Diheterocyclic derivative for inhibiting WRN helicase, pharmaceutical composition containing diheterocyclic derivative and application of diheterocyclic derivative

Abstract

The invention provides a diheterocyclic derivative for inhibiting WRN helicase, a pharmaceutical composition containing the diheterocyclic derivative and application of the diheterocyclic derivative. Specifically, the invention provides a compound as shown in a formula (I), a stereoisomer thereof, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a crystal form thereof, an isotope derivative thereof, a prodrug thereof, a metabolite thereof, a solvate thereof or a hydrate thereof. The invention also provides a composition containing the compound and application of the composition as a WRN helicase inhibitor, and the composition can be used for preparing medicines for treating WRN helicase mediated diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and in particular relates to a biheterocyclic derivative for inhibiting WRN helicase, a pharmaceutical composition containing the biheterocyclic derivative, and application thereof. Background Art

[0002] Microsatellite instability (MSI) refers to the phenomenon that base errors are inserted, deleted, and merged during DNA replication and recombination due to DNA mismatch repair deficiency (dMMR), resulting in changes in microsatellite sequences (Cell 155, 858–868 (2013)). MSI is present in many tumors, especially in colon cancer, gastric cancer, endometrial cancer, and ovarian cancer (JCO Precis Oncol 2017; 2017: 17.00073-17.00073). There is a significant correlation between MSI and immune checkpoint blockade response, and this feature is often used clinically to treat MSI tumors. However, 45-60% of MSI tumors still do not respond to immune checkpoint blockade (Science 357, 409–413, (2017).), so it is urgent to find new treatments for MSI tumors.

[0003] Werner syndrome RecQ helicase (WRN) is a member of the RecQ family of DNA helicases. It can unwind non-canonical secondary DNA structures that may be encountered during replication and recombination. Its mutation can cause chromosomal instability diseases such as Werner syndrome. Studies have shown that WRN is an important synthetic lethal target in MSI tumor cells. In MSI tumor cells, the loss of WRN helicase activity can cause DNA double-strand breaks and promote cell apoptosis and cell cycle arrest (Nature 568, 551–556, 2019). Studies on the WRN-dependent mechanism in MSI tumor cells have shown (Nature 586, 292–298, 2020) that TA-dinucleotide repeat sequences are highly unstable and undergo large-scale amplification in MSI tumor cells. When WRN is lost, this repeat sequence is susceptible to cleavage by the MUS81 nuclease, resulting in a large number of chromosome breaks. This study provides strong support for the development of targeted WRN therapeutics for MSI cancers. The research group of Professor Zhang Lin / Yu Jian from the University of Pittsburgh reported (PNAS.2022;119(51):e2211775119.) the molecular mechanism of WRN deficiency causing apoptosis in microsatellite unstable colorectal cancer cells. In microsatellite unstable colorectal cancer cells, WRN deficiency produces a synthetic lethal effect through the p53 / PUMA-mediated apoptosis signaling pathway, while p53 mutations cause the synthetic lethal effect to be significantly weakened or even disappear. In addition, Dr. Mathew J Garnett of the Wellcome Sanger Institute reported (Cancer Discov.2021Aug;11(8):1923-1937) that WRN gene knockout was performed in 60 experimental models of drug-resistant colorectal cancer, and it was found that more than 90% of the models were dependent on WRN. Targeting the WRN gene is expected to help develop new targeted therapies for the treatment of drug-resistant colorectal cancer. Therefore, the development of WRN inhibitors as a single therapy or in combination with targeted drugs, chemotherapy or immunotherapy for dMMR / MSI tumors has great value and prospects. Summary of the invention

[0004] The invention provides a biheterocyclic derivative for inhibiting WRN helicase, a pharmaceutical composition containing the same and application thereof.

[0005] In a first aspect of the present invention, there is provided a compound represented by formula (I), a stereoisomer thereof, an optical isomer thereof, a pharmaceutically acceptable salt thereof, a crystalline form thereof, an isotopic derivative thereof, a prodrug thereof, a metabolite thereof, a solvate thereof or a hydrate thereof,

[0006]

[0007] in,

[0008] X is selected from CH or N; and when X is CH, the CH may be R 5 or R 6 Substitution (ie, X is CR 5 or CR 6 );

[0009] m, n, e and q are each independently selected from: 0, 1, 2 or 3;

[0010] R 1 Select from the following group: C 3-6 Cycloalkyl, C 4-6 Alkynyl, C 4-10 Cycloalkenyl, C 6 -C 10 Aryl, 5-10 membered heteroaryl, saturated or partially unsaturated 5-10 membered heterocyclic group; wherein said R 1 can be optionally replaced by 0, 1, 2 or 3 R 1a , R 1b , R 1c , R 1d , R 1e , R 1f , R 1g , R i or R h replace;

[0011] R 2 Select from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl;

[0012] R 3 and R 4 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Alkoxy, C 6-10 Alkylamino, C 1-4 Haloalkoxy; R 3 and R 4 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 7 replace;

[0013] R 5 and R 6 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, -SO 2 CF 3 、-SF 5 , C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R 5 and R 6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, each independently optionally replaced by 0, 1, 2 or 3 R 8 replace;

[0014] R 7 and R 8 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, or two R 7 or R 8 co-form = O;

[0015] Cy A Select from the following group:

[0016] Among them, the Cy A Can be optionally replaced by one or more R 12 Substituents are substituted; d is selected from 0, 1, 2 or 3; each R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, cyano substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl, C 1-2 Haloalkoxy substituted C 1-4 Alkyl, or two R 12 Together they form =O; B is selected from O or S;

[0017] Cy B is selected from the group consisting of 6-9 membered heteroaryl; R3 and R 4 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 6-10 Alkylamino, C 1-4 Haloalkoxy, or two R 3 or R 4 Together = O; R 3 and R 4 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 42 replace;

[0018] R 42 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, or two R 42 co-form = O;

[0019] Wherein, the heteroaryl or heterocyclic group includes 1, 2, 3, 4 or 5 heteroatoms selected from the following group as ring atoms: N, O, S or Se; the carbocyclic ring, heterocyclic ring, heterocyclic group may be a monocyclic ring, a fused ring, a spirocyclic ring or a bridged ring structure, which may be saturated or partially unsaturated, but not aromatic;

[0020] R 1a and R 1b Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1b and R 1c Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0021] R 1c and R 1dEach independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino;

[0022] R 1e and R 1f Each independently selected from the group consisting of H, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 alkyl;

[0023] Each R 1g Each independently selected from the group consisting of H, D, hydroxyl, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy;

[0024] R i C 3-6 Cycloalkoxy;

[0025] Each R h Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

[0026] In another preferred embodiment, R 1 Select from the following group:

[0027] (1)C 3-6 Cycloalkyl, the C 3-6 The cycloalkyl group may be optionally substituted with 0, 1, 2 or 3 R 1a or R 1b Replacement; R 1a and R 1b Each independently selected from the group consisting of H, D, oxo, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1b and R 1c Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0028] (2)C 4-6 Alkynyl, C 4-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino;

[0029] (3) R is selected from CH, CD, CR 1e or N; Y, Z, W, U are independently selected from CR 1e R 1f NR 1e or oxygen atom, R 1e and R 1f Each independently selected from the group consisting of H, deuterium, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl; R 1e and R 1f Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0030] (4) R is selected from CH, CD, CR 1e or N; K, F, G are independently selected from CR 1e R 1f NR 1e or oxygen atom, R 1e and R 1fEach independently selected from the group consisting of H, deuterium, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl; R 1e and R 1f Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0031] (5)

[0032] (6) a selenium-containing aromatic heterocycle, wherein the selenium-containing aromatic heterocycle is independently optionally substituted with 0, 1, 2 or 3 R 1g Replacement; R 1g Each independently selected from the group consisting of H, D, hydroxyl, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy;

[0033] (7) R i Selected from C 3-6 Cycloalkoxy or halogen substituted C 3-6 Cycloalkoxy, and Cy C Selected from the following group: phenyl, 5-12 membered heteroaryl, C 4-10 Cycloalkenyl, C 4-10 heterocyclic group; f is selected from 0, 1, 2 or 3; R h Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

[0034] (8) Cy Dis selected from the following group: benzo five-membered heterocycle, benzo five-membered aromatic heterocycle, benzo six-membered heterocycle, benzo six-membered aromatic heterocycle, pyrido five-membered heterocycle, pyrido five-membered aromatic heterocycle, pyrimido five-membered heterocycle, pyrimido five-membered aromatic heterocycle, pyrido six-membered heterocycle, pyrido six-membered aromatic heterocycle, pyrimido six-membered heterocycle, pyrimido six-membered aromatic heterocycle; the heteroatoms of the above heterocycles and aromatic heterocycles are selected from O, N and S; p and t are each independently selected from 0, 1, 2, 3 or 4; R m and R n Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy; R m and R n Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; R m Can be 0, 1, 2, 3 or 4 R m1 Replacement, R n Can be 0, 1, 2, 3 or 4 R n1 Replacement; R m1 and R n1 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkoxy;

[0035] R 2 Select from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl;

[0036] Cy A Select from the following group:

[0037]

[0038] R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, cyano substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl, C 1-2Haloalkoxy substituted C 1-4 Alkyl; B is selected from O or S; d is selected from 0, 1, 2 or 3;

[0039] in represents a covalent bond to the parent nucleus, and represents a covalent bond connected to the carbonyl group; -* represents a covalent bond connected;

[0040] Cy B is selected from the group consisting of 6-9 membered heteroaryl;

[0041] R 3 and R 4 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 6-10 Alkylamino, C 1-4 Haloalkoxy; R 3 and R 4 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 7 replace;

[0042] R 7 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy;

[0043] X is selected from CH or N; and when X is CH, the CH may be R 5 or R 6 Substitution (ie, X is CR 5 or CR 6 );

[0044] R 5 and R 6 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, -SO 2 CF 3 、-SF 5 , C 3-6 Cycloalkyl, C6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R 5 and R 6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, each independently optionally replaced by 0, 1, 2 or 3 R 8 replace;

[0045] R 8 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

[0046] In another preferred embodiment, R 1 Preferably from the following group:

[0047]

[0048]

[0049] In another preferred embodiment, Cy A Select from the following group:

[0050] Among them, R 12 and d are as defined above.

[0051] In another preferred embodiment, Cy A Select from the following group:

[0052]

[0053] in represents a covalent bond to the parent nucleus, and represents a covalent bond to the carbonyl group;

[0054] R 1 Select from the following group:

[0055] (1)C 3-6 Cycloalkyl, C 3-6 The cycloalkyl group is optionally substituted with 0, 1, 2 or 3 R 1a and R 1b Replacement; R 1a and R 1bEach independently selected from the group consisting of H, D, oxo, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1a and R 1b Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0056] (2)C 4-6 Alkynyl, C 2-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino;

[0057] (3) Cycloalkenyl, wherein the cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl is unsubstituted or substituted with 1, 2, 3 or 4, preferably 1 or 2 R 33 and R 34 Replacement; R 33 and R 34 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 33 and R 34 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 35 Replacement; R 35 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-4 Alkoxy, C 1-4 Haloalkoxy;

[0058] (4) heterocyclyl, wherein the heterocyclyl 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 is unbridged or bridged and the bridge is 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or substituted with 1, 2, 3 or 4 R 36 or R 37 Replacement; R 36 and R 37 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 36 and R 37 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 38 Replacement; R 38 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy;

[0059] (5) phenyl, heteroaryl; wherein the heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O, S and Se, 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, wherein the phenyl and heteroaryl are unsubstituted or replaced by 1, 2 or 3 independently selected from R 39 and R 40 Substituents, where R 39 and R 40 are independently selected from H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 39 and R 40 Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-6 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 41 Replacement; R 41 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy;

[0060] Cy B Selected from the group consisting of 6-membered heteroaryl; R 3 and R 4 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 6-10 Alkylamino, C 1-4 Haloalkoxy; R 3 and R 4 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 42 Replacement; R 42 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy;

[0061] R 2 Select from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl;

[0062] R 3 Selected from the following group: halogen, cyano, C 3-6Alkoxy, C 1-4 Haloalkyl;

[0063] X is selected from CH or N; and when X is CH, the CH may be R 5 or R 6 Substitution (ie, X is CR 5 or CR 6 );

[0064] R 5 and R 6 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, -SO 2 CF 3 、-SF 5 , C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R 5 and R 6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, each independently optionally replaced by 0, 1, 2 or 3 R 8 replace;

[0065] R 8 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

[0066] In another preferred embodiment, Cy A Select from the following group:

[0067]

[0068] In another preferred embodiment, Cy B Preferably from the following group:

[0069]

[0070] Among them, R 44 , R 45 and R 46Each independently selected from H, D, halogen, CN, OH, C 1-6 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkylamino, C 2-4 Alkenyl, C 1-4 Halogenated alkenyl, C 2-4 Alkynyl; R as described above 45 and R 46 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 47 Replacement; R 47 Independently selected from the group consisting of H, D, halogen, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated cycloalkyl, C 1-4 Halogenated alkoxy.

[0071] In another preferred embodiment, R 1 Preferably from the following group:

[0072]

[0073]

[0074] In another preferred embodiment, Cy B Select from the following group:

[0075]

[0076] wherein h is selected from 0, 1, 2 or 3; R 44 , R 45 and R 46 Each independently selected from H, D, halogen, CN, OH, C 1-6 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkylamino, C 2-4 Alkenyl, C1-4 Halogenated alkenyl, C 2-4 Alkynyl; R as described above 45 and R 46 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 47 Replacement; R 47 Independently selected from the group consisting of H, D, halogen, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated cycloalkyl, C 1-4 Haloalkoxy;

[0077] R 1 Select from the following group: C 3-6 Cycloalkyl, C 3-6 Cycloalkyl, C 4-6 Alkynyl, cycloalkenyl, heterocyclyl, phenyl, heteroaryl;

[0078] Among them, the C 3-6 The cycloalkyl group is optionally substituted with 0, 1, 2 or 3 R 1a and R 1b Replacement; R 1a and R 1b Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1a and R 1b Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S;

[0079] The C 4-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino;

[0080] The cycloalkenyl group is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl group is unsubstituted or substituted by 1, 2, 3 or 4, preferably 1 or 2 R 33 and R 34 Replacement; R 33 and R 34 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 33 and R 34 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 35 Replacement; R 35 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-4 Alkoxy, C 1-4 Haloalkoxy;

[0081] The heterocyclyl is a 5- or 6-membered fully saturated or partially unsaturated group comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, and wherein the heterocyclyl is unbridged or bridged and the bridge is 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or substituted by 1, 2, 3 or 4 R 36 or R 37 Replacement; R 36 and R 37 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 36 and R 37Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 38 Replacement; R 38 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy;

[0082] The heteroaryl group is a 5- or 6-membered fully unsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O, S and Se, 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, wherein the phenyl and heteroaryl groups are unsubstituted or replaced by 1, 2 or 3 groups independently selected from R 39 and R 40 Substituents, where R 39 and R 40 are independently selected from H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 39 and R 40 Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-6 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 41 Replacement; R 41 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy;

[0083] Cy A Select from the following group:

[0084]

[0085]

[0086] R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, cyano substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl, C 1-2 Haloalkoxy substituted C 1-4 Alkyl; B is selected from O or S;

[0087] in represents a covalent bond to the parent nucleus, and represents a covalent bond to the carbonyl group;

[0088] R 2 Select from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl;

[0089] R 3 Selected from the group consisting of halogen, cyano, C 3-6 Alkoxy, C 1-4 Haloalkyl;

[0090] X is selected from CH or N; and when X is CH, the CH may be R 5 or R 6 Substitution (ie, X is CR 5 or CR 6 );

[0091] R 5 and R 6 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, -SO 2 CF 3 、-SF 5 , C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R 5 and R 6can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, each independently optionally replaced by 0, 1, 2 or 3 R 8 replace;

[0092] R 8 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

[0093] In another preferred embodiment, R 1 Preferably from the following group:

[0094]

[0095]

[0096] In another preferred embodiment, Cy A Select from the following group:

[0097]

[0098] In some embodiments, the compound is selected from the group consisting of:

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109]

[0110] The second aspect of the present invention provides a pharmaceutical composition comprising the compound as described in the first aspect of the present invention, its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates or its hydrates; and a pharmaceutically acceptable carrier.

[0111] In another preferred embodiment, the pharmaceutical composition further comprises other therapeutic agents.

[0112] In another preferred embodiment, the other therapeutic agents are chemotherapeutic drugs, immunotherapeutic drugs, and targeted drugs.

[0113] In another preferred embodiment, the chemotherapy drug is selected from: anastrozole, bicalutamide, bleomycin sulfate, busulfan, busulfan injection, capecitabine, N4-pentyloxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, cytosine arabinoside, cytarabine liposome injection, dacarbazine, actinomycin, daunorubicin hydrochloride, daunorubicin citrate, liposome injection, dexamethasone, docetaxel, doxorubicin hydrochloride, etoposide , fludarabine phosphate, 5-fluorouracil, flutamide, tezacitabine, gemcitabine, hydroxyurea, idarubicin, ifosfamide, irinotecan, L-asparaginase, folinic acid calcium, melphalan, 6-mercaptopurine, methotrexate, mitoxantrone, gemtuzumab ozogamicin, paclitaxel, pentostatin, tamoxifen citrate, teniposide, 6-thioguanine, thiotepa, tirapazamine, topotecan hydrochloride for injection, vinblastine, vincristine, and vinorelbine; preferably 5-fluorouracil (5-FU) and irinotecan.

[0114] In another preferred embodiment, the immunotherapy drug is selected from: PD-1 inhibitors (including small molecules and antibody drugs), PD-L1 inhibitors (including small molecules and antibody drugs), CTLA-4 inhibitors, PD-1 / CTLA-4 dual antibodies, PD-1 / TIGIT dual antibodies, PD-L1 / 4-1BB dual antibodies, PD1 / VEGF-A dual antibodies, PD-1 / LAG-3 dual antibodies;

[0115] Among them, the PD-1 inhibitor is selected from the group consisting of: nivolumab, pembrolizumab, carrelizumab, toripalimab, sintilimab, tislelizumab, penampalimab, sepalizumab, serotonin, dostarlimab, and PDR001;

[0116] Among them, the PD-L1 inhibitor is selected from: durvalumab, atezolizumab, envolizumab, and sugemalimab;

[0117] Wherein, the CTLA-4 inhibitor is selected from ipilimumab;

[0118] Among them, the PD-1 / CTLA-4 dual antibody is selected from cardunilizumab.

[0119] In another preferred embodiment, the targeted drugs include but are not limited to VEGFR inhibitors (lenvatinib), NTRK inhibitors (entrectinib), BRAFV600 inhibitors (dabrafenib), MEK inhibitors (trametinib), EGFR monoclonal antibodies (cetuximab), PARP inhibitors (olaparib, niraparib, fluzoparib and pamiparib, etc.) and ATR inhibitors.

[0120] The third aspect of the present invention provides a method for preparing a pharmaceutical composition, comprising the steps of: mixing a pharmaceutically acceptable carrier with the compound described in the present invention, its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates or its hydrates, to form the pharmaceutical composition as described in the second aspect of the present invention.

[0121] The fourth aspect of the present invention provides a compound as described in the first aspect of the present invention, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystalline form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, or the pharmaceutical composition as described in the second aspect of the present invention for the preparation of a medicament for treating WRN helicase-mediated diseases.

[0122] In another preferred embodiment, the disease is a tumor.

[0123] In another preferred embodiment, the disease is a microsatellite unstable tumor.

[0124] In another preferred embodiment, the tumor is characterized by microsatellite instability-high (MSI-H), or a tumor with defective mismatch repair (dMMR) or "dMMR characteristics";

[0125] Wherein, the microsatellite unstable tumors include, but are not limited to, uterine body tumors, endometrial cancer, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenal cortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, endocervical adenocarcinoma, esophageal cancer, breast cancer, renal clear cell carcinoma and ovarian serous cystic carcinoma;

[0126] Among them, tumors with "defective mismatch repair" (dMMR) or "dMMR characteristics" include but are not limited to lung cancer, breast cancer, kidney cancer, colorectal cancer, ovarian cancer, prostate cancer, upper aerodigestive tract cancer, gastric cancer, endometrial cancer, liver cancer, pancreatic cancer, hematopoietic and lymphoid tissue cancer, skin cancer, thyroid cancer, pleural cancer, autonomic nervous system tumors, soft tissue tumors, rhabdoid sarcoma, melanoma and other tumors.

[0127] In another preferred embodiment, the drug is administered orally, by injection, inhalation or cavity administration.

[0128] In another preferred embodiment, the dosage form of the drug is selected from the group consisting of tablets, capsules, dispersants, suspensions, granules, sprays, gels, sustained-release agents, oral solutions, pills and nanoformulations.

[0129] In a fifth aspect, the present invention provides a method for treating a WRN helicase-mediated disease, comprising the steps of administering to a subject in need of treatment an effective amount of the compound as described in the first aspect, its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystalline forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates or its hydrates, or the pharmaceutical composition as described in the second aspect.

[0130] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0131] After extensive and in-depth research, the inventors discovered for the first time a bicyclic heterocyclic derivative with a novel structure and good WRN helicase inhibitory activity. On this basis, the inventors completed the present invention.

[0132] the term

[0133] Unless otherwise stated, the following terms used in this application (including the specification and claims) have the definitions given below.

[0134] "Alkyl" (alone or as part of another group) refers to a monovalent straight or branched saturated hydrocarbon group consisting only of carbon and hydrogen atoms. Among them, "C1-C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms, preferably C1-C4 alkyl. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl, n-hexyl, octyl, dodecyl, etc. In the present invention, alkyl is also intended to include deuterated alkyl, examples of which include, but are not limited to, CD 3 , CD 2 CD 3 , CD 2 CD 2 CD 3 .

[0135] "Alkoxy" (alone or as part of another group) refers to a radical of the formula -OR or -R'-OR, wherein R is an alkyl radical as defined herein and R' is an alkylene radical. Examples of alkoxy radicals include, but are not limited to, methoxy, ethoxy, isopropoxy, tert-butoxy, -CH 2 O-CH 3 、-CH 2 CH 2 -O-CH 3 、-CH 2 -O-CH 2 CH 3 wait.

[0136] "Halogen" (alone or as part of another group) refers to fluorine, chlorine, bromine or iodine.

[0137] "Haloalkyl" (alone or as part of another group) refers to a group obtained by replacing one or more hydrogen atoms in the alkyl group as described above with the same or different halogen atoms. 1 -C 4 Examples of alkyl and haloalkyl groups include, but are not limited to: -CH 2 Cl, -CH 2 CF 3 、-CH 2 CCl 3 , perfluoroalkyl (e.g., -CF 3 -、-CF 2 CF 3 )wait.

[0138] "Haloalkoxy" (alone or as part of another group) refers to a radical of the formula -OR, wherein R is a haloalkyl radical as defined herein. Examples of haloalkoxy radicals include, but are not limited to, trifluoromethoxy, difluoromethoxy, 2,2,2-trifluoroethoxy, and the like.

