Thickened polycyclic compound and preparation method and pharmaceutical application thereof

Abstract

Relates to a fused polycyclic compound and a preparation method and pharmaceutical application thereof. The invention provides a compound with a specific general formula, a preparation method of the compound, a pharmaceutical composition comprising the compound, and a pharmaceutical application of the compound in treating related diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and relates to a fused polycyclic compound and a preparation method thereof, a pharmaceutical composition containing the compound and the medical use thereof. Background Art

[0002] Cells in any organism possess multiple DNA repair mechanisms to maintain genome stability and integrity, which is essential for survival. DNA mismatch repair (MMR) is a highly conserved pathway that plays a key role in maintaining genome stability by correcting errors that occur during DNA replication, recombination, and repair. (Pecina-Slaus, N., Kafka, A., Salamon, I. & Bukovac, A. Mismatch Repair Pathway, Genome Stability and Cancer. Front Mol. Biosci. 7, 122 (2020)). Defects in the MMR mechanism can cause genome hypermutation and instability, manifested as high-frequency insertions and deletions of short repetitive DNA sequences (microsatellites) throughout the genome, a phenomenon known as microsatellite instability (MSI).

[0003] (Kim, TM, Laird, PW & Park, PJ The landscape of microsatellite instability in colorectal and endometrial cancer genomes. Cell 155, 858–868 (2013), Vernole, P. et al. Common fragile sites in colon cancer cell lines: role of mismatch repair, RAD51 and poly (ADP-ribose) polymerase-1. Mutat. Res 712, 40–48 (2011)). Microsatellites are generated by DNA polymerase slippage during replication or mismatch repair, and are prone to misplacement and framework shift mutations, which can be recognized and corrected by the MMR process in normal cells. However, in cancer cells that lack normal mismatch repair mechanisms, errors that occur during replication or repair accumulate and the genomic mutation rate increases (Lower, SS, McGurk, MP, Clark, AG, et al, Satellite DNA evolution: old ideas, new approaches. Curr. Opin. Genet. Dev., 2018; 49: 70-78). High-frequency deletions and insertions of microsatellites can lead to MSI, resulting in 10-30% of ovarian, colon, gastric, and endometrial cancers (Aaltonen, LA et al. Clues to the pathogenesis of familial colorectal cancer, Science 260, 812-816 (1993), Bonneville R et al., Landscape of Microsatellite Instability Across 39 Cancer Types. JCO Precis Oncol. 1: PO. 17.00073 (2017)).Compared with MSS cancers, patients with high MSI (MSI-H) cancers show better overall prognosis and reduced metastatic potential, higher tumor mutation burden and immunogenicity (Kang, S. et al. The significance of microsatellite instability in colorectal cancer after controlling for clinicopathological factors. Med. (Baltim.) 97, e0019 (2018)), and MSI-H tumors tend to be resistant to immunotherapy and chemotherapy (Le, DT et al. Phase II Open-Label Study of Pembrolizumab in Treatment-Refractory, Microsatellite Instability-High / Mismatch Repair-Deficient Metastatic Colorectal Cancer: KEYNOTE-164. J. Clin. Oncol. 38, 11–19 (2020), Overman, MJ et al. Nivolumab in patients with metastatic DNA mismatch repair-deficient or microsatellite instability-high colorectal cancer (CheckMate 142): an open-label, multicentre, phase2 study. Lancet Oncol. 18, 1182–1191 (2017), Fuca, G. et al. Ascites and resistance to immune checkpoint inhibition in dMMR / MSI-H metastatic colorectal and gastric cancers. J. Immunother. Cancer 10, 4001 (2022)).

[0004] Recently, multiple independent large-scale functional genomic screens using more than 300 human cancer cell lines found that Werner syndrome RecQ helicase (WRN) is selectively required for MSI-H cell survival (Behan, FM et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511–516 (2019), McDonald ER et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3): 577-592 (2017), Chan, EM et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551–556 (2019)). WRN is one of the five RecQ-like helicases in humans. It is a multifunctional enzyme with helicase and exonuclease activities and plays an important role in multiple pathways.

[0005] DNA repair and maintenance of genome integrity, including DNA replication, transcription, DNA repair and telomere maintenance (Bohr, VARising from the RecQ-age: the role of human RecQ helicases in genomemaintenance. Trends Biochem Sci. 33, 609–620 (2008), Singh, DK, Ahn, B. & Bohr, V A Roles of RECQ helicases in recombination based DNA repair, genomic stability and aging. Biogerontology 10, 235–252 (2009), Rossi, ML, Ghosh, AK & Bohr, V A Roles of Werner syndrome protein in protection of genome integrity. DNA Repair (Amst.) 9, 331–344 (2010)).

[0006] Loss of WRN leads to various defects in cellular and genomic structure, including cell cycle arrest, DNA breakage, mitotic defects, chromosome fragmentation, and apoptosis in MSI (but not MSS cell lines) cells (Behan, FM et al. Prioritization of cancer therapeutic targets using CRISPR-Cas9 screens. Nature 568, 511–516 (2019), McDonald ER et al., Project DRIVE: A Compendium of Cancer Dependencies and Synthetic Lethal Relationships Uncovered by Large-Scale, Deep RNAi Screening. Cell 170(3): 577-592 (2017), Chan, EM et al. WRN helicase is a synthetic lethal target in microsatellite unstable cancers. Nature 568, 551–556 (2019)). In addition, studies on loss-of-function mutations and deletions of WRN further identified that the helicase function of WRN is indispensable for the survival of MSI cells (Lieb, S. et al. Werner syndrome helicase is a selective vulnerability of microsatellite instability-high tumor cells. Elife 8, e43333 (2019)). These results clearly indicate that targeting WRN, especially the helicase domain, may be a promising strategy for treating high MSI cancers. Summary of the invention

[0007] In one aspect, the present invention provides a compound of formula (I)-(XI), (I') or its tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof:

[0008]

[0009]

[0010] in,

[0011] Ring A and Ring B are each independently selected from an aryl group or a heteroaryl group;

[0012] Y1 is CH, NH, N, O or S;

[0013] Y2 is CH, NH, N, O or S;

[0014] Y3 is CH, NH, N, O or S;

[0015] X1 and X2 are each independently CH or N;

[0016] M1, M2, M5, M6, each independently represents CH, NH, N, O or S;

[0017] M3 and M4 are each independently -NR a ,-CR a R b ;

[0018] R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0019] Y2 is CH, NH, N, O or S;

[0020] Y3 is CH, NH, N, O or S;

[0021] is a single bond or a double bond, provided that Not at the same time a double bond;

[0022] R1, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0023] Two R2 together with the atoms to which they are bound form a C3-C10 cycloalkyl or a C3-C10 heterocyclic group, wherein the cycloalkyl or heterocyclic group is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, carbonyl, and oxo;

[0024] R5 and R6 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0025] or, R5 and R6 together with the atoms to which they are attached form a C3-C10 cycloalkyl group, a C3-C10 heterocyclyl group, a C6-C14 aryl group, or a C5-C12 heteroaryl group containing 1 to 4 heteroatoms selected from O, S and N;

[0026] R7 is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0027] Alternatively, two R7 together with the atoms to which they are attached form a C3-C10 cycloalkyl or C3-C10 heterocyclyl; L1 is a bond, O, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl, -C3-C10 cycloalkyl-CONH-, -C1-C6 alkyl-C3-C10 cycloalkyl-CONH, -C1-C6 haloalkyl-NH-, -C3-C10-heterocyclyl-NH-alkyl, -C3-C10 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C10 heteroaryl-C1-C6 alkyl, -S(O)2-NH- and -C1-C6 alkyl-S(O)2-NH-;

[0028] L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(R c )-;

[0029] R c is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl;

[0030] Preferably, ring A is a C6-C14 aryl group or a C5-C12 heteroaryl group containing 1 to 4 heteroatoms selected from O, S and N;

[0031] R 10 is a cycloalkenyl radical, wherein the cycloalkenyl radical is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms; or a heterocyclyl radical, wherein the heterocyclyl radical is a 5- or 6-membered fully saturated or partially unsaturated radical comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, and wherein the heterocyclyl radical is unbridged or bridged and the bridge is 1 or 2 carbon atoms; or a heteroaryl radical, wherein the heteroaryl radical is a 5- or 6-membered fully unsaturated monocyclic radical comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1; or a phenyl radical; wherein the cycloalkenyl radical, heterocyclyl radical, heteroaryl radical, phenyl radical is unsubstituted or substituted by one or more substituents selected from halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0032] Or, R10 is C2-C4 alkenyl, optionally substituted by R4 or -O-R4;

[0033] m is 0, 1, 2, 3 or 4;

[0034] p is 0, 1, 2 or 3;

[0035] q is 0, 1, 2, or 3;

[0036] r is 0, 1, 2, or 3;

[0037] s is 2 or 3;

[0038] t is 0, 1, 2, or 3;

[0039] w is 0, 1, 2, or 3; and

[0040] n is 0, 1, 2 or 3.

[0041] In one embodiment, the compound has the formula (XII)-(XIII), (XIX)-(XXI), (XIIB)

[0042]

[0043] R8 is a C3-C12 fused cycloalkyl, a C2-C10 fused heterocyclyl, a spiroheterocyclyl or a spirocycloalkyl, wherein each of the fused cycloalkyl, the fused heterocyclyl, the spiroheterocyclyl or the spirocycloalkyl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, and oxo;

[0044] R9 is a C3-C10 cycloalkyl group or a C3-C10 heterocyclic group, wherein the cycloalkyl group and the heterocyclic group are each optionally substituted by one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, oxo, C1-6 haloalkyl, and C3-5 cycloalkyl; L6 is a C3-C10 cycloalkyl group or a C2-C10 heterocyclic group, wherein the cycloalkyl group and the heterocyclic group are each optionally substituted by one or more substituents selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, oxo, C1-6 haloalkyl, and C3-5 cycloalkyl;

[0045] Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R4, m, p, q, r, t are defined as described above.

[0046] In one embodiment, R1, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 hydroxyalkyl and C2-C6 alkenyl;

[0047] In a preferred embodiment, R1, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C3 alkyl and -CF3.

[0048] In one embodiment, the compound has the formula (XXII) or (XXIII),

[0049]

[0050] Each X is independently selected from F, Cl, Br, I;

[0051] Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R7, m, n, p, r, t, w are as defined above.

