), (V), and (VI),

[0091] Y is CH2;

[0092] R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being replaced by one or more substituents selected from group B; the substituents in group B are: deuterium;

[0093] R 2 For substituted ethyl, and substituted The substitution refers to being replaced by one or more substituents selected from group B; the substituents in group B are F, methyl, or cyclopropyl.

[0094] In specific embodiments, the compounds of formula (I) described in this invention are selected from the following compounds:

[0095] This invention also provides methods for preparing compounds of formula (I), pharmaceutically acceptable salts thereof, isotope-labeled compounds thereof, solvates thereof, solvates of pharmaceutically acceptable salts thereof, or prodrugs thereof. Specifically, these methods include the following preparation methods:

[0096] Method 1:

[0097] Step 1: Compound a-1 reacts with carboxylic acid R 2COOH undergoes a Minisci reaction to give compound a-2; in which X 1 X 2 and R 2 The definition is the same as described above; Y 1 and Y 2 Each is independently selected from Cl, Br, I, OTs, OMs, OTf; Y 3 Selected from C1-C6 alkyl and benzyl groups;

[0098] Step 2: Compound a-2 reacts with R-containing compounds under the catalysis of a metal reagent. 1 Boron reagents, containing R 1 Tin reagents, containing R 1 The zinc reagent undergoes a coupling reaction to give compound a-3; wherein, R 1 The definition is the same as described above; the metal reagent can be a Pd or Ni-containing reagent, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, NiCl2; the R-containing reagent... 1 Boron reagents can be R 1 B(OH)2, R 1 B(Pin); the one containing R 1 The tin reagent can be R 1 Sn(Bu)3; the R-containing 1 The zinc reagent can be (R) 1 )2Zn、R 1 ZnCl, R 1 ZnBr;

[0099] Step 3: Compound a-3 undergoes a coupling reaction with a methyl-containing boron reagent and a methyl-containing zinc reagent under the catalysis of a metal reagent to obtain compound a-4; wherein, the metal reagent can be a Pd or Ni-containing reagent, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, NiCl2; the methyl-containing boron reagent can be MeB(OH)2, MeB(Pin); the methyl-containing zinc reagent can be (Me)2Zn, MeZnCl, MeZnBr;

[0100] Step 4: Compound a-4 undergoes a halogenation reaction to give compound a-5; wherein, Y 4 Selected from Cl, Br, I;

[0101] Step 5: Compound a-5 undergoes a cyclization reaction with amine a-6 to give compound (I-1); where R 4 R 5 R 6 R 7 and R 8 The definition is the same as described above.

[0102] Method 2:

[0103] Step 1: Compound a-7 and R 2 H undergoes a nucleophilic substitution reaction to give compound a-8; in which X 1 X 2 and R 2 The definition is the same as described above; Y 3 Selected from C1-C6 alkyl and benzyl groups; Y 5 Selected from F, Cl, Br, OTs, OMs, OTf; Y 6 Selected from Cl, Br, I, OTs, OMs, OTf, and Me;

[0104] Step 2: Compound a-8 undergoes a halogenation reaction to give compound a-9; where Y 4 Selected from Cl, Br, I;

[0105] Step 3: Compound a-9 reacts with R-containing compounds under the catalysis of a metal reagent. 1 Boron reagents, containing R 1 Tin reagents, containing R 1 The zinc reagent undergoes a coupling reaction to give compound a-10; wherein, R 1 The definition is the same as described above; the metal reagent can be a Pd or Ni-containing reagent, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, NiCl2; the R-containing reagent... 1 Boron reagents can be R 1 B(OH)2, R 1 B(Pin); the one containing R 1 The tin reagent can be R 1 Sn(Bu)3; the R-containing 1 The zinc reagent can be (R) 1 )2Zn、R 1 ZnCl, R 1 ZnBr;

[0106] Step 4: Compound a-10 undergoes a coupling reaction with a methyl-containing boron reagent and a methyl-containing zinc reagent under the catalysis of a metal reagent, and the resulting compound then undergoes a halogenation reaction to give a-5; or compound a-10 directly undergoes a halogenation reaction to give a-5; wherein, Y 4 The reagents are selected from Cl, Br, and I; the metal reagents can be reagents containing Pd or Ni, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, and NiCl2; the methyl-containing boron reagents can be MeB(OH)2 or MeB(Pin); the methyl-containing zinc reagents can be (Me)2Zn, MeZnCl, or MeZnBr.

[0107] Step 5: Compound a-5 undergoes a cyclization reaction with amine a-6 to give compound (I-1); where R 4 R 5 R 6 R 7 and R 8 The definition is the same as described above.

[0108] Method 3:

[0109] Step 1: Compound a-7 reacts with carboxylic acid R 1 COOH or R 1 B(OH)₂ undergoes a Minisci reaction to give compound a-11; wherein, X 1 X 2 and R 1 The definition is the same as described above; Y 3 Selected from C1-C6 alkyl and benzyl groups; Y 5 Selected from F, Cl, Br, OTs, OMs, OTf; Y 6 Selected from Cl, Br, I, OTs, OMs, OTf, and Me;

[0110] Step 2: Compound a-11 and compound R 2 H undergoes a nucleophilic reaction or a metal reagent-catalyzed coupling reaction to give a-10; or compound a-11 reacts with R-containing compounds under metal reagent catalysis. 2 Boron reagents, containing R 2 Tin reagents, containing R 2 Zinc reagents, containing R 2 Grignard reagents or reagents containing R 2 The carboxylic acid undergoes a coupling reaction to give compound a-10; or compound a-11 undergoes a coupling reaction with a boron reagent (such as isopropenyl borate pinacol ester, 2-(1-cyclopropylvinyl)borate pinacol ester, etc.) or a tin reagent (such as tributyl(1-ethoxyethylene)tin, etc.) under metal reagent catalysis, followed by acidic hydrolysis or oxidative treatment. The resulting ketone carbonyl intermediate reacts with a fluorinating reagent to give compound a-10; wherein, R 2 The definition is the same as described above; the metal reagent can be a reagent containing Pd, Cu, Ni, or Fe, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, CuI, NiCl2, FeCl3, Fe(acac)3; the R-containing reagent... 2 Boron reagents can be R 2 B(OH)2, R 2 B(Pin); the one containing R 2 The tin reagent can be R 2Sn(Bu)3; the R-containing 2 The zinc reagent can be (R) 2 )2Zn、R 2 ZnCl, R 2 ZnBr; the R-containing 2 Grignard reagents can be R 2 MgCl, R 2 MgBr; the R-containing 2 Carboxylic acids can be R 2 COOH; the acidic conditions can be carried out in the presence of trifluoroacetic acid, p-toluenesulfonic acid, or hydrochloric acid; the oxidation conditions can be carried out in the presence of K2OsO4 / NaIO4, RuCl3 / Oxone, or O3; the fluorinating agent can be DAST, BAST, or SF4.

[0111] Step 3: Compound a-10 undergoes a coupling reaction with a methyl-containing boron reagent and a methyl-containing zinc reagent under the catalysis of a metal reagent, and the resulting compound then undergoes a halogenation reaction to obtain a-5; or compound a-10 directly undergoes a halogenation reaction to obtain a-5; wherein, Y 4 The reagents are selected from Cl, Br, and I; the metal reagents can be reagents containing Pd or Ni, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, and NiCl2; the methyl-containing boron reagents can be MeB(OH)2 or MeB(Pin); the methyl-containing zinc reagents can be (Me)2Zn, MeZnCl, or MeZnBr.

[0112] Step 4: Compound a-5 undergoes a cyclization reaction with amine a-6 to give compound (I-1); where R 4 R 5 R 6 R 7 and R 8 The definition is the same as described above.

[0113] Method 4:

[0114] Step 1: Compound a-12 reacts with carboxylic acid R 2 COOH undergoes a Minisci reaction to give compound a-13; in which X 1 X 2 and R 2 The definition is the same as described above; Y 1 Selected from Cl, Br, I, OTs, OMs, OTf;

[0115] Step 2: Compound a-13 undergoes a carbonylation reaction catalyzed by a metal reagent to yield compound a-14; wherein, Y... 3Selected from C1-C6 alkyl and benzyl groups; the metal reagent may be a Pd-containing reagent, such as Pd(dppf)Cl2 or Pd(PPh3)Cl2;

[0116] Step 3: Compound a-14 reacts with R-containing compounds under the catalysis of a metal reagent. 1 Boron reagents, containing R 1 Tin reagents, containing R 1 The zinc reagent undergoes a coupling reaction to give compound a-4; wherein, R 1 The definition is the same as described above; the metal reagent can be a Pd or Ni-containing reagent, such as Pd(dppf)Cl2, Pd(PPh3)Cl2, Pd(PPh3)4, NiCl2; the R-containing reagent... 1 Boron reagents can be R 1 B(OH)2, R 1 B(Pin); the one containing R 1 The tin reagent can be R 1 Sn(Bu)3; the R-containing 1 The zinc reagent can be (R) 1 )2Zn、R 1 ZnCl, R 1 ZnBr;

[0117] Step 4: Compound a-4 undergoes a halogenation reaction to give compound a-5; wherein, Y 4 Selected from Cl, Br, I;

[0118] Step 5: Compound a-5 undergoes a cyclization reaction with amine a-6 to give compound (I-1); where R 4 R 5 R 6 R 7 and R 8 The definition is the same as described above.

[0119] The present invention also provides a pharmaceutical composition comprising the above-described compound as shown in formula (I), a pharmaceutically acceptable salt thereof, an isotope label thereof, a solvate thereof, a solvate of a pharmaceutically acceptable salt thereof, or a prodrug thereof, and pharmaceutical excipients.

[0120] In the composition described herein, the amount of the compound represented by formula (I), its pharmaceutically acceptable salt, its isotopic label, its solvate, its pharmaceutically acceptable salt solvate, or its prodrug may be an effective therapeutic amount.

[0121] The present invention also provides the use of the above-described compound of formula (I), its pharmaceutically acceptable salt, its isotopic label, its solvate, its pharmaceutically acceptable salt solvate, or its prodrug in the preparation of a WRN inhibitor or medicament; said medicament being a medicament for treating and / or preventing disease by inhibiting WRN.

[0122] In one embodiment, the treatment and / or prevention of solid tumors with microsatellite high instability (MSI-high, MSI-H) or mismatch repair deficiency (dMMR) by inhibiting WRN is preferred for colorectal cancer.

[0123] The present invention also provides the use of the above-described compound of formula (I), its pharmaceutically acceptable salt, its isotopic label, its solvate, its pharmaceutically acceptable salt solvate, or its prodrug in the preparation of a medicament for treating and / or preventing cancer.

[0124] The cancer may include solid tumors with microsatellite instability (MSI-high, MSI-H) or mismatch repair deficiency (dMMR), such as endometrial cancer, colorectal cancer, gastric cancer, ovarian cancer, etc.; preferably colorectal cancer.

[0125] The present invention also provides a method for treating and / or preventing cancer, which involves administering to a patient a therapeutically effective amount of the above-described compound as shown in formula (I), its pharmaceutically acceptable salt, its isotopic label, its solvate, its pharmaceutically acceptable salt solvate, or its prodrug.