[0139] "Cycloalkyl" (alone or as part of another group) refers to a monovalent saturated carbocyclic group consisting of a mono- or bicyclic ring consisting only of carbon and hydrogen atoms, wherein "C3-C8 cycloalkyl" refers to a saturated carbocyclic group containing 3 to 8 carbon atoms, preferably a C3-C6 cycloalkyl. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc. Cycloalkyl may be optionally substituted with one or more substituents, wherein each substituent is independently hydroxyl, alkyl, alkoxy, halogen, haloalkyl, amino, monoalkylamino, or dialkylamino. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

[0140] "Cycloalkyloxy" (alone or as part of another group) refers to a radical of the formula -OR, wherein R is a cycloalkyl radical as defined herein. Exemplary cycloalkyloxy radicals include cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0141] "Heterocyclyl" (alone or as part of another group) refers to a fully saturated or partially unsaturated cyclic group (including but not limited to 3-7 membered monocyclic, 6-11 membered bicyclic, or 8-16 membered tricyclic system), wherein at least one heteroatom is present in a ring having at least one carbon atom. Each heterocyclic ring containing heteroatoms carries 1, 2, 3 or 4 heteroatoms selected from nitrogen atoms, oxygen atoms or sulfur atoms, wherein the nitrogen atoms or sulfur atoms may be oxidized and the nitrogen atoms may be quaternized. Heterocycloalkane (base) refers to a fully saturated heterocyclic ring (base). The heterocyclic group may be attached to any heteroatom or carbon atom residue of the ring or ring system molecule. Among them, "3-8 membered heterocyclyl" refers to a group having 3-8 ring members. Typical monocyclic heterocycles include, but are not limited to, azetidinyl, pyrrolidinyl, oxetanyl, pyrazolinyl, imidazolinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, hexahydroazepinyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxanyl and tetrahydro-1,1-dioxythiophene, etc. Polycyclic heterocyclic groups include spirocyclic, condensed and bridged heterocyclic groups; wherein the spirocyclic, condensed and bridged heterocyclic groups are optionally connected to other groups through single bonds, or further connected to other cycloalkyl, heterocyclic, aryl and heteroaryl groups through any two or more atoms on the ring.

[0142] "Aryl" (alone or as part of other groups) refers to an aromatic cyclic hydrocarbon compound group with 1-5 rings, especially monocyclic and bicyclic groups. Where there are two or more aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group can be connected by a single bond (such as biphenyl) or fused (such as naphthalene, anthracene, etc.). Among them, "C6-C12 aryl" refers to an aromatic cyclic hydrocarbon compound group containing 6, 7, 8, 9, 10, 11 or 12 ring carbon atoms. Examples of aryl (especially monocyclic and bicyclic groups) include, but are not limited to, phenyl, biphenyl or naphthyl. Aryl can be fused to heterocyclic groups through a single bond or any two adjacent ring C atoms, for example: benzotetrahydrofuranyl, chromanyl, benzodioxanyl, wait.

[0143] "Heteroaryl" (alone or as part of another group) refers to an aromatic group with heteroatoms in the backbone, wherein "5-12 membered heterocyclyl" refers to a monocyclic, bicyclic or tricyclic group of 5 to 12 ring atoms, which contains at least 1 (such as 1, 2 or 3) ring heteroatoms selected from N, O or S, and the remaining ring atoms are C aromatic rings, it should be clear that the point of attachment of the heteroaryl group should be located on the aromatic ring. Heteroaryl preferably has 5-8 ring atoms (5-8 members), more preferably 5-6 ring atoms (5-6 members). Examples of heteroaryl groups include, but are not limited to, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, furanyl, pyranyl, pyridinyl, pyrrolyl, pyrazolyl, pyrimidinyl, quinolyl, isoquinolyl, benzofuranyl, benzothiophenyl, benzothiopyranyl, benzimidazolyl, benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranyl, indolyl, isoindolyl, triazolyl, triazine, quinoxalinyl, purinyl, quinazolinyl, quinolizinyl, naphthyridinyl, pteridinyl, carbazolyl, aza Base, diazepine Base, acridinium, etc.

[0144] "Polysubstituted" means involving two or more substitutions.

[0145] In the present invention, the alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl and other groups include substituted alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl and the like unless otherwise specified, and the substituents include (but are not limited to): halogen, hydroxyl, cyano, alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl and the like.

[0146] "Deuterated compound" refers to a compound in which one or more hydrogen atoms (H) are replaced by deuterium atoms (D). 1-4 Deuterated alkyl refers to a compound in which at least three hydrogen atoms (H) or more hydrogen atoms (H) are replaced by deuterium atoms (D), and the substituents include (but are not limited to) deuterated methyl, deuterated ethyl, and the like.

[0147] Active ingredients

[0148] As used herein, "compound of the present invention" refers to the compound represented by formula (I), and also includes its stereoisomers, optical isomers, pharmaceutically acceptable salts, crystalline forms, isotopic derivatives, prodrugs, metabolites, solvates or hydrates thereof.

[0149] Unless otherwise specified, the structural formulas described in the present invention are intended to include all stereoisomers (such as cis-trans isomers, enantiomers, diastereomers and conformational isomers): R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, cis-trans isomers of cycloalkanes, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers or conformational isomers are all within the scope of the present invention.

[0150] The compounds of the present invention may contain cis-trans isomers, one or more chiral carbon atoms, and thus may produce stereoisomeric forms of cis-trans isomers, chiral isomers, enantiomers, diastereomers and other combinations. Cis-trans isomerism refers to the diastereoisomerism phenomenon that occurs due to the restriction of free rotation in the compound molecule, which causes the different arrangements of various groups in space. This restriction is generally caused by the presence of functional groups that cannot rotate freely in the structure of organic compounds, such as C=C double bonds, C=N double bonds, C=S double bonds, N=N double bonds, heterocycles or cycloalkanes. Organic molecules containing such isomers, such as olefins, azo compounds, alicyclic hydrocarbons, etc., are considered cis-trans isomers. The cis form refers to the same ligands in adjacent positions, generally represented by "cis" or "cis-"; the trans form refers to the same ligands in diagonal positions, generally represented by "trans" or "trans-". Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. The present invention is intended to include all possible isomers, as well as racemates and optically pure forms thereof. The compounds of the present invention may be prepared using racemates, cis-trans isomers, chiral isomers, diastereomers or enantiomers as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0151] Conventional techniques for preparing / isolating individual optical isomers (i.e., cis-trans isomers and chiral isomers) include chiral synthesis from appropriate cis-trans precursors or optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high performance liquid chromatography, see, for example, Gerald Gübitz and Martin G. Schmid (Eds.), Chiral Separations, Methods and Protocols, Methods in Molecular Biology, Vol. 243, 2004; A. M. Stalcup, Chiral Separations, Annu. Rev. Anal. Chem. 3:341-63, 2010; Fumiss et al. (eds.), VOGEL'S ENCYCLOPEDIA OF PRACTICAL ORGANIC CHEMISTRY 5.sup.THED., Longman Scientific and Technical Ltd., Essex, 1991, 809-816; Heller, Acc. Chem. Res. 1990, 23, 128.

[0152] If a synthesis of a specific stereoisomer of the compound of the present invention is to be designed, it can be prepared by asymmetric synthesis, or derivatized with a chiral auxiliary, the resulting stereo mixture can be separated, and then the chiral auxiliary can be removed to obtain a pure cis-trans monomer, a chiral monomer or a mixed stereo monomer. If the molecule contains a cis-trans isomeric center, it can be purified by column chromatography (normal phase silica gel column or reverse phase high performance liquid preparation) to obtain a pure cis product or trans product; in addition, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, it can be used with a suitable optically active acid or base to form a diastereomeric salt, and then separated by conventional means such as fractional crystallization or chromatography, and then a pure enantiomer is obtained.

[0153] The present invention also includes isotope-labeled compounds (i.e., isotope derivatives) that are equivalent to the original compounds disclosed herein. However, in practice, it is common for one or more atoms to be replaced by atoms with different atomic weights or mass numbers. Examples of isotopes in the isotope derivatives of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes, such as 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F and 36Cl, respectively. The isotope derivatives of the compounds of the present invention are all within the scope of protection of the present invention. Herein, 3H-labeled compounds and 14C-labeled compounds are useful in tissue distribution experiments of drugs and substrates. The preparation and detection of tritium (i.e., 3H) and carbon-14 (i.e., 14C) labeled compounds are relatively easy and are the first choice among isotopes. In addition, heavier isotope substitutions such as deuterium, i.e., 2H, may be preferred in certain cases due to their good metabolic stability, which may have advantages in certain therapeutics, such as increasing half-life in vivo or reducing dosage. Isotopically labeled compounds can be prepared by general methods by replacing readily available isotopically labeled reagents with non-isotopic reagents using the protocols disclosed in the examples.

[0154] As used herein, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0155] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobromides, sulfates, nitrates, phosphates, and the like; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetates, trifluoroacetates, propionates, caproates, caprylates, decanoates, undecylenates, glycolates, gluconates, lactates, sebacates, adipates, glutarates, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamate, pyroglutamate, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginate, ascorbate, salicylates, 4-aminosalicylates, naphthalene disulfonates, and the like. These salts may be prepared by methods known in the art.

[0156] "Pharmaceutically acceptable base addition salt" refers to a salt formed with an inorganic base or an organic base that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. The salt derived from organic base includes but is not limited to the following salt: primary amines, secondary amines and tertiary amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. These salts can be prepared by methods known to the profession.

[0157] The metabolites of the compound of formula (I) and its pharmaceutically acceptable salts, and the prodrugs that can be converted into the compound of formula (I) and its pharmaceutically acceptable salts in vivo are also included in the protection scope of the present invention.

[0158] As used herein, the term "solvate" refers to a complex in which the compound represented by formula (I) is coordinated with solvent molecules to form a specific ratio.

[0159] As used herein, the term "hydrate" refers to a complex in which the compound represented by formula (I) is coordinated with water molecules to form a specific ratio.

[0160] As described herein, the compounds of the present invention can be substituted with any number of substituents or functional groups to expand their scope. In general, the term "substituted" refers to replacing hydrogen radicals with specified structural substituents. When multiple positions in a specific structure are substituted with multiple specific substituents, the substituents can be the same or different at each position. The term "substituted" used herein includes all allowed organic group substitutions. In a broad sense, allowed substituents include non-cyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic groups. Herein, heteroatom nitrogen can have hydrogen substituents or any allowed organic groups described above to supplement its valence. In addition, the present invention is not intended to limit the allowed substitution of organic groups in any way. The present invention believes that the combination of substituents and variable groups is good in the treatment of diseases in the form of stable compounds. The term "stable" here refers to a stable compound that is tested for a long enough time to maintain the integrity of the compound structure, preferably for a long enough time, and is used for the above purposes herein.

[0161] Pharmaceutical compositions and methods of administration

[0162] Since the compounds of the present invention can inhibit WRN helicase and are used to treat diseases such as tumors, the compounds of the present invention and their stereoisomers, optical isomers, pharmaceutically acceptable salts, crystal forms, isotopic derivatives, prodrugs, metabolites, solvates or hydrates thereof, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to prevent and / or treat (stabilize, alleviate or cure) WRN helicase-related diseases, preferably tumors; more preferably, microsatellite unstable tumors; further preferably, tumors characterized by high microsatellite instability (MSI-H) or having "defective mismatch repair deficiency (dMMR)" or "dMMR characteristics".

[0163] Microsatellite unstable tumors are particularly present in colorectal cancer, gastric cancer and endometrial cancer, and also in adrenocortical carcinoma, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, prostate cancer and ovarian cancer. Microsatellite unstable tumors include but are not limited to uterine body tumors, endometrial cancer, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenocortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, endocervical adenocarcinoma, esophageal cancer, breast cancer, kidney renal clear cell carcinoma and ovarian serous cystic carcinoma.

[0164] Tumors with "defective mismatch repair" (dMMR) or a "dMMR signature" include cancer types associated with documented MLH1, PMS2, MSH2, MSH3, MSH6, MLH3, and PMS1 mutations or epigenetic silencing, microsatellite fragile sites, or other gene inactivation mechanisms, including but not limited to lung cancer, breast cancer, kidney cancer, colorectal cancer, ovarian cancer, prostate cancer, upper aerodigestive tract cancer, gastric cancer, endometrial cancer, liver cancer, pancreatic cancer, hematopoietic and lymphoid tissue cancer, skin cancer, thyroid cancer, pleural cancer, autonomic nervous system tumors, soft tissue tumors, rhabdoid sarcoma, melanoma, and other tumors.

[0165] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention and a pharmaceutically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 10-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0166] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0167] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administration methods include, but are not limited to, oral administration and parenteral administration (intravenous, intramuscular or subcutaneous administration).

[0168] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0169] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0170] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0171] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0172] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0173] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0174] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable therapeutic agents. Drugs co-administered with WRN inhibitors include but are not limited to chemotherapeutic drugs, immunotherapy drugs, and targeted drugs. When co-administered, the pharmaceutical composition also includes one or more (2, 3, 4 or more) other pharmaceutically acceptable therapeutic agents. One or more (2, 3, 4 or more) of the other pharmaceutically acceptable therapeutic agents can be used simultaneously with the compounds of the present invention to treat microsatellite instability (MSI) type or DNA mismatch repair defective (dMMR) tumors.

[0175] Chemotherapy drugs

[0176] Chemotherapy drugs include, but are not limited to, anastrozole, bicalutamide, bleomycin sulfate, busulfan, busulfan injection, capecitabine, N4-pentyloxycarbonyl-5-deoxy-5-fluorocytidine, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, cyclophosphamide, cytarabine, cytosine arabinoside, cytarabine liposome injection, dacarbazine, actinomycin, daunorubicin hydrochloride, daunorubicin citrate, liposome injection, dexamethasone, docetaxel, doxorubicin hydrochloride, etoposide, fluorophosphate dabrabine, 5-fluorouracil, flutamide, tezacitabine, gemcitabine, hydroxyurea, idarubicin, ifosfamide, irinotecan, L-asparaginase, leucovorin, melphalan, 6-mercaptopurine, methotrexate, mitoxantrone, gemtuzumab ozogamicin, paclitaxel, pentostatin, tamoxifen citrate, teniposide, 6-thioguanine, thiotepa, tirapazamine, topotecan hydrochloride for injection, vinblastine, vincristine and vinorelbine; especially 5-fluorouracil (5-FU) and irinotecan.

[0177] Immunotherapy drugs

[0178] Immunotherapy drugs include but are not limited to PD-1 inhibitors (including small molecules and antibody drugs), PD-L1 inhibitors (including small molecules and antibody drugs), CTLA-4 inhibitors, PD-1 / CTLA-4 dual antibodies, PD-1 / TIGIT dual antibodies, PD-L1 / 4-1BB dual antibodies, PD1 / VEGF-A dual antibodies, PD-1 / LAG-3 dual antibodies and other immunotherapy drugs.

[0179] Among them, PD-1 inhibitors include but are not limited to nivolumab, pembrolizumab, carrelizumab, toripalimab, sintilimab, tilelizumab, penampalimab, sepalizumab, slulizumab, dostarlimab and PDR001.

[0180] PD-L1 inhibitors include, but are not limited to, durvalumab, atezolizumab, envoiriumab, and sugemalimab.

[0181] CTLA-4 inhibitors include, but are not limited to, ipilimumab.

[0182] PD-1 / CTLA-4 dual antibodies include but are not limited to cardunilimab.

[0183] Targeted drugs

[0184] Targeted drugs include but are not limited to VEGFR inhibitors (lenvatinib), NTRK inhibitors (entrectinib), BRAFV600 inhibitors (dabrafenib), MEK inhibitors (trametinib), EGFR monoclonal antibodies (cetuximab), PARP inhibitors (olaparib, niraparib, fluzoparib and pamiparib, etc.) and ATR inhibitors.

[0185] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1-2000 mg, preferably 20-500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0186] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually carried out under conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts. The compounds of the present invention are all determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 The solvents for NMR measurement are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS).

[0187] The abbreviations of the present invention are defined as follows:

[0188] M: molar concentration, such as 1M hydrochloric acid means 1 mol / L hydrochloric acid solution

[0189] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate

[0190] T3P: 1-propylphosphoric anhydride

[0191] DIPEA: also written as DIEA, diisopropylethylamine, also known as N,N-diisopropylethylamine

[0192] EA: Ethyl acetate

[0193] TFA: trifluoroacetic acid

[0194] DMF: N,N-dimethylformamide

[0195] THF: Tetrahydrofuran

[0196] PE: Petroleum ether

[0197] LC-MS: Liquid chromatography-mass spectrometry

[0198] DMSO: dimethyl sulfoxide

[0199] CbzCl: benzyl chloroformate

[0200] Xantphos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0201] TLC: Thin layer chromatography

[0202] IC 50 : Half inhibitory concentration refers to the concentration at which half of the maximum inhibitory effect is achieved.

[0203] Unless indicated to the contrary, compounds exemplified herein are named and numbered using ChemBioDraw Ultra 14.0.

[0204] Preparative HPLC separation conditions: Preparative column: Agilent 10Prep-C18 250*21.2 mm, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 45%-55% acetonitrile, 30 mL / min.

[0205] Intermediate 8b: 2-Bromo-N-(2-methoxy-4-(trifluoromethyl)phenyl)acetamide

[0206]

[0207] 2-Methoxy-4-(trifluoromethyl)aniline (8b-1) (200 mg, 1.05 mmol) and 4-dimethylaminopyridine (128 mg, 1.05 mmol) were dissolved in dry dichloromethane (2 mL), cooled to about 0°C, and bromoacetyl bromide (252 mg, 1.25 mmol) was added dropwise. After the addition, the mixture was returned to room temperature and reacted for 18 h. The reaction solution was concentrated and the crude product was flash separated (PE: EA = 20:1) to obtain compound 8b (320 mg) as a white solid.

[0208] MS (ESI) m / z 312 [M+H] + .

[0209] Intermediate 16a: 4-Hydroxy-1,5-dimethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0210]

[0211] First step 5-methyl-2H-1,3-oxazine-2,6(3H)-dione 16a-2

[0212] At room temperature, compound 16a-1 (5.6 g, 50 mmol) was dissolved in THF (10 mL) and TMSN was added. 3(6.0 g 52 mmol), in N 2 The mixture was stirred at 70°C for 2 hours. After the reaction was completed as monitored by TLC, the mixture was cooled to room temperature and the organic phase was concentrated under reduced pressure to obtain a crude product 16a-2 (1.2 g).

[0213] MS (ESI) m / z 128.10 [M+H] + .

[0214] Step 2 3,5-dimethyl-2H-1,3-oxazine-2,6(3H)-dione 16a-3

[0215] Compound 16a-2 (1.0 g, 7.8 mmol) was dissolved in acetonitrile (10 mL) and K 2 CO 3 (1.3 g, 9.45 mmol), then MeI (1.88 g, 13.4 mmol) was added. The mixture was reacted at 70°C for 16 h. After the reaction was completed by TLC monitoring, ethyl acetate (30 mL) was added and the insoluble solid was removed by filtration. The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (PE / EtOAc=2 / 1) to obtain compound 16a-3 (320 mg).

[0216] MS (ESI) m / z 142.10 [M+H] + .

[0217] Step 3 4-Hydroxy-1,5-dimethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid ethyl ester 16a-4

[0218] Dissolve diethyl malonate (312 mg, 1.95 mmol) in anhydrous THF (20 mL), add NaH (78 mg, 60% in mineral oil) under ice bath, and react at room temperature for 20 min. Add dropwise a THF solution of compound 16a-3 (250 mg of 16a-3 dissolved in 1 mL of THF). React at room temperature for 2 hours. After the reaction is completed by TLC monitoring, add saturated NH 4 Cl solution (20 mL), extracted with ethyl acetate (30 mL × 3), combined the organic phases and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (PE / EtOAc=2 / 1) to obtain compound 16a-4 (130 mg).

[0219] MS (ESI) m / z 212.10 [M+H] + .

[0220] Step 4: 4-Hydroxy-1,5-dimethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid 16a

[0221] Compound 16a-4 (130 mg, 0.61 mmol) was dissolved in HBr (2 mL, 33 wt.% in HOAc), and then reacted at 70°C for 3 h. The mixture was concentrated under reduced pressure to obtain a crude product, Compound 16a (130 mg).

[0222] MS (ESI) m / z 182.20 [MH] - .

[0223] Intermediate 23a 6-(Benzyloxy)-7-methylpyrazolo[1,5-a]pyrimidine-5-carboxylic acid

[0224]

[0225] The first step is diethyl 2-(benzyloxy)-3-hydroxyfumarate 23a-1

[0226] Ethyl 2-(benzyloxy)acetate (19a) (70 g, 360.66 mmol) and diethyl oxalate (63.25 g, 432.79 mmol) were dissolved in tetrahydrofuran (700 mL), the reaction solution was cooled to -78 ° C, 2.0 M lithium diisopropylamide (216.4 ml, 432.79 mmol) was slowly added dropwise, and the reaction was carried out at -78 ° C for 1 h. Saturated aqueous ammonium chloride solution (300 mL) was added to quench the reaction, and the pH was adjusted to 6 with 85% aqueous phosphoric acid solution, and extracted with ethyl acetate (300 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 23a-1 (106 g) as an orange-yellow oil.

[0227] MS (ESI) m / z 295.3 [M+H] + .

[0228] Step 2 6-(Benzyloxy)-7-hydroxypyrazolo[1,5-a]pyrimidine-5-carboxylic acid ethyl ester 23a-2

[0229] 2H-pyrazole-3-amine (3 g, 36.123 mmol) was dissolved in DMF (30 mL), and compound 23a-1 (15.94 g, 54.184 mmol) and cesium carbonate (17.65 g, 54.164 mmol) were added in sequence, and the mixture was reacted at 110°C for 18 h. After cooling to room temperature, ice water (30 mL) was added, and 2.0 M hydrochloric acid solution was added dropwise to adjust the pH to 5-6, and the mixture was extracted with dichloromethane (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by column chromatography (PE: EA = 1:2) to obtain compound 23a-2 (1 g) as a reddish brown oil.

[0230] MS (ESI) m / z 314.3 [M+H] + .

[0231] Step 3 6-(Benzyloxy)-7-chloropyrazolo[1,5-a]pyrimidine-5-carboxylic acid ethyl ester 23a-3

[0232] Compound 23a-2 and N,N-diethylaniline (47.69 mg, 0.3198 mmol) were dissolved in phosphorus oxychloride (1.47 g, 9.5814 mmol) and reacted at 120°C for 3 h. The mixture was concentrated under reduced pressure and the crude product was separated by column chromatography (PE:EA=4:1) to obtain compound 23a-3 (420 mg) as a light yellow solid.

[0233] MS (ESI) m / z 332.3 [M+H] + .

[0234] Step 4: 6-(Benzyloxy)-7-methylpyrazolo[1,5-a]pyrimidine-5-carboxylic acid ethyl ester 23a-4

[0235] Compound 23a-3 was dissolved in DMSO (5 mL), and tricyclohexylphosphine (71.23 mg, 0.254 mmol), tris(dibenzylideneacetone)palladium (116.30 mg, 0.127 mmol) and cesium carbonate (826.93 mg, 2.538 mmol) were added in sequence. The atmosphere was replaced with nitrogen three times, and the reaction was carried out at 130°C for 3 h. The mixture was cooled to room temperature, diluted with water (5 mL), extracted with ethyl acetate (5 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was separated by column chromatography (PE:EA=1:1) to obtain compound 23a-4 (200 mg) as a light yellow solid.

[0236] MS (ESI) m / z 312.0 [M+H] + .

[0237] Step 5: 6-(Benzyloxy)-7-methylpyrazolo[1,5-a]pyrimidine-5-carboxylic acid 23a

[0238] Compound 23a-4 (200 mg, 0.643 mol) was dissolved in tetrahydrofuran (2 mL) and water (2 mL), and lithium hydroxide (46.19 mg, 1.928 mmol) was added, and the mixture was reacted at room temperature for 2 h. The reaction solution was diluted with water (2 mL), and the pH was adjusted to 6 with 2.0 M dilute hydrochloric acid, and extracted with dichloromethane (5 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 23a (160 mg) as a light yellow solid.