[0052] In one embodiment, the compound has the formula (XXIV)-(XXVI),

[0053]

[0054] Ring C is C6-10 aryl or C2-C9 heteroaryl;

[0055] R 10 is a cycloalkenyl group, wherein the cycloalkenyl group is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms; or a heterocyclyl group, wherein the heterocyclyl group is a 5- or 6-membered fully saturated or partially unsaturated group, including ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, wherein the heterocyclyl group is non-bridged or bridged, and the bridge is 1 or 2 carbon atoms; or a heteroaryl group, wherein the heteroaryl group is a 5- or 6-membered fully unsaturated monocyclic group, including ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1; or a phenyl group; wherein the cycloalkenyl group, heterocyclyl group, heteroaryl group, phenyl group is unsubstituted or selected from halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0056] Or, R 10 is C2-C4 alkenyl, optionally substituted by R4 or -O-R4;

[0057] u is 2,3,4;

[0058] Two R's 11connected to form a C1-C3 alkylene bridge or a -CH=CH- bridge or a -CH=CH-CH2- bridge or a C1-C3 heteroalkylene bridge, wherein the C1-C3 alkylene bridge or the -CH=CH-CH- bridge or the C1-C3 heteroalkylene bridge is optionally substituted by one or more halogens, and the heteroatom is selected from one heteroatom among N, O and S; the other R 11 are independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0059] R 12 are independently selected from hydrogen, deuterium, halogen, amino, cyano, carbonyl, hydroxy, alkoxy, alkoxy, haloalkyl and hydroxyalkyl, -O-haloalkyl;

[0060] R 13 is selected from halogen, amino, cyano, oxo, hydroxy, alkoxy, haloalkyl and hydroxyalkyl, -O-haloalkyl;

[0061] v is 1,2,3,4.

[0062] In one embodiment, the compound has the formula (XXVII)-(XXXIII), (XXXV)-(XXXVII), (XXVII-A), (XXVII-B), (XXXVI-A),

[0063]

[0064] Among them, R 14 Selected from the group consisting of: Each X is independently selected from F, Cl, Br, I;

[0065] R 10a is a 5-6 membered heteroaryl group, wherein the 6 membered heteroaryl group has 2 or more heteroatoms, wherein the 5-6 membered heteroaryl group is unsubstituted or substituted by one of a plurality of substituents selected from halogen, amino, cyano, carbonyl, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0066] Ring D is a condensed C6-C12 aryl group, a condensed C1-C12 heteroaryl group, or a condensed heterocyclic group;

[0067] Ring E is C6-C12 aryl or C1-C12 heteroaryl;

[0068] Ring F is C6-C10 aryl or C2-C9 heteroaryl;

[0069] L3 is selected from O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(Rc)-;

[0070] L7 is selected from -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(Rc)-;

[0071] R 15 is a heterocyclyl or spiroheterocyclyl or spirocycloalkyl group, wherein the heterocyclyl, spiroheterocyclyl and spirocycloalkyl group are unsubstituted or substituted by one of a plurality of substituents selected from halogen, amino, cyano, carbonyl, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0072] R 16 A group selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl, -O-haloalkyl.

[0073] In one embodiment, ring A is a C6-C14 aryl group or a C5-C12 heteroaryl group containing 1-4 heteroatoms selected from O, S and N.

[0074] In one embodiment, the compound has the formula (XXXVIII)-(XXXIX),

[0075]

[0076] R 10b Selected from the group consisting of:

[0077]

[0078] Among them, the R 4a is selected from the group consisting of halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0079] R 4b is selected from the group consisting of amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl;

[0080] R 4c is selected from the group consisting of halogen, amino, cyano, carbonyl, hydroxy, alkyl, haloalkyl and hydroxyalkyl;

[0081] m1 is 1 or 2;

[0082] m2 is 0, 1 or 2;

[0083] R1, R3, R4, R7, Y1, Y2, Y3, r, t, w, p, q, L2, X1, X2, M1, and M2 are as defined in the above general formula.

[0084] In an embodiment, two R2 and the atoms to which they are attached form a C3-C10 bridged cycloalkyl or a C3-C10 bridged heterocyclyl;

[0085] In a preferred embodiment represent

[0086] In one embodiment represent Represents the site of attachment to L2.

[0087] In one embodiment, R5 and R6 are independently selected from hydrogen, deuterium, halogen, amino, cyano, carbonyl, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0088] Alternatively, R5 and R6 together with the atoms to which they are attached are in the form

[0089] In an embodiment, R7 is selected from hydrogen, deuterium, halogen, amino, cyano, carbonyl, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl; or, two of R7 and the atoms to which they are attached form a bridged C3-C10 cycloalkyl or a bridged C3-C10 heterocyclyl, preferably, represent

[0090] In an embodiment, R a and R b are independently selected from hydrogen, deuterium, halogen, amino, cyano, carbonyl, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0091] In an embodiment, L1 is a bond, O, C1-C3 alkyl, C2-C3 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, -C3-C6 cycloalkyl-CONH-, -C1-C3 alkyl-C3-C6 cycloalkyl-CONH, -C1-C3 haloalkyl-NH-, -C3-C6-heterocyclo-NH-alkyl, -C3-C6 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C10 heteroaryl-C1-C6 alkyl, -S(O)2-NH-, and -C1-C6 alkyl-S(O)2-NH-;

[0092] In embodiments, L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, and -CON(R c )-;

[0093] R c is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl;

[0094] In one embodiment, Ring A is In one embodiment, Ring B is selected from the group consisting of phenyl, pyridyl, pyridinyl or pyrimidinyl.

[0095] In one embodiment, yes In one embodiment, yes In one embodiment, yes In one embodiment, in Formula (XIIB), yes or yes In one embodiment, R8 is selected from the following groups:

[0096] In one embodiment, R9 is R 10 It is C 1-5 Alkyl, C 3-5 Cycloalkyl.

[0097] In one embodiment, L6 is or L6-L2 is In one embodiment, in the general formula (XXII), X connected to the phenyl group is Cl, and / or CH n X 3-n It's CF3.

[0098] In one embodiment, in the general formula (XXIII), CH n X 3-n It's CF3.

[0099] In one embodiment, is selected from the following groups:

[0100] yes

[0101] yes

[0102] * represents the site of attachment to L2.

[0103] In one embodiment, in the general formula (XXVII), yes and / or, R 10 yes

[0104] In one embodiment, in the general formula (XXIX), R 10a yes Preferably, In one embodiment, in Formula (XXX), Ring D is

[0105] and Ring D is optionally substituted with (R12)q;

[0106] In one embodiment, in Formula (XXXI), Ring E is and ring E is optionally (R7) w replace;

[0107] In one embodiment, in the general formula (XXXII), L3 is a C1-C6 alkyl group, preferably, -CH(CH3)-; In one embodiment, in the general formula (XXXIII), R1 is H; L3 is a C1-C6 alkyl group, preferably, -CH2-, -CH(CH3)-; R 15 is a spiroheterocyclic group or

[0108] In the general formula (XXXVI), ring C is or thick and C1-C 12 heteroaryl; and Ring C is optionally substituted with (R12)q;

[0109] Or, in general formula (XXXVI), yes In general formula (XXXVII), yes and / or yes Preferably,

[0110] * represents the site of attachment to L2.

[0111] In the general formula (XXXVI-A), yes In one embodiment, in each of the above general formulas, R 10 Preferably, it is a 5-6-membered heterocyclic ring having an unsaturated double bond, containing 1-2 heteroatoms selected from O, N and S, and optionally substituted by deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 10 Preferably, R 10 More preferably, In one embodiment, R 10b yes Preferably,

[0112] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of any general formula of the present invention, or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt, solvate or prodrug thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0113] On the other hand, the present invention relates to a method for treating a disease mediated by WRN, comprising administering to a subject in need thereof an effective amount of a compound of any general formula of the present invention or a tautomer, cis or trans isomer, mesomorph, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt solvate thereof, or a prodrug thereof, or a pharmaceutical composition comprising the same.

[0114] Preferably, the disease mediated by WRN is selected from cancers characterized by high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), more preferably selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer and ovarian cancer.

[0115] On the other hand, the present invention relates to a method for treating cancer, comprising administering to a subject in need thereof an effective amount of a compound of the general formula of the present invention, or a tautomer, a cis isomer or a trans isomer, a mesomer, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt solvate thereof, or a prodrug thereof, or a pharmaceutical composition comprising the same.

[0116] In a preferred embodiment, the cancer is selected from cancers characterized by high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), more preferably selected from colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, renal cancer and ovarian cancer. DETAILED DESCRIPTION OF THE INVENTION

[0118] The following are definitions of terms used in this application. Any term not defined herein is given the normal meaning of the term as understood by those skilled in the art.

[0119] "Alkyl" refers to a saturated aliphatic hydrocarbon group including C1-C20 straight chain and branched groups. Preferably, the alkyl group is an alkyl group having 1 to 12, sometimes preferably 1 to 6, and sometimes more preferably 1 to 3 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, 2-methylhexyl, n-heptyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl , 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, N-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl and branched chain isomers thereof. More preferably, the alkyl group is a lower alkyl group having 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment, preferably the substituent is one or more substituents independently selected from the group consisting of alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonyl, alkylamino, thiol, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyl, hetero, cycloalkylthio, heteroalkylthio, heteroalkylthio and carbonyl.

[0120] "Alkenyl" refers to an alkyl group having at least two carbon atoms and at least one carbon-carbon double bond as defined above, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc., preferably C2-20 alkenyl, more preferably C2-12 alkenyl, and most preferably C2-6 alkenyl. The alkenyl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, independently selected from alkyl, halogen, alkoxy, alkenyl, alkynyl, alkylsulfonyl, alkylamino, thiol, hydroxyl, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, hetero, cycloalkylthio, heterothio, heteroalkylthio and carbonyl.

[0121] "Alkynyl" refers to an alkyl group having at least two carbon atoms and at least one carbon-carbon triple bond as defined above, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc., preferably C2-20 alkynyl, more preferably C2-12 alkynyl, most preferably C2-6 alkynyl. The alkynyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkyl sulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, aryl, heteroaryl, cycloalkyloxy, heteroalkyloxy, cycloalkylthio and heteroalkylthio.