[0126] The term "pharmaceutical acceptable" means that something is relatively non-toxic, safe, and suitable for patient use.

[0127] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. See Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, Camille G. Wermuth, 2011, 2nd Revised Edition) for details.

[0128] The term "solvate" refers to a substance formed by the combination of a compound and a solvent (including but not limited to water, methanol, ethanol, etc.). Solvates are classified into stoichiometric solvates and non-stoichiometric solvates.

[0129] The term "solvate of a pharmaceutically acceptable salt" refers to a substance formed by the combination of a compound with a pharmaceutically acceptable acid or base and a solvent (including but not limited to water, methanol, ethanol, etc.). The amount of solvent can be stoichiometric or non-stoichiometric.

[0130] The "-" at the end of a group indicates that the group is attached to the rest of the molecule through that site. For example, CH3-C(=O)- refers to an acetyl group.

[0131] exist In the diagram, the wavy line indicates the connection point between the group and other parts of the molecule.

[0132] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0133] The term "oxo" refers to the =O group, where an oxygen atom replaces two hydrogen atoms on the same carbon atom; that is, a carbonyl group replaces a methylene group.

[0134] The term "alkyl" refers to a straight-chain or branched, saturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6 or C2-C6). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, etc.

[0135] The term "alkoxy" refers to the group R. X -O-,R X The definition is the same as the term "alkyl". Alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, etc.

[0136] The term "alkenyl" refers to a straight-chain or branched, unsaturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C2-C6) and having one or more (e.g., 1, 2, or 3) carbon-carbon sp groups. 2 Double bond. Alkenyl groups include, but are not limited to: vinyl groups, wait.

[0137] The term "alkynyl" refers to a straight-chain or branched, unsaturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C2-C6) and possessing one or more (e.g., 1, 2, or 3) carbon-carbon triple bonds. Alkenyl groups include, but are not limited to: ethynyl, wait.

[0138] The term "cycloalkyl" refers to a cyclic, saturated monovalent hydrocarbon group having a specified number of carbon atoms (e.g., C3-C6), which can be monocyclic, bridged, or spirocyclic. Cycloalkyl groups include, but are not limited to: wait.

[0139] The term "cycloalkenyl" refers to an alicyclic hydrocarbon group containing one, two, or three double bonds, having a specified number of carbon atoms (e.g., C3-C12, C3-C8, C3-C6, C5-C8, C5-C6). Cycloalkenyl groups include, but are not limited to: wait.

[0140] The term "cycloalkoxy" refers to the group R Y -O-,R Y The definition is the same as the term "cycloalkyl". Cycloalkoxy groups include, but are not limited to, cyclopropoxy groups.

[0141] The term "aryl" refers to an aryl group having a specified number of carbon atoms (e.g., C6-C). 10 Aryl groups are cyclic, unsaturated monovalent hydrocarbon groups, which can be monocyclic or polycyclic (e.g., two or three). When polycyclic, the monocyclic rings share two atoms and one bond, and each ring is aromatic. Aryl groups include, but are not limited to, phenyl and naphthyl groups.

[0142] In this invention, the term "heterocyclic group" refers to a non-aromatic cyclic group comprising at least one carbon atom and at least one (e.g., 1-3) cyclic heteroatoms selected from N, O, and S, wherein the sulfur atom may optionally be oxidized or amination. Examples of "heterocyclic groups" specifically include cycloalkyl groups in which one or more cyclic carbons are selected from -O-, -N=, -NR-, -S-, -S(=O)-, and -S(=O)2-, as defined in this invention. The group formed by partial substitution, wherein R is hydrogen, C is C 1-4 Alkyl, C 3-6 Cycloalkyl or nitrogen-protected groups (e.g., benzyloxycarbonyl, p-methoxybenzylcarbonyl, tert-butoxycarbonyl, acetyl, benzoyl, benzyl, p-methoxy-benzyl, p-methoxy-phenyl, 3,4-dimethoxybenzyl, etc.). "Heterocyclic groups" include monocyclic and fused rings, bridged rings, spirocyclic, and other bicyclic structures, and may be partially or completely saturated, such as 4-10 membered saturated or unsaturated heterocyclic groups, 4-6 membered saturated or unsaturated heterocyclic groups, 3-8 membered heterocyclic groups, 4-8 membered heterocyclic groups, 3-6 membered heterocyclic groups, etc.; such as tetrahydrofuranyl, pyrrolidinyl, oxetyl, oxetylhexyl, aziridine, thioheterocyclic, 1,2-dithioheterocyclic, 1,3-dithioheterocyclic, azirheptanyl, oxetylheptanyl, etc. For example, the heterocyclic groups described in this invention may preferably be selected from the following groups:

[0143] In this invention, the term "heteroaryl" refers to a monocyclic, bicyclic, or fused polycyclic cyclic aromatic hydrocarbon group having a specified number of ring atoms (e.g., 5-10), which contains at least one (e.g., 1-3) cyclic heteroatoms independently selected from N, O, and S (e.g., N), with the remaining ring atoms being carbon atoms; such as imidazolyl, pyridinyl, pyrrololyl, thiazolyl, furanyl, oxazolyl, isoxazolyl, pyrazolyl, thiophene, pyrimidinyl, 1,2,4-triazolyl, benzoxazolyl, imidazopyridyl, triazolylpyridinyl, benzofuranyl, pyrazolylpyrimidinyl, benzo-m-dioxacyclopentenyl, indolyl, quinolinyl, isoquinolinyl, etc.

[0144] The term “heteroary ring” satisfies at least one of the following conditions, and the rest are defined the same as the term “heteroaryl”: 1. It is connected to the rest of the molecule by two or more single bonds; 2. It shares two atoms and one bond with the rest of the molecule.

[0145] The term "isotope-labeled compound" refers to an isotope-labeled compound in which one or more atoms, compared to a compound of formula (I), are replaced by atoms with atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of H, C, N, O, S, F, and Cl, such as... 2 H, 3 H, 13 C 11 C 14 C 15 N、 18 O、 17 O、 32 P, 35 S, 18 F and 36 Cl. The compounds of the present invention containing the aforementioned isotopes and / or other isotopes, pharmaceutically acceptable salts thereof, solvates thereof, solvates of pharmaceutically acceptable salts thereof, or prodrugs thereof are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as those doped with radioactive isotopes (e.g.,... 3 H and 14 Compounds in (C) can be used for drug and / or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, heavier isotopes (such as deuterium, i.e., 2Hydrogen (H or D) substitution can provide certain therapeutic advantages derived from greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirement), and may therefore be preferred in certain circumstances. The compounds of the present invention may be specifically defined as being substituted with deuterium or tritium. Furthermore, the presence of hydrogen in the substituents without a separate mention of the terms deuterium or tritium does not exclude deuterium or tritium, but may also include deuterium or tritium.

[0146] The term "therapeutic effective dose" refers to the amount given to a patient that is sufficient to effectively treat the disease. Therapeutic effective doses will vary depending on the type of compound, the type of disease, the severity of the disease, the patient's age, etc., but may be adjusted as appropriate by those skilled in the art.

[0147] The term "pharmaceutical excipients" refers to all substances contained in a pharmaceutical preparation other than the active pharmaceutical ingredient, and are generally divided into two main categories: excipients and additives. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 Edition) and Handbook of Pharmaceutical Excipients (Paul J Sheskey, Bruno C Hancock, Gary P Moss, David J Goldfarb, 2020, 9th Edition).

[0148] The term "treatment" refers to eliminating the cause of an illness or relieving symptoms.

[0149] The term "prevention" refers to reducing the risk of developing a disease.

[0150] The term "patient" refers to any animal, typically a mammal such as a human, that requires treatment or prevention of disease. Mammals include, but are not limited to: cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans.

[0151] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0152] The reagents and raw materials used in this invention are all commercially available.

[0153] The positive and progressive effects of the present invention are as follows: the compounds of the present invention have novel structures, and further, the compounds of the present invention satisfy one or more of the following advantages: (1) good inhibitory activity against WRN; (2) good inhibitory activity against the proliferation of SW48 cells; (3) excellent pharmacokinetic properties (e.g., low clearance rate and high oral exposure); (4) excellent antitumor activity. Attached Figure Description

[0154] Figure 1 shows the tumor growth curves of the control group and each experimental group in the pharmacodynamic evaluation experiment of the mouse SW48 human colon cancer xenograft model.

[0155] Figure 2 shows the relative body weight changes of mice in the control group and each experimental group during the pharmacodynamic evaluation experiment of the SW48 human colon cancer xenograft model. Detailed Implementation

[0156] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0157] In each embodiment, 1 H NMR was recorded using a BRUKER AVANCE NEO 400MHz JNM-ECZ400s NMR spectrometer, and chemical shifts are expressed as δ (ppm). Liquid chromatography-mass spectrometry (LC-MS) was recorded using a Shimadzu LC-20AD Agilent 1260 mass spectrometer. Preparative HPLC separation was performed using a WATERS2545 Shimadzu LP-20AP liquid chromatograph.

[0158] The following abbreviations may be used in the following experimental descriptions:

[0159] Table 1

[0160] Preparation of intermediates

[0161] Preparation of intermediate INT-1:

[0162] Step 1: (S)-(1-Cyclopropyl-2-hydroxyethyl) tert-butyl carbamate

[0163] (S)-2-((tert-Butoxycarbonyl)amino)-2-cyclopropylacetic acid INT-1-1 (10.0 g, 46.5 mmol) was dissolved in tetrahydrofuran (100 mL), and borane (66.6 mL, 66.6 mmol, 1 M THF solution) was slowly added dropwise at 0 °C. The system was stirred at room temperature for 2 hours. Then, methanol (60 mL) was slowly added dropwise to quench the reaction. The mixture was concentrated under vacuum to obtain a crude product, which was purified by column chromatography (PE:EA = 3:1) to obtain compound INT-1-2 (6.80 g, 72.7%, colorless oil).

[0164] Step 2: (S)-(1-Cyclopropyl-2-aldehyde)tert-butyl carbamate

[0165] (S)-(1-cyclopropyl-2-hydroxyethyl)carbamate tert-butyl ester INT-1-2 (6.30 g, 31.3 mmol) was dissolved in dichloromethane (70 mL). The system was stirred at 0 °C for 10 minutes, and then Dys-Martin oxidant (15.9 g, 37.6 mmol) was slowly added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, saturated sodium bicarbonate solution (50 mL) was slowly added dropwise, and the mixture was extracted with ethyl acetate (100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (PE:EA = 4:1) to give compound INT-1-3 (1.60 g, crude product, colorless oil).

[0166] Step 3: (S,E)-(1-Cyclopropyl-3-(Methylsulfonyl)allyl)tert-butyl carbamate

[0167] Compound (S)-(1-cyclopropyl-2-aldehyde)carbamate tert-butyl ester INT-1-3 (1.40 g, 7.03 mmol) and diethyl methanesulfonyl methylphosphonate ester INT-1-4 (1.62 g, 7.03 mmol) were dissolved in tetrahydrofuran (15 mL), and potassium carbonate (2.43 g, 17.6 mmol) was added. The mixture was stirred at 60 °C for 4 hours under nitrogen protection. After the reaction was complete, the mixture was filtered and concentrated. The residue was purified by column chromatography (PE:EA = 2:1) to give product INT-1-5 (1.30 g, crude product, white solid). The crude product was dissolved in ethyl acetate (10 mL), and petroleum ether was slowly added until a solid precipitated. Filtering yielded compound INT-1-5 (850 mg, yield 43.9%, white solid). LCMS calc. for C8H 14 NO4S[M+H-C4H8] + :m / z=222.0; Found: 222.0; Chiral HPLC Ee%=97.2.