[0239] MS (ESI) m / z 284 [M+H] + .

[0240] Intermediate 28a 4,4,5,5-tetramethyl-2-(4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-1,3,2-dioxaborolane

[0241]

[0242] First step 4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]28a-2

[0243] The synthesis of (1-(thiophen-2-yl)cyclopropyl)methanol (28a-1) refers to patent WO2022 / 206706. Compound (28a-1) (1.3 g, 8.44 mmol) was dissolved in dry acetonitrile (100 mL), and paraformaldehyde (329 mg, 10.96 mmol) and indium trichloride (187 mg, 0.84 mmol) were added in sequence. Under nitrogen protection, the temperature was raised to 70 ° C for reaction for 2 h, cooled to room temperature, and the reaction solution was concentrated under reduced pressure. The crude product was purified by column chromatography (petroleum ether) to obtain compound 28a-2 (600 mg) as a colorless oil.

[0244] Step 2 4,4,5,5-Tetramethyl-2-(4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-1,3,2-dioxaborolane 28a

[0245] Under nitrogen protection, compound 28a-2 (600 mg, 3.61 mmol) was dissolved in dry tetrahydrofuran (10 mL), cooled to -70 ° C, 2.5 M n-butyl lithium n-hexane solution (1.52 mL, 3.8 mmol) was added dropwise, and the temperature was restored to room temperature for 1 h. The reaction solution was cooled to -70 ° C, 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (740 mg, 3.98 mmol) was added dropwise, and the temperature was raised to room temperature for 2 h. Saturated ammonium chloride solution (30 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and concentrated, and the crude product was separated and purified by column chromatography (PE / EA=3 / 2) to obtain compound 28a (520 mg) as a yellow oil.

[0246] MS (ESI) m / z 293 [M+H] + .

[0247] Intermediate 29a 2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl-5,5-d2)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0248]

[0249] The first step is 2-(thiophen-3-yl)ethyl-1,1-d 2 -1-ol 29a-2

[0250] LiAlD 4 (1.0 g, 25.61 mmol) was added to a single-mouth bottle, THF (20 mL) was added at 0°C, stirred for 10 min, 20 mL of THF solution of compound 29a-1 (4.0 g, 25.61 mmol) was slowly added, and the mixture was reacted at 0°C for 1 h. The mixture was heated to room temperature and stirred for 1 h. The reaction solution was cooled to 0°C, potassium sodium tartrate was added until no obvious gas was released, stirred for 1 h, petroleum ether (50 mL) was added, stirred and filtered, and the filtrate was concentrated to obtain compound 29a-2 (2.5 g), a yellow oil.

[0251] 1 H NMR (400 MHz, CDCl 3 )δ7.29(dd,J=4.9,3.0Hz,1H),7.05(d,J=2.2Hz,1H),6.98(dd,J=4.9,0.9Hz,1H),2.89(s,2H).

[0252] Step 2 4,7-dihydro-5H-thieno[2,3-c]pyran-5,5-d 2 29a-3

[0253] Compound 29a-2 (2.5 g, 19.20 mmol) was dissolved in acetonitrile, and paraformaldehyde (724 mg, 24.96 mmol) and InCl were added. 3 (425 mg, 1.92 mmol), N 2 Replace three times, react at 85°C for 2.5h. The reaction solution was cooled to room temperature, water (200mL) was added, and extracted with a mixed solvent of PE / TBME (PE / TBME=1 / 1, 70mL x 2), the organic phases were combined, concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether / DCM=10 / 1 to 2 / 1) to obtain compound 29a-3 (1.7g), a yellow oil.

[0254] 1 H NMR (400 MHz, CDCl 3 )δ7.14(d,J=5.0Hz,1H),6.82(d,J=5.0Hz,1H),4.82(s,2H),2.74(s,2H).

[0255] Step 3 2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl-5,5-d 2 )-4,4,5,5-Tetramethyl-1,3,2-dioxaborolane 29a

[0256] Compound 29a-3 (1.7 g, 11.95 mmol) was dissolved in THF (20 mL), cooled to -70 °C, and n-BuLi (2.5 M in THF, 5.2 mL, 13.15 mmol) was added. Stir at -70 °C for 15 min, warm to room temperature, and after about 1 h, the reaction solution changed from yellow-green to brown-green. The reaction solution was cooled to -70 °C, and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.45 g, 13.15 mmol) was added. The reaction solution was slowly warmed to room temperature and reacted overnight. The reaction solution was cooled to -70 °C, and saturated NH 4 The reaction was quenched with aqueous Cl solution (20 mL), extracted with ethyl acetate (25 mL x 2), and the organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to give compound 29a (2.1 g) as a yellow solid.

[0257] 1 H NMR (400 MHz, CDCl 3 )δ7.36(s,1H),4.85(t,J=1.4Hz,2H),4.16-4.07(m,1H),3.94-3.84(m,1H),2.75(s,2H),1.33(s,12H).

[0258] Intermediate 30a Trimethyl((3-methyloxetan-3-yl)ethynyl)silane

[0259]

[0260] The first step is 3-methyloxetane-3-carboxaldehyde 30a-2

[0261] Pyridinium chlorochromate (10.1 g, 25.61 mmol) and silica gel (100-200 mesh, 10.0 g) were dissolved in DCM (100 mL), and compound 30a-1 (4.0 g, 39.16 mmol) was added at 0°C, and the mixture was reacted at room temperature overnight. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain compound 30a-2 (2.8 g) as a colorless oil.

[0262] 1 H NMR (400 MHz, CDCl 3 )δ9.95(s,1H),4.87(d,J=6.4Hz,2H),4.50(d,J=6.4Hz,2H),1.48(s,3H).

[0263] Step 2 3-(2,2-dibromovinyl)-3-methyloxetane 30a-3

[0264] Dissolve triphenylphosphine (10.48 g, 39.95 mmol) in DCM (50.0 mL), cool to 0 °C, and add CBr 4 (6.62 g, 19.98 mmol), react at 0°C for 0.5 h, then add compound 30a-2 (1.0 g, 9.99 mmol), warm to room temperature and react for 1 h. After the reaction is completed, add petroleum ether (150 mL), stir for 0.5 h, a large amount of solid precipitates, filter. Concentrate the filtrate under reduced pressure, and purify the crude product by column chromatography to obtain compound 30a-3 (1.7 g), a light yellow oil.

[0265] 1 H NMR (400 MHz, CDCl 3 )δ6.69(s,1H),4.81(d,J=5.8Hz,2H),4.38(d,J=6.0Hz,2H),1.60(s,3H).

[0266] Step 3: Trimethyl((3-methyloxetane-3-yl)ethynyl)silane 30a

[0267] Compound 30a-3 (1.0 g, 3.91 mmol) was dissolved in anhydrous THF (15 mL). The reaction solution was cooled to -70 °C, n-BuLi (2.5 M in THF, 3.6 mL, 8.99 mmol) was added, and the mixture was reacted at -70 °C for 1 h. Then TMSCl (1.49 g, 13.68 mmol) was added at -70 °C, and the mixture was reacted at -70 °C for 0.5 h, and then the mixture was heated to room temperature for 0.5 h. The reaction solution was concentrated under reduced pressure, petroleum ether (15 mL) was added, and the mixture was filtered through silica gel. The filtrate was concentrated to obtain compound 30a (590 mg) as a light yellow oil.

[0268] 1 H NMR (400 MHz, CDCl 3 )δ4.82(d,J=5.3Hz,2H),4.37(d,J=5.4Hz,2H),1.61(s,3H),0.18-0.15(s,9H).

[0269] Intermediate 37a 4,4,5,5-tetramethyl-2-(5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-1,3,2-dioxaborolane

[0270]

[0271] First step 5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]37a-2

[0272] The synthesis of compound 37a-1 refers to patent PCT / CN / 2022 / 083485. Compound 37a-1 (1.9 g, 12.33 mmol), paraformaldehyde (0.4 g, 13.33 mmol) and indium trichloride (0.13 g, 0.58 mmol) were dissolved in acetonitrile (20 mL), purged with nitrogen, reacted at 45 ° C for 0.5 hours, then heated to 65 ° C for 0.5 hours, and then heated to 85 ° C for 2 hours, cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate, combined organic phases, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography to obtain compound 37a-2 (0.8 g) as an oil.

[0273] 1 H NMR (400 MHz, CDCl 3 )δ7.11(dd,J=5.1,0.9Hz,1H),6.48(d,J=5.1Hz,1H),4.92(s,2H),3.70(s,2H),0.99–0.94(m,2H),0.93–0.87(m,2H).

[0274] Step 2 4,4,5,5-Tetramethyl-2-(5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-1,3,2-dioxaborolane 37a

[0275] Compound 37a-2 (0.8 g, 4.81 mmol) was dissolved in THF (10 mL), the reaction solution was cooled to -78 ° C, 2.5 M n-butyl lithium n-hexane solution (1.9 mL, 5.0 mmol) was added dropwise, the reaction was continued for 10 minutes, the temperature was slowly raised to room temperature for 0.5 hour, the temperature was again lowered to -78 ° C, isopropanol pinacol borate (0.98 g, 5.26 mmol) was added dropwise, the reaction was continued for 10 minutes, water (20 mL) was added at low temperature to quench the reaction, the reaction was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography to obtain compound 37a (0.5 g) as a white solid.

[0276] MS (ESI) m / z 293.1 [M+H] +

[0277] Intermediate 41a tert-Butyl 2-(methoxymethyl)piperazine-1-carboxylate

[0278]

[0279] The first step is 1-tert-butyl-2-hydroxymethylpiperazine-1,4-dicarboxylic acid-4-benzyl ester 41a-2

[0280] Compound 41a-1 (5.0 g, 23.12 mmol) was dissolved in THF (100 mL) and water (20 mL), and NaHCO 3 (7.77 g, 92.47 mmol) and CbzCl (5.9 g, 34.68 mmol). Warm to room temperature and react for 12 h. Concentrate under reduced pressure, add EtOAc (150 mL) and water (50 mL). Separate the organic phase, concentrate under reduced pressure to obtain a crude product, and purify the crude product by column chromatography to obtain compound 41a-2 (9.7 g), a yellow oil.

[0281] MS (ESI) m / z 251.1 [M-Boc+H] +

[0282] Step 2 1-tert-butyl-2-(methoxymethyl)piperazine-1,4-dicarboxylic acid 4-benzyl ester 41a-3

[0283] Compound 41a-2 (9.2 g, 26.25 mmol) was dissolved in THF (100 mL), cooled to -30 °C, added with NaH (2.1 g, 52.51 mmol, 60% wt), stirred for 10 min, and added with CH 3 I (11.18 g, 5.0 mL, 78.76 mmol), warm to room temperature and react overnight. The reaction solution was cooled to 0°C and saturated NH 4 Cl aqueous solution (20 mL) was added until no obvious gas was generated, water (300 mL) was added, and EtOAc was extracted (100 mL x 2). The organic phases were combined and concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain compound 41a-3 (5.5 g) as a colorless oil.

[0284] MS (ESI) m / z 387.1 [M+Na] +

[0285] Step 3 tert-Butyl 2-(methoxymethyl)piperazine-1-carboxylate 41a

[0286] Compound 41a-3 (5.5 g, 15.09 mmol) was dissolved in THF (100 mL), and 10% Pd / C (1.0 g, 844.37 μmol) was added. The reaction mixture was deoxygenated three times by displacement, and the reaction mixture was reacted at room temperature overnight. The reaction mixture was filtered through diatomaceous earth and washed with MeOH (200 mL). The filtrate was concentrated under reduced pressure to obtain compound 41a (3.5 g) as a yellow-brown oil.

[0287] MS (ESI) m / z 231.1 [M+H] +

[0288] Intermediate 42a 4-Hydroxy-1,5,6-trimethyl-2-oxo-1,2-dihydropyridine-3-carboxylic acid

[0289]

[0290] The synthesis of compound 42a-1 was based on patent WO2007007903. Compound 42a-1 (210 mg, 0.93 mmol) was dissolved in HBr (2 mL, 33 wt.% in HOAc) and reacted at 65°C for 3 h. The reaction solution was concentrated under reduced pressure to obtain compound 42a (190 mg), which was used directly in the next step.

[0291] MS (ESI) m / z 198.2 [M+H] + .

[0292] Intermediate 43a 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

[0293]

[0294] First step 2-(2-bromo-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 43a-1

[0295] Compound 1g (2.26 g, 3.4 mmol) was dissolved in ethyl acetate (20 mL), and hydrogen chloride-ethyl acetate (20 mL, 80 mmol, 4 M in EtOAc) was added, and the mixture was reacted at room temperature for 6 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product 43a-1 (2.2 g). The crude product was used directly in the next step.

[0296] MS (ESI) m / z 562.1 [M+H] +

[0297] Step 2 2-(6-(4-(5-(Benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 43a

[0298] The crude product 43a-1 (2.2 g, 3.4 mmol) was dissolved in 1,4-dioxane (30 mL), compound 1e (0.91 g, 3.75 mmol), triethylamine (1.02 g, 10.2 mmol) and 1-propylphosphoric acid cyclic anhydride (1.6 g, 5.1 mmol, 50% in EtOAc) and reacted at room temperature for 3 hours. After TLC monitoring, the reaction was completed, saturated sodium bicarbonate solution was added, and ethyl acetate was extracted. The organic phase was concentrated to obtain a crude product, which was purified by ethyl acetate beating to obtain compound 43a (1.8 g).

[0299] MS (ESI) m / z 788.1 [M+H] +

[0300] Intermediate 43b 6-methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine

[0301]

[0302] First step 6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridine 43b-2

[0303] 4,5,6,7-Tetrahydrothiophene[2,3-c]pyridine hydrochloride (43b-1) (1.37 g, 10 mmol) was dissolved in tetrahydrofuran (40 mL), potassium carbonate (2.07 g, 15 mmol) and methyl p-toluenesulfonate (1.48 g, 8 mmol) were added, and the mixture was reacted at room temperature for 16 hours. After the reaction, water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL). The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (PE / EA=1:3) to obtain compound 43b-2 (0.25 g).

[0304] MS (ESI) m / z 154.0 [M+H] + .

[0305] Step 2 6-Methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine 43b

[0306] Compound 43b-2 (250 mg, 1.63 mmol) was dissolved in tetrahydrofuran (10 mL), and n-butyl lithium (0.72 mL, 1.8 mmol, 2.5 M in THF) was added dropwise at -78 °C. After reacting at -78 °C for 1 hour, isopropanol pinacol borate (334 mg, 1.8 mmol) was added and the reaction was continued at -78 °C. After the reaction was completed by TLC monitoring, saturated ammonium chloride solution (30 mL) was added, and the mixture was extracted with ethyl acetate. The organic phase was concentrated to obtain a crude product, which was separated and purified by column chromatography (DCM / MeOH=10:1) to obtain compound 43b (220 mg).

[0307] MS (ESI) m / z 280.1 [M+H] + .

[0308] Intermediate 44a 5-Methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine

[0309]

[0310] The first step is 5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridine 44a-2

[0311] 4,5,6,7-Tetrahydrothieno[3,2-c]pyridine hydrochloride (44a-1) (500 mg, 3.60 mmol) was dissolved in dry methanol (10 mL), acetic acid (2 mL) and 37% formaldehyde (3.17 g, 36.00 mmol) were added, and the mixture was stirred at room temperature for 1 h. Sodium triacetoxyborohydride (3.05 g, 7.2 mmol) was then added in batches, and the mixture was stirred at room temperature for 48 h. The reaction was complete when detected by TLC, and the reaction solution was concentrated under reduced pressure. The crude product was separated by column chromatography (DCM / MeOH=20:1) to obtain compound 44a-2 (350 mg) as a light yellow oil.

[0312] MS (ESI) m / z 154.0 [M+H] + .

[0313] Step 2 5-Methyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine 44a

[0314] Compound 44a-2 (350 mg, 2.29 mmol) was dissolved in dry tetrahydrofuran (10 mL), cooled to -70 ° C, 2.5 M n-butyl lithium (0.92 ml, 2.29 mmol) was slowly added dropwise, and the reaction was continued at -70 ° C for 30 minutes, followed by dropwise addition of isopropanol pinacol borate (469.07 mg, 2.52 mmol), and the temperature was naturally raised to room temperature for 2 hours. After TLC detection, the reaction was complete, saturated ammonium chloride aqueous solution was added for quenching (5 mL), and ethyl acetate was extracted (10 mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The crude product was separated by column chromatography (DCM / MeOH=10:1) to obtain compound 44a (180 mg) as a light yellow solid.

[0315] MS (ESI) m / z 280.1 [M+H] + .

[0316] Example 1 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0317]

[0318]

[0319] Step 1 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 1c

[0320] The synthesis of intermediate 1a refers to patent WO2022249060A1, and the synthesis of intermediate 1b refers to patent WO2022268230A1. Compound 1b (115 mg, 0.68 mmol) and 1a (133 mg, 0.23 mmol) were dissolved in dry dimethyl sulfoxide (2 mL), potassium acetate (124 mg, 1.36 mmol) was added, the temperature was raised to 120 ° C for reaction for 18 h, compound 1b (115 mg, 0.68 mmol) and potassium acetate (124 mg, 1.36 mmol) were added, and the reaction was continued at 120 ° C for 18 h. Cool to room temperature, add water (20 mL) to dilute, extract with ethyl acetate (20 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and separate the crude product on a silica gel plate (PE: EA = 2: 1) to obtain compound 1c (57 mg) as a white solid.

[0321] MS (ESI) m / z 715.2 [M+H] + .

[0322] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 1d

[0323] Add dry dichloromethane (1 mL) to compound 1c (57 mg, 0.08 mmol) to dissolve, add trifluoroacetic acid (1 mL), stir at room temperature for 1 h, concentrate, and feed the crude product directly to the next step.

[0324] MS (ESI) m / z 615 [M+H] + .

[0325] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 1f

[0326] Compound 1d was dissolved in DMF (2 mL), and intermediate 1e (synthesis reference patent WO2022249060A1) (25.3 mg, 0.1 mmol) and DIEA (103 mg, 0.8 mmol) were added in sequence. The reaction solution was cooled to about 0°C, and a 50% ethyl acetate solution of T3P (76 mg, 0.12 mmol) was added dropwise. After addition, the mixture was returned to room temperature and stirred for 3 h. Water (20 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated and purified on a silica gel plate (PE: EA = 1:3) to obtain compound 1f (20 mg) as a white solid.

[0327] MS (ESI) m / z 841.2 [M+H] + .

[0328] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 1

[0329] Trifluoroacetic acid (2 mL) was added to compound 1f (20 mg, 0.042 mmol), and the temperature was raised to 50 °C for 18 h. The reaction solution was concentrated and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) to give compound 1 (3.0 mg) as a white solid.

[0330] MS (ESI) m / z 751.2 [M+H] + .

[0331] 1 H NMR (400 MHz, DMSO-d 6 )δ10.34(br s,1H),8.53(s,1H),8.03(d,J=8.6Hz,1H),7.96(s,1H),7.72(d,J=8.7Hz,1H),5.21(s,2H),4.51(d,J=11.3Hz,1H),3.50(m, 7H),3.23(m,2H),2.94(m,3H),2.77(d,J=9.8Hz,1H),2.60(d,J=9.9Hz,1H),2.43(s,3H),2.17(m,4H),1.16(t,J=7.4Hz,3H).

[0332] Example 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0333]

[0334] Step 1 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 2b

[0335] Compound 1a (0.10 g, 0.151 mmol) was dissolved in dioxane (5 mL) and water (1 mL), and compound 2a (synthesis reference patent WO2019023468 A1) (0.06 g, 0.20 mmol), potassium phosphate (0.11 g, 0.500 mmol) and Pd(dppf)Cl2 (11.0 mg, 0015 mmol) were added.2 The reaction was carried out at 85°C for 1 h. After the reaction was completed, the mixture was extracted with ethyl acetate (30 mL × 3), and the organic phases were combined and concentrated under reduced pressure to obtain a crude product, compound 2b (67 mg).

[0336] MS (ESI) m / z 720.10 [M+H] + .

[0337] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 2c

[0338] Compound 2b (67.0 mg, 0.093 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, compound 2c (67.0 mg)

[0339] MS (ESI) m / z 620.10 [M+H] + .

[0340] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 2d

[0341] Compound 2c (0.067 g, 0.153 mmol) was dissolved in THF (10 mL), and compound 1e (49.0 mg, 0.200 mmol), triethylamine (170 mg, 1.53 mmol) and T 3 P (204 mg, 0.3 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, it was extracted with ethyl acetate (50 mL × 3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc = 1 / 1) to obtain compound 2d (0.050 g)

[0342] MS (ESI) m / z 846.10 [M+H] + .

[0343] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 2

[0344] Compound 2d (0.050 g, 0.061 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50°C for 3 h. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia water in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) and lyophilized to obtain compound 2 (17.0 mg).

[0345] MS (ESI) m / z 756.20 [M+H] + .

[0346] 1 H NMR (400 MHz, CDCl 3 )δ11.79(s,1H),9.10(s,1H),8.58(s,1H),8.48(d,J=8.8Hz,1H),7.63(d,J=2.0H z,1H),7.53(d,J=8.8Hz,1H),6.94(s,1H),5.13(d,J=15.6Hz,1H),5.03(d,J=15. 6Hz,1H),3.94-3.75(m,5H),3.49-3.41(m,1H),3.23-3.06(m,5H),2.87-2.64(m, 6H), 2.56 (s, 3H), 2.45-2.28 (m, 3H), 2.04 (q, J = 7.5Hz, 2H), 1.33 (t, J = 7.5Hz, 3H).

[0347] Example 3 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-cyclopropyl-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0348]

[0349] Step 1 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-cyclopropyl-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 3a

[0350] Compound 1a (0.100 g, 0.151 mmol) was dissolved in DMF (5 mL), cyclopropyltributyltin (66.2 mg, 0.20 mmol) and tetrakis(triphenylphosphine)palladium (8.90 mg, 0.0151 mmol) were added, and N 2 The reaction was carried out at 135°C for 8 h. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EA=1 / 1) to obtain compound 3a (80.0 mg).

[0351] MS (ESI) m / z 624.10 [M+H] + .

[0352] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-cyclopropyl-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 3b

[0353] Compound 3a (80.0 mg, 0.128 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, compound 3b (80.0 mg).

[0354] MS (ESI) m / z 524.10 [M+H] + .

[0355] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-cyclopropyl-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 3c

[0356] Compound 3b (0.080 g, 0.153 mmol) was dissolved in THF (10 mL), and compound 1e (49.0 mg, 0.200 mmol), triethylamine (170 mg, 1.53 mmol) and T 3P (204 mg, 0.3 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, it was extracted with ethyl acetate (50 mL × 3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc = 1 / 1) to obtain compound 3c (0.056 g).

[0357] MS (ESI) m / z 750.10 [M+H] + .

[0358] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-cyclopropyl-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 3

[0359] Compound 4 (0.056 g, 0.075 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50 °C for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) and lyophilized to obtain compound 3 (17.0 mg).

[0360] MS (ESI) m / z 660.20 [M+H] + .