[0122] "Alkylene" refers to a saturated straight or branched aliphatic hydrocarbon group having 2 residues, derived by removing two hydrogen atoms from the same carbon atom or two different carbon atoms of the parent alkane. The straight or branched group contains 1-20 carbon atoms, preferably 1-12 carbon atoms, and more preferably 1-6 carbon atoms. Non-limiting examples of alkylene include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butyl (-CH2CH2CH2CH2-), etc. The alkylene group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterooxyalkoxy, cycloalkylthio and heteroalkylthio.

[0123] "Alkenylene" refers to an alkylene group having at least two carbon atoms and at least one carbon-carbon double bond as defined above, preferably C2-20 alkenylene, more preferably C2-12 alkenylene, and most preferably C2-6 alkenylene. Non-limiting examples of alkenylene include, but are not limited to, -CH=CH-, -CH=CHCH2-, -CH=CHCH2CH2-, -CH2CH=CHCH2-, etc. The alkenylene group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, and these groups are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterooxyalkoxy, cycloalkylthio, and heteroalkylthio.

[0124] "Alkynylidene" refers to an alkynyl group having at least two carbon atoms and at least one carbon-carbon triple bond as defined above, preferably C2-20 alkynylidene, more preferably C2-12 alkynylidene, most preferably C2-6 alkynylidene. Non-limiting examples of alkenylidene include but are not limited to -CH≡CH-, -CH≡CHCH2-, -CH≡CHCH2CH2-, -CH2CH≡CHCH2-, etc. Alkynylidene may be substituted or unsubstituted. When substituted, the substituent is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, and these groups are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkyl sulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterooxyalkoxy, cycloalkylthio and heteroalkylthio.

[0125] "Cycloalkyl" refers to a saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon radical having 3-20 carbon atoms, preferably 3-12 carbon atoms, more preferably 3-10 carbon atoms, most preferably 3-8 carbon atoms or 3-6 carbon atoms. Representative examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyls include cycloalkyls having spiral rings, fused rings, or bridged rings.

[0126] "Spirocycloalkyl" refers to a 5-20-membered polycyclic group whose rings are connected by a common carbon atom (called a spiro atom), wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, the spirocycloalkyl is 6 to 14 members, more preferably 6 to 12 members, and most preferably 7 to 8 members. According to the number of common spiro atoms, the spirocycloalkyl is divided into a monospirocycloalkyl, a dispirocycloalkyl or a polyspirocycloalkyl, preferably a monospirocycloalkyl or a dispirocycloalkyl, more preferably a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or a 5-membered monospirocycloalkyl. Representative examples of spirocycloalkyl include, but are not limited to, the following substituents:

[0127]

[0128] "Fused cycloalkyl" refers to a 5-20-membered polycyclic hydrocarbon group, in which each ring in the system shares a pair of adjacent carbon atoms with another ring, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, the fused cycloalkyl is 6 to 14 members, more preferably 6 to 12 members, more preferably 7 to 10 members, and most preferably 0.7 to 8 members. Depending on the number of rings, the fused cycloalkyl is divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused cycloalkyl, preferably a bicyclic or tricyclic fused cycloalkyl, more preferably a 5-membered / 5-membered, or a 5-membered / 6-membered bicyclic fused cycloalkyl. Representative examples of fused cycloalkyls include, but are not limited to, the following substituents:

[0129] "Bridged cycloalkyl" refers to a 5-20-membered polycyclic hydrocarbon group in which every two rings in the system share two unconnected carbon atoms. The ring may have one or more double bonds, but there is no completely conjugated π electron system. Preferably, the bridged cycloalkyl is 5 to 14 members, more preferably 5 to 10 members, and most preferably 5 to 8 members. Depending on the number of rings, the bridged cycloalkyl is divided into a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, and preferably refers to a bicyclic, tricyclic or tetracyclic bridged cycloalkyl, more preferably a bicyclic or tricyclic bridged cycloalkyl. Representative examples of bridged cycloalkyl include, but are not limited to, the following substituents:

[0130] The cycloalkyl group may be fused to the ring of an aryl, heteroaryl or heterocycloalkyl group, wherein the ring bound to the parent structure is a cycloalkyl group. Representative examples include, but are not limited to, indaneacetic acid, tetralin, benzocycloheptyl, etc. The cycloalkyl group is optionally substituted or unsubstituted. When a substituent group is substituted, the substituent group is preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, and the substituent group is independently selected from alkyl, halogen, alkoxy, alkenyl, alkynyl, alkyl sulfonyl, alkylamino, thiol, hydroxyl, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, assorted groups, cycloalkylthio, assorted thio groups and oxo groups. Representative examples include, but are not limited to, the following substituent groups: and

[0131] "Heterocyclyl" refers to a 3-20 membered saturated and / or partially unsaturated monocyclic or polycyclic hydrocarbon radical having one or more, sometimes preferably one to five, sometimes more preferably one to three heteroatoms selected from the group consisting of N, O and S(O)m (wherein m is 0, 1 or 2), but excluding -OO-, -OS- or -SS- in the ring, the remaining ring atoms being C. Preferably, the heterocyclyl is 3-12 membered with 1 to 4 heteroatoms; more preferably 3-10 membered with 1 to 3 heteroatoms; more preferably 3-7 membered with 1 to 3 heteroatoms; most preferably 5-6 membered with 1 to 2 heteroatoms. Representative examples of monocyclic heterocyclyls include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, sulfomorpholinyl, homopiperazinyl, and the like. Polycyclic heterocyclyls include heterocyclyls having spiral rings, fused rings, or bridged rings.

[0132] "Spiro heterocyclyl" refers to a 5-20-membered polycyclic heterocyclyl, whose rings are connected by a common carbon atom (called a spiro atom), wherein the ring has one or more, sometimes preferably one to five, sometimes more preferably one to three heteroatoms selected from the group consisting of N, O and S(O)m (wherein m is 0, 1 or 2) as ring atoms, and the remaining ring atoms are C, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, the spiro heterocyclyl is 6 to 14 members, more preferably 7 to 10 members, and most preferably 7 to 8 members. According to the common number of spiro atoms, the spiro heterocycle is divided into a monospiro heterocyclyl, a dispiro heterocyclyl or a polyspiro heterocyclyl, preferably a monospiro heterocyclyl or a dispiro heterocyclyl, more preferably a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or a 5-membered or a 5-membered or a 5-membered / 6-membered monospiro heterocycle. Representative examples of spiro heterocyclyls include, but are not limited to, the following substituents:

[0133]

[0134] "Fused heterocyclyl" refers to a 5-20 membered polycyclic heterocyclyl, wherein each ring in the system shares a pair of adjacent carbon atoms with another ring, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, and wherein the ring has one or more, sometimes preferably one to five, sometimes more preferably one to three, heteroatoms selected from the group consisting of N, O and S(O)p (wherein p is 0, 1 or 2) as ring atoms, and the remaining ring atoms are C. Preferably, the fused heterocyclyl is 6 to 14 members, more preferably 6 to 12 members, more preferably 7 to 10 members, and most preferably 7 to 8 members. According to the number of rings, the fused heterocyclyl is divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclyl, preferably a bicyclic or tricyclic fused heterocyclyl, more preferably a 5-membered / 5-membered, or a 5-membered / 6-membered bicyclic fused heterocyclyl. Representative examples of fused heterocyclyls include, but are not limited to, the following substituents:

[0135]

[0136] "Bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic alkyl group, wherein each two rings in the system share two unconnected atoms, the ring may have one or more double bonds, but there is no completely conjugated π electron system, and the ring has one or more selected from N, O, and S (O) m (wherein m is 0, 1 or 2) as ring atoms, and the remaining ring atoms are C. Preferably, the bridged heterocyclic group is 6 to 14 members, more preferably 6 to 12 members, more preferably 7 to 10 members, and most preferably 7 to 8 members. Depending on the number of rings, the bridged heterocyclic group is divided into a bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic group, and preferably refers to a bicyclic, tricyclic or tetracyclic bridged heterocyclic group, more preferably a bicyclic or tricyclic bridged heterocyclic group. Representative examples of bridged heterocyclic groups include, but are not limited to, the following substituents:

[0137] The heterocyclic ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring bound to the parent structure is a heterocyclic ring. Representative examples include, but are not limited to, the following substituents: wait.

[0138] The heterocyclic group is optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heteroalkyloxy, cycloalkylthio, heteroalkyl and alkylthio.

[0139] "Aryl" refers to a 6-14-membered all-carbon monocyclic or polycyclic fused aromatic ring (a "fused" ring system means that each ring in the system shares a pair of adjacent carbon atoms with another ring in the system) group, and has a completely conjugated π electron system. Preferably, the aryl group is 6-10 members, such as phenyl and naphthyl, and phenyl is most preferred. The aryl group can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring bound to the parent structure is an aryl group. Representative examples include, but are not limited to, the following substituents:

[0140]

[0141] The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, and sometimes more preferably one to three, and the substituents are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterooxyalkoxy, cycloalkylthio, heteroalkyl and alkylthio.

[0142] "Heteroaryl" refers to heteroaromatic ring systems, including monocyclic, bicyclic and tricyclic fused heteroaryl rings. Exemplary heteroaryl groups include 5- to 14-membered heteroaryl systems, 1 to 4 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen, preferably heteroaryl groups are heteroatoms with 1 to 10 heteroatoms selected from the group consisting of oxygen, sulfur and nitrogen. The heteroaryl group is preferably a 5-membered or 6-membered monocyclic heteroaryl group or an 8-12-membered bicyclic heteroaryl group, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazolyl, isoxazolyl, oxadiazolyl, pyrazinyl and the like, preferably pyridyl, oxazolyl, isoxazolyl, oxadiazolyl, tetrazolyl, triazolyl, thienyl, imidazolyl, pyrazolyl, pyrimidinyl and thiazolyl, more preferably pyrazolyl, furyl, pyridyl, oxazolyl, isoxazole, furanyl and pyrimidinyl.

[0143] The bicyclic heteroaryl is preferably a 5-membered / 5-membered bicyclic heteroaryl, or a 5-membered / 6-membered bicyclic heteroaryl, or a 6-membered / 5-membered bicyclic heteroaryl. Representative examples include, but are not limited to, the following substituents:

[0144] The heteroaryl group may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring bound to the parent structure is the heteroaryl group. Representative examples include, but are not limited to, the following substituents:

[0145]

[0146] The heteroaryl group may be substituted or unsubstituted. The substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, and the substituents are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterooxyalkoxy, cycloalkylthio, cycloalkylthio, heteroalkylthio.