[0168] Step 4: (S,E)-1-Cyclopropyl-3-(Methanesulfonyl)propyl-2-en-1-amine hydrochloride

[0169] (S,E)-(1-cyclopropyl-3-(methanesulfonyl)allyl)carbamate tert-butyl ester INT-1-5 (200 mg, 727 μmol) was dissolved in hydrochloric acid (2 mL, 4N EA solution), and the system was stirred at room temperature for 0.5 hours. After the reaction was completed, the solution was concentrated under reduced pressure to give compound INT-1 (127 mg, crude product, white solid). 1H NMR(400MHz, DMSO-d6)δ8.73(s,3H),7.07(dd,J=15.4,1.2Hz,1H),6.80(dd,J=15.4,6.2Hz,1 H),3.35-3.27(m,1H),3.06(s,3H),1.23-0.95(m,1H),0.75-0.48(m,3H),0.46-0.32(m,1H).

[0170] Example 1: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(1,1-difluoroethyl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0171] Step 1: Methyl 5-bromo-4-chloro-6-(1,1-difluoroethyl)nicotinic acid

[0172] A mixture of methyl 5-bromo-4-chloronicotinate Cpd-1-1 (1.00 g, 3.99 mmol), 2,2-difluoropropionic acid (880 mg, 7.98 mmol), silver nitrate (680 mg, 3.99 mmol), and potassium persulfate (5.40 g, 20.0 mmol) in acetonitrile (10 mL) and water (10 mL) was stirred at 50 °C for 16 hours. After the reaction was complete, the mixture was extracted with ethyl acetate (20 mL × 2), the organic layers were combined, washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-1-2 (480 mg, yield 38.6%, colorless oil). LCMS calc. for C9H8BrClF2NO2[M+H) + :m / z=313.9 / 315.9; Found:313.8 / 315.8.

[0173] Step 2: Methyl 4-chloro-6-(1,1-difluoroethyl)-5-phenylnicotinic acid

[0174] Under an argon atmosphere, compound Cpd-1-2 (480 mg, 1.54 mmol), phenylboronic acid (210 mg, 1.70 mmol), Pd(dppf)Cl2 (130 mg, 150 μmol), and potassium phosphate (650 mg, 3.08 mmol) were stirred at 80 °C for 3 hours in a mixture of toluene (10 mL) and water (2.5 mL). After cooling to room temperature, the mixture was extracted with ethyl acetate (10 mL × 2). The combined organic layers were washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-1-3 (390 mg, yield 81.1%, yellow solid). LCMS calc. for C 15 H 13 ClF2NO2[M+H] + :m / z=312.1;Found:312.0.

[0175] Step 3: Methyl 6-(1,1-difluoroethyl)-4-methyl-5-phenylnicotinic acid

[0176] Under an argon atmosphere, compound Cpd-1-3 (300 mg, 960 μmol), methylboric acid (115 mg, 1.92 mmol), Pd(dppf)Cl2 (78.6 mg, 100 μmol), and potassium phosphate (511 mg, 2.41 mmol) were stirred at 100 °C for 3 hours in a mixture of toluene (8 mL) and water (2 mL). After cooling to room temperature, the mixture was extracted with ethyl acetate (10 mL × 2). The combined organic layers were washed with saturated brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-1-4 (300 mg, crude product, colorless oil). LCMS calc. for C 16 H 16 F2NO2[M+H] + :m / z=292.1;Found:292.1.

[0177] Step 4: Methyl 4-(bromomethyl)-6-(1,1-difluoroethyl)-5-phenylnicotinic acid

[0178] At room temperature, azobisisobutyronitrile (16.9 mg, 100 μmol) was added to a carbon tetrachloride (8 mL) solution of compound Cpd-1-4 (300 mg, 1.03 mmol) and N-bromosuccinimide (275 mg, 1.54 mmol). The mixture was stirred at 80 °C for 16 hours. The mixture was then cooled and concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-1-5 (220 mg, yield 57.7%, white solid). LCMS calc. for C 16 H 15 BrF₂NO₂[M+H] + :m / z=370.0 / 372.0; Found:369.9 / 371.7.

[0179] Step 5: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(1,1-difluoroethyl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0180] Compound Cpd-1-5 (220 mg, 590 μmol), INT-1 (189 mg, 890 μmol), and diisopropylethylamine (384 mg, 2.97 mmol) were stirred at 80 °C for 16 hours in a mixture of tetrahydrofuran (8 mL). The mixture was then cooled and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:1) and SFC to give compound Cpd-1 (12.9 mg, yield 4.99%, white solid). LCMS calc. for C 22 H 23 F2N2O3S[M+H] + :m / z=433.1;Found:433.0; 1 H NMR(400MHz, DMSO-d6)δ9.01(s,1H),7.51-7.42(m,5H),6.87(d,J=16.4Hz,1H),6.80(dd,J=15.2,4.4Hz,1H),4.41-4.30(m,2H),4 .16-4.12(m,1H),2.99(s,3H),1.99(t,J=19.2Hz,3H),1.33-1.23(m,1H),0.65-0.59(m,1H),0.56-0.46(m,2H),0.29-0.23(m,1H).

[0181] Example 2: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(3-fluoro-3-methylazacyclobutan-1-yl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0182] Step 1: Methyl 6-chloro-4-methylpyridine-3-carboxylic acid ester

[0183] At room temperature, iodomethane (710 μL, 11.5 mmol) was added dropwise to a mixture of Cpd-2-1 (1.71 g, 9.97 mmol) and potassium carbonate (2.75 g, 19.9 mmol) in N,N-dimethylformamide (15 mL). The system was stirred at 40 °C for 2 hours, cooled to room temperature, diluted with ethyl acetate (100 mL), washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-2-2 (1.74 g, 94.1% yield, white solid). LCMS calc. for C8H9ClNO2[M+H) + :m / z=186.0;Found:186.1.

[0184] Step 2: 4-(bromomethyl)-6-chloropyridine-3-carboxylic acid methyl ester

[0185] Under argon atmosphere, a mixture of Cpd-2-2 (1.50 g, 8.08 mmol), N-bromosuccinimide (1.58 g, 8.89 mmol), and azobisisobutyronitrile (270 mg, 1.62 mmol) in 1,2-dichloroethane (25 mL) was stirred at 70 °C for 16 hours. After cooling to room temperature, the mixture was quenched with saturated sodium thiosulfate solution (50 mL) and extracted with dichloromethane (30 mL × 3). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-2-3 (770 mg, yield 36.0%, white solid). LCMS calc. for C8H8BrClNO2[M+H] + :m / z=263.9 / 265.9; Found:263.8 / 265.7.

[0186] Step 3: (S)-6-chloro-2-(1-cyclopropyl-2-hydroxyethyl)-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0187] In a sealed tube, a solution of Cpd-2-3 (700 mg, 2.65 mmol), INT-1-6 (364 mg, 2.65 mmol), and N,N-diisopropylethylamine (1.38 mL, 7.94 mmol) in n-butanol (7 mL) was stirred at 140 °C for 90 min. The mixture was cooled and concentrated, and the residue was purified by silica gel column chromatography (EA 100%) to give compound Cpd-2-4 (413 mg, yield 61.8%, pale yellow solid). LCMS calc. for C 12 H 14 ClN2O2[M+H] + :m / z=253.1;Found:252.9.

[0188] Step 4: (S)-2-(1-cyclopropyl-2-hydroxyethyl)-6-(3-fluoro-3-methylazacyclobutane-1-yl)-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0189] In a sealed tube, a mixture of Cpd-2-4 (403 mg, 1.59 mmol), 3-fluoro-3-methylazacyclobutane hydrochloride (600 mg, 4.78 mmol), and N,N-diisopropylethylamine (1.10 mL, 6.38 mmol) in N-methylpyrrolidone (8 mL) was stirred at 140 °C for 16 hours. After cooling and concentration, the residue was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give compound Cpd-2-5 (290 mg, 82.1% yield, white solid). LCMS calc. for C 16 H 21 FN3O2[M+H] + :m / z=306.2;Found:306.8.

[0190] Step 5: (S)-7-bromo-2-(1-cyclopropyl-2-hydroxyethyl)-6-(3-fluoro-3-methylazacyclobutane-1-yl)-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0191] At room temperature, N-bromosuccinimide (247 mg, 1.39 mmol) was added to a solution of Cpd-2-5 (283 mg, 930 μmol) in N,N-dimethylformamide (3 mL). The mixture was stirred at room temperature for 10 minutes and diluted with ethyl acetate (50 mL). The organic compound was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound Cpd-2-6 (356 mg, crude product, white solid), which was used directly in the next reaction. LCMS calc. for C 16 H 20BrFN3O2[M+H] + :m / z=384.1 / 386.1; Found:383.9 / 385.9.

[0192] Step Six: (S)-2-(1-Cyclopropyl-2-hydroxyethyl)-6-(3-fluoro-3-methylazacyclobutane-1-yl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0193] Under argon protection, a mixture of Cpd-2-6 (356 mg, 930 μmol), phenylboronic acid (169 mg, 1.39 mmol), potassium carbonate (384 mg, 2.78 mmol), and Pd(dppf)Cl2 (75.7 mg, 90.0 μmol) in acetonitrile (10 mL) and water (2.5 mL) was stirred at 80 °C for 30 min. After cooling to room temperature, the mixture was diluted with ethyl acetate (100 mL) and washed with water (50 mL) and saturated brine (50 mL). The solution was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give compound Cpd-2-7 (290 mg, yield 82.1%, white solid). LCMS calc. for C 22 H 25 FN3O2[M+H] + :m / z=382.2;Found:381.5.

[0194] Step 7: (S)-2-cyclopropyl-2-(6-(3-fluoro-3-methylazacyclobutane-1-yl)-3-oxo-7-phenyl-1,3-dihydro-2H-pyrrolo[3,4-c]pyridin-2-yl)acetaldehyde

[0195] A solution of Cpd-2-7 (290 mg, 760 μmol) and Dietrich Martin oxidant (484 mg, 1.14 mmol) in dichloromethane (10 mL) was stirred at room temperature for 1 hour. The reaction mixture was diluted with dichloromethane (50 mL) and quenched with saturated sodium bicarbonate solution (25 mL) and saturated sodium thiosulfate solution (25 mL). The mixture was stirred at room temperature for 10 minutes. The separated organic phase was washed with saturated sodium bicarbonate solution (25 mL), saturated sodium thiosulfate solution (25 mL), and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound Cpd-2-8 (288 mg, crude, white solid), which was used directly in the next reaction. LCMS calc. for C 22 H 23 FN3O2[M+H] + m / z = 380.2; Found: 379.7.