[0361] 1 H NMR (400 MHz, CDCl 3 )δ11.79(br s,1H),8.96(s,1H),8.58(s,1H),8.47(d,J=8.6Hz,1H),7.66(d,J=1.9Hz,1H) ,7.54(d,J=8.8Hz,1H),5.64(d,J=12.8Hz,1H),5.06(s,2H),4.78(d,J=12.8Hz ,1H),3.83-3.78(m,2H),3.52-3.46(m,1H),3.22-2.98(m,3H),2.86-2.70(m, 2H), 2.56 (s, 3H), 2.15-2.08 (m, 1H), 1.31 (t, J = 7.5Hz, 3H), 1.13-1.05 (m, 4H).

[0362] Example 4 N-(1-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-5-hydroxy-N,6-dimethylpyrimidine-4-carboxamide

[0363]

[0364] First step (1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)(methyl)carbamic acid tert-butyl ester 4b

[0365] Compound 4a (synthesis reference patent WO2022249060A1) (0.100 g, 0.178 mmol) was dissolved in DMSO (5 mL), and tert-butyl (pyrrolidin-3-yl)carbamate (35.6 mg, 1.78 mmol) and silver tetrafluoroborate (34.5 mg, 0.178 mmol) were added. 2 After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EA=1 / 1) to obtain compound 4b (95 mg)

[0366] MS (ESI) m / z 680.10 [M+H] + .

[0367] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-(methylamino)pyrrolidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 4c

[0368] Compound 4b (95.0 mg, 0.139 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, compound 4c (95.0 mg).

[0369] MS (ESI) m / z 580.10 [M+H] + .

[0370] Step 3 5-(Benzyloxy)-N-(1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-N,6-dimethylpyrimidine-4-carboxamide 4d

[0371] Compound 4c (0.095 g, 0.153 mmol) was dissolved in THF (10 mL), and compound 1e (49 mg, 0.200 mmol), triethylamine (170 mg, 1.53 mmol) and T 3 P (204 mg, 0.3 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, ethyl acetate was extracted (50 mL × 3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc = 1 / 1) to obtain compound 4d (52 mg)

[0372] MS (ESI) m / z 805.80 [M+H] + .

[0373] Step 4: N-(1-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-5-hydroxy-N,6-dimethylpyrimidine-4-carboxamide

[0374] Compound 4d (0.052 g, 0.068 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50°C for 3 h. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) and lyophilized to obtain compound 4 (17 mg).

[0375] MS (ESI) m / z 716.20 [M+H] + .

[0376] 1 H NMR (400 MHz, CDCl 3)δ11.58(brs,1H),9.02(s,1H),8.63(s,1H),8.49(d,J=8.8Hz,1H),7.64(d,J= 2.1Hz,1H),7.53(d,J=8.8Hz,1H),6.98(s,1H),5.10(s,2H),4.37(d,J=3.1Hz,2 H),3.92(t,J=5.4Hz,2H),3.57(m,2H),3.34(m,2H),3.22(s,3H),3.15-3.10(m, 3H), 2.73 (m, 2H), 2.55 (s, 3H), 2.38 (m, 1H), 2.24 (m, 1H), 1.33 (t, J = 7.5Hz, 3H).

[0377] Example 5 N-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0378]

[0379]

[0380] Step 1: 2-Bromo-N-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetamide 5b

[0381] Compound 5a (2.07 g, 10 mmol) was dissolved in DCM (25 mL) and DMAP (1.48 g, 12 mmol) was added. Under an ice bath, bromoacetyl bromide (2.02 g, 10 mmol) was added dropwise and the reaction was carried out at room temperature for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was diluted with dichloromethane (50 mL), and washed with water (20 mL) and saturated brine (20 mL) respectively. The organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EA=3 / 1) to obtain compound 5b (1.6 g).

[0382] MS (ESI) m / z 327.90 [M+H] + .

[0383] Step 2 4-(4-(2-((6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 5d

[0384] Compound 5b (129 mg, 0.39 mmol) and 5c (synthesis reference patent WO2022249060A1) (137 mg, 0.32 mmol) were dissolved in DMF (3 mL), and DIPEA (126 mg, 0.98 mmol) was added, and the mixture was reacted at 80°C for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10 / 1) to obtain compound 5d (160 mg).

[0385] MS (ESI) m / z 678.20 [M+H] + .

[0386] Step 3 N-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 5e

[0387] Compound 5d (160 mg, 0.23 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure to obtain a crude product 5e (160 mg).

[0388] MS (ESI) m / z 578.20 [M+H] + .

[0389] Step 4 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-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-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)acetamide 5f

[0390] Compound 5e (60 mg, 0.1 mmol) was dissolved in DMF (3 mL), and EDCI (19 mg, 0.1 mmol), HOBt (13 mg, 0.1 mmol), and DIEA (39 mg, 0.3 mmol) were added at room temperature. Compound 1e (15 mg, 0.1 mmol) was then added and reacted at room temperature for 16 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 5f (30 mg).

[0391] MS (ESI) m / z 804.20 [M+H] + .

[0392] Step 5: N-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 5

[0393] Compound 5f (60 mg, 0.03 mmol) was dissolved in TFA (3 mL) and reacted at 50 °C for 16 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) to obtain compound 5 (4.5 mg).

[0394] MS (ESI) m / z 714.2 [M+H] + .

[0395] 1 H NMR (400 MHz, CDCl 3 )δ11.79(s,1H),8.87(s,1H),8.59(s,1H),8.16(s,1H),7.11(s,1H),6. 96(s,1H),5.67(d,J=12.0Hz,1H),5.08(s,2H),4.80(d,J=12.0Hz,1H), 4.30(d,J=2.8Hz,2H),3.92(m,2H),3.78(m,2H),3.50(m,1H),3.18(m,2 H), 3.07 (m, 1H), 2.85-2.73 (m, 4H), 2.57 (s, 3H), 1.32 (t, J = 7.5Hz, 3H).

[0396] Example 6 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0397]

[0398] Step 1 Methyl 2-(Methylamino)cyclopent-1-ene-1-carboxylate 6b

[0399] Compound 6a (0.71 g, 5 mmol) was dissolved in THF (10 mL) and MeNH 2 -THF (12.5 mL, 25 mmol, 2M in THF), in N 2 The mixture was stirred for 2 hours under protection. After the reaction was completed by TLC monitoring, ethyl acetate (20 mL) was added, and the organic phase was washed with water (10 mL) and brine (10 mL) respectively. The organic phase was concentrated under reduced pressure to obtain the crude product 6b (560 mg).

[0400] MS (ESI) m / z 156.10 [M+H] + .

[0401] Step 2 2-(3-ethoxy-N-methyl-3-oxopropionylamino)cyclopent-1-ene-1-carboxylic acid methyl ester 6c

[0402] Compound 6b (560 mg, 3.6 mmol) was dissolved in toluene (10 mL), and NaOAc (590 mg, 7.2 mmol) was added, followed by ethyl 3-chloro-3-oxopropanoate (538 mg, 3.6 mmol). The mixture was reacted at room temperature for 6 h. After the reaction was completed by TLC monitoring, ethyl acetate (30 mL) was added, and the mixture was washed with water (10 mL) and brine (10 mL) respectively. The organic phase was concentrated under reduced pressure to obtain a crude product, compound 6c (460 mg).

[0403] MS (ESI) m / z 270.10 [M+H] + .

[0404] Step 3 4-Hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridine-3-carboxylic acid methyl ester 6d

[0405] Compound 6c (460 mg, 0.153 mmol) was dissolved in anhydrous methanol (30 mL), and MeONa solution (0.5 mL, 30 wt% in MeOH) was added dropwise, and the mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was dried to about 10 mL, 30 mL of water was added, and the pH was adjusted to about 7 with dilute HCl under ice bath, and extracted with dichloromethane (30 mL × 3), and the organic phases were combined and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (PE / EtOAc = 2 / 1) to obtain compound 6d (390 mg).

[0406] MS (ESI) m / z 224.10 [M+H] + .

[0407] Step 4: 4-Hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridine-3-carboxylic acid 6e

[0408] Compound 6d (38 mg, 0.17 mmol) was dissolved in anhydrous THF (1 mL), potassium tert-butoxide (1 mL, 1 Min THF) was added at room temperature, and then reacted at 80°C for 3 h. The pH was adjusted to about 7 with dilute HCl under ice bath, extracted with dichloromethane (20 mL × 3), the organic phases were combined, and concentrated under reduced pressure to obtain crude product 6e (24 mg).

[0409] MS (ESI) m / z 210.10 [M+H] + .

[0410] Step 5: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 6

[0411] Compound 6e (24 mg, 0.11 mmol) was dissolved in DMF (2 mL), and EDCI (22 mg, 0.11 mmol), HOBt (15 mg, 0.11 mmol) and DIEA (36 mg, 0.28 mmol) were added at room temperature. Compound 6f (synthesis reference patent WO2022249060A1) was then added and reacted at room temperature for 16 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography (DCM / MeOH=10:1) to obtain compound 6 (1.4 mg). MS (ESI) m / z757.20[M+H] + .

[0412] 1 H NMR (400 MHz, CDCl 3 )δ9.00(s,1H),8.49(d,J=8.8Hz,1H),7.64(s,1H),7.54(d,J=8.8Hz,1H),7.36(s,1H),6.97(s,1H),5.09(s,2H),4.37(m,2H),3.92( m,2H),3.74-3.65(m,4H),3.43(s,3H),3.16(m,2H),2.95-2.84(m,4H),2.73(m,4H),2.16(m,2H),2.03(m,2H),1.32(t,J=7.6Hz,3H)

[0413] Example 7 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(4,6-dimethyl-3-oxo-3,4-dihydropyrazine-2-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0414]

[0415] Step 1 3-Chloro-1,5-methylpyrazin-2(1H)-one 7b

[0416] Compound 7a (100 mg, 0.69 mmol) and potassium carbonate (191 mg, 1.38 mmol) were dissolved in DMF (5 mL). Methyl iodide (118 mg, 0.83 mmol) was added to the reaction solution and stirred at room temperature for 4 h. After the reaction was completed, the solvent was removed by concentration under reduced pressure. Ethyl acetate / H 2 The organic layers were combined and washed with saturated brine, and the organic layer was dried over anhydrous sodium sulfate to obtain crude compound 7b (100 mg) as a yellow oil, which was used directly in the next step.

[0417] MS (ESI) m / z 159.2 [M+H] + .

[0418] Step 2 4,6-Dimethyl-3-oxo-3,4-dihydropyrazine-2-carboxylic acid phenyl ester 7c

[0419] Compound 7b (100 mg, 0.63 mmol), phenyl formate (231 mg, 1.89 mmol), Xantphos (36 mg, 0.063 mmol), palladium acetate (7 mg, 0.031 mmol) and TEA (159 mg, 1.57 mmol) were dissolved in anhydrous acetonitrile (2 mL), replaced with nitrogen three times, and refluxed at 80 ° C for 4 h. The reaction solution was cooled to room temperature and filtered with celite, and the solvent was removed by concentration under reduced pressure, and DCM / H 2 The organic layer was combined and washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography (PE / EA=1 / 1) to obtain compound 7c (40 mg) as an off-white solid.

[0420] MS (ESI) m / z 245.1 [M+H] + .

[0421] 1 H NMR (400 MHz, CDCl 3 )δ7.42(m,3H),7.30-7.22(m,3H),3.61(s,3H),2.39(s,3H).

[0422] Step 3 4,6-Dimethyl-3-oxo-3,4-dihydropyrazine-2-carboxylic acid 7d

[0423] Compound 7c (25 mg, 0.10 mmol) was dissolved in THF / MeOH / H 2 O (1 / 1 / 1), lithium hydroxide (7 mg, 0.30 mmol) was added to the system and reacted at room temperature for 1 h. After the reaction, the pH was adjusted to 6 with 1N hydrochloric acid (1.0 M), and the solvent was removed by concentration under reduced pressure to obtain crude compound 7d (20 mg) as a yellow solid.

[0424] MS (ESI) m / z 169.1 [M+H] + .

[0425] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-6-(4-(4,6-dimethyl-3-oxo-3,4-dihydropyrazine-2-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 7

[0426] Compound 7d (20 mg, 0.12 mmol) and compound 6f (68 mg, 0.12 mmol) were dissolved in DMF (2 mL), TEA (48 mg, 0.48 mmol) was added, and T 3P / EA solution (57 mg, 0.18 mmol) was reacted at room temperature for 2 h. 2 O for three times, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia water in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) to obtain compound 7 (3 mg) as a white solid.

[0427] MS (ESI) m / z 716.2 [M+H] + .

[0428] 1 H NMR (400 MHz, CDCl 3 )δ8.99(s,1H),8.48(d,J=8.7Hz,1H),7.64(s,1H),7.53(d,J=8.5Hz,1H),7.00(s,1H) ,6.96(s,1H),5.38-5.32(m,1H),5.11(d,J=15.2Hz,1H),5.06(d,J=15.2Hz,1H),4.74- 4.67(m,1H),4.36(m,2H),3.92(m,2H),3.76(m,2H),3.53(s,3H),3.48(m,1H),3.18-3 .12(m,3H),2.78(m,1H),2.72(m,2H),2.63(m,1H),2.30(s,3H),1.30(t,J=7.6Hz,3H).

[0429] Example 8 2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methoxy-4-(trifluoromethyl)phenyl)acetamide

[0430]

[0431]

[0432] Step 1 4-(5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 8a

[0433] Compound 5c (200 mg, 0.47 mmol) and compound 2a (185 mg, 0.70 mmol) were dissolved in 1,4-dioxane (5 mL) and water (1.5 mL), and Pd(dppf)Cl was added. 2 (19 mg, 0.023 mmol) and sodium carbonate (99.2 mg, 0.94 mmol), replaced with nitrogen three times, heated to 100 ° C for 16 h, cooled to room temperature, diluted with water (20 mL), extracted with dichloromethane (30 mL x 3), the organic phase was dried over anhydrous sodium sulfate, concentrated, and the crude product was flash separated (DCM / MeOH=30 / 1) to obtain compound 8a (120 mg) as a brown solid.

[0434] Step 2 4-(5-ethyl-4-(2-(2-methoxy-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 8c

[0435] Compound 8a (120 mg, 0.25 mmol) and compound 8b (85 mg, 0.27 mmol) were dissolved in dry acetonitrile (2 mL), and N,N-diisopropylethylamine (96 mg, 0.74 mmol) was added. The mixture was reacted at room temperature for 18 h. The reaction solution was concentrated and separated on a silica gel plate (PE:EA=2:1) ​​to obtain compound 8c (40 mg) as a white solid.

[0436] MS (ESI) m / z 716.4 [M+H] + .

[0437] Step 3 2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methoxy-4-(trifluoromethyl)phenyl)acetamide 8d

[0438] Add dry dichloromethane (1 mL) to compound 8c (40 mg, 0.056 mmol) to dissolve, add trifluoroacetic acid (1 mL), stir at room temperature for 1 h, concentrate, add saturated sodium bicarbonate aqueous solution (10 mL) to the residue, extract with dichloromethane (20 mL x 3), combine the organic phases, dry over anhydrous sodium sulfate, concentrate, and the crude product 8d is directly fed to the next step.

[0439] MS (ESI) m / z 616 [M+H] + .

[0440] Step 4: 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methoxy-4-(trifluoromethyl)phenyl)acetamide 8e

[0441] Compound 8d was dissolved in DMF (2 mL), and compound 1e (17.7 mg, 0.073 mmol) and N,N-diisopropylethylamine (72 mg, 0.56 mmol) were added in sequence. The reaction solution was cooled to about 0°C, and a 50% ethyl acetate solution of T3P (53.4 mg, 0.084 mmol) was added dropwise. After addition, the mixture was returned to room temperature and stirred for 3 h. Water (20 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated and purified on a silica gel plate (PE: EA = 1:3) to obtain compound 8e (19 mg) as a white solid.

[0442] MS (ESI) m / z 842.5 [M+H] + .

[0443] Step 5 2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methoxy-4-(trifluoromethyl)phenyl)acetamide 8

[0444] Trifluoroacetic acid (1 mL) was added to compound 8e (19 mg, 0.023 mmol), and the temperature was raised to 50 °C for 18 h. The reaction solution was concentrated and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) to give compound 8 (10 mg) as a white solid.

[0445] MS (ESI) m / z 752.4 [M+H] + .

[0446] 1 H NMR (400 MHz, DMSO-d 6)δ10.11(s,1H),8.55(s,1H),8.20(d,J=8.4Hz,1H),7.36(s,1H),7.27(d,J=8.4Hz,1H), 6.71(s,1H),5.30(s,2H),4.52(d,J=12.1Hz,1H),3.98(s,3H),3.73(t,J=6.8Hz,2H),3. 55-3.38(m,4H),3.00-2.88(m,3H),2.81(m,1H),2.66-2.54(m,2H),2.44(s,3H),2.27(m ,2H),2.17(t,J=7.2Hz,1H),1.90-1.85(m,2H),1.76-1.60(m,4H),1.16(t,J=7.4Hz,3H).

[0447] Example 9 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(6,7-dihydro-4H-thieno[3,2-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide

[0448]

[0449]

[0450] Step 1 6,7-Dihydro-4H-thieno[3,2-c]pyran 9b

[0451] Compound 9a (12.8 g, 0.1 mol), paraformaldehyde (3.9 g, 0.13 mol), indium trichloride (1.1 g, 5 mmol), and acetonitrile (600 mL) were added to a reaction flask, and the reaction solution was heated to 80°C and stirred for 2 h under argon protection. The crude product was concentrated under reduced pressure and purified by column chromatography (PE / EtOAc=20 / 1) to obtain compound 9b (6.5 g) as a light yellow solid.

[0452] MS (EI) m / z 140 [M].

[0453] Step 2 2-(6,7-Dihydro-4H-thieno[3,2-c]pyran-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane 9c

[0454] Compound 9b (4.2 g, 30 mmol) was dissolved in anhydrous tetrahydrofuran (60 mL), and n-butyl lithium solution (20 mL, 1.6 M in hexane) was slowly added dropwise at -70 °C. After addition, the mixture was heated to room temperature and stirred for 1 h. The mixture was cooled to -70 °C and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane was slowly added dropwise. After addition, the mixture was heated to room temperature and stirred for 2 h. Saturated NH 4 The reaction was quenched with Cl, and ethyl acetate (200 mL) and saturated brine (10 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (PE / EtOAc=20 / 1) to give compound 9c (4.2 g) as a light yellow solid.

[0455] MS (ESI) m / z 267 [M+H] + .

[0456] Step 3 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(6,7-dihydro-4H-thieno[3,2-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 9d

[0457] Compound 1a (226 mg, 0.5 mmol), compound 9c (133 mg, 0.5 mmol), Pd(dppf)Cl 2 (41 mg, 0.05 mmol) and sodium carbonate (159 mg, 1.5 mmol) were added to the sealed tube, and the argon atmosphere was replaced three times. 1,4-dioxane (5 mL) and H 2 O (1 mL), sealed, heated to 100 ° C and stirred for 16 h. Ethyl acetate (50 mL) and saturated brine (10 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=3 / 1) to obtain compound 9d (62 mg) as a light yellow solid.

[0458] MS (ESI) m / z 744 [M+Na] + .

[0459] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(6,7-dihydro-4H-thieno[3,2-c]pyran-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 9e

[0460] Compound 9d (60 mg, 0.08 mmol) was dissolved in DCM (2 mL), TFA (2 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated, ethyl acetate (30 mL) was added, and saturated sodium bicarbonate was adjusted to alkaline, and the organic phase was washed with saturated brine and concentrated under reduced pressure to obtain crude compound 9e (60 mg), which was directly used in the next step.

[0461] MS (ESI) m / z 622 [M+H] + .

[0462] Step 5 2-(6-(4-(5-benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(6,7-dihydro-4H-thieno[3,2-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 9f

[0463] Compound 9e (60 mg, 0.08 mmol) was dissolved in DMF (2 mL), and compound 1e (24 mg, 0.1 mmol), DIPEA (54 mg, 0.4 mmol) and T were added. 3 P (80 mg, 0.4 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 2 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction, and ethyl acetate was extracted (50 mL). The organic phase was washed three times with saturated brine and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (PE / EtOAc=1 / 5) to obtain compound 9f (50 mg) as a light yellow solid.

[0464] MS (ESI) m / z 848 [M+H] + .

[0465] Step 6: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(6,7-dihydro-4H-thieno[3,2-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyridin-4(7H)-yl)acetamide 9

[0466] Compound 9f (50 mg, 0.06 mmol) was dissolved in trifluoroacetic acid (3 mL), and the reaction mixture was reacted at 50°C for 3 h. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated under reduced pressure and ethyl acetate (30 mL) was added, and the saturated sodium bicarbonate aqueous solution was adjusted to Ph = 8, extracted with ethyl acetate, and the organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc = 1 / 5) to obtain compound 9 (26 mg) as a white solid.

[0467] MS (ESI) m / z 758 [M+H] + .

[0468] 1 H NMR (400 MHz, CDCl 3 )δ11.78(s,1H),9.04(s,1H),8.58(s,1H),8.47(d,J=8.8Hz,1H),7.61(s,1H),7.5 2(d,J=8.8Hz,1H),7.51(s,1H),5.66(d,J=12.8Hz,1H),5.15(s,2H),4.79(d,J=12 .8Hz,1H),4.71(s,2H),4.00(m,2H),3.81(m,2H),3.49(m,1H),3.19(m,2H),3.07( m,1H),2.92(m,2H),2.84(m,1H),2.77(m,1H),2.56(s,3H),1.32(t,J=7.5Hz,3H).

[0469] Example 10 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyanocyclohex-1-en-1-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0470]

[0471]

[0472] Step 1 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-cyanocyclohex-1-en-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 10b

[0473] The synthesis of intermediate 10a refers to patent WO2015144001 A1. Compound 1a (200 mg, 0.30 mmol) and 10a (105 mg, 0.45 mmol) were dissolved in 1,4-dioxane (4 mL) and water (0.4 mL), 1,1-bis(diphenylphosphino)ferrocenepalladium dichloromethane complex (24.5 mg, 0.03 mmol) and sodium carbonate (95.4 mg, 0.90 mmol) were added, nitrogen was replaced three times, the temperature was raised to 100 ° C for 16 h, cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by silica gel column (PE: EA = 1: 4) to obtain compound 10b (47 mg) as a yellow-brown solid.

[0474] MS (ESI) m / z 689 [M+H] + .

[0475] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyanocyclohex-1-en-1-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 10c

[0476] Compound 10b (47 mg, 0.068 mmol) was dissolved in dry dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (5 mL), and the mixture was concentrated under reduced pressure. The operation was repeated at least 3 times, and the crude product was directly used for the next reaction to obtain compound 10c (54 mg) as a yellow-brown solid.

[0477] MS (ESI) m / z 589 [M+H] + .

[0478] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4-cyanocyclohex-1-en-1-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 10d

[0479] Compound 10c (54 mg, crude product) was dissolved in DMF (2 mL), and compound 1e (26.85 mg, 0.110 mmol) and triethylamine (46.55 mg, 0.46 mmol) were added in sequence. The reaction solution was cooled to about 0°C, and a 50% ethyl acetate solution of 1-propylphosphoric acid cyclic anhydride (43.9 mg, 0.138 mmol) was added dropwise. After addition, the mixture was returned to room temperature and reacted for 2 h. Water (10 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified on a silica gel plate (PE / EA=1 / 2) to obtain compound 10d (20 mg) as a light yellow solid.

[0480] MS (ESI) m / z 815 [M+H] + .