[0147] "Alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl) groups, wherein alkyl is defined as above. Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, etc. Alkoxy can be substituted or unsubstituted. When substituted, the substituents are preferably one or more, sometimes preferably one to five, sometimes more preferably one to three, and the substituents are independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylsulfonyl, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heteroalkyloxy, cycloalkylthio and heterothio.

[0148] "Fused" refers to any ring structure described herein that is fused to an existing ring structure. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure that becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be substituted with a nitrogen atom, an oxygen atom, and a sulfur atom.

[0149] "Haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above. The hydrogen atoms of the present invention may be substituted with deuterium, its isotope.

[0150] Any hydrogen atom in the compounds of the embodiments of the present invention may also be replaced by a deuterium atom.

[0151] "Bond" refers to a covalent bond using the symbol "—".

[0152] "Hydroxyalkyl" means an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0153] "Hydroxy" refers to an -OH group.

[0154] "Halogen" refers to a fluorine, chlorine, bromine or iodine atom.

[0155] "Amino" refers to the -NH2 group.

[0156] "Cyano" refers to a -CN group.

[0157] "Nitro" refers to a -NO2 group.

[0158] "Oxo" refers to a =0 group.

[0159] "Carboxy" refers to a -C(O)OH group.

[0160] "Alkoxycarbonyl" means a -C(O)O(alkyl) or (cycloalkyl) group where alkyl and cycloalkyl are as defined above.

[0161] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance may or may not occur. For example, "heterocyclyl optionally substituted with alkyl" means that alkyl may be present but need not be, and that the description includes instances where the heterocyclyl is substituted with alkyl and instances where the heterocyclyl is not substituted.

[0162] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are independently replaced by a corresponding number of substituents. It goes without saying that the substituent is present in its only possible chemical position. A person skilled in the art will be able to determine whether a substitution is possible or not, experimentally or theoretically, without undue effort. For example, the combination of an amino or hydroxyl group with free hydrogen and a carbon atom with an unsaturated bond (e.g. olefinic) may be unstable.

[0163] Unless otherwise defined, a subunit can be connected to a group from either end. For example, in the general formula (I), L2 can be –CONH-, in which case it includes two connection methods. One is that M2 is directly connected to C(O) to form M2-CONH-. The other is that M2 is directly connected to NH to form M2-NH-CO-.

[0164] "Pharmaceutical composition" refers to a mixture of one or more compounds described in the present invention or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to an organism, facilitate the absorption of the active ingredient, and thus exhibit biological activity.

[0165] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have corresponding biological activities.

[0166] Dosage Form: The pharmaceutical composition or combination of the present invention may be, for example, in a unit dose of about 1-1000 mg of active ingredient for a subject of about 50-70 kg.

[0167] In some embodiments, in the above-mentioned pharmaceutical composition, the amount of the compound, tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof is about 0.1-95% by weight (calculated based on the free base); preferably, about 5-70%, for example 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%.

[0168] In some embodiments, the above-mentioned pharmaceutical composition is formulated into tablets, capsules, liquid forms or injection forms.

[0169] In some embodiments, in the above-mentioned pharmaceutical composition, the amount of the compound, tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof is about 1-1000 mg (calculated based on the free base); preferably, about 1-500 mg, more preferably, about 1 mg, 2 mg, 3 mg, 5 mg, 10 mg, 20 mg, 40 mg, 50 mg, 60 mg, 80 mg, 100 mg, 200 mg, 300 mg, 400 mg or 500 mg.

[0170] In some embodiments, the compound, tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer or mixture thereof or a pharmaceutically acceptable salt thereof can be administered by any suitable route of administration, such as oral, parenteral, buccal, sublingual, nasal, rectal, intrathecal or transdermal administration, and pharmaceutical compositions adjusted accordingly.

[0171] In some embodiments, the compound, tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt is formulated in a solid or liquid form, such as a solid or liquid form, such as a syrup, suspension, emulsion, tablet, capsule, powder, granule or lozenge.

[0172] In the compounds of the present invention, atoms may exhibit their natural isotopic abundance, or one or more atoms may be artificially enriched with the same atomic number but with an atomic mass or mass number different from the atomic mass or mass number that is predominantly present in nature. All suitable isotopic variants of the general formula of the present invention and its embodiments are described and claimed herein. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H, also represented herein as D). Protium is the major hydrogen isotope found in nature. Enrichment with deuterium may provide certain therapeutic advantages, such as increasing half-life in vivo or reducing dosage requirements, or may provide compounds that can be used as standards for characterization of biological samples. Isotopically enriched compounds may be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by methods similar to those described in the processes and examples herein, using appropriate isotopically enriched reagents and / or intermediates. DETAILED DESCRIPTION

[0173] Example

[0174] Synthesis steps

[0175] All contents involved in WO2024120378A2 are added to the present invention by reference. The compounds of the present application can also be synthesized by referring to the preparation methods of the intermediates and compounds of the patent.

[0176] Preparation Example 1

[0177] 2-(2-(3,6-dihydro-2H-pyran-4-yl)-7-ethyl-6-(2-(3-hydroxypicolinoyl)-2,5-diazabicyclo[5.1.0]octan-5-yl)-5-oxoimidazolo[1,2-a]pyrimidin-8(5H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 228

[0178]

[0179] Step 1: 4-(tert-butyl) 5-oxo-1,4-diazacyclohexane-1,4-dicarboxylic acid 1-benzyl ester, 228-2

[0180] 5-Oxo-1,4-diazepane-1-carboxylic acid benzyl ester (4g, 16.11mmol), di-tert-butyl dicarbonate (5.27g, 24.17mmol), DMAP (3.94g, 32.22mmol) in THF (30mL) solution and the mixture were stirred at room temperature overnight. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA, the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. Heptane (100mL) was added, the resulting precipitate was collected by filtration, the filtrate was concentrated under reduced pressure, and then heptane (20mL) was added, the resulting precipitate was collected by filtration, and the filter cake was dried in vacuo to give 228-2 (24.9g, 66.33% yield). 1 H NMR(400MHz, DMSO-d6)δ:6.58(br,1H),3.62-3.60(m,4H),3.04-3.28(m,2H),2.63-2.61(m,2H),1.49(s,9H).

[0181] Step 2: 4-(tert-butyl) 5-hydroxy-1,4-diazepane-1,4-dicarboxylic acid 1-benzyl ester, 228-3 DIBAL (1M, 28.85 mL) was added dropwise to a solution of 228-2 (6.7 g, 19.23 mmol) in dry DCM (30 mL) and stirred at -78 °C under N2 atmosphere for 2 hours. The mixture was quenched with potassium sodium tartrate, filtered through silica ore, the filtrate was extracted with DCM, the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography to give 228-3 (4.6 g, 68.26% yield). Mass calc C 18 H 26 N2O5 for 350.18found:251.2(M-Boc+H) + ESI.

[0182] Step 3: 4-Benzyl-1-tert-butyl-2,3-dihydro-1H-1,4-diazepine-1,4(5H)-dicarboxylate, 228-4 A mixture of 228-3 (2.6 g, 7.42 mmol), DIEA (5.75 g, 44.52 mmol) and acetyl chloride (2.91 g, 37.1 mmol) in Tol (20 mL) was stirred at 120 °C overnight. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography to give 228-4 (2 g, 81.09% yield).

[0183] Mass calc C 18 H 24N2O4 for 332.17found:233.1(M-Boc+H) + ESI.

[0184] Step 4: Benzyl 2,5-diazabicyclo[5.1.0]octane-5-carboxylate, 228-5

[0185] 228-4 (500 mg, 1.50 mmol) was suspended in Et2O (5 mL), ZnEt2 (929.02 mg, 7.52 mmol) was added, and stirred at room temperature for 10 min under N2 atmosphere. Then CH2I2 (2.42 g, 9.03 mmol) was added, and the mixture was stirred at room temperature overnight. Potassium sodium tartrate was added to quench the mixture at 0°C, and the mixture was filtered through silica ore and washed with EA. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography to obtain 228-5 (250 mg, 67.48% yield).

[0186] Mass calc C 14 H 18 N2O2 for 246.14found:247.2(M+H) + ESI.

[0187] Step 5: 2-(tert-butyl) 2,5-diazabicyclo[5.1.0]octane-2,5-dicarboxylic acid benzyl ester, 228-6 A solution of 228-5 (500 mg, 2.03 mmol), di-tert-butyl dicarbonate (1.33 g, 6.09 mmol) in MeOH (5 mL) and the mixture were stirred overnight at RT. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA, and the organic layer was washed with brine, dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by column chromatography to give 228-6 (660 mg, 93.85% yield).

[0188] Mass calc C 19 H 26 N2O4 for 346.19found:247.26(M-Boc+H) + ESI.

[0189] Step 6: tert-butyl 2,5-diazabicyclo[5.1.0]octane-2-carboxylate, 228-7

[0190] 228-6 (660 mg, 1.91 mmol) was dissolved in MeOH (2 mL) and Pd / C was added in the presence of H2. The reaction mixture was stirred at room temperature until the substrate disappeared. The reaction mixture was filtered and the filter was used directly in the next step.

[0191] Mass calc C 11 H 20 N2O2 for 212.15found:213.2(M+H) + ESI.

[0192] Step 7: 5-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-(2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2,5-diazabicyclo[5.1.0]octane-2-carboxylic acid tert-butyl ester, 228-8

[0193] Int-1 (76.36 mg, 0.14 mmol) was suspended in DMSO (0.3 mL), CsF (33.73 mg, 0.22 mmol) and 228-7 (300 mg, 1.41 mmol) were added, and then stirred at 100 ° C for 1 hour. RM was concentrated under reduced pressure and purified with a reverse phase column (C18 column) to give the product 228-8 (30 mg, 31.61% yield). Mass calc C 33 H 40 F3N7O5 for671.3found:616.3(M-tBu+H) + ESI.

[0194] Step 8: 2-(6-(2,5-diazabicyclo[5.1.0]octan-5-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 228-9

[0195] 228-8 (30 mg, 0.04 mmol) was suspended in DCM (1 mL), TFA (0.5 mL) was added, and the mixture was stirred at room temperature for 2 hours. The mixture was washed with aqueous NaHCO3, dried over Na2SO4, and concentrated under reduced pressure. The RM was concentrated under reduced pressure and used without purification.