[0196] Step 8: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(3-fluoro-3-methylazacyclobutan-1-yl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0197] At room temperature, a solution of INT-1-4 (262 mg, 1.14 mmol), lithium chloride (48.3 mg, 1.14 mmol), and DBU (170 μL, 1.14 mmol) in anhydrous acetonitrile (5 mL) was stirred for 15 min. The system was cooled to 0 °C, and a solution of Cpd-2-8 (288 mg, 760 μmol) in acetonitrile (10 mL) was added dropwise. The mixture was then stirred at 25 °C for 30 min. The system was quenched with ice water (50 mL) and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (EA:MeOH = 20:1) and prep-HPLC (FA / MeCN) to give compound Cpd-2 (100 mg, yield 28.9%, white solid). LCMS calc. for C 24 H 27 FN3O3S[M+H] + :m / z=456.2;Found:456.2. 1 H NMR (400MHz, CDCl3) δ8.65 (s, 1H), 7.53-7.39 (m, 3H), 7.35 (d, J = 6.9Hz, 2H), 6 .94(dd,J=15.2,4.3Hz,1H),6.45(dd,J=15.2,1.8Hz,1H),4.30-4.20(m,2H), 4.11(d,J=17.4Hz,1H),4.00-3.82(m,2H),3.75-3.54(m,2H),2.93(s,3H),1. 51(d,J=21.5Hz,3H),1.12-0.99(m,1H),0.82-0.70(m,1H),0.63-0.35(m,3H).

[0198] Step Nine: (S)-2-Amino-2-cyclopropyl ethanol-1-ol hydrochloride

[0199] INT-1-2 (11.0 g, 54.4 mmol) was added to hydrochloric acid (110 mL, 2 M ethyl acetate solution) in an ice bath, and the mixture was stirred at room temperature for 6 hours. LCMS analysis showed complete consumption of the starting material and detection of the desired compound. The reaction mixture was filtered to give INT-1-6 (5.15 g, 69.1% yield, white solid). LCMS calc. for C5H12 NO[M+H] + :m / z=102.1;Found:102.2.

[0200] Example 3: (S,E)-6-(1-cyclopropyl-3-(methanesulfonyl)allyl)-3-(3-fluoro-3-methylazacyclobutane-1-yl)-4-phenyl-5,6-dihydro-7H-pyrrolo[3,4-b]pyridin-7-one

[0201] Step 1: 3-Methylpyridine nitride-2-carboxylic acid methyl ester

[0202] 3-chloroperoxybenzoic acid (10.1 g, 49.6 mmol, 85%) was added to a solution of Cpd-3-1 (5.00 g, 33.1 mmol) in dichloromethane (100 mL) under ice bath conditions. The system was stirred at room temperature for 16 hours, then water (100 mL) was added and the mixture was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated sodium bicarbonate solution (50 mL × 3) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give compound Cpd-3-2 (3.85 g, yield 69.2%, pale yellow oil). 1 H NMR (400MHz, CDCl3) δ8.12 (d, J = 6.4Hz, 1H), 7.23-7.18 (m, 1H), 7.15 (d, 1H), 4.01 (s, 3H), 2.29 (s, 3H).

[0203] Step 2: 3-Methyl-4-nitropyridine nitroxide-2-carboxylic acid methyl ester

[0204] Cpd-3-2 (3.85 g, 22.9 mmol) was added to concentrated sulfuric acid (25 mL) at 0 °C. Fuming nitric acid (6.7 mL) was slowly added dropwise to the solution over 20 minutes. The system was slowly heated to 95 °C and stirred for 5 hours. After cooling, the mixture was poured into ice water (100 mL) and neutralized with saturated sodium carbonate solution. It was extracted with dichloromethane (100 mL × 2), the organic layers were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was slurried (DCM:PE = 1:10) and purified to give compound Cpd-3-3 (2.74 g, yield 56.2%, pale yellow solid). 1 H NMR (400MHz, CDCl3) δ8.15 (d, J = 7.2Hz, 1H), 8.01 (d, J = 7.2Hz, 1H), 4.05 (s, 3H), 2.54 (s, 3H).

[0205] Step 3: Methyl 3-methyl-4-aminopyridine-2-carboxylic acid ester

[0206] At room temperature, a system of Cpd-3-3 (2.74 g, 12.9 mmol) and palladium on carbon (680 mg, 10%) in methanol (100 mL) was stirred for 18 hours under a hydrogen atmosphere (1 atm). After filtration and concentration, compound Cpd-3-4 (2.12 g, crude product, pale yellow solid) was obtained. 1 H NMR (400MHz, CDCl3) δ8.15 (d, J = 5.3Hz, 1H), 6.62 (d, J = 5.4Hz, 1H), 4.31 (s, 2H), 3.94 (s, 3H), 2.29 (s, 3H).

[0207] Step 4: Methyl 3-methyl-4-amino-5-bromopyridine-2-carboxylic acid ester

[0208] N-bromosuccinimide (2.38 g, 13.4 mmol) was added to a solution of Cpd-3-4 (2.02 g, 12.2 mmol) and ammonium acetate (100 mg, 1.34 mmol) in acetonitrile (40 mL). The mixture was stirred at room temperature for 3 hours, then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 20:1) to give compound Cpd-3-5 (3.03 g, 91.5% yield, white solid). 1 H NMR (400MHz, CDCl3) δ8.37(s,1H),4.79(s,2H),3.95(s,3H),2.39(s,3H).

[0209] Step 5: Methyl 3-methyl-4-iodo-5-bromopyridine-2-carboxylic acid ester

[0210] A mixture of cuprous iodide (1.55 g, 8.16 mmol) and acetonitrile (20 mL) was heated to 50 °C, and then tert-butyl nitrite (2.43 mL, 20.4 mmol) was added. The system was stirred at 50 °C for 0.5 h, and then a solution of Cpd-3-5 (1.00 g, 4.08 mmol) in acetonitrile (30 mL) was added. The reaction mixture was heated to 80 °C and stirred for 10 min. After cooling, the reaction mixture was quenched with saturated sodium bicarbonate aqueous solution (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-3-6 (810 mg, yield 55.8%, white solid). 1 H NMR (400MHz, CDCl3) δ8.55(s,1H),3.99(s,3H),2.76(s,3H).

[0211] Step Six: Methyl 5-bromo-3-methyl-4-phenylpyridine-2-carboxylic acid

[0212] Under argon protection, a mixture of 1,4-dioxane (50 mL) and water (12.5 mL) of Cpd-3-6 (2.17 g, 6.10 mmol), phenylboronic acid (740 mg, 6.10 mmol), cesium carbonate (3.97 g, 12.2 mmol), and Pd(dppf)Cl2 (500 mg, 610 μmol) was stirred at 50 °C for 6 hours. After cooling, the mixture was extracted with ethyl acetate (50 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-3-7 (1.42 g, yield 76.1%, white solid). LCMS calc. for C 14 H 13 BrNO2[M+H] + :m / z=306.0 / 308.0; Found:306.2 / 308.2.

[0213] Step 7: Methyl 5-bromo-3-(bromomethyl)-4-phenylpyridine-2-carboxylic acid

[0214] A mixture of Cpd-3-7 (1.42 g, 4.64 mmol), N-bromosuccinimide (990 mg, 5.57 mmol), and azobisisobutyronitrile (380 mg, 2.32 mmol) in carbon tetrachloride (50 mL) was stirred at 90 °C for 3 hours. After cooling and filtration, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-3-8 (2.03 g, 88.7% yield, pale yellow solid). LCMS calc. for C 14 H 12 Br2NO2[M+H] + :m / z=383.9 / 385.9 / 387.9; Found:383.9 / 385.9 / 387.8.

[0215] Step 8: (S)-3-bromo-6-(2-((tert-butyldiphenylsilyl)oxo)-1-cyclopropylethyl)-4-phenyl-5,6-dihydro-7H-pyrrole[3,4-b]pyridin-7-one

[0216] A methanol (40 mL) solution of Cpd-3-8 (2.00 g, 4.05 mmol), INT-1-7 (1.38 g, 4.05 mmol), and triethylamine (1.76 mL, 12.2 mmol) was stirred at 70 °C for 3 hours. The system was concentrated by cooling and purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-3-9 (1.76 g, yield 71.0%, white solid). LCMS calc. for C 34 H 36 BrN2O2Si[M+H] + :m / z=611.2 / 613.2; Found:611.2 / 613.2.

[0217] Step Nine: (S)-6-(2-((tert-butyldiphenylsilyl)oxo)-1-cyclopropylethyl)-3-(3-fluoro-3-methylazacyclobutane-1-yl)-4-phenyl-5,6-dihydro-7H-pyrrole[3,4-b]pyridin-7-one

[0218] In a sealed tube, a solution of Cpd-3-9 (1.48 g, 2.42 mmol), 3-fluoro-3-methylazacyclobutane hydrochloride (460 mg, 3.63 mmol), Xantphos-Pd G2 (430 mg, 480 μmol), and cesium carbonate (2.37 g, 7.26 mmol) in 1,4-dioxane (30 mL) was stirred at 100 °C for 16 hours under argon protection. After cooling and filtration, the filter cake was washed with dichloromethane (50 mL), the filtrates were combined and concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EA 100%) to give compound Cpd-3-10 (1.54 g, 92.4% yield, white solid). LCMS calc. for C 38 H 43 FN3O2Si[M+H] + :m / z = 620.3; Found: 620.0

[0219] Step 10: (S)-6-(1-cyclopropyl-2-hydroxyethyl)-3-(3-fluoro-3-methylazacyclobutane-1-yl)-4-phenyl-5,6-dihydro-7H-pyrrole[3,4-b]pyridin-7-one

[0220] At room temperature, tetrabutylammonium fluoride (7.26 mL, 7.26 mmol, 1 M tetrahydrofuran solution) was added to a solution of Cpd-3-10 (1.50 g, 2.42 mmol) in tetrahydrofuran (25 mL), and stirred for 1 hour. The system was concentrated under reduced pressure, diluted with ethyl acetate (50 mL), washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (EA:EtOH = 8:1) to give compound Cpd-3-11 (480 mg, yield 52.0%, pale yellow solid). LCMS calc. for C 22 H 25 FN3O2[M+H] + :m / z=382.2;Found:381.7.

[0221] Step 11: (S)-2-cyclopropyl-2-(3-(3-fluoro-3-methylazacyclobutane-1-yl)-7-oxo-4-phenyl-5,7-dihydro-6H-pyrrolo[3,4-b]pyridin-6-yl)acetaldehyde

[0222] A solution of Cpd-3-11 (480 mg, 1.26 mmol) and Dietrich Martin oxidant (800 mg, 1.89 mmol) in dichloromethane (25 mL) was stirred at room temperature for 1 hour. The system was diluted with dichloromethane (50 mL) and quenched with saturated sodium bicarbonate solution (25 mL) and saturated sodium thiosulfate solution (25 mL). The mixture was stirred at room temperature for 10 minutes, separated, and the organic phase was washed with saturated sodium bicarbonate solution (25 mL), saturated sodium thiosulfate solution (25 mL), and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound Cpd-3-12 (500 mg, crude, white solid), which was used directly in the next reaction. LCMS calc. for C 22 H 23 FN3O2[M+H] + :m / z=380.2;Found:380.2.