[0481] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyanocyclohex-1-en-1-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 10

[0482] Compound 10d (20 mg, 0.025 mmol) was dissolved in trifluoroacetic acid (1 mL), and the temperature was raised to 50°C for reaction for 3 h. The reaction solution was concentrated under reduced pressure and purified by silica gel plate separation (DCM / MeOH=30 / 1) to obtain compound 10 (12.2 mg) as a light yellow solid.

[0483] MS (ESI) m / z 725 [M+H] + .

[0484] 1 H NMR (400 MHz, DMSO-d 6 )δ10.35(s,1H),10.23(s,1H),8.57(s,1H),8.05(d,J=8.0Hz,1H),7.96(s,1H ),7.71(d,J=8.0Hz,1H),6.79(s,1H),5.31(s,2H),5.08-4.98(m,1H),4.52(d, J=12.8Hz,1H),3.54-3.43(m,4H),3.16(m,1H),2.99(m,2H),2.81(d,J=11.2Hz ,1H),2.67-2.54(m,3H),2.44(s,3H),2.02-1.93(m,4H),1.26(t,J=7.5Hz,3H)

[0485] Example 11 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-(difluoromethylene)piperidin-1-yl)-5-ethyl-6-(4-(4-hydroxy-1-methyl)-2-oxo-2,5,6,7-tetrahydro-1H-cyclopenta[b]pyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0486]

[0487] Compound 6e (38 mg, 0.18 mmol) was dissolved in DMF (2 mL), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (34.8 mg, 0.18 mmol), 1-hydroxybenzotriazole (24.5 mg, 0.18 mmol) and N,N-diisopropylethylamine (58.6 mg, 0.45 mmol) were added in sequence. The mixture was stirred at room temperature for 10 minutes, and compound 1d (55.9 mg, 0.09 mmol) was added. The mixture was reacted at room temperature for 48 h, and water (20 mL) was added to quench the mixture. The mixture was extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia water in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) to give compound 11 (10.9 mg) as a light yellow solid.

[0488] MS (ESI) m / z 806.4 [M+H] + .

[0489] 1 H NMR (400 MHz, DMSO-d 6 )δ10.33(s,1H),8.03(d,J=8.6Hz,1H),7.96(s,1H),7.72(d,J=8.3Hz,1H),5.21(s,2H),3.59-3.3 9(m,8H),3.30(s,3H),2.92(m,6H),2.69(m,4H),2.18(m,4H),2.00(m,2H),1.16(t,J=7.4Hz,3H).

[0490] Example 12 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(1,1-difluorospiro[2.5]oct-5-en-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0491]

[0492]

[0493] Step 1 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(1,1-difluorospiro[2.5]oct-5-en-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 12b

[0494] The synthesis of compound 12a refers to patent WO2021141041A1. Compound 1a (150 mg, 0.23 mmol) was dissolved in dioxane (4 mL) and water (0.5 mL), and compound 12a (124 mg, 0.46 mmol), sodium carbonate (48 mg, 0.45 mmol) and Pd(dppf)Cl were added. 2 (17 mg, 0.023 mmol), N 2 The reaction was carried out at 100°C for 1 h. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate (30 mL×3), and the organic phases were combined and concentrated under reduced pressure to obtain a crude product, Compound 12b (80 mg).

[0495] MS (ESI) m / z 671.1 [M–C 4 H 7 ] +

[0496] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(1,1-difluorospiro[2.5]oct-5-en-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 12c

[0497] Compound 12b (80 mg, 0.11 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, compound 12c (80 mg).

[0498] MS (ESI) m / z 625.10 [M+H] + .

[0499] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(1,1-difluorospiro[2.5]oct-5-en-6-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 12d

[0500] Compound 12c (80 mg, 0.128 mmol) was dissolved in THF (10 mL), and compound 1e (38 mg, 0.154 mmol), triethylamine (130 mg, 1.28 mmol) and T 3 P (61 mg, 0.192 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, it was extracted with ethyl acetate (20 mL × 3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc = 1 / 1) to obtain compound 12d (70 mg).

[0501] MS (ESI) m / z 852.20 [M+H] + .

[0502] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(1,1-difluorospiro[2.5]oct-5-en-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 12

[0503] Compound 12d (50 mg, 0.059 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50°C for 3 h. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) and lyophilized to obtain compound 12 as a white solid (35 mg).

[0504] MS (ESI) m / z 762.20 [M+H] + .

[0505] 1 H NMR (400 MHz, CDCl 3)δ11.78(s,1H),9.07(s,1H),8.58(s,1H),8.49(d,J=8.7Hz,1H),7.64(s,1H),7.53(d ,J=8.7Hz,1H),7.01(s,1H),5.65(m,1H),5.10(s,2H),4.78(m,1H),3.80(m,2H),3.49 (m,1H),3.25-3.00(m,3H),2.90-2.65(m,4H),2.56(s,3H),2.54-2.44(m,1H),2.30-2 .18(m,1H),1.82(m,2H),1.32(t,J=7.6Hz,3H),1.19-1.08(m,1H),0.90-0.80(m,1H).

[0506] Example 13 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(2-oxaspiro[3.5]non-6-en-7-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0507]

[0508] The first step is 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(2-oxaspiro[3.5]non-6-ene-7-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 13b. The synthesis of compound 13a refers to patent WO2021055744 A1. Compound 1a (100 mg, 0.15 mmol) was dissolved in dioxane (4 mL) and water (0.5 mL), and compound 13a (75 mg, 0.30 mmol), potassium phosphate (65 mg, 0.30 mmol) and Pd(dppf)Cl were added. 2 (10 mg, 0.015 mmol), N 2 The reaction was carried out at 85° C. for 1 h. After the reaction was completed, the mixture was extracted with ethyl acetate (30 mL×3), and the organic phases were combined and concentrated under reduced pressure to obtain a crude product, compound 13b (70 mg).

[0509] MS (ESI) m / z 651.1 [M–C 4 H 7 ] + .

[0510] Step 2: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(2-oxaspiro[3.5]non-6-ene)-7-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 13c

[0511] Compound 13b (70 mg, 0.10 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain a crude product, Compound 13c (50 mg).

[0512] MS (ESI) m / z 606.10 [M+H] + .

[0513] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(2-oxaspiro[3.5]non-6-en-7-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 13d

[0514] Compound 13c (50 mg, 0.083 mmol) was dissolved in THF (5 mL), and compound 7 (24 mg, 0.100 mmol), triethylamine (84 mg, 0.83 mmol) and THF were added. 3 P (40 mg, 0.125 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed, it was extracted with ethyl acetate (20 mL × 3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (PE / EtOAc = 1 / 1) to obtain compound 13d (35 mg).

[0515] MS (ESI) m / z 832.20 [M+H] + .

[0516] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(2-oxaspiro[3.5]non-6-en-7-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 13

[0517] Compound 13d (35 mg, 0.042 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50°C for 3 h. After the reaction was completed under TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) and lyophilized to obtain compound 13 as a white solid (10 mg).

[0518] MS (ESI) m / z 742.20 [M+H] + .

[0519] 1H NMR (400MHz, CDCl 3 )δ9.06(s,1H),8.58(s,1H),8.49(d,J=8.7Hz,1H),7.64(s,1H),7.53(d,J=8.7Hz,1 H),6.94(s,1H),5.65(m,1H),5.09(s,2H),4.78(m,1H),4.52(d,J=6.0Hz,1H),4.43( d,J=6.0Hz,1H),4.28-4.53(m,1H),3.80(m,2H),3.49(m,1H),3.18(m,2H),3.06(m,1 H),2.85-2.56(m,5H),2.56(s,3H),2.05(m,2H),1.61(m,2H),1.33(t,J=7.5Hz,3H).

[0520] Example 14 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3-(methylsulfonyl)azetidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0521]

[0522] Step 1: 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(3-(methylsulfonyl)azetidin-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 14a

[0523] 1a (200 mg, 0.302 mmol), AcOK (178 mg, 1.81 mmol) and 3-methylsulfonyl-azetidine hydrochloride (156 mg, 0.906 mmol) were dissolved in DMSO (4 mL) and reacted at 120 °C for 16 h. After the reaction was complete, H 2 O (30 mL x 3), extracted with ethyl acetate (30 mL), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 14a (100 mg) as a yellow solid.

[0524] MS (ESI) m / z 717.3 [M+H] + .

[0525] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(3-(methylsulfonyl)azetidin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 14b

[0526] Compound 14a (100 mg, 0.139 mmol) was dissolved in TFA (2 mL) and reacted at room temperature for 1 h. The mixture was concentrated under reduced pressure, saturated sodium carbonate solution was added, and the mixture was extracted with dichloromethane (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 14b (41 mg).

[0527] MS (ESI) m / z 617.2 [M+H] + .

[0528] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-(3-(methylsulfonyl)azetidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 14c

[0529] Compound 14b (41 mg, 0.0655 mmol), compound 1e (21 mg, 0.0865 mmol) and DIPEA (43 mg, 0.333 mmol) were dissolved in DMF (2 mL), and T3P (50% wt, 64 mg, 0.0998 mmol) was added and reacted at room temperature for 16 h. After the reaction was completed, H 2 The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 14c (45 mg).

[0530] MS (ESI) m / z 843.4 [M+H] + .

[0531] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3-(methylsulfonyl)azetidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 14

[0532] Compound 14c (45 mg, 0.0534 mmol) was dissolved in TFA (2 mL) and reacted at 50°C for 3 h. After concentration under reduced pressure, the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 20%-30% acetonitrile, 50 mL / min) was purified by reverse phase to obtain compound 14 (3 mg) as a white solid.

[0533] MS (ESI) m / z 753.3 [M+H] + .

[0534] 1 H NMR (400 MHz, DMSO-d 6 )δ10.79–9.80(m,1H),8.50(s,1H),7.99(d,J=8.5Hz,1H),7.89(s,1H),7.64(d,J=7.2Hz,1H),5.16(s,2H),4.52–4.33(m,2H),4.21(t, J=8.7Hz,2H),4.10(dd,J=9.2,5.6Hz,2H),3.42(d,J=11.0Hz,3H),2.96(s,3H),2.92–2.50(m,6H),2.37(s,3H),1.09(t,J=7.4Hz,3H).

[0535] Example 15 2-(2-(3-oxabicyclo[3.1.0]hexane-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide

[0536]

[0537]

[0538] Step 1: 4-(2-(3-oxabicyclo[3.1.0]hexane-6-yl)-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 15b

[0539] The synthesis of intermediate 15a refers to patent WO2020072695A1. Compound 1a (530 mg, 0.8 mmol), compound 15a (264 mg, 1.2 mmol), Pd(dppf)Cl 2 (59 mg, 0.08 mmol) and potassium phosphate (424 mg, 2.0 mmol) were added to the sealed tube, the argon atmosphere was replaced three times, 1,4-dioxane (10 mL) and H 2 O (2 mL), sealed, heated to 80 ° C and stirred for 16 h. Ethyl acetate (100 mL) and saturated brine (10 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 2) to obtain compound 15b (31 mg) as a light yellow solid.

[0540] MS (ESI) m / z 666 [M+H] + .

[0541] Step 2 2-(2-(3-oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 15c

[0542] Compound 15b (31 mg, 0.047 mmol) was dissolved in DCM (2 mL), TFA (2 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated, ethyl acetate (30 mL) was added, and saturated sodium bicarbonate solution was adjusted to pH = 8, and the organic phase was washed with saturated brine and concentrated under reduced pressure to obtain crude compound 15c (33 mg), which was used directly in the next step without purification.

[0543] MS (ESI) m / z 566 [M+H] + .

[0544] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(3-oxabicyclo[3.1.0]hexan-6-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 15d

[0545] Compound 15c (33 mg, 0.047 mmol) was dissolved in THF (2 mL), and compound 1e (14 mg, 0.056 mmol), DIPEA (30 mg, 0.23 mmol) and THF were added. 3 P (44 mg, 0.07 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 3 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction, and ethyl acetate was extracted (50 mL). The organic phase was washed three times with saturated brine and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (EtOAc) to obtain compound 15d (21 mg) as a light yellow solid.

[0546] MS (ESI) m / z 792 [M+H] + .

[0547] Step 4: 2-(2-(3-oxabicyclo[3.1.0]hexane-6-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 15

[0548] Compound 15d (21 mg, 0.026 mmol) was dissolved in trifluoroacetic acid (2 mL), and the reaction mixture was reacted at 50°C for 3 h. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated under reduced pressure, ethyl acetate (30 mL) was added, and saturated sodium bicarbonate was adjusted to alkalinity for extraction. The organic phase was washed once with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (MeOH / EtOAc=1 / 10) to obtain compound 15 (11 mg).

[0549] MS (ESI) m / z 702 [M+H] + .

[0550] Example 16 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1,5-dimethyl-2-oxo-1,2-dihydropyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0551]

[0552] Compound 16a (64 mg, 0.35 mmol) was dissolved in DMF (3 mL), and EDCI.HCl (86 mg, 0.45 mmol), HOBt (60 mg, 0.45 mmol), and DIEA (117 mg, 0.9 mmol) were added at room temperature. Compound 6f was then added and reacted at 50°C for 5 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was separated and purified by silica gel column chromatography (DCM / MeOH=10:1) to obtain compound 16 (31.2 mg).

[0553] MS (ESI) m / z 731.40 [M+H] + .

[0554] 1 H NMR (400 MHz, CDCl 3 )δ9.03(s,1H),8.48(d,J=8.6Hz,1H),7.63(s,1H),7.54(d,J=8.6Hz,1H),7.14(s,1H),6.95(s,1H),5.12(s,2H),4.36(m,2H),4.18(m,1H),3 .91(t,J=5.6Hz,2H),3.66(m,2H),3.46(s,3H),3.34(m,2H),3.20-3.1 0(m,2H),2.71(m,4H),2.04(s,3H),2.00(m,1H),1.30(t,J=7.6Hz,3H)

[0555] Example 17 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(cyclopropylethynyl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0556]

[0557]

[0558] First step 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(cyclopropylethynyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 17a

[0559] Compound 1a (500 mg, 0.76 mmol) was dissolved in DMF (4 mL), and cyclopropylacetylene (75 mg, 1.13 mmol), Xphos-Pd-G2 (70 mg, 0.076 mmol) and triethylamine (1 mL) were added. 2 After the reaction was completed, water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 17a (340 mg).

[0560] MS (ESI) m / z 592.1 [M–C 4 H 7 ] + .

[0561] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(cyclopropylethynyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 17b

[0562] Compound 17a (340 mg, 0.53 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain a crude product, Compound 17b (340 mg).

[0563] MS (ESI) m / z 548.20 [M+H] + .

[0564] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(cyclopropylethynyl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 17c

[0565] Compound 17b (150 mg, 0.27 mmol) was dissolved in THF (5 mL), and compound 1e (80 mg, 0.33 mmol), triethylamine (273 mg, 2.7 mmol) and THF were added. 3 P (129 mg, 0.41 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 1 h. After the reaction was completed by TLC monitoring, water (20 mL) was added, and ethyl acetate was extracted (20 mL × 3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (PE / EtOAc = 1 / 1) to obtain compound 17c (90 mg).

[0566] MS (ESI) m / z 774.20 [M+H] + .

[0567] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(cyclopropylethynyl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 17

[0568] Compound 17c (90 mg, 0.12 mmol) was dissolved in trifluoroacetic acid (3 mL) and reacted at 50°C for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase purification (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 5% to 20% acetonitrile, 30 mL / min) to give compound 17 as a white solid (20 mg).

[0569] MS (ESI) m / z 684.20 [M+H] + .

[0570] 1 H NMR (400 MHz, CDCl 3 )δ9.13(s,1H),8.58(s,1H),8.49(d,J=8.7Hz,1H),7.66(s,1H),7.53(d,J=8.4Hz,1H),5.67(m,1H),5.07(s,2H),4.79(m,1H), 4.12(m,1H),3.80(m,2H),3.48(m,1H),317(m,2H),3.05(m,1H),2.81(m,2H),2.56(s,3H),1.33(t,J=7.5Hz,3H),0.94(m,4H).

[0571] Example 18 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyclopropyloxyphenyl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopentadienyl[b]pyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0572]

[0573] First step 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-cyclopropyloxyphenyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 18a

[0574] Compound 1a (100 mg, 0.15 mmol) was dissolved in 1,4-dioxane (4 mL), and compounds 2-(4-cyclopropyloxyphenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (60 mg, 0.23 mmol), Pd(PPh 3 ) 4 (18 mg, 0.015 mmol), potassium phosphate (70 mg, 0.30 mmol) and water (1 mL). 2 After the reaction was completed, water (20 mL) was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 18a (70 mg).

[0575] MS (ESI) m / z 660.1 [M–C 4 H 7 ] + .

[0576] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyclopropyloxyphenyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 18b

[0577] Compound 18a (70 mg, 0.098 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain compound 18b (60 mg).

[0578] MS (ESI) m / z 616.20 [M+H] + .

[0579] Step 3 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-cyclopropyloxyphenyl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-2-oxo-2,5,6,7-tetrahydro-1H-cyclopentadienyl[b]pyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 18

[0580] Compound 18b (60 mg, 0.097 mmol) was dissolved in DMF (3 mL), and compound 6e (24 mg, 0.117 mmol), DIPEA (75 mg, 0.58 mmol), EDCI.HCl (44 mg, 0.23 mmol) and HOBT (31 mg, 0.23 mmol) were added, and the reaction mixture was reacted at 60°C for 8 h. After the reaction was completed as monitored by TLC, the mixture was extracted with ethyl acetate (20 mL×3), the organic phases were combined, and the mixture was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10 / 1) to obtain compound 18 (5 mg).

[0581] MS (ESI) m / z 807.20 [M+H] + .

[0582] 1 H NMR (400 MHz, CDCl 3 )δ9.19(s,1H),8.49(d,J=8.6Hz,1H),8.20(d,J=8.5Hz,2H),7.57(s,1H), 7.51(d,J=8.0Hz,1H),7.11(d,J=8.6Hz,2H),5.35(m,1H),5.15(s,2H),3.8 0(m,1H),3.66(m,4H),3.43(s,3H),3.35(m,2H),2.93(m,2H),2.85(m,2H), 2.73(m,2H),2.35(m,1H),2.15(m,2H),1.31(t,J=7.5Hz,3H),0.84(m,4H).

[0583] Example 19 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(6-hydroxypyrazole)[1,5-a]pyrimidine-5-carbonyl)piperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0584]

[0585]

[0586] Step 1: Sodium (Z)-2-(Benzyloxy)-3-ethoxy-3-oxoprop-1-en-1-ol 19b

[0587] Dissolve (benzyloxy)ethyl acetate (19a) (5g, 25.7mmol) in tetrahydrofuran (30mL), cool the reaction solution to 0°C, add sodium hydride (1.13g, 60% in mineral oil), and return to room temperature for 1h. Cool the reaction solution to 0°C again, add ethyl formate (2.14g, 28.8mmol) dropwise, and return to room temperature for 3h after addition. The reaction solution is concentrated under reduced pressure, and the crude product is directly fed to the next step. Compound 19b (6.3g) is a gray solid.

[0588] MS (ESI) m / z 245 [M+H] + .

[0589] Step 2 (Z)-2-(Benzyloxy)-3-ethoxyethyl acrylate 19c

[0590] Compound 19b (6.3 g, 25.8 mmol) was dissolved in DMF (30 mL), and iodoethane (4 g, 25.8 mmol) was added. After the addition, the temperature was raised to 35°C and stirred for 1 h. Water (30 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified on a silica gel plate (PE / EA=2 / 1) to obtain compound 19c (6 g) as a light yellow oil.

[0591] MS (ESI) m / z 251 [M+H] + .

[0592] Step 3 6-(Benzyloxy)pyrazolo[1,5-a]pyrimidin-5-ol 19d

[0593] Dissolve 2H-pyrazole-3-amine (1g, 12.04mmol) in DMF (30mL), add compound 19c (4.52g, 18.06mmol) and cesium carbonate (5.88g, 18.06mmol), and heat to 110℃ for 18h after addition. After cooling to room temperature, add ice water (100mL), and add 2.0M hydrochloric acid dropwise to adjust the pH value to about 5-6. Filter the mixture under reduced pressure, wash the filter cake with water (10mL x 3), and dry to obtain compound 19d (670mg) as a yellow-brown solid.

[0594] MS (ESI) m / z 242 [M+H] + .

[0595] Step 4: 6-(Benzyloxy)-5-chloropyrazolo[1,5-a]pyrimidine 19e

[0596] Compound 19d (670 mg, 2.78 mmol) was dissolved in dry acetonitrile (7 mL), phosphorus oxychloride (1.29 g, 8.34 mmol) was added, and the temperature was raised to 35°C and stirred for 5 h. The mixture was concentrated under reduced pressure, and the crude product was separated and purified on a silica gel plate (PE / EA=4 / 1) to obtain compound 19e (200 mg) as a light yellow solid.

[0597] MS (ESI) m / z 260 [M+H] + .

[0598] Step 5: 6-(Benzyloxy)pyrazolo[1,5-a]pyrimidine-5-carboxylic acid phenyl ester 19f

[0599] Compound 19e (200 mg, 0.78 mmol) was dissolved in acetonitrile (2 mL), and tri-tert-butylphosphine tetrafluoroborate (22.68 mg, 0.078 mmol), palladium acetate (8.76 mg, 0.039 mmol) and triethylamine (394.64 mg, 3.9 mmol) were added in sequence. The atmosphere was replaced with nitrogen three times, the temperature was raised to 80 ° C and the reaction was carried out for 16 h. The mixture was cooled to room temperature, diluted with water (4 mL), extracted with ethyl acetate (5 mL x 3), the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE / EA=1 / 1) to obtain compound 19g (140 mg) as a white solid.

[0600] MS (ESI) m / z 345 [M+H] + .

[0601] Step 6: 6-(Benzyloxy)pyrazolo[1,5-a]pyrimidine-5-carboxylic acid 19h

[0602] Compound 19g (140 mg, 0.406 mol) was dissolved in tetrahydrofuran (1 mL) and water (1 mL), and lithium hydroxide (29.6 mg, 1.218 mmol) was added. After the addition, the mixture was stirred at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia water in H 2 O, mobile phase B: acetonitrile, 5% to 20% acetonitrile, 30 mL / min) to give compound 19h (70 mg) as a white solid.

[0603] MS (ESI) m / z 270 [M+H] + .

[0604] Step 7 2-(6-(4-(6-(benzyloxy)pyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-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-chloro-4-(trifluoromethyl)phenyl)acetamide 19i

[0605] Compound 1e (50 mg, 0.088 mmol) was dissolved in DMF (2 mL), and compound 19h (28.61 mg, 0.106 mmol) and N,N-diisopropylethylamine (44.5 mg, 0.44 mmol) were added. The reaction solution was cooled to about 0°C, and a 50% ethyl acetate solution of 1-propylphosphoric acid cyclic anhydride (84.6 mg, 0.133 mmol) was added dropwise. After addition, the mixture was returned to room temperature and reacted for 2 hours. Water (5 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (PE: EA = 1:2) to obtain compound 19i (30 mg) as a white solid.

[0606] MS (ESI) m / z 817 [M+H] + .

[0607] Step 8 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(6-hydroxypyrazolyl)[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 19

[0608] Compound 19i (30 mg, 0.037 mmol) was dissolved in trifluoroacetic acid (1 mL), and the temperature was raised to 50 °C for 18 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 30%-50% acetonitrile, 30 mL / min) to give compound 19 (3.8 mg) as a white solid.

[0609] MS (ESI) m / z 727 [M+H] + .