[0196] Mass calc C 28 H 32F3N7O3 for 571.25found:572.27(M+H) + ESI.

[0197] Step 9: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-7-ethyl-6-(2-(3-hydroxypyridinyl)-2,5-diazabicyclo[5.1.0]octan-5-yl)-5-oxoimidazolo[1,2-a]pyrimidin-8(5H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 228

[0198] 228-9 (30.00 mg, 0.05 mmol), 3-hydroxypicolinic acid (7.30 mg, 0.05 mmol), EDCI (20.10 mg, 0.1 mmol), HOBT (14.18 mg, 0.1 mmol) and pyridine (12.45 mg, 0.16 mmol) were dissolved in DCM (0.5 mL) and then stirred at room temperature for 2 hours. RM was purified by reverse phase column (C18 column) to provide the product (8 mg, 22% yield).

[0199] Mass calc C 35 H 36 F3N7O5 for 691.27found:692.31(M+H) + ESI.

[0200] 1 H NMR(400MHz,DMSO-d6)δ:10.30(br,1H),10.01(s,1H),8.05-8.03(m,1H),d,7.32(J=8.4Hz,1H), 7.63(s,1H),7.53(d,J=8.4Hz,1H),7.29-7.26(m,2H),6.83(s,1H),5.23-5.22(m,2H),4.41-4.3 8(m,2H),3.82-3.79(m,2H),3.53-3.50(m,2H),3.30-3.06(m,4H),2.92-2.86(m,2H),2.52-2.51 (m,2H),2.35(s,3H),2.00-1.98(m,1H),1.26-1.23(m,4H),0.85-0.84(m,1H),0.37-0.32(m,1H).

[0201] Preparation Example 2

[0202] 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-fluoro-8-(3-hydroxypicolinyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 236

[0203]

[0204] Step 1: (3S,4R)-3,4-dihydroxypyrrolidine-1-carboxylic acid tert-butyl ester, 236-1

[0205] Tert-butyl 2,5-dihydro-1H-pyrrole-1-carboxylate (27.08 g, 160 mmol) was dissolved in EtOH (100 mL) and water (40 mL), and MgSO4 (38.51 g, 320 mmol) and KMnO4 (50.57 g, 320 mmol) solutions were added at 0°C and stirred at 0°C for 0.5 hours. The mixture was then stirred overnight at room temperature. The mixture was added to celite and stirred for 20 minutes, filtered and quenched with aqueous Na2SO3, extracted with DCM, and the organic layer was washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The crude product was purified by column chromatography to give 236-1 (9 g, 27.67% yield). Mass calc C9H 17 NO4 for203.14found:104.1(M-Boc+H) + ESI.

[0206] Step 2: tert-Butyl bis(2-oxoethyl)carbamate, 236-2

[0207] PhI(OAc)2 (19.97 g, 62 mmol) was added to a solution of 236-1 (9 g, 44.28 mmol) in DCM (50 mL) and stirred at 0°C for 3 h. The mixture was then quenched with aqueous NaHCO3, extracted with DCM, and the organic layer was washed with brine and dried over Na2SO4. The RM was concentrated under reduced pressure and used without purification. Mass calc C7H 15 NO4 for 201.1found:102.1(M-Boc+H) + ESI.

[0208] Step 3: tert-Butyl bis(2-hydroxybut-3-en-1-yl)carbamate, 236-3

[0209] At -78 ° C, (vinyl) magnesium bromide (19.57 g, 149.09 mmol) was added to a solution of 236-2 (10 g, 49.70 mmol) in THF (30 mL). The mixture was then stirred at room temperature overnight. The mixture was dissolved in NH4Cl water, extracted with EA, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain 236-3 (3.8 g, yield 29.71%). Mass calc C 12 H 23 NO4for 257.16found:158.1(M-Boc+H) + ESI.

[0210] Step 4: ((tert-Butyloxycarbonyl)azadiyl)bis(but-3-ene-1,2-diyl)bis(2,2,2-trichloroacetimidate), 236-4

[0211] DBU (1.33 g, 8.74 mmol) and trichloroacetonitrile (15.15 g, 104.93 mmol) were added to a solution of 236-3 (4.5 g, 17.49 mmol) in DCM (80.06 mL) and stirred at room temperature for 16 hours at 0°C. The mixture was concentrated under reduced pressure. The crude product was diluted with EA and water, extracted once with EA, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography to give 236-4 (8.1 g, 84.82% yield). 1 H NMR(400MHz,DMSO-d6)δ:8.36(s,2H),5.86-5.76(m,2H),5.67-5.61(m,2H),5.44-5.37(m,2 H),5.28-5.24(m,2H),3.84-3.80(m,1H),3.70-3.66(m,1H),3.49-3.39(m,2H),1.46(s,9H).

[0212] Step 5: tert-Butyl 4-(2-phenylpropan-2-yl)-3,5-divinylpiperazine-1-carboxylate, 236-5 [Ir(cod)Cl]2 (498.15 mg, 0.74 mmol) was added to a solution of CUMYLAMINE (2.41 g, 17.8 mmol) in DCE (60 mL) at 0°C, and then the mixture was added to a solution of 236-4 (8.1 g, 14.83 mmol) in DCE (60 mL) under N2 atmosphere and stirred at room temperature overnight. The mixture was filtered and concentrated under reduced pressure to produce a residue. The residue was diluted with EA and water, extracted once with EA, and the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The crude product was purified by column chromatography to give 236-5 (3.2 g, 60.52% yield). Mass calc C 22 H 32 N2O2 for 356.25found:357.2(M+H) + ESI.

[0213] Step 6: (1R,5S)-8-(2-phenylpropan-2-yl)-3,8-diazabicyclo[3.2.1]oct-6-ene-3-carboxylic acid tert-butyl ester, 236-6

[0214] Grubbs Gen 2nd (381.02 mg, 0.45 mmol) was added to a solution of 236-5 (3.2 g, 8.98 mmol) in Tol (80 mL). The reaction mixture was stirred at 125 °C for 16 hours. The mixture was filtered and concentrated under reduced pressure to produce a residue. The residue was diluted with EA and water, extracted once with EA, and the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by column chromatography to obtain 236-6 (2.6 g, 88.19% yield). Mass calcC 20 H 28 N2O2 for 328.22found:329.2(M+H) + ESI.

[0215] 1 H NMR(400MHz,DMSO-d6)δ:7.59-7.57(m,2H),7.32-7.26(m,2H),7.23-7.21(m,1H),6.0 0-5.94(m,2H),3.64-3.60(m,2H),3.53-3.46(m,2H),1.41(s,9H),1.26-1.24(m,6H).

[0216] Step 7: (1S,5S,6S)-6-hydroxy-8-(2-phenylpropan-2-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester, 236-7

[0217] 236-6 (1.5 g, 4.57 mmol) was suspended in THF (8 mL), (-)-diisopinenylartemisinylborane (2.29 g, 7.99 mmol) was added, and the mixture was stirred at room temperature overnight under N2 atmosphere. Then NaOH (3 M, 13.71 mmol, 4.6 mL) was added, and 30% H2O2 (2.8 mL) was added after degassing, and stirred at 50 ° C for 2 h. The mixture was quenched with Na2S2CO3 aqueous solution, extracted with EA, and the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by column chromatography to obtain 236-7 (1.3 g, 82.16% yield). Mass calc C 20 H 30 N2O3 for 346.23found:347.2(M+H) + ESI.

[0218] Step 8: (1S,5S,6S)-6-fluoro-8-(2-phenylpropan-2-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylic acid tert-butyl ester, 236-8

[0219] DAST (488.51 mg, 3.03 mmol) was added to a solution of 236-7 (200 mg, 0.577 mmol) in DCM (2 mL) and stirred at 0°C for 2 hours under N2 atmosphere. The mixture was diluted with aqueous NaHCO3, extracted with DCM, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a residue. The crude product was purified by column chromatography to obtain 236-8 (360 mg, 51.13% yield). Masscalc C 20 H 29 FN2O2 for348.22found:349.2(M+H) + ESI.

[0220] Step 9: (1R, 5R, 6R)-6-fluoro-8-(2-phenylpropan-2-yl)-3,8-diazabicyclo[3.2.1]octane, 236-9

[0221] TMSOTf (344.43 mg, 1.55 mmol), lutidine (249.08 mg, 2.32 mmol) were added to a solution of 236-8 (135 mg, 0.39 mmol) in DCM (2 mL) and stirred at 0 °C for 2 h under N2 atmosphere. The RM was concentrated under reduced pressure and used without purification. Mass calc C 15 H 21 FN2 for 248.17found:249.2(M+H) + ESI.

[0222] Step 10: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-fluoro-8-(2-phenylpropan-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 236-10

[0223] Int-1 (43.52 mg, 0.08 mmol) was suspended in DMSO (0.2 mL), and CsF (24.47 mg, 0.16 mmol) and 236-9 (40 mg, 0.16 mmol) were added, and then stirred at 100 ° C overnight. The RM was concentrated under reduced pressure and purified with a reverse phase column (C18 column) to give 236-10 (5 mg, 8.74% yield). Mass calc C 37 H 41 F4N7O3 for707.32found:708.4(M+H) + ESI.

[0224] Step 11: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-fluoro-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 236-11

[0225] 236-10 (5 mg, 7.04 μmol) was suspended in TFA (1 mL), and the mixture was stirred at 85°C for 3 hours. The mixture was washed with aqueous NaHCO3, extracted with DCM, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The RM was concentrated under reduced pressure and used without purification. Mass calc C 28 H 31 F4N7O3 for 589.24found:590.5(M+H)+ ESI.

[0226] Step 12: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-fluoro-8-(3-hydroxypicolinyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 236

[0227] 236-11 (4.5 mg, 7.63 μmol), 3-hydroxypyridine-2-carboxylic acid (1.38 mg, 9.92 μmol),

[0228] HOBT (2.06 mg, 15.27 μmol), EDCI (2.92 mg, 15.27 μmol) and pyridine (1.81 mg, 22.90 μmol) were dissolved in DCM (1 mL) and then stirred at room temperature for 2 hours. The RM was purified by reverse phase column (C18 column) to give the product (1.6 mg, 29.55% yield). Mass calc C 34 H 34 F4N8O5 for 710.26found:711.3(M+H) + ESI.