[0223] Step 12: (S,E)-6-(1-cyclopropyl-3-(methanesulfonyl)allyl)-3-(3-fluoro-3-methylazacyclobutane-1-yl)-4-phenyl-5,6-dihydro-7H-pyrrole[3,4-b]pyridin-7-one

[0224] At room temperature, a solution of INT-1-4 (437 mg, 1.90 mmol), lithium chloride (80.4 mg, 1.14 mmol), and DBU (280 μL, 1.90 mmol) in anhydrous acetonitrile (15 mL) was stirred for 15 minutes, then cooled to 0 °C. A solution of Cpd-3-12 (500 mg, 1.26 nmol) in acetonitrile (10 mL) was added dropwise. The resulting system was stirred at 25 °C for 30 minutes. The reactants were quenched with ice water (50 mL) and extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated brine (50 mL), dried over sodium sulfate pentahydrate, filtered, and concentrated. The residue was purified sequentially by silica gel column chromatography (EA:EtOH = 10:1), prep-HPLC (FA / MeCN), and SFC to give compound Cpd-3 (91.6 mg, yield 15.5%, white solid). LCMS calc. for C 24 H 27 FN3O3S[M+H] + m / z = 456.2; Found: 455.9; 1 H NMR (400MHz, DMSO-d6) δ8.07(s,1H),7.55-7.43(m,5H),6.87-6.76(m,2H),4.38-4.23(m,2H),4.13(dd,J=10.0,3.1Hz,1H),3.7 9-3.59(m,4H),3.00(s,3H),1.46(d,J=22.0Hz,3H),1.34-1.24(m,1H),0.70-0.58(m,1H),0.56-0.45(m,2H),0.31-0.18(m,1H).

[0225] Step Thirteen: (S)-2-((tert-butyldiphenylsilyl)oxy)-1-cyclopropylethane-1-amine

[0226] To a solution of INT-1-6 (5.15 g, 37.6 mmol) in dichloromethane (60 mL), tert-butyldiphenylchlorosilane (10.3 g, 39.5 mmol) and imidazole (6.09 g, 89.5 mmol) were added, and the mixture was stirred at 40 °C for 12 hours. The reaction solution was diluted with dichloromethane (100 mL), washed with saturated brine (40 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM:MeOH = 10:1) to give compound INT-1-7 (8.25 g, yield 64.5%, pale yellow oil). LCMS calc. for C 21 H 30 NOSi[M+H] + :m / z=340.2;Found:340.2.

[0227] Example 4: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(cyclopropyldifluoromethyl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0228] Step 1: 5-Bromo-4-methylnicotinic acid methyl ester

[0229] Under ice bath conditions, sulfoxide (21.5 g, 181 mmol) was slowly added dropwise to a methanol (130 mL) solution of Cpd-4-1 (13.0 g, 60.2 mmol). The system was stirred at 80 °C for 12 hours. After cooling and concentration, the crude product was dissolved in ethyl acetate (100 mL), washed with saturated sodium bicarbonate solution (50 mL × 2) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-4-2 (11.7 g, yield 84.5%, white solid). LCMS calc. for C8H9BrNO2[M+H] + :m / z=230.0 / 232.0; Found:230.0 / 232.1.

[0230] Step 2: 5-Bromo-6-(cyclopropylcarbonyl)-4-methylnicotinic acid methyl ester

[0231] At room temperature, silver nitrate (8.64 g, 50.7 mmol) and potassium persulfate (68.7 g, 254 mmol) were added to a mixture of Cpd-4-2 (11.7 g, 50.9 mmol) and 2-cyclopropyl-2-carbonylacetic acid (11.6 g, 102 mmol) in acetonitrile (110 mL) and water (30 mL). The system was stirred at 50 °C for 12 hours under nitrogen protection. After cooling and filtration, the filtrate was extracted with ethyl acetate (100 mL × 2), the combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-4-3 (12.7 g, yield 83.7%, white solid). LCMS calc. for C 12 H 13 BrNO3[M+H] + :m / z=298.0 / 300.0; Found:298.1 / 300.1.

[0232] Step 3: 5-Bromo-6-(2-cyclopropyl-1,3-dithiapentan-2-yl)-4-methylnicotinic acid methyl ester

[0233] Under ice bath conditions, boron trifluoride diethyl ether (3.82 g, 26.9 mmol) was added dropwise to a methanol (60 mL) solution of Cpd-4-3 (6.69 g, 22.4 mmol) and 1,2-ethanedithiol (3.80 g, 40.4 mmol), and the system was stirred at 40 °C for 16 hours. After cooling, the system was poured into a saturated sodium carbonate solution (100 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-4-4 (3.27 g, yield 38.9%, white solid). LCMS calc. for C 14 H 17 BrNO2S2[M+H] + :m / z=374.0 / 376.0; Found:374.2 / 376.2.

[0234] Step 4: 5-Bromo-6-(cyclopropyldifluoromethyl)-4-methylnicotinic acid methyl ester

[0235] At -60°C, pyridine hydrofluoric acid (4.66 g, 47.0 mmol) and cooled Cpd-4-4 (4.40 g, 11.7 mmol) in dichloromethane (50 mL) were added dropwise to a solution of N-iodosuccinimide (5.29 g, 23.5 mmol) in dichloromethane (50 mL). The reaction mixture was brought to room temperature over 1 hour, then poured into a saturated sodium carbonate solution (50 mL) and extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-4-5 (588 mg, yield 15.6%, white solid). LCMS calc. for C 12 H 13 BrF₂NO₂[M+H] + :m / z=320.0 / 322.0; Found:320.1 / 322.2.

[0236] Step 5: Methyl 6-(cyclopropyldifluoromethyl)-4-methyl-5-phenylnicotinic acid ester

[0237] At room temperature, Pd(dppf)Cl2 (182 mg, 223 μmol) was added to a mixture of 1,4-dioxane (10 mL) of Cpd-4-5 (714 mg, 2.23 mmol), phenylboronic acid (407 mg, 3.35 mmol), and cesium fluoride (1.01 g, 6.69 mmol) in 1,4-dioxane (2 mL) and water. The reaction mixture was stirred at 80 °C for 12 hours under nitrogen protection. After cooling, the mixture was diluted with ethyl acetate (100 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-4-6 (586 mg, yield 82.8%, yellow oil). LCMS calc. for C 18 H 18 F2NO2[M+H] + :m / z=318.1;Found:318.2.

[0238] Step Six: 4-(bromomethyl)-6-(cyclopropyldifluoromethyl)-5-phenylnicotinic acid methyl ester

[0239] At room temperature, N-bromosuccinimide (493 mg, 2.77 mmol) and azobisisobutyronitrile (30.2 mg, 185 μmol) were added to a solution of Cpd-4-6 (586 mg, 1.85 mmol) in carbon tetrachloride (10 mL), and the mixture was stirred at 80 °C for 16 hours. After cooling and filtration, the filtrate was concentrated, and the residue was purified by reverse (Prep-C18) to give compound Cpd-4-7 (357 mg, yield 48.7%, yellow oil). LCMS calc. for C 18 H 17 BrF₂NO₂[M+H] + :m / z=396.0 / 398.0; Found:396.2 / 398.2.

[0240] Step 7: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(cyclopropyldifluoromethyl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0241] At room temperature, N,N-diisopropylethylamine (546 mg, 4.23 mmol) was added to a solution of Cpd-4-7 (335 mg, 845 μmol) and INT-1 (441 mg, 1.27 mmol) in tetrahydrofuran (10 mL), and the mixture was stirred at 40 °C for 16 hours. The mixture was concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 1:1) and prep-HPLC (FA / MeCN) to give compound Cpd-4 (210 mg, yield 54.6%, white solid). LCMS calc. for C 24 H 25 F2N2O3S[M+H] + m / z = 459.2; Found: 458.8; 1 H NMR (400MHz, DMSO-d6) δ9.04 (s, 1H), 7.53-7.35 (m, 5H), 6.93-6.74 (m, 2H), 4.41-4.24 (m, 2H), 4.14 (dd, J= 9.7,4.5Hz,1H),2.99(s,3H),2.01-1.83(m,1H),1.33-1.22(m,1H),0.67-0.45(m,7H),0.31-0.22(m,1H).

[0242] Example 5: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(3-methylazacyclobutane-1-yl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0243] Step 1: 4-Methyl-6-(3-methylazacyclobutan-1-yl)nicotinic acid methyl ester

[0244] A mixture of Cpd-2-2 (3.00 g, 16.2 mmol), 3-methylazacyclobutane (1.73 g, 24.3 mmol), and N,N-diisopropylethylamine (4.19 g, 32.4 mmol) in N,N-dimethylformamide (20 mL) was stirred at 60 °C for 3 hours. After cooling, the mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL × 3). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-5-1 (2.00 g, yield 53.1%, yellow solid). LCMS calc. for C 12 H 17 N₂O₂[M+H] + :m / z=221.1;Found:221.2.

[0245] Step 2: methyl 5-bromo-4-methyl-6-(3-methylazonobutan-1-yl)nicotinic acid

[0246] A solution of Cpd-5-1 (2.00 g, 9.10 mmol) and N-bromosuccinimide (1.78 g, 10.0 mmol) in N,N-dimethylformamide (20 mL) was stirred at 25 °C for 15 hours. The system was diluted with water (10 mL) and extracted with ethyl acetate (20 mL × 3). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 3:1) to give compound Cpd-5-2 (1.16 g, yield 47.2%, yellow solid). LCMS calc. for C 12 H 16 BrN2O2[M+H] + :m / z=299.0 / 301.0; Found:299.1 / 301.1.

[0247] Step 3: 4-Methyl-6-(3-methylazacyclobutan-1-yl)-5-phenylnicotinic acid methyl ester

[0248] Under argon protection, a mixture of Cpd-5-2 (1.00 g, 3.34 mmol), phenylboronic acid (611 mg, 5.01 mmol), potassium carbonate (924 mg, 6.68 mmol), and Pd(dppf)Cl2 (242 mg, 334 μmol) in dioxane (10 mL) and water (1 mL) was stirred at 80 °C for 3 hours. After cooling, the mixture was diluted with ethyl acetate (100 mL) and washed with water (50 mL) and saturated brine (50 mL). The solution was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 2:1) to give compound Cpd-5-3 (500 mg, yield 40.4%, yellow solid). LCMS calc. for C 18 H 21 N₂O₂[M+H] + :m / z=297.2;Found:297.2.

[0249] Step 4: 4-(chloromethyl)-6-(3-methylazonobutan-1-yl)-5-phenylnicotinic acid methyl ester

[0250] Under argon protection, a mixture of Cpd-5-3 (500 mg, 1.67 mmol), N-chlorosuccinimide (248 mg, 1.86 mmol), and azobisisobutyronitrile (55.4 mg, 337 μmol) in carbon tetrachloride (10 mL) was stirred at 40 °C for 16 hours. After cooling and filtration, the filtrate was concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 1:1) to give compound Cpd-5-4 (300 mg, yield 51.1%, yellow solid). LCMS calc. for C 18 H 20 ClN2O2[M+H] + :m / z=331.1;Found:331.1.