[0610] 1 H NMR (400 MHz, DMSO-d 6)δ10.39(s,1H),8.34(s,1H),8.04(d,J=8.6Hz,1H),7.94(d,J=8.6Hz,1 H),7.90(s,1H),7.69(d,J=8.6Hz,1H),6.82(s,1H),6.57(s,1H),5.30(s ,2H),4.52(d,J=12.4Hz,1H),4.24(m,2H),3.79(m,3H),3.49(m,3H),3. 17(m,2H),2.99(m,3H),2.81(m,1H),2.63(m,1H),1.17(t,J=7.6Hz,3H).

[0611] Example 20 (S)-N-(1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4)-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-5-hydroxy-N,6-dimethylpyrimidine-4-carboxamide

[0612]

[0613] First step (S)-(1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)(methyl)carbamic acid tert-butyl ester 20a

[0614] Compound 4a (80 mg, 0.14 mmol) was dissolved in dimethyl sulfoxide (2 mL), and (S)-methyl(pyrrolidin-3-yl)carbamic acid tert-butyl ester (286.2 mg, 1.43 mmol) and silver tetrafluoroborate (29.2 mg, 0.15 mmol) were added. The atmosphere was replaced with nitrogen three times, and the temperature was raised to 130 °C for reaction for 5 h. The mixture was cooled to room temperature, diluted with water (5 mL), extracted with ethyl acetate (5 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was separated by silica gel column chromatography (PE:EA=1:1) to give compound 20a (28 mg) as a white solid.

[0615] MS (ESI) m / z 680 [M+H] + .

[0616] Step 2 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-(methylamino)pyrrolidin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 20b

[0617] Compound 20a (28 mg, 0.041 mmol) was dissolved in dry dichloromethane (0.5 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was stirred at room temperature for 1 h. The mixture was concentrated under reduced pressure, and the crude product was dissolved in dichloromethane (3 mL), and the mixture was concentrated under reduced pressure. The operation was repeated at least 3 times to obtain compound 20b (30 mg) as a yellow-brown solid. The crude product was directly fed to the next step.

[0618] MS (ESI) m / z 580 [M+H] + .

[0619] Step 3 (S)-5-(Benzyloxy)-N-(1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro)-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-N,6-dimethylpyrimidine-4-carboxamide 20c

[0620] Compound 20b (30 mg, 0.052 mmol) was dissolved in DMF (1 mL), and compound 1e (15.23 mg, 0.062 mmol) and N,N-diisopropylethylamine (33.6 mg, 0.26 mmol) were added in sequence. The reaction solution was cooled to about 0°C, and a 50% ethyl acetate solution of 1-propylphosphoric acid cyclic anhydride (49.6 mg, 0.078 mmol) was added dropwise. After addition, the mixture was returned to room temperature and stirred for 2 h. Water (5 mL) was added to quench the mixture, and the mixture was extracted with dichloromethane (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified on a silica gel plate (PE / EA=1 / 2) to obtain compound 20c (8 mg) as a white solid.

[0621] MS (ESI) m / z 806 [M+H] + .

[0622] Step 4 (S)-N-(1-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4)-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)pyrrolidin-3-yl)-5-hydroxy-N,6-dimethylpyrimidine-4-carboxamide 20

[0623] Compound 20c (20 mg, 0.025 mmol) was dissolved in trifluoroacetic acid (1 mL), and the temperature was raised to 50 °C for 18 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated by HPLC (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 30%-50% acetonitrile, 30 mL / min) to give compound 20 (1.7 mg) as a white solid.

[0624] MS (ESI) m / z 715 [M+H] + .

[0625] 1 H NMR (400 MHz, DMSO-d 6 )δ8.06(d,J=8.4Hz,1H),7.91(s,2H),7.67(d,J=8.0Hz,1H),6.82(s,1H),5.32(m,1H),5.26(s,2H),4.39(m ,1H),4.25(m,2H),3.80(m,2H),2.99(s,3H),2.88(m,3H),2.22(m,3H),2.07(m,3H),1.27(t,J=7.5Hz,3H).

[0626] Example 21 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-((R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0627]

[0628] First step (2R)-4-(1-methoxy-1,3-dioxolan-2-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 21b

[0629] Compound 21a (4.74 g 22.7 mmol) was dissolved in acetonitrile (40 mL), and (R)-tert-butyl 2-methylpiperazine-1-carboxylate (5 g 24.97 mmol) and potassium carbonate (6.27 g, 45.39 mmol) were added. The mixture was reacted at room temperature for 4 h. Water was added and extracted with ethyl acetate (100 mL×3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 21b (6.69 g).

[0630] MS (ESI) m / z 328 [M+H] +.

[0631] Step 2 (R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 21d

[0632] The synthesis of compound 21c refers to WO2022249060A1. Compound 21b (3.5 g, 10.66 mmol), compound 21c (1.58 g 9.69 mmol) and anhydrous phosphoric acid (1.04 g 10.66 mmol) were added to a reaction bottle, dissolved with ethanol (20 mL), heated to 80 ° C, reacted for 16 hours, concentrated the reaction solution, added water, extracted with ethyl acetate (50 mL × 3), combined the organic phases, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound 21d (1.65 g).

[0633] MS (ESI) m / z 441 [M+H] + .

[0634] Step 3 (R)-4-(2-bromo-4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 21e

[0635] Compound 21d (1.65 g, 3.74 mmol) was dissolved in acetonitrile (10 mL), and 2-bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (1.3 g 4.11 mmol) and DIPEA (1.45 g 11.22 mmol) were added. The mixture was reacted at room temperature for 3 h. Water was added, and the mixture was extracted with ethyl acetate (30 mL×3). The organic phases were combined and concentrated under reduced pressure to give a crude product. The crude product was purified by silica gel column chromatography to give compound 21e (1.13 g).

[0636] MS (ESI) m / z 677 [M+H] + .

[0637] Step 4: (2R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 21f

[0638] Compound 21e (0.50 g, 0.74 mmol) was dissolved in 1,4-dioxane (10 mL) and water (2 mL), and compound 2a (0.23 g, 0.89 mmol), potassium phosphate (0.47 g, 2.22 mmol) and Pd(dppf)Cl were added. 2 (54.0 mg, 0.074 mmol), N 2 The mixture was heated to 85°C under protection and reacted for 16 h. Water was added and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain compound 21f (243 mg).

[0639] MS (ESI) m / z 734 [M+H] + .

[0640] Step 5: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-((R)-3-methylpiperazine-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 21 g

[0641] Compound 21f (243 mg, 0.38 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (3 mL) was added, and the mixture was reacted at room temperature for 1 h and then concentrated under reduced pressure to obtain a crude product 21 g (210.0 mg). The crude product was used directly in the next step without purification.

[0642] MS (ESI) m / z 634 [M+H] + .

[0643] Step 6: 2-(6-((R)-4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 21h

[0644] Compound 21g (210 mg, 0.33 mmol) was dissolved in THF (10 mL), and compound 1e (97 mg, 0.39 mmol), triethylamine (330 mg, 3.3 mmol) and T 3P (300 mg, 0.49 mmol, 50% wt in EA), the reaction solution was reacted at room temperature for 3 h, and then saturated sodium bicarbonate solution was added to quench, extracted with ethyl acetate (50 mL×3), the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (PE / EtOAc=1 / 1) to obtain compound 21h (103 mg).

[0645] MS (ESI) m / z 860.5 [M+H] + .

[0646] Step 7: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-((R)-4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 21

[0647] Compound 21h (103 mg, 0.15 mmol) was dissolved in trifluoroacetic acid (3 mL), heated to 50 °C for 3 h, and then concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative liquid chromatography (preparative column: YMCC18, mobile phase A: 0.1% ammonia in H 2 O, mobile phase B: acetonitrile, 30%-50% acetonitrile, 30 mL / min) to obtain compound 21 (53 mg) as a white solid.

[0648] MS (ESI) m / z 770.4 [M+H] + .

[0649] 1 H NMR (400 MHz, DMSO-d 6)δ10.35(s,1H),10.25(br s,1H),8.56(d,J=4.4Hz,1H),8.06(d,J=8.6Hz,1H),7.96(d,J=2.1Hz,1H),7.71(dd,J=8.7,2.1Hz,1H) ,6.74(s,1H),5.31(s,2H),4.78(m,0.5H),4.39(d,J=12.4Hz,0.5H),3.74(m,3.5H),3.49(m,1.5H),3. 12(m,1H),2.92(m,1H),2.80(d,J=12.0Hz,0.5H),2.68(d,J=12.0Hz,0.5H),2.58(m,2H),2.44(d,J=3. 8Hz,2H),2.27(m,2H),1.89(m,2H),1.78-1.61(m,4H),1.40(m,3H),1.23(s,3H),1.20(t,J=7.5Hz,3H).

[0650] Example 22 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0651]

[0652]

[0653] Step 1 2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane 22b

[0654] Compound 22a (2.1 g, 15 mmol) was dissolved in anhydrous tetrahydrofuran (30 mL), and n-butyl lithium solution (10 mL, 1.6 M in hexane) was slowly added dropwise at -70 °C. After the addition was completed, the mixture was heated to room temperature and stirred for 1 h. The temperature was lowered to -70 °C and 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3.1 g, 16.5 mmol) was slowly added dropwise. After the addition was completed, the mixture was heated to room temperature and stirred for 2 h. Saturated NH 4 The reaction was quenched with Cl, and ethyl acetate (200 mL) and saturated brine (10 mL) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product. The crude product was purified by column chromatography (PE / EtOAc=20 / 1) to give compound 22b (1.0 g) as an off-white solid.

[0655] MS (ESI) m / z 267 [M+H] + .

[0656] Step 2 tert-Butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate 22c

[0657] Compound 1a (954 mg, 1.4 mmol), compound 22b (460 mg, 1.7 mmol), Pd(dppf)Cl 2 (105 mg, 0.14 mmol) and sodium carbonate (459 mg, 4.3 mmol) were added to the sealed tube, and the argon atmosphere was replaced three times. 1,4-dioxane (20 mL) and H 2 O (4 mL), sealed, heated to 100 ° C and stirred for 16 h. Ethyl acetate (100 mL) and saturated brine (10 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=3 / 1) to obtain compound 22c (350 mg) as a light yellow solid.

[0658] MS (ESI) m / z 722 [M+H] + .

[0659] Step 3 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 22d

[0660] Compound 22c (350 mg, 0.48 mmol) was dissolved in DCM (5 mL), TFA (5 mL) was added, and the mixture was reacted at room temperature for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated, ethyl acetate (100 mL) was added, saturated sodium bicarbonate was adjusted to alkalinity, and extraction was performed. The organic phase was washed with saturated brine and concentrated under reduced pressure to obtain crude compound 22d (310 mg), which was used directly in the next step without purification.

[0661] MS (ESI) m / z 622 [M+H] + .

[0662] Step 4: 2-(6-(4-(5-benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 22e

[0663] Compound 22d (310 mg, 0.48 mmol) was dissolved in THF (10 mL), and compound 1e (129 mg, 0.53 mmol), DIPEA (310 mg, 2.4 mmol) and T 3 P (458 mg, 0.72 mmol, 50% in EA), the reaction mixture was reacted at room temperature for 3 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction, and ethyl acetate was extracted (100 mL). The organic phase was washed three times with saturated brine and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (EtOAc) to obtain compound 22e (175 mg) as a white solid.

[0664] MS (ESI) m / z 848 [M+H] + .

[0665] Step 5: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 22

[0666] Compound 22e (175 mg, 0.2 mmol) was dissolved in trifluoroacetic acid (10 mL), and the reaction mixture was reacted at 50°C for 3 h. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated under reduced pressure, ethyl acetate (100 mL) was added, and saturated sodium bicarbonate was adjusted to alkalinity for extraction. The organic phase was washed once with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (EtOAc) to obtain compound 22 (86 mg).

[0667] MS (ESI) m / z 758 [M+H] + .

[0668] 1 H NMR (400 MHz, CDCl 3)δ11.78(s,1H),9.04(s,1H),8.58(s,1H),8.47(d,J=8.7Hz,1H),7.61(s ,2H),7.52(d,J=8.8Hz,1H),5.66(m,1H),5.16(s,2H),4.83(s,2H),4.79 (m,1H),3.97(t,J=5.6Hz,2H),3.81(m,2H),3.49(m,1H),3.18(m,2H),3. 10(m,1H),2.84(m,1H),2.76(m,3H),2.56(s,3H),1.33(t,J=7.5Hz,3H).

[0669] Example 23 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(6-hydroxy-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0670]

[0671] Step 1 2-(6-(4-(6-(Benzyloxy)-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-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-chloro-4-(trifluoromethyl))phenyl)acetamide 23b

[0672] Compound 6f (150 mg, 0.265 mmol) was dissolved in DMF (2 mL), and compound 23a (90.03 mg, 0.318 mmol) and DIEA (171.24 mg, 1.325 mmol) were added. The reaction solution was cooled to 0°C, and a 50% ethyl acetate solution of 1-propylphosphoric acid cyclic anhydride (252.95 mg, 0.3975 mmol) was added dropwise. After addition, the mixture was returned to room temperature and reacted for 2 h. Water (5 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 23b (110 mg) as a reddish brown solid. The crude product was directly used in the next step.

[0673] MS (ESI) m / z 831.2 [M+H] + .

[0674] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(6-hydroxy-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 23

[0675] Compound 23b (110 mg, 0.132 mmol) was dissolved in trifluoroacetic acid (1 mL), reacted at 50° C. for 3 h, and the reaction solution was concentrated under reduced pressure. The crude product was purified by preparative liquid chromatography to give compound 23 (32.9 mg) as a light yellow solid.

[0676] MS (ESI) m / z 741.2 [M+H] + .

[0677] 1 H NMR (400 MHz, DMSO-d 6 )δ10.33(s,1H),8.12(s,1H),8.05(d,J=8.0Hz,1H),7.96(s,1H),7.71(d,J=8.0H z,1H),6.83(s,1H),6.69(d,J=8Hz,1H),5.31(s,2H),4.53(d,J=8Hz,1H),4.25(d, J=8Hz,2H),3.80(t,J=8Hz,2H),3.54-3.51(m,4H),3.27-3.21(m,2H),3.03-2.96 (m,3H),2.83(d,J=8Hz,1H),2.69(s,3H),2.63(d,J=8Hz,2H),1.18(t,J=8Hz,3H).

[0678] Example 24 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(6-hydroxy-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0679]

[0680] Step 1 2-(6-(4-(6-(Benzyloxy)-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 24a

[0681] Compound 22d (150 mg, 0.255 mmol) was dissolved in DMF (2 mL), and compound 23a (74.69 mg, 0.306 mmol) and DIEA (165.09 mg, 1.277 mmol) were added. The reaction solution was cooled to 0°C, and a 50% ethyl acetate solution of 1-propylphosphoric acid cyclic anhydride (121.7 mg, 0.3825 mmol) was added dropwise, and the reaction was allowed to react at room temperature for 2 h. Water (5 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 24a (120 mg) as a reddish brown solid.

[0682] MS (ESI) m / z 887.2 [M+H] + .

[0683] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(6-hydroxy-7-methylpyrazolo[1,5-a]pyrimidine-5-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 24

[0684] Compound 24a (120 mg, 0.135 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50° C. for 3 h. The reaction solution was concentrated under reduced pressure and the crude product was purified by preparative liquid chromatography to give compound 24 (6.3 mg) as a light yellow solid.

[0685] MS (ESI) m / z 797.2 [M+H] + .

[0686] 1 H NMR (400 MHz, DMSO-d 6)δ10.38(s,1H),8.12(s,1H),8.05(d,J=8.0Hz,1H),7.96(s,1H),7.71(d,J=8 .0Hz,1H),7.52(s,1H),6.69(d,J=8Hz,1H),5.31(s,2H),4.77(2,2H),4.54(d, J=8Hz,2H),3.38(t,J=8Hz,2H),3.53-3.42(m,3H),3.03-2.98(m,2H),2.85(d, J=8Hz,1H),2.71(s,1H),2.69(s,3H),2.62(d,J=8Hz,2H),1.19(t,J=8Hz,3H).

[0687] Example 25 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)benzofuran-7-yl)acetamide

[0688]

[0689] Step 1-Bromo-2-(2,2-diethoxyethoxy)-4-(trifluoromethyl)benzene 25b

[0690] 2-Bromo-5-(trifluoromethyl)phenol (25a) (20 g, 83.35 mmol) and 2-bromo-1,1-diethoxyethane (24.51 g, 125.025 mmol) were dissolved in DMF (40 mL), cesium carbonate (40.74 g, 125.025 mmol) was added, and the mixture was reacted at 120°C for 18 h. After the reaction was completed as monitored by TLC, water (50 mL) was added for dilution, and ethyl acetate was extracted (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography (PE: EA = 20: 1) to obtain compound 25b (16 g) as a colorless transparent oil.

[0691] MS (ESI) m / z 379.1 [M+Na] + .

[0692] Step 2 7-Bromo-4-(trifluoromethyl)benzofuran 25c

[0693] Compound 25b (10 g, 28.088 mmol) was dissolved in toluene (150 mL), 85% polyphosphoric acid (104.4 g, 309 mmol) was added, a 10 mL water separator was added, nitrogen was replaced 3 times, and the reaction was carried out at 120°C for 24 h. After the reaction was completed by TLC monitoring, it was cooled to room temperature, the upper toluene organic phase was separated, and the lower liquid was extracted with petroleum ether (100 mL x 3), the upper and lower organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain a crude compound 25c (2 g) as a yellow oil.

[0694] Step 3 1,1-Diphenyl-N-(4-(trifluoromethyl)benzofuran-7-yl)methanamine 25d

[0695] Compound 25c (2 g, 7.577 mmol) and diphenylmethaneimine (1.51 g, 8.335 mmol) were dissolved in toluene (20 mL), and 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (282.69 mg, 0.454 mmol), tris(dibenzylideneacetone)dipalladium (207.87 mg, 0.227 mmol) and sodium tert-butoxide (2.18 g, 22.731 mmol) were added. The atmosphere was replaced with nitrogen three times, and the reaction was carried out at 110 °C for 18 h. The mixture was cooled to room temperature and filtered through celite. The filter cake was washed with ethyl acetate (10 mL x 3). The filtrate was concentrated under reduced pressure to obtain compound 25d (2 g) as a yellow-brown oil.

[0696] MS (ESI) m / z 366.3 [M+H] + .

[0697] Step 4: 4-(Trifluoromethyl)benzofuran-7-amine 25e

[0698] Compound 25d (2 g, 182.55 mmol) was dissolved in 2.0 M ethanolic hydrochloric acid solution (20 mL), reacted at 50° C. for 3 h, cooled to room temperature, and concentrated under reduced pressure. The concentrate was separated by column chromatography (PE:EA=1:1) to give compound 25e (90 mg) as a yellow oil.

[0699] MS (ESI) m / z 202.2 [M+H] + .

[0700] Step 5: 2-Bromo-N-(4-(trifluoromethyl)benzofuran-7-yl)acetamide 25f

[0701] Compound 25e (90 mg, 0.4477 mmol) and DMAP (54.70 mg, 0.4477 mmol) were dissolved in dry dichloromethane (2 mL). The reaction solution was cooled to 0°C, and bromoacetyl bromide (108.44 mg, 0.5372 mmol) was added dropwise. The reaction was allowed to react at room temperature for 2 h. The reaction solution was concentrated, and the crude product was separated by column chromatography (PE / EA=20 / 1) to give compound 25f (70 mg) as a light yellow solid.

[0702] MS (ESI) m / z 322.1 [M+H] + .

[0703] Step 6: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4-(2-oxo-2-((4-(trifluoromethyl)benzofuran-7-yl)amino)ethyl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 25g

[0704] Compound 5c (100.54 mg, 0.2337 mmol) and compound 25f (90 mg, 0.2804 mmol) were dissolved in dry acetonitrile (3 mL), DIEA (90.54 mg, 0.7011 mmol) was added, and the mixture was reacted at room temperature for 18 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated on a silica gel plate (PE / EA=1 / 1) to obtain compound 25g (32 mg) as a light yellow solid.

[0705] MS (ESI) m / z 672.5 [M+H] +

[0706] Step 7: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)benzofuran-7-yl)acetamide 25h

[0707] Compound 25g (32 mg, 0.0477 mmol) was dissolved in dry dichloromethane (0.5 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, saturated sodium bicarbonate (2 mL) was added, and dichloromethane was extracted (3 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 25h (30 mg) as a reddish brown solid.

[0708] MS (ESI) m / z 572.4 [M+H] + .

[0709] Step 8 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-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-(4-(trifluoromethyl)benzofuran-7-yl)acetamide 25i

[0710] Compound 25h (30 mg, 0.0525 mmol) was dissolved in DMF (1 mL), compound 1e (15.38 mg, 0.063 mmol) and DIEA (33.93 mg, 0.2625 mmol) were added, T3P (50.11 mg, 0.07875 mmol, 50% in EA) was added dropwise, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, water (5 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 25i (20 mg) as a reddish brown solid.

[0711] MS (ESI) m / z 798.6 [M+H] + .

[0712] Step 9 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(4-(trifluoromethyl)benzofuran-7-yl)acetamide 25

[0713] Compound 25i (20 mg, 0.025 mmol) was dissolved in trifluoroacetic acid (1 mL) and reacted at 50° C. for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the crude product was purified by preparative liquid chromatography to obtain compound 25 (5.1 mg) as a light yellow solid.

[0714] MS (ESI) m / z 708.5 [M+H] + .

[0715] 1 H NMR (400 MHz, DMSO-d 6)δ11.07(s,1H),8.56(s,1H),8.35(d,J=8.0Hz,1H),8.05(d,J=8.0Hz,1H),7 .60(d,J=8.0Hz,1H),7.13(s,1H),6.81(s,1H),5.36(s,2H),4.52(d,J=8.0H z,2H),4.23(s,2H),3.79(t,J=8Hz,2H),3.66-3.48(m,4H),3.05-2.94(m,3H ),2.82(d,J=8.0Hz,1H),2.64-2.55(m,2H),2.44(s,3H),1.19(t,J=8Hz,3H).

[0716] Example 26 N-(2,4-dichlorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0717]

[0718] Step 1 2-Bromo-N-(2,4-dichlorophenyl)acetamide 26b

[0719] 2,4-Dichloroaniline (26a) (1 g, 6.212 mmol) and DMAP (758.9 mg, 6.212 mmol) were dissolved in dry dichloromethane (10 mL), and bromoacetyl bromide (1.49 g, 7.454 mmol) was added dropwise at 0°C. The mixture was reacted at room temperature for 18 h. The reaction solution was concentrated and the crude product was purified by column chromatography (PE / EA=20 / 1) to give compound 26b (980 mg) as a white solid.

[0720] MS (ESI) m / z 282.1 [M+H] + .

[0721] Step 2 tert-Butyl 4-(4-(2-((2,4-dichlorophenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate 26d

[0722] Compound 26c (300 mg, 0.617 mmol) and compound 26b (260 mg, 0.9256 mmol) were dissolved in dry acetonitrile (3 mL), DIEA (239 mg, 1.851 mmol) was added, and the mixture was reacted at room temperature for 18 h. The reaction solution was concentrated and the crude product was separated by column chromatography (PE / EA=2 / 1) to give compound 26d (230 mg) as a light yellow solid.

[0723] MS (ESI) m / z 688.1 [M+H] + .