[0229] 1 H NMR(400MHz, DMSO-d6)δ:11.79(br,1H),10.02(s,1H),8.16-8.14(m,1H),7.72(d,J=8.4Hz ,1H),7.63(s,1H),7.53(d,J=8.4Hz,1H),7.46-7.45(m,1H),7.43-7.41(m,2H),6.82(s,1H ),5.62-5.47(m,1H),5.38-5.31(m,3H),5.08-4.83(m,1H),4.25(s,2H),3.81-3.65(m,3H) ,2.96-2.90(m,3H),2.67-2.51(m,3H),2.35(s,3H),2.01-1.99(m,3H),1.24-1.23(m,3H).

[0230] Preparation Example 3

[0231] 2-(6-(6,6-difluoro-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-1,4-diazepan-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 360

[0232]

[0233] Step 1: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-hydroxy-1,4-diazepan-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 360-2

[0234] 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (360-1,4 g, 7.40 mmol) was suspended in DMSO (10 ml), 1,4-diazepan-6-ol (8.60 g, 74.03 mmol) and AgBF4 (2.88 g, 14.81 mmol) were added, and then stirred at 120° C. for 4 hours. The mixture was diluted with EA and water, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by reverse phase column (C18 column) to give 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-hydroxy-1,4-diazepan-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (360-2, 2.5 g, 58% yield).

[0235] Mass calc C 27 H 32 F3N7O4 for 575.25found:576.34(M+H) + ESI.

[0236] Step 2: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6-hydroxy-1,4-diazepane-1-carboxylic acid tert-butyl ester, 360-3

[0237] A solution of 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(6-hydroxy-1,4-diazepam-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (360-2, 10 mg, 17.37 μmol), (Boc)2O (4.55 mg, 20.85 μmol) and DIEA (3.37 mg, 26.06 μmol, 4.54 μL) in MeOH (1 mL) was stirred at room temperature overnight. The mixture was concentrated under reduced pressure and used in the next step without purification.

[0238] Mass calc C 32 H 40 N7O6F3 for 675.30found:674.3(MH) - ESI.

[0239] Step 3: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6-oxo-1,4-diazepane-1-carboxylic acid tert-butyl ester, 360-4

[0240] Dess-Martin reagent (82.86 mg, 195.35 μmol) was added to a solution of 4-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-[2-methyl-4-(trifluoromethyl)aniline]-2-oxoethyl]-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl]-6-hydroxy-1,4-diazepane-1-carboxylic acid tert-butyl ester (360-3.66 mg, 97.68 μmol) in DCM (6 mL) in three portions. The resulting mixture was stirred for 16 hours. The mixture was filtered through silica ore and the solvent was removed in vacuo. The crude material was purified by silica gel chromatography using a gradient elution of 40-100% ethyl acetate / hexanes to afford tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6-oxo-1,4-diazepane-1-carboxylate (360-4,60 mg, 89.06 μmol, 91.18% yield)

[0241] Mass calc C32 H 38 N7O6F3 for 673.28found:672.3(MH) - ESI.

[0242] Step 4: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6,6-difluoro-1,4-diazepane-1-carboxylic acid tert-butyl ester, 360-5

[0243] Tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6-oxo-1,4-diazacyclopentane-1-carboxylate (100 mg, 148.44 μmol) was suspended in DCM (5 mL), DAST (400.00 mg, 2.48 mmol) was added, and the RM was stirred at room temperature for 20 h. The mixture was dissolved with aqueous NaHCO3, the mixture was diluted with DCM and water, extracted once with DCM, the organic layer was washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by reverse phase column (C18 column) to give tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6,6-difluoro-1,4-diazepane-1-carboxylate (360-5.12 mg, 17.25 μmol, 11.62% yield). Mass calc C 32 H 38 F5N7O5 for 695.23,found:596.3(M-Boc+H) + ESI.

[0244] 1H NMR(400MHz, CDCl3-d6)δ:9.982(s,1H),7.712-7.691(d,1H,J=8.4),7.631(s,1H),7.551 -7.530(d,1H,J=8.4),6.838(s,1H),5.218(s,2H),4.273-4.267(d,2H,J=2.4Hz),3.987-3 .829(m,2H),3.816-3.780(t,2H,J=14.4Hz),3.761-3.672(m,2H),3.476-3.423(m,2H),3. 204-2.926(m,4H),2.602-2.555(m,2H),2.360(s,3H),1.435(s,9H),1.355-1.295(m,3H).

[0245] Step 5: 2-(6-(6,6-difluoro-1,4-diazepam-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 360-6

[0246] TFA (500.00 mg, 4.39 mmol) was added to a solution of tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-6,6-difluoro-1,4-diazepane-1-carboxylate (360-5,30 mg, 43.12 μmol) in DCM (4 mL) and the RM was stirred at room temperature for 1 h. The RM was concentrated under reduced pressure and used without purification. Masscalc C 27 H 30 N7O3F5 for 595.23,found:596.3(M+H) + ESI.

[0247] Step 6: 2-(6-(6,6-difluoro-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-1,4-diazepam-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide 4-Pyrimidinecarboxylic acid, 5-hydroxy-6-methyl-(12.94 mg, 83.95 μmol), pyridine (9.96 mg, 125.93 μmol, 10.14 μL), EDCI (16.08 mg, 83.95 μmol) and HOBT (11.34 mg, 83.95 μmol) were dissolved in DCM (993.22 μL). 2-[6-(6,6-difluoro-1,4-diazepam-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]-N-[2-methyl-4-(trifluoromethyl)phenyl]acetamide (25.00 mg, 41.98 μmol) was added to the solution at 0°C. After stirring for 1 h, RM was purified by reverse phase column (C18 column) to give the product 2-(6-(6,6-difluoro-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-1,4-diazepam-1-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (3.2 mg, 4.37 μmol, 10.42% yield). Mass calcC 33 H 34 F5N9O5 for 731.26,found:732.3(M+H) + ESI.

[0248] 1 H NMR(400MHz, CDCl3-d6)δ:10.010(s,1H),8.568(s,1H),7.715-7.685(m,1H),7.63 0(s,1H),7.547-7.526(d,1H,J=8.4Hz),6.834(s,1H),5.221(s,2H),4.675-4.208 (m, 3H), 4.147-3.944 (m, 2H), 3.812 (s, 2H), 3.673-3.499 (m, 4H), 3.143-3.004 (m, 4H), 2.602-2.555 (m, 2H), 2.44 (s, 3H), 2.360 (s, 3H), 1.355-1.295 (m, 3H). Preparation Example 4

[0249] 2-(6-(1-acetyl-3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, Compound 437

[0250]

[0251] Step 1: tert-Butyl 3,3-difluoro-4-(((perfluorobutyl)sulfonyl)oxy)-3,6-dihydropyridine-1(2H)-carboxylate, 340-1

[0252] DBU (19.4 g, 127.43 mmol) was added to a solution of tert-butyl 3,3-difluoro-4-oxopiperidine-1-carboxylate (10 g, 42.51 mmol) in THF (100 mL) and stirred at -20 ° C for 10 minutes. Then 1,1,2,2,3,3,4,4,4,4-nonafluorobutane-1-sulfonyl fluoride (38.55 g, 127.60 mmol) in THF (40 mL) was added to the mixture and stirred at -20 ° C for another hour. The mixture was diluted with EA and water, extracted twice with EA, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography to give 340-1 (14 g, 63.66% yield). 1 H NMR (400MHz, DMSO-d6) δ: 6.75 (s, 1H), 4.23-4.21 (m, 2H), 4.04-3.98 (m, 2H), 1.40 (s, 9H).

[0253] Step 2: (2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)boronic acid, int1-B(OH)2

[0254] i-PrMgCl·LiCl (6 g, 6 mmol) was added dropwise to a solution of int-1 (540.33 mg, 1 mmol) in dry THF (2 mL) and stirred for 20 min. Subsequently, trimethyl borate (0.65 mL, 5.8 mmol) was added dropwise to the mixture and stirred at room temperature for 2 h. The mixture was quenched with MeOH, extracted with EA and water, and the organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The RM was concentrated under reduced pressure and used without purification. Mass calc C 22 H 23BF3N5O5 for 505.17found:504.2(MH) - ESI.

[0255] Step 3: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((4-methyl-3-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,3-difluoro-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester, 390-1

[0256] Int1-B(OH)2 (450 mg, 0.77 mmol) was suspended in 1,4-dioxane (5 mL), 340-1 (1.98 g, 3.83 mmol) and K3PO4 (211.43 mg, 1 mmol) were added, the mixture was degassed with argon for 10 minutes, PdCl2(dppf) (310.49 mg, 0.38 mmol) was added, and the mixture was stirred at 80°C for 2 h. The mixture was concentrated under reduced pressure, and the crude product was purified by reverse phase column (C18 column) to give 437-1 (160 mg, 30.77% yield).

[0257] Mass calc C 32 H 35 F5N6O5 for 678.26found:677.36(MH) - ESI.

[0258] 1 H NMR (400MHz, DMSO-d6) δ: 10.06 (s, 1H), 7.71 (d, J = 8.4Hz, 1H), 7.63 (s, 1H), 7.53(d,J=8.4Hz,1H),6.85(s,1H),6.38(s,1H),5.33-5.17(m,2H),4.28-4. 26(m,3H),3.82-3.79(m,3H),3.51-3.48(m,1H),3.27-3.24(m,1H),2.75-2 .70(m,2H),2.51-2.50(m,2H),2.36(s,3H),1.45(s,9H),1.18-1.14(m,3H).

[0259] Step 4: 2-(6-(3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 437-2

[0260] 437-1 (110 mg, 0.16 mmol) was suspended in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was washed with aqueous NaHCO3, dried over Na2SO4, and concentrated under reduced pressure. The RM was concentrated under reduced pressure and used without purification. Mass calc C 27 H 27 F5N6O3 for578.21found:579.38(M+H) + ESI.

[0261] Step 5: 2-(6-(1-acetyl-3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

[0262] Acetic acid (6.23 mg, 103.71 μmol), pyridine (12.31 mg, 155.57 μmol, 12.53 μL), EDCI (19.86 mg, 103.71 μmol) and HOBT (14.01 mg, 103.71 μmol) were dissolved in DCM (2.0 mL). To this solution was added 437-2 (30 mg, 51.86 μmol) at 0°C. After stirring for 1 h, RM was purified by HPLC to give the product 2-(6-(1-acetyl-3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (5 mg, 8.06 μmol, 15.54% yield). Mass calc C 29 H 29 F5N6O4 for 620.22found:621.3(M+H) + ESI.