[0251] Step 5: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(3-methylazacyclobutan-1-yl)-7-phenyl-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0252] A methanol (10 mL) solution of Cpd-5-4 (300 mg, 910 μmol), INT-1 (238 mg, 1.13 mmol), and triethylamine (367 mg, 3.63 mmol) was stirred at 60 °C for 15 hours. The mixture was then cooled and concentrated, and the residue was purified by Prep-HPLC (FA / MeCN) to give compound Cpd-5 (30.5 mg, yield 6.81%, white solid). LCMS calc. for C 24 H 28 N3O3S[M+H] + :m / z=438.2;Found:438.1; 1 H NMR(400MHz, DMSO-d6)δ8.45(d,J=3.9Hz,1H),7.51-7.40(m,5H),6.83-6.78(m,2H),4.30(q,J=17.7Hz,2H),4.13-4.04(m,1H),3.7 1(t,J=8.3Hz,2H),3.23-3.18(m,2H),2.99(s,3H),1.31-1.19(m,1H),1.05(d,J=6.8Hz,3H),0.69-0.40(m,3H),0.30-0.17(m,1H).

[0253] Example 6: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(cyclopropyldifluoromethyl)-7-(phenyl-d5)-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0254] Step 1: 3,5-Dibromo-2-(cyclopropyldifluoromethyl)-4-methylpyridine

[0255] At room temperature, 2-cyclopropyl-2,2-difluoroacetic acid (10.9 g, 79.7 mmol) was added to a mixture of compound Cpd-6-1 (10.0 g, 39.9 mmol), potassium persulfate (53.9 g, 199 mmol), silver nitrate (6.77 g, 39.9 mmol), acetonitrile (100 mL), and water (100 mL). The mixture was stirred at 50 °C for 16 hours. After cooling to room temperature, the mixture was extracted with ethyl acetate (100 mL × 2). The organic layers were combined, washed with saturated brine (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 10:1) to give compound Cpd-6-2 (4.60 g, yield 33.9%, yellow oil). LCMS calc. for C 10 H9Br2FN[MF] + :m / z=319.9 / 321.9 / 323.9; Found:319.9 / 321.9 / 323.9.

[0256] Step 2: Methyl 5-bromo-6-(cyclopropyldifluoromethyl)-4-methylnicotinic acid

[0257] A mixture of Cpd-6-2 (4.60 g, 13.5 mmol), triethylamine (10.9 g, 108 mmol), and Pd(dppf)Cl2 (1.10 g, 1.35 mmol) in methanol (80 mL) was stirred at 70 °C for 16 hours under a carbon monoxide atmosphere. After cooling and concentration, the residue was purified by silica gel column chromatography (PE:EA = 5:1) to give compound Cpd-6-3 (2.80 g, yield 64.8%, yellow oil). LCMS calc. for C 12 H 13 BrF₂NO₂[M+H] + :m / z=320.0 / 322.0; Found:320.0 / 322.0.

[0258] Step 3: Methyl 6-(cyclopropyl difluoromethyl)-4-methyl-5-(phenyl-d5)nicotinic acid

[0259] Under an argon atmosphere, a mixture of Cpd-6-3 (2.80 g, 8.75 mmol), (phenyl-d5)boric acid (1.33 g, 10.5 mmol), Pd(dppf)Cl2 (710 mg, 870 μmol), potassium phosphate (3.71 g, 17.5 mmol), toluene (30 mL), and water (6 mL) was stirred at 100 °C for 4 hours. After cooling to room temperature, the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic layers were washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-6-4 (2.70 g, 95.8% yield, yellow oil). LCMS calc. for C 18 H 13 D5F2NO2[M+H] + :m / z=323.2;Found:323.3.

[0260] Step 4: 4-(bromomethyl)-6-(cyclopropyldifluoromethyl)-5-(phenyl-d5)nicotinic acid methyl ester

[0261] At room temperature, azobisisobutyronitrile (80.0 mg, 470 μmol) was added to a solution of compound Cpd-6-4 (1.00 g, 3.10 mmol) and N-bromosuccinimide (830 mg, 4.65 mmol) in carbon tetrachloride (15 mL), and the mixture was stirred at 80 °C for 16 hours. After cooling and concentration, the residue was purified by silica gel column chromatography (PE:EA = 4:1) to give compound Cpd-6-5 (1.00 g, 80.3% yield, yellow oil). LCMS calc. for C 18 H 12 BrD5F2NO2[M+H] + :m / z=401.1 / 403.1; Found:401.0 / 403.0.

[0262] Step 5: (S,E)-2-(1-cyclopropyl-3-(methanesulfonyl)allyl)-6-(cyclopropyldifluoromethyl)-7-(phenyl-d5)-1,2-dihydro-3H-pyrrole[3,4-c]pyridin-3-one

[0263] At room temperature, Cpd-6-5 (1.00 g, 2.49 mmol) was added to a solution of INT-1 (690 mg, 3.24 mmol) and N,N-diisopropylethylamine (1.61 mg, 12.5 mmol) in tetrahydrofuran (15 mL). The mixture was stirred at 80 °C for 16 hours. After cooling to room temperature, the mixture was extracted with ethyl acetate (20 mL × 2). The organic layers were combined, washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE:EA = 1:2) to give compound Cpd-6 (520 mg, yield 45.0%, yellow solid). LCMS calc. for C 24 H 20 D5F2N2O3S[M+H] + :m / z=464.2;Found:464.0; 1 H NMR(400MHz, DMSO-d6)δ9.04(s,1H),6.88(dd,J=15.6,0.8Hz,1H),6.79(dd,J=15.6,4.8Hz,1H),4.40-4.32(m,2H),4.15- 4.12(m,1H),2.99(s,3H),1.99-1.84(m,1H),1.31-1.23(m,1H),0.64-0.59(m,3H),0.55-0.46(m,4H),0.28-0.22(m,1H).

[0264] Bioevaluation

[0265] I. Biochemical Activity Assay: DNA Unwinding Fluorescence Assay of WRN Helicase

[0266] A fluorescence-based (FI) WRN helicase assay was used to detect the inhibitory effect of compounds on WRN helicases. Recombinant His-labeled WRN (517-1238) used in the assay was expressed in insect cells and purified to 90% purity using a Ni-NTA column. Fluorescently labeled ssDNA was synthesized by GenScript, with the specific sequences 5'-TTT TTT TTT TTT TTT TTT TTT TTT TTT TTT TTT CGT ACC CGA TGT GTT CGT TC-3' (SEQ ID NO:1) and 5'-GAA CGA ACA CAT CGG GTA CG TTT TTT TTT TTT TTT TTT TTT TTT TTT TTT TTT-3' (SEQ ID NO:2). After annealing, dsDNA substrates were formed for the assay. Capturing strand ssDNA was synthesized by GenScript, with the specific sequence 5'-GAACGAACACATCGGGTACG-3' (SEQ ID NO:3).

[0267] The compound was serially diluted using DMSO in a 384PP Plate compound dilution plate. 0.15 μL of the compound was transferred to a 384 reaction microplate (Corning 4514) using an Echo, ensuring a final DMSO concentration of 1% (double replicate). 5 μL of WRN (517-1238) enzyme solution (containing 0.2 mM ATP) was added to each well of the 384 reaction microplate, centrifuged for 1 minute, and incubated at 25°C for 240 minutes. Wells containing 1% DMSO and enzyme served as high controls, and wells containing the same concentration of DMSO and buffer served as low controls. 5 μL of a mixture containing dsDNA and Capture strand ssDNA solution was added to each well, centrifuged for 1 minute, and then 5 μL of ATP solution was added. The plate was incubated at 25°C for 30 minutes. (Final reagent concentrations: 5 nM WRN (517-1238), 100 nM dsDNA, 1 μM Capture strand ssDNA, 4 mM ATP). Fluorescence intensity signals were read on a BMG (CLARIO Star Plusacu) microplate reader (Ex: 620nm and Em: 685nm). The inhibition percentage of the compound-treated wells was normalized between the high and low controls (%inhibition = (FI reading high control - FI reading compound) / (FI reading high control - FI reading low control) * 100). Four-parameter IC50 curves were then fitted and analyzed using XLfit 5.5.0. 50It is the compound concentration corresponding to 50% inhibition on the curve.

[0268] Table 2. Results of DNA unwinding fluorescence assay using WRN helicase

[0269] Note 1: Ref. 1 is selected from WO2024010782A1.

[0270] Note 2: A refers to IC 50 >1.0μM, B refers to 0.5μM <IC 50 ≤1.0μM, where C refers to 0.1μM <IC 50 ≤0.5μM, where D refers to 0.05μM <IC 50 ≤0.1μM, E refers to IC 50 ≤0.05μM.

[0271] The above data show that the compounds in the embodiments of the present invention have good inhibitory activity against WRN helicase function.

[0272] II. Cell viability assay: Proliferation inhibition activity assay of SW48 cells

[0273] The antiproliferative effect of the compound on human colon cancer cell lines was evaluated using ATP quantification, and this activity correlated well with the number of viable cells. Cell line SW48 (ATCC, CCL-231) was cultured in DMEM (Gibco, 11965118) medium supplemented with 10% (v / v) fetal bovine serum (Gibco, 10099141C). Cell lines were cultured according to the standard instructions of the American Center for Type Culture Collection. Cell lines were identified by short tandem repeat profiles.

[0274] In the proliferation assay, 4000 SW48 cells were seeded in a 96-well clear flat-bottom black TC-treated plate (Corning, 3603) with 200 μL of medium per well and cultured overnight in the medium to recover. The compounds were added at increasing concentration gradients, and DMSO treatment was used as a positive control. After 5 days of treatment, the microplate was equilibrated to room temperature for 30 minutes, and the number of live cells was measured by adding CellTiter-Glo reagent (Promega, G7573). The chemiluminescence readings were measured on a Spark (Tecan). The % growth rate (%G) of each well was calculated using the formula: when T > V0, %G = ((T - V0) / V0)) * 100; when T < V0, %G = ((T - V0) / (V - V0)))*100; V0 is the cell viability at the time of adding the compound treatment; V and T represent the cell viabilities of the vehicle control treatment and the compound treatment at the final detection, respectively. 100%, 0%, and -100% represent no cell proliferation inhibition effect, complete cell proliferation arrest, and complete cell killing, respectively. According to the growth rate %G, the half-maximal growth inhibitory concentration GI was then calculated by fitting to a four-parameter nonlinear curve using the program GraphPad 10 software. 50 .

[0275] Table 3. Results of the proliferation inhibition activity assay of SW48 cells

[0276] Note 1: Ref. 1 is selected from WO2024010782A1,

[0277] Note 2: A refers to GI 50 > 5 μM, B refers to 0.5 μM < GI 50 ≤ 5 μM, C refers to 0.05 μM < GI 50 ≤ 0.5 μM, D refers to 0.01 μM < GI 50 ≤ 0.05 μM, E refers to 0.05 μM < GI 50 ≤ 0.01 μM, F refers to GI 50 ≤ 0.005 μM.

[0278] The above data show that the compounds of the embodiments of the present invention have good inhibitory activity against the proliferation of SW48 cells.