[0724] Step 3 N-(2,4-dichlorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 26e

[0725] Compound 26d (230 mg, 0.3347 mmol) was dissolved in dry dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, and the mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, saturated sodium bicarbonate aqueous solution (3 mL) was added to the residue, and the mixture was extracted with dichloromethane (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to obtain compound 26e (200 mg) as a reddish brown solid.

[0726] MS (ESI) m / z 588.1 [M+H] + .

[0727] Step 4 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2,4-dichlorophenyl)acetamide 26f

[0728] Compound 26e (150 mg, 0.255 mmol) was dissolved in DMF (2 mL), compound 1e (74.69 mg, 0.306 mmol) and DIEA (164.78 mg, 1.275 mmol) were added, the reaction solution was cooled to 0°C, T3P (243.4 mg, 0.3825 mmol, 50% in EA) was added dropwise, and the reaction was carried out at room temperature for 2 h. Water (5 mL) was added to quench, and ethyl acetate was extracted (5 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 26f (130 mg) as a reddish brown solid.

[0729] MS (ESI) m / z 814.2 [M+H]+ .

[0730] Step 5: N-(2,4-dichlorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 26

[0731] Compound 26f (130 mg, 0.1599 mmol) was dissolved in trifluoroacetic acid (2 mL), and reacted at 50° C. for 3 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative liquid chromatography to obtain compound 26 (34.2 mg) as a white solid.

[0732] MS (ESI) m / z 724.2 [M+H] + .

[0733] 1 H NMR (400 MHz, DMSO-d 6 )δ10.25(s,1H),8.45(s,1H),7.72(d,J=8.0Hz,1H),7.71(s,1H),7.53( s,1H),7.42(dd,J=8.0Hz,1H),6.67(s,1H),5.26(s,2H),4.77(s,2H),4. 54(d,J=8.0Hz,1H), 3.88(t,J=8Hz,2H),3.61-3.45(m,4H),3.01-2.97(m ,3H),2.83(m,1H),2.71(t,J=8Hz,2H),2.35(s,3H),1.20(t,J=8Hz,3H).

[0734] Example 27 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0735]

[0736] First step (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 27b

[0737] The synthesis of compound 27a refers to patent WO2022249060. Compound 27a (147 mg, 0.22 mmol), compound 22b (69 mg, 0.26 mmol), Pd(dppf)Cl 2 (16 mg, 0.02 mmol), sodium carbonate (69 mg, 0.65 mmol), 1,4-dioxane (5 mL) and H 2 O (1 mL) was added to the reaction flask, argon was replaced three times, and the reaction was carried out at 100°C for 4 h. Ethyl acetate (50 mL) and saturated brine (10 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 27b (155 mg) as a light yellow solid.

[0738] MS (ESI) m / z 736.5 [M+H] + .

[0739] Step 2 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 27c

[0740] Compound 27b (155 mg, 0.21 mmol) was dissolved in DCM (2 mL), TFA (2 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated, ethyl acetate (50 mL) was added, saturated sodium bicarbonate was adjusted to pH = 8, and the organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude compound 27c (123 mg).

[0741] MS (ESI) m / z 636.4 [M+H] + .

[0742] Step 3 (R)-2-(6-(4-(5-benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 27d

[0743] Compound 27c (123 mg, 0.19 mmol) was dissolved in THF (2 mL), and compound 1e (51 mg, 0.21 mmol), DIPEA (129 mg, 1.0 mmol) and T 3 P (191 mg, 0.3 mmol, 50% in EA), reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction, extracted with ethyl acetate (50 mL), the organic phase was washed three times with saturated brine, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=30 / 1) to obtain compound 27d (144 mg) as a light yellow solid.

[0744] MS (ESI) m / z 862.6 [M+H] + .

[0745] Step 4 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 27

[0746] Compound 27d (144 mg, 0.17 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 4 h. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated under reduced pressure, ethyl acetate (50 mL) was added, and saturated sodium bicarbonate aqueous solution was adjusted to pH = 8 for extraction. The organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound 27 (51 mg).

[0747] MS (ESI) m / z 772.6 [M+H] + .

[0748] 1 H NMR (400 MHz, CDCl 3)δ11.78(s,1H),8.96(s,1H),8.58(s,1H),8.45(d,J=8.8Hz,1H),7.61(s,1H),7 .59(s,1H),7.52(d,J=8.8Hz,1H),5.65-5.62(m,1H),5.24(d,J=16.0Hz,1H),5.1 3(d,J=16.0Hz,1H),4.82(s,2H),4.76(m,1H),3.98-3.84(m,3H),3.73(m,1H),3. 55-3.35(m,2H),3.16-2.95(m,2H),2.75(m,3H),2.56(s,3H),1.34-1.27(m,6H).

[0749] Example 28 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0750]

[0751] First step 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 28b

[0752] Compound 28a (160 mg, 0.55 mmol) and compound 1a (300 mg, 0.45 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL), (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (33 mg, 0.045 mmol) and sodium carbonate (144 mg, 1.36 mmol) were added, nitrogen was replaced three times, the reaction was carried out at 100 ° C for 4 h, and the mixture was cooled to room temperature. Water (20 mL) was added for dilution, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA=2 / 1) to obtain compound 28b (180 mg) as a yellow solid.

[0753] MS (ESI) m / z 692[MC 4 H 7 ] + .

[0754] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(4'H,6'H-spiro[cyclopropane]-1,7'-thieno[3,2-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 28c

[0755] Compound 28b (180 mg, 0.24 mmol) was dissolved in dry dichloromethane (2 mL), trifluoroacetic acid (2 mL) was added, the mixture was reacted at room temperature for 1 h, and the mixture was concentrated under reduced pressure. Saturated sodium bicarbonate solution (10 mL) was added, and the mixture was extracted with dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 28c (150 mg) as a yellow solid.

[0756] MS (ESI) m / z 648 [M+H] + .

[0757] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(4'H,6'H-spiro)[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 28d

[0758] Compound 28c (100 mg, 0.154 mmol) and compound 1e (45 mg, 0.184 mmol) were dissolved in dry DMF (3 mL), DIEA (99.5 mg, 0.77 mmol) was added, and the mixture was cooled to 0°C. A 50% ethyl acetate solution (147 mg, 0.23 mmol) of 1-propylphosphoric acid cyclic anhydride was added dropwise. The mixture was reacted at room temperature for 1 h, and water (10 mL) was added to quench the mixture. The mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by column chromatography (PE / EA=1 / 3) to give compound 28d (100 mg) as a white solid.

[0759] MS (ESI) m / z 874 [M+H] + .

[0760] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(4'H,6'H-spiro[cyclopropane-1,7'-thieno[3,2-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 28

[0761] Compound 28d (100 mg, 0.11 mmol) was dissolved in trifluoroacetic acid (5 mL), reacted at 50° C. for 18 h, and concentrated under reduced pressure. The crude product was dissolved in dichloromethane, and saturated aqueous sodium bicarbonate solution was added. The organic phase was concentrated under reduced pressure. The crude product was separated by preparative liquid chromatography to obtain compound 28 (33.0 mg) as a white solid.

[0762] MS (ESI) m / z 784 [M+H] + .

[0763] 1 H NMR (400 MHz, DMSO-d 6 )δ10.37(s,1H),8.57(s,1H),8.05(d,J=8.7Hz,1H),7.97(s,1H),7.72(d,J=8.8Hz, 1H),7.45(s,1H),5.32(s,2H),4.72(s,2H),4.52(d,J=11.4Hz,1H),3.67(s,2H),3. 55–3.45(m,3H),3.27–3.21(m,2H),2.99(s,3H),2.82(d,J=10.4Hz,1H),2.64(d,J= 11.7Hz,1H),2.44(s,3H),1.19(t,J=7.4Hz,3H),1.08(d,J=6.7Hz,2H),0.93(s,2H).

[0764] Example 29 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl-5,5-d 2 )-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0765]

[0766] The first step is 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl-5,5-d 2 tert-Butyl 5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate 29b

[0767] Compound 1a (1.0 g, 1.51 mmol), compound 29a (485 mg, 1.81 mmol), Pd(dppf)Cl 2 (110 mg, 150.85 μmol) and Na 2 CO 3 (480 mg, 4.53 mmol) was dissolved in 1,4-dioxane (20 mL) and water (4 mL), N 2 Replace three times and react at 85°C for 3h. The reaction solution was cooled to room temperature, DCM (50mL) was added, washed with saturated brine (50mL), and the organic phase was concentrated under reduced pressure. The crude product was purified by column chromatography to obtain compound 29b (1.0g) as a yellow solid.

[0768] MS (ESI) m / z 668.1 [MC 4 H 7 ] +

[0769] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5,5-d 2 )-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 29c

[0770] Compound 29b (500 mg, 690.42 μmol) was dissolved in DCM (2 mL), TFA (2 mL) was added, and the mixture was reacted at room temperature for 1.5 h. The mixture was concentrated under reduced pressure, and DCM (50 mL) and saturated NaHCO were added. 3 The aqueous solution was separated and the organic phase was washed with anhydrous Na 2 SO 4 After drying, the residue was concentrated under reduced pressure to give compound 29c (420 mg) as a yellow solid.

[0771] MS (ESI) m / z 624.1 [M+H] +

[0772] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl-5,5-d 2 )-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 29d

[0773] Compound 29c (400 mg, 640.95 μmol) and compound 1e (189 mg, 769.14 μmol) were dissolved in DMF (5.0 mL), and DIPEA (414 mg, 3.20 mmol) and T3P (50% wt in EtOAc, 693 mg, 961.42 μmol) were added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed, the mixture was concentrated under reduced pressure, and DCM (20 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 29d (520 mg) as a yellow solid.

[0774] MS (ESI) m / z 850.3 [M+H] +

[0775] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-(methoxymethyl)-piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 29

[0776] Compound 29d (520 mg, 611.54 μmol) was dissolved in TFA (5 mL) and reacted at 50 °C for 4 h. The mixture was concentrated under reduced pressure, and DCM (10 mL) and saturated NaHCO 3 The solution (5 mL) was added, the organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by preparative liquid chromatography to give compound 29 (15.1 mg) as a light yellow solid.

[0777] MS (ESI) m / z 760.2 [M+H] +

[0778] 1 H NMR (400 MHz, DMSO-d 6 )δ10.40(s,1H),8.54(s,1H),8.05(d,J=8.8Hz,1H),7.97(d,J=1.2Hz,1H),7. 71(dd,J=8.8,1.6Hz,1H),7.52(s,1H),5.34(s,2H),4.76(s,2H),4.53(d,J=1 2.0Hz,1H),3.52-3.46(m,3H),3.26(m,1H),3.05-2.95(m,3H),2.83(d,J=10. 8Hz,1H),2.69(s,2H),2.66-2.62(m,1H),2.43(s,3H),1.19(t,J=7.6Hz,3H).

[0779] Example 30 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-((3-methyloxetane-3-yl)ethynyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0780]

[0781] Step 1 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-((3-methyloxetane-3-yl)ethynyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 30b

[0782] Compound 1a (200 g, 301.71 μmol), compound 30a (203 mg, 1.21 mmol), Xphos-Pd-G2 (24 mg, 30.17 μmol) and CsF (275 mg, 301.71 μmol) were dissolved in DMF (2 mL) and TEA (0.4 mL). 2 Replacement three times, 90 ° C reaction solution for 3h. The reaction solution was concentrated under reduced pressure, and EtOAc (15mL) and saturated NH 4 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 30b (200 mg) as a yellow solid.

[0783] MS (ESI) m / z 622.1 [MC 4 H 7 ] +

[0784] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-((3-methyloxetan-3-yl)ethynyl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4-(7H)-yl)acetamide 30c

[0785] Compound 30b (80 mg, 117.97 μmol) was dissolved in DCM (1.5 mL), TFA (1.5 mL) was added, and the mixture was reacted at room temperature for 1.5 h. After the reaction was completed, the mixture was concentrated under reduced pressure, and DCM (15 mL) and saturated NaHCO were added. 3 The organic phase was separated and concentrated under reduced pressure to give compound 30c (72 mg) as a yellow solid.

[0786] MS (ESI) m / z 578.1 [M+H] +

[0787] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-((3-methyloxetane-3-yl)ethynyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 30d

[0788] Compound 30c (72 mg, 124.57 μmol) and compound 1g (37 mg, 149.48 μmol) were dissolved in DMF (1.0 mL), and DIPEA (81 mg, 622.84 μmol) and T3P (50% wt in EtOAc, 134.63 mg, 186.85 μmol) were added, and the mixture was reacted at room temperature for 1 h. After the reaction was completed, the mixture was concentrated under reduced pressure, and DCM (10 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 30d (86 mg) as a yellow solid.

[0789] MS (ESI) m / z 804.3 [M+H] +

[0790] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-((3-methyloxetane-3-yl)ethynyl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-(7H)-yl)acetamide 30

[0791] Compound 30d (84 mg, 104.45 μmol) was dissolved in TFA (1 mL) and reacted at 50 °C for 3 h. The reaction solution was concentrated under reduced pressure, and DCM (10 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by preparative liquid chromatography to give compound 30 (27.3 mg) as a white solid.

[0792] MS (ESI) m / z 714.2 [M+H] +

[0793] 1 H NMR (400 MHz, DMSO-d 6)δ10.33(s,1H),8.52(s,1H),8.10(d,J=8.6Hz,1H),7.96(d,J=1.3Hz,1H),7.71(dd ,J=8.6,1.3Hz,1H),5.33(s,2H),4.75(d,J=5.6Hz,2H),4.52(d,J=12.0Hz,1H),4.4 5(d,J=5.6Hz,2H),3.51-3.44(m,3H),3.22(m,1H),3.03-2.94(m,3H),2.82(d,J=11 .2Hz,1H),2.64(d,J=11.2Hz,1H),2.43(s,3H),1.64(s,3H),1.18(t,J=7.4Hz,3H).

[0794] Example 31 N-(2-chloro-4-cyanophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0795]

[0796] Step 1 2-Bromo-N-(2-chloro-4-cyanophenyl)acetamide 31b

[0797] Compound 31a (2.0 g, 13.11 mmol) and DMAP (2.5 g, 20.46 mmol) were dissolved in DCM (40 mL). The reaction solution was cooled to 0°C, bromoacetyl bromide (4.68 g, 23 mmol) was added, and the reaction was allowed to react at room temperature for 1 h. After the reaction was completed, the reaction solution was quenched with water (50 mL), extracted with dichloromethane, and the organic phase was concentrated under reduced pressure and purified by silica gel column chromatography to obtain compound 31b (3.0 g) as a light yellow solid.

[0798] MS (ESI) m / z 273.1 [M+H] +

[0799] Step 2 4-(2-bromo-4-(2-((2-chloro-4-cyanophenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 31c

[0800] Compound 5c (500 mg, 1.17 mmol) and compound 31b (400 mg, 1.46 mmol) were dissolved in acetonitrile (10 mL), DIEA (0.42 mL, 2.40 mmol) was added, and the mixture was reacted at room temperature for 3 h. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain compound 31c (680 mg) as a yellow solid.

[0801] MS (ESI) m / z 563.3 [M–C 4 H 7 ] +

[0802] Step 3 4-(4-(2-((2-chloro-4-cyanophenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 31d

[0803] Compound 31c (300 mg, 483.95 μmol) and compound 22b (160 mg, 601.14 μmol) were dissolved in 1,4-dioxane (5 mL) and water (1 mL), and Na 2 CO 3 (160 mg, 1.51 mmol) and Pd(dppf)Cl 2 (35.0 mg, 47.83 μmol). The reaction solution was replaced with nitrogen three times and reacted at 85°C for 16 h. The reaction solution was concentrated under reduced pressure and the crude product was purified by column chromatography to obtain compound 31d (70 mg) as a yellow solid.

[0804] MS (ESI) m / z 623.3 [M–C 4 H 7 ] +

[0805] Step 4: N-(2-chloro-4-cyanophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 31e

[0806] Compound 31d (70 mg, 103.06 μmol) was dissolved in DCM (2 mL) and TFA (1 mL) and reacted at room temperature for 1 h. The reaction solution was concentrated under reduced pressure, and DCM (10 mL) and saturated NaHCO 3 The organic phase was concentrated under reduced pressure and purified by column chromatography to give compound 31e (77 mg) as a yellow solid.

[0807] MS (ESI) m / z 579.4 [M+H] +

[0808] Step 5 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-cyanophenyl)acetamide 31f

[0809] Compound 31e (77 mg, 132.97 μmol) and compound 1e (42 mg, 171.95 μmol) were dissolved in DMF (1.5 mL), and DIPEA (86 mg, 665.39 μmol) and T3P (143 mg, 396.93 μmol) were added, and the mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography to obtain compound 31f (55 mg) as a yellow solid.

[0810] MS (ESI) m / z 805.5 [M+H] +

[0811] Step 6: N-(2-chloro-4-cyanophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 31

[0812] Compound 31f (53 mg, 65.81 μmol) was dissolved in TFA (2.0 mL) and reacted at 50 °C for 3 h. The reaction solution was concentrated, and DCM (15 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure. The crude product was purified by preparative liquid chromatography to give compound 31 (12.7 mg) as a white solid.

[0813] MS (ESI) m / z 715.5 [M+H] +

[0814] 1 H NMR (400 MHz, DMSO-d 6)δ10.29(br s,1H),8.54(s,1H),8.15(d,J=1.8Hz,1H),8.05(d,J=8.4Hz,1H),7.80(dd,J =8.4,1.8Hz,1H),7.51(s,1H),5.34(s,2H),5.32(m,1H),4.76(s,2H),4.53( d,J=12.0Hz,1H),3.87(t,J=5.6Hz,2H),3.50(m,2H),2.99(m,2H),2.82(m,1 H), 2.71-2.63 (m, 3H), 2.43 (s, 3H), 2.04-1.95 (m, 2H), 1.18 (t, J = 7.2Hz, 3H).

[0815] Example 32 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(1-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0816]

[0817] First step 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester 32c

[0818] The synthesis of compounds 32a and 32b is based on WO2022249060. Compound 32a (500 mg, 1.18 mmol) was dissolved in acetonitrile (8 mL), 2-bromo-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (32b) (392 mg, 1.24 mmol) and DIEA (456 mg, 3.54 mmol) were added, and the mixture was reacted at 70°C for 18 h. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA=2 / 1) to obtain compound 32c (420 mg) as a light yellow solid.

[0819] MS (ESI) m / z 604 [MC 4 H 7 ] + .

[0820] Step 2 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester 32d

[0821] Compound 32c (320 mg, 0.486 mmol) was dissolved in DMF (5 mL), and compound 22b (190 mg, 0.714 mmol), X-phosPdG3 (20 mg, 0.02 mmol), potassium phosphate (200 mg, 0.94 mmol) and water (0.5 mL) were added, and the mixture was reacted at 80°C for 1 h under nitrogen protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA=1 / 1) to obtain compound 32d (400 mg) as a white solid.

[0822] MS (ESI) m / z 663.1 [MC 4 H 7 ] + .

[0823] Step 3 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-6-(1,2,3,6-tetrahydropyridin-4-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 32e

[0824] Compound 32d (400 mg, 0.55 mmol) was dissolved in DCM (8 mL) and trifluoroacetic acid (4 mL), reacted at room temperature for 1 h, concentrated under reduced pressure, added with saturated sodium bicarbonate solution, extracted with ethyl acetate (20 mL×3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give compound 32e (250 mg) as a white solid.

[0825] MS (ESI) m / z 619.1 [M+1] + .

[0826] Step 4 2-(6-(1-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 32f

[0827] Compound 32e (250 mg, 0.405 mmol) was dissolved in DMF (10 mL), and compound 1e (125 mg, 0.526 mmol), 1-propylphosphoric anhydride ethyl acetate solution (250 mg, 0.607 mmol) and DIEA (150 mg, 1.16 mmol) were added, and the mixture was reacted at room temperature for 1 h. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain compound 32f (300 mg) as a white solid.

[0828] MS (ESI) m / z 845.2 [M+1] + .

[0829] Step 5: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(1-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-1,2,3,6-tetrahydropyridin-4-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 32

[0830] Compound 32f (30 mg, 0.036 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 1.5 h. Saturated sodium bicarbonate solution was added to the reaction solution to adjust to neutrality, extracted with dichloromethane, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound 32 (13 mg) as a white solid.

[0831] MS (ESI) m / z 755.2 [M+1] + .

[0832] 1 H NMR (400 MHz, DMSO-d 6 )δ8.05(dd,J=8.0,4.0Hz,1H),7.88(s,1H),7.82(d,J=8.0Hz,1H),7.64(d ,J=8.0Hz,1H),7.53(s,1H),6.02–5.63(m,2H),5.41–5.10(m,3H),4.77(s, 2H),4.01(d,J=4.0Hz,1H),3.87(t,J=5.5Hz,2H),2.78–2.64(m,4H),2.36– 2.29(m,1H),2.21(s,3H),1.99(s,1H),1.76(d,J=4.0Hz,1H),1.23(s,3H).

[0833] Example 33 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0834]

[0835] First step (S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 33b

[0836] The synthesis of compound 33a refers to patent WO2022249060. Compound 33a (490 mg, 0.72 mmol), compound 22b (193 mg, 0.72 mmol), Pd(dppf)Cl 2 (32 mg, 0.04 mmol), sodium carbonate (228 mg, 2.16 mmol) were dissolved in 1,4-dioxane (20 mL) and H 2 O (4 mL), replaced with nitrogen three times, reacted at 100 ° C for 4 h. Ethyl acetate (150 mL) and saturated brine (30 mL) were added, the organic phase was separated, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 33b (220 mg) as a light yellow solid.

[0837] MS (ESI) m / z 736.5 [M+H] + .

[0838] Step 2 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 33c

[0839] Compound 33b (220 mg, 0.65 mmol) was dissolved in DCM (10 mL), and hydrogen chloride ethanol solution (2 mL, 4 mol / L) was added and reacted at room temperature for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain crude compound 33c (160 mg).

[0840] MS (ESI) m / z 636.4 [M+H] + .

[0841] Step 3 (S)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 33d

[0842] Compound 33c (100 mg, 0.17 mmol) was dissolved in DMF (2 mL), and compound 1e (41 mg, 0.17 mmol), triethylamine (52 mg, 0.51 mmol), EDCI (33 mg, 0.17 mmol) and HOBt (23 mg, 0.17 mmol) were added, and the reaction was carried out at room temperature for 16 h. After the reaction was completed by TLC monitoring, ethyl acetate (50 mL) was added, and the organic phase was washed three times with saturated brine (20 mL). The organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=30 / 1) to obtain compound 33d (76 mg) as a light yellow solid.

[0843] MS (ESI) m / z 862.6 [M+H] + .

[0844] Step 4 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 33

[0845] Compound 33d (76 mg, 0.1 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 3 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure, ethyl acetate (50 mL) and saturated sodium bicarbonate aqueous solution were added, the organic phase was separated, and the mixture was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10 / 1) to obtain compound 33 (2.5 mg).

[0846] MS (ESI) m / z 772.2 [M+H] + .

[0847] 1 H NMR (400 MHz, CDCl 3)δ11.77(s,0.5H),11.70(s,0.5H),9.04(s,1H),8.60(d,J=6.6Hz,1H),8.49(d,J=8. 6Hz,1H),7.63(m,2H),7.53(m,1H),5.75(m,0.5H),5.48(m,0.5H),5.13(s,2H),5.06( m,0.5H),4.84(s,2H),4.68(m,0.5H),3.98(m,3.5H),3.81(m,1H),3.69(m,0.5H),3. 36-3.16(m,2H),2.86-2.66(m,4H),2.57(s,1.5H),2.56(s,1.5H),1.40-1.32(m,6H).