[0263] 1H NMR (400MHz, DMSO-d6) δ: 10.073 (s, 1H), 7.729-7.707 (d, 1H, J = 8.8Hz), 7.633 (s, 1H),7.552-7.532(d,1H,J=8.0),6.855(s,1H),6.423-6.389,1H,J=13.6Hz),5.3 40-5.176 (m, 2H), 4.523-3.929 (m, 6H), 3.817 (t, 2H, J = 5.6 Hz), 2.729-2.687 (m, 2H), 2.602-2.555 (m, 2H), 2.362 (s, 3H), 2.131 (m, 3H), 1.184-1.154 (m, 3H). Preparation Example 5

[0264] 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(1-ethyl-3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide

[0265]

[0266] Step 1: 2-(6-(3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, 437-2

[0267] 437-1 (110 mg, 0.16 mmol) was suspended in DCM (5 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. The mixture was washed with aqueous NaHCO3, dried over Na2SO4, and concentrated under reduced pressure. The RM was concentrated under reduced pressure and used without purification. Mass calc C 27 H 27 F5N6O3for 578.21found:579.38(M+H) + ESI.

[0268] Step 2: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(1-ethyl-3,3-difluoro-1,2,3,6-tetrahydropyridin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide, compound 438

[0269] Acetaldehyde (53.30 mg, 1.21 mmol) was added to a solution of 437-2 (70 mg, 0.12 mmol) in THF (7 mL) and stirred at room temperature for 10 minutes. Sodium triacetoxyborohydride (76.93 mg, 0.36 mmol) was then added and the mixture was stirred at room temperature for 2 hours. The mixture was adjusted to pH 8 with aqueous NaHCO3, diluted with EA and water, extracted once with EA, the organic layer was washed with brine, dried with Na2SO4, filtered, and concentrated under reduced pressure. The crude oil was purified by reverse phase column (C18 column) to give the product (21 mg, 28.61% yield). Mass calc C 29 H 31 F5N6O3 for 606.24found:605.32(MH) - ESI.

[0270] 1 H NMR(400MHz, DMSO-d6)δ:10.05(s,1H),7.71(d,J=8.4Hz,1H),7.62(s,1H),7.53(d,J=8.4 Hz,1H),6.84(s,1H),6.35(s,1H),5.34-5.16(m,2H),4.27-4.26(m,2H),3.82-3.80(m,2H ),3.35-3.34(m,1H),3.16-3.08(m,2H),2.89-2.82(m,1H),2.75-2.69(m,2H),2.59-2.52(m,4H),2.36(s,3H),1.18-1.13(m,3H),1.09-1.06(m,3H). The compounds in the following table were synthesized by or with reference to the above method.

[0271] Table 1

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279]

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291] Biological experiments

[0292] WRN helicase assay

[0293] In order to evaluate the inhibitory properties of the compounds of the invention on WRN helicase activity, a fluorescence assay using forked DNA was established. In the helicase assay, His-WRN produced in-house was used. 527-1072Fluorescent forked DNA prepared by annealing OLIGOA-BHQ2 (TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTCGTACCCGAT GTGTTCGTTC-BHQ2) and OLIGOB-TAMRA (TAMRA-GAACGAACACA TCGGGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT) was used as a substrate. Test compounds were prepared into 2.5-fold serial dilutions of 7 gradient points with DMSO, and then 1.5 μl of each concentration was added to 48.5 μl of assay buffer (25 mM Tris-HCl (pH 8.0), 50 mM NaCl, 2 mM MgCl2, 1 mM DTT, 0.05% Tween-20, and 2.5 μg / ml BSA) for further dilution. 5 μl of 45 nM WRN protein and 5 μl of each diluted compound solution were transferred to a 384-well assay plate and pre-incubated at room temperature for 30 minutes. Each concentration of compound was added to the plate in triplicate. Controls were included in the test to validate the assay. Instead of compound solution, assay buffer containing 3% DMSO was added to the positive control (no inhibition), and no protein was added to the negative control (maximum inhibition). Forked DNA and ATP were diluted to 300 nM and 6 mM, respectively, with assay buffer. To initiate the reaction, 5 μl of dsDNA and ATP solution were added to the assay plate. After incubation at room temperature for 60 minutes, the plate was transferred to a Victor Nivo multimode plate reader (Perkin Elmer, Waltham, MA) and the fluorescence output was monitored. Dose response curves were plotted using GraphPad Prism 9 (GraphPad Software, San Diego, CA) to derive IC50s.

[0294] WRN ATPase assay

[0295] To evaluate the inhibitory properties of the compounds of the present invention on DNA-dependent WRN ATPase activity, an ATPase assay using the ADP-Glo ​​assay kit (Promega, Madison, WI) was established. In the ATPase assay, the same WRN protein and DNA substrate as described in the helicase assay were used. The serial dilution of the compound in DMSO was the same as for the helicase assay. In further dilutions, 2 μl of each concentration was transferred to 48 μl of assay buffer. 5 μl of 100nM WRN protein and 2.5 μl of each diluted compound solution were preincubated in a 384-well assay plate at room temperature for 30 minutes. All test wells were set up in triplicate. Then, 2.5 μl of assay buffer containing 400nM DNA substrate and 4mM ATP was added to each well and incubated for 60 minutes. A no protein control (maximum inhibition) and a no compound control (no inhibition) were included in the test. In order to stop the reaction and consume excess ATP, 10 μl of the reagent ADP-Glo ​​from the assay kit was added. TM , and incubated for 40 minutes. Then, 20 μl of ATP detection reagent was added. After 30 minutes of incubation, luminescence was measured using a Victor Nivo multi-mode plate reader (Perkin Elmer, Waltham, MA). Dose-response curves spanning IC50s were generated by GraphPad Prism 9 (GraphPad Software, San Diego, CA).

[0296] Cell viability assay

[0297] High microsatellite instability (MSI) cell line SW48 and microsatellite stable MSS cell line SW480 were purchased from ATCC and cultured in DMEM GlutaMax medium (Gibco, 10564-011) consisting of penicillin-streptomycin (Gibco-15140122) and 10% fetal bovine serum (Gibco-10082-147) at 37°C in a humidified 5% CO2 incubator.

[0298] Cells were seeded at 3000 cells / well in a white clear bottom 384-well plate (PerkinElmer). The next day, twelve 3-fold serial dilutions of 10 mM DMSO compound stock solutions were added to each replicate well. Blank wells contained medium without cells, and DMSO control wells contained cells and DMSO only. After 72 hours of treatment, the cells were treated with CellTiter Cell viability was analyzed using the 2.0 assay (Promega, G9243), and after incubation at room temperature for 10 minutes, luminescent signals were detected on a VICTOR NivoTM multimode plate reader (PerkinElmer). For data analysis, all data points were normalized by subtracting the blank value, and the ratio of the DMSO control value was calculated. IC50 values ​​were calculated using GraphPad Prism 9.

[0299] Table 2

[0300]

[0301]

[0302]

[0303]

[0304]

[0305]

[0306]

[0307]

[0308] N / A means not tested.

Claims

1. A compound of formula (I)-(XI), (I'), or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, Ring A and Ring B are each independently selected from an aryl group or a heteroaryl group; Y1 is CH, NH, N, O or S; Y2 is CH, NH, N, O or S; Y3 is CH, NH, N, O or S; X1 and X2 are each independently selected from CH or N; M1, M2, M5, M6 are each independently selected from CH, NH, N, O or S; M3 and M4 are each independently selected from -NR a ,-CR a R b ; R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl; is a single bond or a double bond, provided that they are adjacent Not a double bond at the same time; R1, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; Two R2 together with the atoms to which they are bound form a C3-C10 cycloalkyl or a C3-C10 heterocyclic group, wherein the cycloalkyl or heterocyclic group is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, carbonyl, and oxo; R5 and R6 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; or, R5 and R6 together with the atoms to which they are attached form a C3-C10 cycloalkyl group, a C3-C10 heterocyclyl group, a C6-C14 aryl group, or a C1-C12 heteroaryl group containing 1 to 4 heteroatoms selected from O, S and N; R7 is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl; Or, two R7 together with the atoms to which they are attached form a C3-C10 cycloalkyl group, a C1-C10 heterocyclyl group; L1 is a bond, O, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl, -C3-C10 cycloalkyl-CONH-, -C1-C6 alkyl-C3-C10 cycloalkyl-CONH, -C1-C6 haloalkyl-NH-, -C3-C10-heterocyclo-NH-alkyl, -C3-C10 cycloalkyl-NH-, -C1-C6 alkyl-C(NH)-NH, -C6-C10 heteroaryl-C1-C6 alkyl, -S(O)2-NH-, and -C1-C6 alkyl-S(O)2-NH-; L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(Rc)-; R c is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, and hydroxyalkyl; Preferably, ring A is a C6-C14 aryl group or a C1-C12 heteroaryl group containing 1 to 4 heteroatoms selected from O, S and N; R 10 is a cycloalkenyl radical, wherein the cycloalkenyl radical is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms; or a heterocyclyl radical, wherein the heterocyclyl radical is a 5- or 6-membered fully saturated or partially unsaturated radical comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, and wherein the heterocyclyl radical is unbridged or bridged and the bridge is 1 or 2 carbon atoms; or a heteroaryl radical, wherein the heteroaryl radical is a 5- or 6-membered fully unsaturated monocyclic radical comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O and S, preferably 1 or 2 ring heteroatoms, wherein the total number of ring S atoms does not exceed 1 and the total number of ring O atoms does not exceed 1; or a phenyl radical; wherein the cycloalkenyl radical, heterocyclyl radical, heteroaryl radical, phenyl radical is unsubstituted or substituted by one or more substituents selected from halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; Or, R 10 is C2-C4 alkenyl, optionally substituted by R4 or -O-R4; m is 0, 1, 2, 3, or 4; p is 0, 1, 2, or 3; q is 0, 1, 2, or 3; r is 0, 1, 2, or 3; s is 2 or 3; t is 0, 1, 2, or 3; w is 0, 1, 2, or 3; and n is 0, 1, 2, or 3.