[0279] III. Pharmacokinetic study in mice

[0280] 1. Experimental purpose: To test the pharmacokinetics of the compound in CD-1 mice.

[0281] 2. Experimental instruments and materials:

[0282] Animals: CD-1 mice (male)

[0283] Table 4 Instruments:

[0284] Table 5 Reagents:

[0285] Solvent:

[0286] 10% NMP+15% Kolliphor EL+1% Tween-20+74% (20% HP-β-CD)

[0287] 3. Experimental methods:

[0288] Six male mice were used. They were fasted the evening before administration but allowed free access to water. During the experiment, the mice had free access to food and water. The drugs were administered intravenously or by gavage. The animals' condition was observed and any abnormal behaviors were recorded after administration. Blood samples were collected from the orbital sinus at 0.0833, 0.25, 1, 2, 4, 8, and 24 hours after intravenous injection; and at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after gavage. 20 μL of plasma was collected, and 250 μL of acetonitrile (containing dexamethasone as an internal standard) was added to precipitate proteins. The mixture was centrifuged at 4000 rpm for 20 minutes at 4°C. 180 μL of the supernatant was mixed with 180 μL of an aqueous solution containing 0.1% formic acid in a 96-well plate. Another 10 μL of the sample was then used for LC-MS / MS analysis.

[0289] 4. Data processing methods and results:

[0290] A standard curve was established using the internal standard method, with the theoretical standard curve concentration plotted on the x-axis and the peak area ratio (peak area of ​​the tested compound / peak area of ​​the internal standard) plotted on the y-axis. A linear regression method (weighted factor 1 / X²) was used, and R0 was calculated. 2 >0.9900. Calculations for unknown samples were performed using a standard curve. Pharmacokinetic parameters were calculated using a non-compartmental analysis model in WinNonlin 8.2 software and presented in the report. Parameters include Clint, C... max And AUC, etc.

[0291] Table 6: Pharmacokinetic parameters of the compounds in CD-1 mice

[0292] The data above show that the compounds in the embodiments of the present invention have a lower clearance rate and a higher oral exposure in mice, and are superior to the reference compounds.

[0293] IV. Pharmacokinetic Studies in Rats

[0294] 1. Experimental objective: To test the pharmacokinetics of the compound in SD rats.

[0295] 2. Experimental instruments and materials:

[0296] Animal: Male SD rat

[0297] Table 7 Instruments:

[0298] Table 8 Reagents:

[0299] Solvent:

[0300] Intravenous injection: 40% HP-β-CD aqueous solution; Gavage: 0.2% HPC + 0.5% Tween 80 aqueous solution.

[0301] 3. Experimental methods:

[0302] Six male rats were used. They were fasted the evening before administration but allowed free access to water. During the experiment, the rats were given free access to food and water, and were administered the drug via intravenous injection or gavage. The animals' condition was observed and any abnormal behaviors were recorded after administration. Blood was collected via jugular vein at 0.0833, 0.25, 1, 2, 4, 8, and 24 hours after intravenous injection; and via gavage at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after gavage. 50 μL of plasma was collected, and 500 μL of acetonitrile (containing dexamethasone as an internal standard) was added to precipitate proteins. The mixture was centrifuged at 4000 rpm for 20 minutes at 4°C. 300 μL of the supernatant was mixed with 300 μL of Watson's aqueous solution in a 96-well plate, and then 5 μL of the sample was used for LC-MS / MS analysis.

[0303] 4. Data processing methods and results:

[0304] A standard curve was established using the internal standard method, with the theoretical standard curve concentration plotted on the x-axis and the peak area ratio (peak area of ​​the tested compound / peak area of ​​the internal standard) plotted on the y-axis. A linear regression method (weighted factor 1 / X²) was used, and R0 was calculated. 2 >0.9900. Calculations for unknown samples were performed using a standard curve. Pharmacokinetic parameters were calculated using a non-compartmental analysis model in WinNonlin 8.2 software and presented in the report. Parameters include Clint, C... max And AUC, etc.

[0305] Table 9: Pharmacokinetic parameters of the compounds in SD rats

[0306] The data above show that the compounds in the embodiments of the present invention have a low clearance rate and a high oral exposure in rats, and are significantly superior to the reference compounds.

[0307] V. In vivo efficacy experiment of SW48 human colon cancer xenograft mouse model

[0308] 1. Experimental objective: To test the in vivo pharmacodynamic activity of the compound in a BALB / c Nude female mouse SW48 human subcutaneous xenograft model of colon cancer.

[0309] 2. Experimental materials:

[0310] Animals: 5-6 week old female BALB / c Nude mice;

[0311] Table 10 Solvent Information:

[0312] Solvent: 5% DMSO + 10% Solutol HS15 + 85% PBS buffer (0.01M) Table 11 Main reagent and consumable information:

[0313] Table 12 Main Instrument Information:

[0314] 3. Experimental Design:

[0315] Cell culture: SW48 cells were cultured in a medium containing L-15 + 10% FBS + 1% P / S; the cells were cultured in an incubator at 37°C without carbon dioxide.

[0316] Tumor modeling and grouping: SW48 cells were collected by digestion, counted, and resuspended in a mixture of equal proportions of culture medium and Matrigel (Matrigel, Corning Biocoat 356234) to achieve a concentration of 1*10^7 cells / mL. Cells were then transplanted subcutaneously into the right anterior shoulder and back of mice at a concentration of 1*10^6 cells / mouse (100 μL). Following inoculation, tumor growth was monitored until the average tumor volume reached 137.21 ± 2.59 mm. 3 Students were randomly assigned to groups. The day on which medication began was designated Day 0.

[0317] 4. Experimental Design: The grouping and administration information for this pharmacodynamic experiment are shown in Table 13. Mice in group G1 were administered the solvent control (5% DMSO + 10% Solutol HS15 + 85% PBS buffer (0.01M)) orally once daily. Mice in groups G2, G3, and G4 were administered Ref. 1 orally once daily at doses of 5 mg / kg, 10 mg / kg, and 20 mg / kg, respectively. Mice in groups G5, G6, G7, and G8 were administered Cpd-6 orally once daily at doses of 2.5 mg / kg, 5 mg / kg, 10 mg / kg, and 20 mg / kg, respectively. The administration period was 21 days.

[0318] Table 13. In vivo pharmacodynamic design of SW48 xenograft mouse model

[0319] 5. Experimental observation and result judgment:

[0320] Experimental observation: After cell inoculation, the mice's condition, mobility, food intake, and water consumption were closely monitored daily. Any deaths, abnormal behavior, or disease symptoms were immediately reported and recorded to determine appropriate intervention. After grouping the mice, their body weight was measured twice weekly. Tumor volume was measured twice weekly using calipers. The formula for calculating tumor volume was TV = 0.5 * length * width * width.

[0321] Data processing: Tumor volume inhibition rate (TGI) (%): The formula for calculating TGI (%) is as follows: TGI (%) = [1 – (mean tumor volume at the end of treatment - mean tumor volume at the beginning of treatment) / (mean tumor volume at the end of treatment in the solvent control group - mean tumor volume at the beginning of treatment in the solvent control group)] × 100%.

[0322] Statistical analysis: Tumor volume and animal body weight are expressed as Mean ± SEM (mean standard error). Tumor volumes were statistically compared between different groups. All statistical analyses were performed in GraphPad Prism 9.0. ANOVA was used to analyze significant differences in tumor volume between different groups, with the Vehicle group as a control. p ≥ 0.05 was considered no significant difference, p < 0.05 was considered a significant difference, and p < 0.001 was considered a highly significant difference.

[0323] 6. Experimental Results:

[0324] Figure 1 shows the tumor growth curves of the control group and each experimental group in the pharmacodynamic evaluation experiment of the SW48 human colon cancer xenograft model in mice, and Figure 2 shows the changes in relative body weight of mice. Table 14 shows the average tumor volume, tumor volume inhibition rate (TGI%), and comparison results of each group at the end of the experiment.

[0325] At the experimental endpoint (day 20 after grouping), the mean tumor volume of mice in the solvent control group (G1) was 1532.25 mm. 3 The mean tumor volume in mice treated with 5 mg / kg Ref.1 (G2) was 315.19 mm. 3 (TGI 87.24%, p < 0.001), the mean tumor volume in mice in the 10 mg / kg Ref.1 treatment group (G3) was 104.06 mm. 3(TGI 102.38%, p < 0.001), the mean tumor volume in mice in the 20 mg / kg Ref.1 treatment group (G4) was 75.97 mm. 3 (TGI 104.39%, p < 0.001), the mean tumor volume in the 2.5 mg / kg Cpd-6 treatment group (G5) was 816.18 mm. 3 (TGI was 51.33%, p < 0.001), the mean tumor volume in mice treated with 5 mg / kg Cpd-6 (G6) was 137.91 mm. 3 (TGI 99.95%, p < 0.001), the mean tumor volume in mice in the 10 mg / kg Cpd-6 treatment group (G7) was 85.74 mm. 3 (TGI was 103.69%, p < 0.001), the mean tumor volume in the 20 mg / kg Cpd-6 treatment group (G8) was 78.99 mm. 3 (TGI: 104.18%, p: <0.001). These results indicate that Cpd-6 and Ref.1 exhibited good inhibitory activity against SW48 xenograft growth in mice at all tested doses, showing a dose-related relationship; and Cpd-6 showed superior antitumor activity compared to Ref.1. Mice in all groups were in good condition with relatively stable body weight, and the test substances Cpd-6 and Ref.1 had no effect on mouse body weight.

[0326] Table 14. In vivo efficacy results of the SW48 xenograft mouse model.