2. A compound according to claim 1 or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the compound has formula (XII)-(XIII), (XIX)-(XXI), (XIIB) R8 is C3-C12 fused cycloalkyl, C2-C10 fused heterocyclyl, C2-C10 spiroheterocyclyl or C3-C12 spirocycloalkyl, wherein each of the fused cycloalkyl, fused heterocyclyl, spiroheterocyclyl or spirocycloalkyl is optionally substituted with one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy and oxo; R9 is a C3-C10 bridged cycloalkyl or a C3-C10 bridged heterocyclic group, wherein the bridged cycloalkyl and the bridged heterocyclic group are each optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-5 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, oxo, C1-6 haloalkyl, C3-5 cycloalkyl; L6 is a C3-C10 bridged cycloalkyl or a bridged C2-C10 bridged heterocyclic group, wherein the bridged cycloalkyl and the bridged heterocyclic group are each optionally substituted by one or more substituents selected from hydrogen, deuterium, halogen, amino, cyano, hydroxyl, C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, oxo, C1-6 haloalkyl, C3-5 cycloalkyl; Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R4, m, p, q, r, t are as defined in claim 1.

3. The compound of claim 1 or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the compound has the formula (XXII)-(XXIII): Each X is independently selected from F, Cl, Br, I; Y1, Y2, Y3, M1, M2, L2, X1, X2, R1, R3, R7, m, n, p, r, t, w are as defined in claim 1.

4. The compound of claim 1 or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the compound has the formula (XXIV)-(XXVI), Ring C is C6-C 10 Aryl or C2-C9 heteroaryl; Two R's 11 connected to form a C1-C3 alkylene bridge or a -CH=CH- bridge or a -CH=CH-CH2- bridge or a C1-C3 heteroalkylene bridge, wherein the C1-C3 alkylene bridge or the -CH=CH-CH- bridge or the C1-C3 heteroalkylene bridge is optionally substituted by one or more halogens, and the heteroatom is selected from one heteroatom among N, O and S; the other R 11 are independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 12 are independently selected from hydrogen, deuterium, halogen, amino, cyano, carbonyl, hydroxy, alkoxy, alkoxy, haloalkyl, hydroxyalkyl and -O-haloalkyl; R 13 is halogen, amino, cyano, oxo, hydroxy, alkoxy, haloalkyl, hydroxyalkyl and -O-haloalkyl; u is 2, 3 or 4; v is 1, 2, 3, or 4; R 10 ,L2,X1,X2,R1,R3,R4,R5,R7,m,p,q,r as defined in claim 1.

5. A compound of claim 1 or a tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the compound has the formula (XXVII)-(XXXIII), (XXXV)-(XXXVII), (XXVII-A), (XXVII-B), (XXXVI-A), Among them, R 14 is selected from the following groups: Each X is independently selected from F, Cl, Br, I; R 10a is a 5-6 membered heteroaryl group, wherein the 6 membered heteroaryl group has 2 or more heteroatoms, wherein the 5-6 membered heteroaryl group is unsubstituted or substituted by one or more selected from halogen, amino, cyano, carbonyl, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; Ring D is a C6-C12 fused aryl group, a C1-C12 fused heteroaryl group, or a C1-C12 fused heterocyclic group; Ring E is C6-C12 aryl or C1-C12 heteroaryl; Ring F is C6-C10 aryl or C2-C9 heteroaryl; L3 is O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(Rc)-; L7 is -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C6 alkyl-NH-, -C2-C6 alkenyl-NH-, C2-C6 alkynyl, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6 haloalkyl and -CON(Rc)-; R 15 is a heterocyclyl or spiroheterocyclyl or spirocycloalkyl group, wherein the heterocyclyl, spiroheterocyclyl and spirocycloalkyl group are unsubstituted or substituted by one or more groups selected from halogen, amino, cyano, carbonyl, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 16 is selected from the group consisting of hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl, hydroxyalkyl, and -O-haloalkyl; Ring C,Y1,Y2,Y3,M1,M2,L2,X1,X2,R1,R3,R4,R5,R7,R 10 ,R 12 ,R c ,m,n,p,r,t,w,q as defined in claims 1-4.

6. The compound of claim 1 or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer or mixture, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein the compound has the formula (XXXVIII)-(XXXIX), R 10b is selected from the following groups: Wherein the R 4a is selected from the group consisting of halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 4b is selected from the group consisting of amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 4c is selected from the group consisting of halogen, amino, cyano, carbonyl, hydroxy, alkyl, haloalkyl and hydroxyalkyl; m1 is 1 or 2; m2 is 0, 1 or 2; R1, R3, R4, R7, Y1, Y2, Y3, r, t, w, p, q, L2, X1, X2, M1, M2 are as defined in claim 1.

7. The compound of claim 1, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein: Ring A is a C6-C14 aryl group or a C1-C12 heteroaryl group containing 1 to 4 heteroatoms selected from O, S and N.

8. The compound according to any one of claims 1 to 4, or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein: R1, R3 and R4 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl, C1-C6 hydroxyalkyl and C2-C6 alkenyl; Preferably, R1, R3 and R4 are each independently selected from the following groups: hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, C1-C3 hydroxyalkyl and -CF3.

9. The compound of claim 1, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein two R2 together with the atoms to which they are attached form a bridged C3-C10 cycloalkyl or a bridged C3-C10 heterocyclyl; Preferably, represent or represent * represents the site of attachment to L2.

10. The compound of claim 1, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein R5 and R6 are each independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl, and C1-C3 hydroxyalkyl; Or, R5 and R6 together with the atoms to which they are attached form 11. The compound of claim 1 or 3 or 4, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein R7 is selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl; Or, two of R7 together with the atoms to which they are attached form a bridged C3-C10 cycloalkyl or C3-C10 bridged heterocyclic group, preferably, represent 12. The compound of claim 1, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein R a and R b Each is independently selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxyl, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl.

13. The compound of claim 1, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein L1 is a bond, O, C1-C3 alkyl, C2-C3 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl, -C3-C6 cycloalkyl-CONH-, -C1-C3 alkyl-C3-C6 cycloalkyl-CONH, -C1-C3 haloalkyl-NH-, -C3-C6-heterocyclic-NH-alkyl, -C3-C 6-cycloalkyl-NH-, -C1-C6-alkyl-C(NH)-NH, -C6-C10-heteroaryl-C1-C6-alkyl, -S(O)2-NH- and -C1-C6-alkyl-S(O)2-NH-.

14. The compound of any one of claims 1 to 4, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein L2 is a bond, O, -C(O), -CONH-, C1-C6 alkyl, C2-C6 alkenyl, -C1-C3 alkyl-NH-, -C2-C3 alkenyl-NH-, C2-C3 alkynyl, C1-C3 alkoxy, C1-C3 alkylthio, C1-C3 haloalkyl and -CON(R c)-; R c Selected from hydrogen, deuterium, halogen, amino, cyano, oxo, hydroxy, C1-C3 alkyl, C1-C3 alkoxy, C1-C3 haloalkyl and C1-C3 hydroxyalkyl.

15. The compound of claim 1, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Ring A is selected from the following groups:

16. The compound of claim 1, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof, wherein Ring B is selected from the group consisting of phenyl, pyridyl, pyrrolyl or pyrimidinyl.

17. The compound of claim 2, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein: yes or, yes or, yes In general formula (XIIB), yes yes 18. The compound of claim 2, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, in the general formula (XIII), R8 is selected from the following groups:

19. The compound of claim 2, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein R9 is R 10 It is C 1-5 Alkyl, C 3-5 Cycloalkyl.

20. The compound of claim 2, or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein L6 is or L6-L2 is 21. The compound of claim 3, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, in the general formula (XXII), X attached to the phenyl group is Cl, and / or CH n X 3-n is CF3; in the general formula (XXIII), CH n X 3-n It's CF3.

22. The compound of claim 4, or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein: is selected from the following groups: yes In general formula (XXVI), yes * represents the site of attachment to L2.

23. The compound of claim 5, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein, in the general formula (XXVII), yes and / or, R 10 yes In the general formula (XXIX), R 10a yes Preferably, In the general formula (XXX), ring D is and Ring D is optionally substituted with (R12)q; In the general formula (XXXI), ring E is and ring E is optionally replaced by (R7) w replace; In the general formula (XXXII), L3 is a C1-C6 alkyl group, preferably, -CH(CH3)-; In the general formula (XXXIII), R1 is H; L3 is C1-C6 alkyl, preferably, -CH2-, -CH(CH3)-; R 15 is a spiroheterocyclic group, preferably, In the general formula (XXXVI), ring C is or thick and C1-C 12 heteroaryl; and Ring C is optionally substituted with (R12)q; Or, in general formula (XXXVI), yes In general formula (XXXVII), yes yes Preferably, In the general formula (XXXVI-A), yes In each general formula, R 10 is a 5-6 membered heterocyclic ring having an unsaturated double bond, containing 1-2 heteroatoms selected from O, N and S, and optionally substituted by deuterium, halogen, amino, cyano, oxo, hydroxy, alkyl, alkoxy, haloalkyl and hydroxyalkyl; R 10 It is preferred that R 10 It is more preferred that * represents the site of attachment to L2.

24. The compound of claim 6, or its tautomer, cis or trans isomer, meso-isomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, solvate or prodrug, wherein R 10b yes Preferably, 25. A compound selected from the group consisting of: or a tautomer, cis or trans isomer, meso racemate, racemate, enantiomer, diastereomer or a mixture thereof, or a pharmaceutically acceptable salt, solvate or prodrug thereof.

26. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1 to 25 or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer or mixture, or a pharmaceutically acceptable salt solvate or prodrug thereof, and a pharmaceutically acceptable carrier.

27. A method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of claims 1 to 25 or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, or a prodrug thereof, or a pharmaceutical composition of claim 26.

28. A method of treating a condition or disease in a subject that is treatable by WRN inhibition, comprising administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 25, or a tautomer, cis or trans isomer, meso, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt or solvate thereof, or a prodrug thereof or a pharmaceutical composition of claim 26.

29. A method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of claims 1 to 25 or its tautomer, cis or trans isomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt solvate or prodrug thereof, or a pharmaceutical composition of claim 26, wherein the cancer is selected from the group consisting of high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR), preferably, high microsatellite instability (MSI-H) or mismatch repair deficiency (dMMR) is selected from colorectal, gastric, prostate, endometrial cancer, adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer and ovarian cancer.

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