Claims

1. A compound of formula (I), its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof: in, X 1 is N or CR 0 , R 0 selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, Ci-C6alkyl, haloCi-C6alkyl, deuterated Ci-C6alkyl, Ci-C6alkoxy, haloCi-C6alkoxy, deuterated Ci-C6alkoxy; X 2 is N or CR 3 , R 3 Selected from hydrogen, halogen, cyano, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, unsubstituted or substituted C3-C6 cycloalkyl, unsubstituted or substituted 3-8 membered heterocyclic groups, -NR 3a R 3b ;R 3 The substitution mentioned herein refers to substitution by one or more substituents selected from group A; group A substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR A1 R A2 -CONR A1 R A2 -NR A1 C(O)R A2 ; Y represents O, CR 9 R 10 or R 9 R 10 R 11 R 12 R 13 R 14 Each group is independently selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, C1-C6 alkyl, halo-C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, and deuterated C1-C6 alkoxy. R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted C3-C8 cycloalkyl groups, unsubstituted or substituted C6-C10 aryl groups, unsubstituted or substituted 3-8 membered heterocyclic groups, unsubstituted or substituted 5-10 membered heteroaryl groups, and unsubstituted or substituted C3-C8 cycloalkenyl groups; R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ; R 2 For -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 -OR 17 ;R 16 and R 17 Selected from unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl, unsubstituted or substituted C2-C6 ynyl, unsubstituted or substituted C3-C8 cycloalkyl, unsubstituted or substituted C6-C10 aryl, unsubstituted or substituted 3-8 membered heterocyclic, and unsubstituted or substituted 5-10 membered heteroaryl; R 16 and R 17 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR D1 R D2 -CONR D1 R D2 -NR D1 C(O)R D2 ; R 4 It is hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C3-C8 cycloalkyl, or unsubstituted or substituted 5-6 membered heterocyclic group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR E1 R E2 -CONR E1 R E2 -NR E1 C(O)R E2 ; R 5 It is hydrogen or deuterium; R 6 It is hydrogen or C1-C6 alkyl; R 7 It is hydrogen, C1-C6 alkyl, or halogen; R 8 It is hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C3-C8 cycloalkyl, or unsubstituted or substituted 4-6 membered heterocyclic group; R 8 The substitution mentioned herein refers to substitution by one or more substituents selected from group F; group F substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR F1 R F2 -CONR F1 R F2 -NR F1 C(O)R F2 ; R 2a R 2b R 3a R 3b R A1 R A2 R B1 R B2 R C1 R C2 R D1 R D2 R E1 R E2 R F1 R F2 Each is independently selected from hydrogen, C1-C6 alkyl, halo-C1-C6 alkyl, and C3-C6 cycloalkyl; With S atoms and R 7 Connected separately express It can be a geometric isomer of (Z) or (E); This indicates the connection point of the group with other locations; The *-marked C atom can be a chiral carbon atom or a non-chiral carbon atom; when the *-marked C atom is a chiral carbon atom, the configuration of the chiral carbon atom is S configuration or R configuration; The heteroatoms in the 3-8 membered heterocyclic group, 5-6 membered heterocyclic group, 4-6 membered heterocyclic group, and 5-10 membered heteroaryl group are independently selected from 1, 2, 3, or 4 of N, O, and S.

2. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It satisfies one or more of the following conditions: (1)X 1 For N or CR 0 , (2)R 0 Selected from hydrogen, deuterium, halogen, carboxyl, hydroxyl, cyano, amino, C1-C6 alkyl, halo-C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, and deuterated C1-C6 alkoxy. (3)X 2 For N or CR 3 , (4)R 3 Selected from hydrogen, halogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C1-C6 alkoxy, -NR 3a R 3b ;R 3 The substitution mentioned herein refers to substitution by one or more substituents selected from group A; group A substituents include: deuterium, halogen, C1-C6 alkoxy, halo-C1-C6 alkoxy, -NR A1 R A2 -CONR A1 R A2 -NR A1 C(O)R A2 ;R 3a R 3b R A1 R A2 Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; (5)R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted cyclobutyl, unsubstituted or substituted cyclopentyl, unsubstituted or substituted cyclohexyl, unsubstituted or substituted phenyl, unsubstituted or substituted... No substitution or substitution No substitution or substitution No substitution or substitution R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halogenated C1-C6 alkyl, and C3-C6 cycloalkyl; R 1s It is a C1-C6 alkyl or a halo-C1-C6 alkyl; Preferably, R 1 For R 15 ;R 15 It is an unsubstituted or substituted C6-C10 aryl group; R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; the substituents in group B are deuterium; (6)R 2 Selected from -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; Preferably, R 2 -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups and unsubstituted or substituted 3-8 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups, or C3-C6 cycloalkyl groups; (7)R 4 It is an unsubstituted or substituted C1-C6 alkyl group, an unsubstituted or substituted C3-C6 cycloalkyl group, or an unsubstituted or substituted 4-6 membered heterocyclic group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR E1 R E2 -CONR E1 R E2 -NR E1 C(O)R E2 ;R E1 R E2 Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; Preferably, R 4 It is an unsubstituted C3-C8 cycloalkyl group; (8)R 5 For hydrogen, R 6 For hydrogen, R 7 It is hydrogen; (9)R 8 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 8 The substitution mentioned herein refers to substitution by one or more substituents selected from group F; group F substituents include: deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, amino, -N(CH3)2; Preferably, R 8 It is an unsubstituted C1-C6 alkyl group; (10) Y is CR 9 R 10 or and (11)R 9 R 10 R 11 R 12 R 13 R 14 Each is independently selected from hydrogen.

3. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It satisfies one or more of the following conditions: (1)X 1 For N or CH; (2)X 2 For N or CR 3 R 3 Selected from hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, deuterated C1-C6 alkyl, C1-C6 alkoxy, amino, -NH-(C1-C6 alkyl), -NH-(C3-C6 cycloalkyl); (3)R 1 The phenyl group is either unsubstituted or substituted; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, and C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halo-C1-C6 alkyl, and C3-C6 cycloalkyl; (4)R 2 For -NR 2a R 2b or -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; (5)R 4 It is an unsubstituted or substituted C1-C6 alkyl group, an unsubstituted or substituted cyclopropyl group, or an unsubstituted or substituted cyclobutyl group; R 4 The substitution mentioned herein refers to substitution by one or more substituents selected from group E; group E substituents include: halogen, hydroxyl, cyano, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkoxy; (6)R 8 Selected from methyl, ethyl, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, And (7)Y is -CH2- or -(CH2)2-.

4. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It satisfies one or more of the following conditions: (1)X 1 For N or CH; (2)X 2 For N or CR 3 R 3 Selected from hydrogen, amino, -NH- (C1-C6 alkyl), -NH- (C3-C6 cycloalkyl); (3)R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3; and (4)R 2 Selected from methyl, ethyl, tert-butyl, 5. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It satisfies one or more of the following conditions: (1)X 1 For N or CH; (2)X 2 For N or CR 3 R 3 Selected from hydrogen, amino, -NHCH3, -NH-cyclopropyl; and (3)R 1 It is a phenyl group.

6. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It satisfies one or more of the following conditions: (1)X 1 For N or CH; (2)X 2 For N or CH; (3) Y is -CH2-; (4)R 1 for (5)R 2 for (6)R 4 for and (7)R 8 It is a methyl group.

7. The compound of formula (I) as claimed in any one of claims 1-6, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) has the structure shown in formula (II): in, X 1 X 2 Y, R 1 R 2 and R 8 The definition is as described in any one of claims 1-6.

8. The compound of formula (I) as claimed in any one of claims 1-6, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) has a structure as shown in formula (III), (IV), (V) or (VI): in, Y, R 1 R 2 and R 8 The definition is as described in any one of claims 1-6.

9. The compound of formula (I) as claimed in claim 8, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, It is either Option 1 or Option 2 as follows: Option 1: In equations (III), (IV), (V), and (VI), Y is O, CH2, or -(CH2)2; R 1 For -OR 15 or R 15 ;R 15 Selected from unsubstituted or substituted cyclobutyl, unsubstituted or substituted cyclopentyl, unsubstituted or substituted cyclohexyl, unsubstituted or substituted phenyl, unsubstituted or substituted... No substitution or substitution No substitution or substitution No substitution or substitution R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; group B substituents include: oxo (=O), deuterium, halogen, hydroxyl, cyano, carboxyl, unsubstituted or substituted by one or more substituents selected from group C, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, -NR B1 R B2 -CONR B1 R B2 -NR B1 C(O)R B2 Group C substituents include: deuterium, halogen, C1-C6 alkoxy, halogenated C1-C6 alkoxy, C3-C6 cycloalkyl, cyano, hydroxyl, carboxyl, amino, and -NR. C1 R C2 -CONR C1 R C2 -NR C1 C(O)R C2 ;R B1 R B2 R C1 R C2 Each is independently selected from hydrogen, C1-C6 alkyl, halogenated C1-C6 alkyl, and C3-C6 cycloalkyl; R 1s It is a C1-C6 alkyl or a halo-C1-C6 alkyl; R 2 Selected from -NR 2a R 2b -CONR 2a R 2b -NR 2a C(O)R 2b -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; R 8 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted 4-6 membered heterocyclic groups; R 8 The substitution mentioned herein refers to substitution by one or more substituents selected from group F; group F substituents include: deuterium, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, amino, -N(CH3)2; Option 2: In equations (III), (IV), (V), and (VI), Y is CH2 or -(CH2)2-; R 1 For R 15 ;R 15 It is an unsubstituted or substituted C6-C10 aryl group; R 15 The substitution mentioned herein refers to substitution by one or more substituents selected from group B; the substituents in group B are deuterium; R 2 -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups and unsubstituted or substituted 3-8 membered heterocyclic groups; R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups, or C3-C6 cycloalkyl groups; R 8 It is an unsubstituted C1-C6 alkyl group.

10. The compound of formula (I) as claimed in claim 8, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, Y is O, CH2, or -(CH2)2; R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3; R 2 For -NR 2a R 2b or -R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, unsubstituted or substituted cyclopropyl groups, unsubstituted or substituted cyclobutyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl; R 2a R 2b Each is independently selected from hydrogen or C1-C6 alkyl or C3-C6 cycloalkyl; R 8 Selected from methyl, ethyl, -CH2N(CH3)2, -CH2CH2N(CH3)2, -CH2CH2CH2N(CH3)2, Preferably, in formulas (III), (IV), (V), and (VI), Y is O, CH2, or -(CH2)2; R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being replaced by one or more substituents selected from group B; the substituents in group B are: deuterium; R 2 For R 16 ;R 16 Selected from unsubstituted or substituted C1-C6 alkyl groups, and unsubstituted or substituted... R 16 The substitution mentioned herein refers to substitution by one or more substituents selected from group D; group D substituents are: halogens, C1-C6 alkyl groups, or C3-C6 cycloalkyl groups; R 8 Selected from methyl.

11. The compound of formula (I) as claimed in claim 8, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, Y is CH2; R 1 It is an unsubstituted or substituted phenyl group; the substitution refers to being substituted by one or more substituents selected from group B; group B substituents include: deuterium, halogen, hydroxyl, cyano, carboxyl, C1-C6 alkyl, deuterated C1-C6 alkyl, halo-C1-C6 alkyl, C1-C6 alkoxy-substituted C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkoxy, C1-C6 alkoxy, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 cycloalkoxy, NH2, -CONH2, -NHC(O)CH3; R 2 Selected from methyl, ethyl, tert-butyl, R 8 It is a methyl group.

12. The compound of formula (I) as claimed in claim 1, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug, or pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) is selected from the following compounds:

13. A pharmaceutical composition comprising a compound of formula (I) as claimed in any one of claims 1-12, an isotopic label thereof, an enantiomer, a diastereomer, a transisomer, a solvate, a prodrug or a pharmaceutically acceptable salt thereof, and a pharmaceutical excipient.

14. The use of a compound of formula (I) as described in any one of claims 1-12, an isotopic label thereof, an enantiomer, a diastereomer, a transisomer, a solvate, a prodrug or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 13, in the preparation of a WRN inhibitor or a medicament; said medicament being a medicament for treating and / or preventing disease by inhibiting WRN; or said medicament being a medicament for treating and / or preventing cancer; Preferably, the cancer includes solid tumors with high microsatellite instability or mismatch repair defects; more preferably, the cancer includes endometrial cancer, colorectal cancer, gastric cancer, and ovarian cancer.

15. A method for treating and / or preventing cancer, characterized in that, The patient is given a therapeutically effective amount of the compound of formula (I) as described in any one of claims 1-13, its isotopic label, enantiomer, diastereomer, transisomer, solvate, prodrug or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 13.

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