[0848] Example 34 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0849]

[0850] First step (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester 34a

[0851] Compound 21e (500 mg, 0.74 mmol), compound 22b (236 mg, 0.89 mmol), Pd(dppf)Cl 2 (32 mg, 0.04 mmol) and sodium carbonate (229 mg, 2.22 mmol) were dissolved in 1,4-dioxane (20 mL) and H 2 O (4 mL), replaced with nitrogen three times, reacted at 100° C. for 4 h, added ethyl acetate (150 mL) and saturated brine (30 mL), separated the organic phase, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 34a (480 mg) as a light yellow solid.

[0852] MS (ESI) m / z 736.5 [M+H] + .

[0853] Step 2 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 34b

[0854] Compound 34a (480 mg, 0.65 mmol) was dissolved in DCM (10 mL), and HCl-EtOH (2 mL, 4 M) was added and reacted at room temperature for 3 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain a crude compound 34b (180 mg).

[0855] MS (ESI) m / z 636.4 [M+H] + .

[0856] Step 3 (R)-2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidine-4(7H)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 34c

[0857] Compound 34b (110 mg, 0.17 mmol) was dissolved in DMF (2 mL), and compound 1e (42 mg, 0.17 mmol), triethylamine (52 mg, 0.51 mmol), EDCI (33 mg, 0.17 mmol) and HOBt (23 mg, 0.17 mmol) were added, and the mixture was reacted at room temperature for 16 h. After the reaction was completed by TLC monitoring, ethyl acetate (50 mL) was added, and the organic phase was washed three times with saturated brine (20 mL x 3). The organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=30 / 1) to obtain compound 34c (86 mg) as a light yellow solid.

[0858] MS (ESI) m / z 862.6 [M+H] + .

[0859] Step 4 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 34

[0860] Compound 34c (86 mg, 0.1 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 4 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, ethyl acetate (50 mL) and saturated sodium bicarbonate aqueous solution were added, the organic phase was separated, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10 / 1) to obtain compound 34 (25.2 mg).

[0861] MS (ESI) m / z 772.5 [M+H] + .

[0862] 1 H NMR (400 MHz, DMSO-d 6 )δ10.39(s,1H),8.53(s,1H),8.15(s,1H),8.05(d,J=8.6Hz,1H),7.96(s,1H),7.71(d,J=8.0Hz,1H),7.52(s,1H),5.33(s, 2H),4.76(s,2H),3.87(m,2H),3.69(m,2H),3.53(m,3H),3.20(m,3H),2.96(m,2H),2.70(m,2H),2.42(m,2H),1.23(m,6H).

[0863] Example 35 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide KTH051106

[0864]

[0865] First step (S)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-3-methylpiperazine-1-carboxylic acid tert-butyl ester 35b

[0866] Compound 35a was prepared according to patent WO2022249060. Compound 35a (235 mg, 0.35 mmol), compound 22b (111.1 mg, 0.42 mmol), Pd(dppf)Cl 2(25.5 mg, 0.03 mmol), sodium carbonate (110.7 mg, 1.05 mmol), 1,4-dioxane (5 mL) and H 2 O (1 mL) was added to the reaction flask, argon was replaced three times, and the reaction was carried out at 100°C for 4 h. Ethyl acetate (50 mL) and saturated brine (10 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 35b (100 mg) as a light yellow solid.

[0867] MS (ESI) m / z 680.2 [M–C 4 H 7 ] + .

[0868] Step 2 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 35c

[0869] Compound 35b (100 mg, 0.13 mmol) was dissolved in DCM (4 mL), TFA (3 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated, ethyl acetate (50 mL) was added, saturated sodium bicarbonate was adjusted to alkalinity, and the organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude compound 35c (60 mg).

[0870] MS (ESI) m / z 636.1 [M+H] + .

[0871] Step 3 (S)-2-(6-(4-(5-benzyloxy)-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 35d

[0872] Compound 35c (60 mg, 0.09 mmol) was dissolved in THF (2 mL), and compound 1e (25.32 mg, 0.10 mmol), DIPEA (61 mg, 0.47 mmol) and THF were added. 3P (45 mg, 0.14 mmol, 50% in EA), react at room temperature for 4 h. After the reaction was completed by TLC monitoring, saturated sodium bicarbonate solution was added to quench the reaction, extracted with ethyl acetate (50 mL), and the organic phase was washed three times with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=30 / 1) to obtain compound 35d (68 mg) as a light yellow solid.

[0873] MS (ESI) m / z 862.3 [M+H] + .

[0874] Step 4 (S)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-2-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 35

[0875] Compound 35d (68 mg, 0.07 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 4 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, ethyl acetate (50 mL) was added, and the pH was adjusted to 8 with saturated sodium bicarbonate aqueous solution, and extracted. The organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (EtOAc) to obtain compound 35 (27.3 mg).

[0876] MS (ESI) m / z 772.2 [M+H] + .

[0877] 1 H NMR (400 MHz, CDCl 3 )δ11.77(s,1H),9.02(s,1H),8.58(d,J=9.4Hz,1H),8.44(d,J=8.8Hz,1H),7.61(s,1 H),7.57(s,1H),7.51(dd,J=8.8,2.0Hz,1H),5.65-5.56(m,1H),5.27(d,J=16.0Hz,1 H),5.15(d,J=16.0Hz,1H),4.81(s,2H),4.75(m,1H),3.97-3.85(m,3H),3.72(m,1H) ,3.55-3.35(m,2H),3.09-2.92(m,2H),2.74(m,3H),2.56(s,3H),1.34-1.27(m,6H).

[0878] Example 36 N-(2-chloro-4-fluorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0879]

[0880] Step 1 2-Bromo-N-(2-chloro-4-fluorophenyl)acetamide 36b

[0881] Compound 36a (2 g, 13.7 mmol), 2-bromoacetyl bromide (3.0 g, 15 mmol) and DMAP (2.5 g, 20.4 mmol) were dissolved in DCM (40 mL), replaced with argon three times, and the reaction solution was stirred for 4 h in an ice bath. Ethyl acetate (150 mL) and saturated brine (50 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=5 / 1) to obtain compound 36b (2.87 g) as a white solid.

[0882] MS (ESI) m / z 266.1 [M+H] + .

[0883] Step 2 4-(2-bromo-4-(2-(2-chloro-4-fluorophenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 36c

[0884] Compound 36b (2.05 g, 4.8 mmol), compound 5c (1.4 g, 5.28 mmol), DIEA (1.86 g, 14.4 mmol) and acetonitrile (100 mL) were added to a reaction bottle, argon was replaced three times, and the reaction was carried out at room temperature for 3 h. After the reaction was completed by TLC monitoring, ethyl acetate (150 mL) and saturated brine (50 mL) were added for extraction, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 36c (1.1 g) as a white solid.

[0885] MS (ESI) m / z 556.2 [M–C 4 H 7 ] + .

[0886] Step 3 4-(2-bromo-4-(2-(2-chloro-4-fluorophenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 36d

[0887] Compound 36c (1 g, 3.7 mmol), compound 22b (326 mg, 1.2 mmol), Pd(dppf)Cl 2 (99 mg, 0.13 mmol) and sodium carbonate (433 mg, 4 mmol) were dissolved in 1,4-dioxane (50 mL) and H 2 O (10 mL), argon replacement three times, reaction at 100 ° C for 4 h. Ethyl acetate (150 mL) and saturated brine (50 mL) were added for extraction, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (PE / EtOAc=1 / 1) to obtain compound 36d (408 mg) as a white solid.

[0888] MS (ESI) m / z 616.4 [M–C 4 H 7 ] + .

[0889] Step 4: N-(2-chloro-4-fluorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 36e

[0890] Compound 36d (200 mg, 0.29 mmol) was dissolved in DCM (4 mL), TFA (2.5 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, the mixture was concentrated under reduced pressure, ethyl acetate (50 mL) and saturated sodium bicarbonate aqueous solution were added, and the organic phase was washed with saturated brine and concentrated under reduced pressure to give a crude compound 36e (170 mg).

[0891] MS (ESI) m / z 572.4 [M+H] + .

[0892] Step 5 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-fluorophenyl)acetamide 36f

[0893] Compound 36e (120 mg, 0.20 mmol) was dissolved in THF (10 mL), and compound 1e (56.3 mg, 0.23 mmol), DIPEA (135 mg, 1.04 mmol) and THF were added. 3 P (100 mg, 0.31 mmol, 50% in EA) was added and reacted at room temperature for 4 h. After the reaction was completed by TLC monitoring, a saturated sodium bicarbonate solution was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL). The organic phase was washed three times with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=30 / 1) to obtain compound 36f (30 mg) as a light yellow solid.

[0894] MS (ESI) m / z 798.6 [M+H] + .

[0895] Step 6: N-(2-chloro-4-fluorophenyl)-2-(2-(4,7-dihydro-5H-thieno[2,3-c]pyran-2-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 36

[0896] Compound 36f (30 mg, 0.03 mmol) was dissolved in trifluoroacetic acid (2 mL) and reacted at 50°C for 4 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure, ethyl acetate (50 mL) was added, and the pH was adjusted to 8 with saturated sodium bicarbonate aqueous solution, and extracted. The organic phase was washed with saturated brine and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (EtOAc) to obtain compound 36 (8.8 mg).

[0897] MS (ESI) m / z 708.5 [M+H] +

[0898] 1 H NMR (400 MHz, CDCl 3)δ11.80(s,1H),8.74(s,1H),8.59(s,1H),8.23(m,1H),7.62(s,1H),7.11(d, J=7.6Hz,1H),7.00(m,1H),5.66(m,1H),5.35(m,1H),5.10(s,2H),4.84(s,2H ),3.98(m,2H),3.82(m,2H),3.49(m,1H),3.25-3.15(m,2H),3.07(m,1H),2.8 5(m,1H),2.78(m,2H),2.56(s,3H),2.24-2.19(m,1H),1.33(t,J=7.6Hz,3H).

[0899] Example 37 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0900]

[0901] First step 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 37b

[0902] Compound 37a (0.5 g, 1.71 mmol), 1 g (0.94 g, 1.42 mmol), Pd(dppf)Cl 2 DCM (0.09 g, 0.11 mmol) and sodium carbonate (0.35 g, 3.3 mmol) were dissolved in 1,4-dioxane (20 mL) and water (4 mL), replaced with nitrogen three times, reacted at 100 °C for 6 hours, diluted with water (20 mL), extracted with ethyl acetate, combined organic phases, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography to obtain compound 37b (0.64 g) as a white solid.

[0903] MS (ESI) m / z 748.2 [M+H] +

[0904] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(5'H,7'H-spiro[cyclopropane]-1,4'-thieno[2,3-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 37c

[0905] Compound 37b (0.2 g, 0.26 mmol) was dissolved in dry dichloromethane (5 mL), trifluoroacetic acid (5 mL) was added, the reaction was carried out at room temperature for 3 h, and the mixture was concentrated under reduced pressure. Ethyl acetate and saturated sodium bicarbonate were added to adjust the aqueous solution, and the mixture was extracted with ethyl acetate three times. The organic phases were combined and concentrated to give compound 37c (173 mg).

[0906] MS (ESI) m / z 648.1 [M+H] +

[0907] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(5'H,7'H-spiro)[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 37d

[0908] Compound 37c (173 mg, 0.26 mmol) was dissolved in THF (10 mL), and compound 1e (78 mg, 0.32 mmol), triethylamine (270 mg, 2.67 mmol) and T 3 P (250 mg, 0.39 mmol, 50% wt in EA) was reacted at room temperature for 3 h. Saturated sodium bicarbonate solution was added to quench the mixture, and the mixture was extracted with ethyl acetate (50 mL×3). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography to obtain compound 37d (39.8 mg).

[0909] MS (ESI) m / z 874.2 [M+H] +

[0910] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(5'H,7'H-spiro[cyclopropane-1,4'-thieno[2,3-c]pyran]-2'-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 37

[0911] Compound 37d (39.8 mg, 0.045 mmol) was dissolved in trifluoroacetic acid (10 mL) and reacted at 50°C for 4 h. The crude product was obtained by concentration under reduced pressure, and ethyl acetate and saturated sodium bicarbonate aqueous solution were added, and the product was extracted with ethyl acetate (20 mL) three times. The organic phases were combined, concentrated under reduced pressure, and purified by column chromatography to obtain compound 37 (10 mg).

[0912] MS (ESI) m / z 784.2 [M+H] +

[0913] 1 H NMR (400 MHz, CDCl 3 )δ11.79(s,1H),9.04(s,1H),8.59(s,1H),8.48(d,J=8.6Hz,1H),7.62(s,1H),7.5 3(d,J=8.6Hz,1H),7.32(s,1H),5.67(d,J=13.2Hz,1H),5.13(s,2H),4.93(s,2H), 4.79(d,J=13.2Hz,1H),3.81(t,J=11.7Hz,2H),3.73(s,2H),3.49(m,1H),3.20(m, 2H),3.07(m,1H),2.80(m,2H),2.56(s,3H),1.34(m,3H),1.01(m,2H),0.91(m,2H).

[0914] Example 38 2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

[0915]

[0916] Step 1 4-(5-ethyl-4-(2-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 38b

[0917] Compound 38a (200 mg, 0.31 mmol) and compound 2a (123 mg, 0.465 mmol) were dissolved in 1,4-dioxane (4 mL) and water (0.4 mL), and 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride dichloromethane complex (24.5 mg, 0.03 mmol) and sodium carbonate (98.5 mg, 0.93 mmol) were added. The atmosphere was replaced with nitrogen three times, and the mixture was reacted at 100 °C for 16 h. The mixture was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was separated by column chromatography to obtain compound 38b (53 mg) as a yellow-brown solid.

[0918] MS (ESI) m / z 647.3 [M–C 4 H 7 ] + .

[0919] Step 2 2-(5-ethyl-7-oxo-6-(piperazin-1-yl)-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide 38c

[0920] Compound 38b (53 mg, 0.075 mmol) was dissolved in dry dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the mixture was reacted at room temperature for 1 h. The mixture was concentrated under reduced pressure, and the residue was dissolved in dichloromethane (5 mL), and the mixture was concentrated under reduced pressure. The operation was repeated 3 times to obtain compound 38c (50 mg) as a yellow-brown solid.

[0921] MS (ESI) m / z 604.4 [M+H] + .

[0922] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-7-oxo-2-(1-oxaspiro[4.5]dec-7-en-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide 38d

[0923] Compound 38c (50 mg, 0.082 mmol) was dissolved in DMF (2 mL), compound 1e (23.92 mg, 0.098 mmol) and triethylamine (52.99 mg, 0.41 mmol) were added, the reaction solution was cooled to 0°C, 1-propylphosphoric acid cyclic anhydride 50% ethyl acetate solution (78.27 mg, 0.123 mmol) was added dropwise, and the reaction was allowed to react at room temperature for 2 h. Water (10 mL) was added to quench, and the mixture was extracted with dichloromethane (10 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel plate (PE / EA=1 / 2) to obtain compound 38d (30 mg) as a light yellow solid.

[0924] MS (ESI) m / z 830.6 [M+H] + .

[0925] Step 4: 2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-2-(1-oxaspiro[4.5]dec-7-ene-8-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-fluoro-4-(trifluoromethyl)phenyl)acetamide

[0926] Compound 38d (30 mg, 0.036 mmol) was dissolved in trifluoroacetic acid (1 mL), and reacted at 50° C. for 3 h. The reaction solution was concentrated under reduced pressure and purified by preparative liquid chromatography to give compound 38 (15.7 mg) as a white solid.

[0927] MS (ESI) m / z 740.5 [M+H] + .

[0928] 1 H NMR (400 MHz, DMSO-d 6 )δ10.68(s,1H),8.43(s,1H),8.22(s,1H),7.79(dd,J=8.0Hz,1H),7.56(d,J=8 .0Hz,1H),6.71(s,1H),5.29(s,2H),4.52(d,J=8Hz,1H),3.73(t,J=8Hz,2H),3. 60-3.43(m,4H),3.02-2.93(m,3H),2.82-2.79(m,2H),2.68-2.59(m,2H),2.40 (s,3H),2.27(s,2H),1.92-1.85(m,2H),1.79-1.63(m,4H),1.17(t,J=8Hz,3H).

[0929] Example 39 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((3aR,6aS)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0930]

[0931] First step (3aR,6aS)-5-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran)-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylic acid tert-butyl ester 39a

[0932] Compound 4a (95 mg, 0.17 mmol) was dissolved in DMSO (1 mL), and cis-2-Boc-hexahydropyrrolo[3,4-c]pyrrole (360 mg, 1.7 mmol) and AgBF were added. 4 (33 mg, 0.17 mmol), in N 2 Under protection, the reaction was carried out at 130° C. for 3 hours. After the reaction was completed by TLC monitoring, the organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10 / 1) to obtain compound 39a (35 mg).

[0933] MS (ESI) m / z 692.2 [M+H] + .

[0934] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 39b

[0935] Compound 39a (35 mg, 0.05 mmol) was dissolved in DCM (5 mL), TFA (1 mL) was added, and the mixture was reacted at room temperature for 4 h. After the reaction was completed as monitored by TLC, the mixture was concentrated under reduced pressure to obtain compound 39b (35 mg).

[0936] MS (ESI) m / z 592.2 [M+H] + .

[0937] Step 3 2-(6-((3aR,6aS)-5-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-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-chloro-4-(trifluoromethyl)phenyl)acetamide 39c

[0938] Compound 1e (13.4 mg, 0.55 mmol), EDCI (10.6 mg, 0.55 mmol), HOBt (2.9 mg, 0.55 mmol) and DIEA (19 mg, 150 mmol) were dissolved in DMF (3 mL), and compound 39b (35 mg, 0.5 mmol) was added, and the mixture was reacted at room temperature for 16 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 39c (22 mg).

[0939] MS (ESI) m / z 818.3 [M+H] + .

[0940] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-((3aR,6aS)-5-(5-hydroxy-6-methylpyrimidine-4-carbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 39

[0941] Compound 39c (22 mg, 0.03 mmol) was dissolved in TFA (2 mL) and reacted at 50°C for 16 h. After the reaction was completed as monitored by TLC, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (DCM / MeOH=10:1) to obtain compound 39 (6.4 mg).

[0942] MS (ESI) m / z 728.4 [M+H] + .

[0943] 1 H NMR (400 MHz, CDCl 3)δ12.73(s,1H),8.98(s,1H),8.61(s,1H),8.47(d,J=8.8Hz,1H),7.63(s,1H),7.53 (d,J=8.4Hz,1H),6.96(s,1H),5.35(m,1H),5.07(s,2H),4.53(m,2H),4.40(m,1H), 4.36(m,2H),4.06-4.00(m,1H),3.91(t,J=5.2Hz,2H),3.68(m,1H),3.18(m,1H),3. 04(m,3H),2.72(m,2H),2.55(s,3H),2.22(t,J=7.6Hz,1H),2.02(m,1H),1.31(t,3H)

[0944] Example 40 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)benzofuran-7-yl)acetamide

[0945]

[0946] Step 1 N-(2-methyl-4-(trifluoromethyl)benzofuran-7-yl)-1,1-diphenylmethanamine 40b

[0947] The synthesis of compound 40a is based on WO2023000834 A1. Compound 40a (1.1 g, 3.957 mmol) and diphenylmethaneimine (788.89 mg, 4.353 mmol) were dissolved in toluene (10 mL), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (108.97 mg, 0.119 mmol), tris(dibenzylideneacetone)dipalladium (147.57 mg, 0.237 mmol) and sodium tert-butoxide (1.14 g, 11.871 mmol) were added, nitrogen was replaced three times, the reaction was carried out at 110°C for 18 hours, cooled to room temperature, filtered through diatomaceous earth, the filter cake was washed with ethyl acetate (10 mL x 3), all the filtrates were combined, and concentrated under reduced pressure to obtain a crude compound 40b (1.3 g) as a yellow-brown oil.

[0948] MS (ESI) m / z 380.1 [M+H] +

[0949] Step 2 2-Methyl-4-(trifluoromethyl)benzofuran-7-amine 40c

[0950] Compound 40b (1.3 g, 3.428 mmol) was dissolved in 2M hydrochloric acid-ethanol solution (20 mL), reacted at 50°C for 3 h, cooled to room temperature, concentrated under reduced pressure, and the crude product was purified by column chromatography (PE / EA=1 / 1) to give a crude compound 40c (420 mg) as a yellow oil.

[0951] MS (ESI) m / z 216.1 [M+H] +

[0952] Step 3 2-Bromo-N-(2-methyl-4-(trifluoromethyl)benzofuran-7-yl)acetamide 40d

[0953] Compound 40c (420 mg, 1.953 mmol) and DMAP (238.60 mg, 1.95 mmol) were dissolved in dry dichloromethane (5 mL), cooled to about 0°C, bromoacetyl bromide (473.11 mg, 2.344 mmol) was added dropwise, and the mixture was returned to room temperature for 2 h. Water (5 mL) was added for dilution, and the mixture was extracted with dichloromethane (5 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated on a silica gel plate (PE / EA=20 / 1) to give compound 40d (96 mg) as a light yellow solid.

[0954] MS (ESI) m / z 336.8 [M+H] +

[0955] Step 4: 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((2-methyl-4-(trifluoromethyl)benzofuran-7-yl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester 40e

[0956] Compound 40d (98.65 mg, 0.2293 mmol) and compound 5c (96 mg, 0.2866 mmol) were dissolved in dry acetonitrile (3 mL), DIEA (88.83 mg, 0.6879 mmol) was added, and the mixture was reacted at room temperature for 18 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated on a silica gel plate (PE / EA=1 / 1) to obtain compound 40e (80 mg) as a light yellow solid.

[0957] MS (ESI) m / z 686.3 [M+H] +

[0958] Step 5 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)benzofuran-7-yl)acetamide 40f

[0959] Compound 40e (80 mg, 0.1167 mmol) was dissolved in dry dichloromethane (1 mL), trifluoroacetic acid (1 mL) was added, and the mixture was reacted at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, saturated sodium bicarbonate (2 mL) was added to the residue, and the mixture was extracted with dichloromethane (3 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give a crude product of compound 40f (70 mg) as a reddish brown solid.

[0960] MS (ESI) m / z 586.3 [M+H] + .

[0961] Step 6: 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazine-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)benzofuran-7-yl)acetamide 40 g

[0962] Compound 40f (70 mg, 0.1196 mmol), compound 1e (43.79 mg, 0.1794 mmol) and DIEA (77.29 mg, 0.598 mmol) were dissolved in DMF (1 mL), the reaction solution was cooled to 0°C, 1-propylphosphoric acid cyclic anhydride 50% ethyl acetate solution (76.11 mg, 0.1196 mmol) was added dropwise, and the mixture was returned to room temperature for 2 h. Water (2 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (3 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 40g (70 mg) as a reddish brown solid, which was used directly in the next step.

[0963] MS (ESI) m / z 812.3 [M+H] + .

[0964] Step 9: 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)benzofuran-7-yl)acetamide 40

[0965] Compound 40g (70 mg, 0.025 mmol) was dissolved in trifluoroacetic acid (1 mL), and the mixture was reacted at 50°C for 1 h. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative liquid chromatography to obtain compound 40 (10.7 mg) as a white solid.

[0966] MS (ESI) m / z 722.3 [M+H] + .

[0967] 1 H NMR (400 MHz, DMSO-d 6 )δ11.02(s,1H),8.40(s,1H),7.96(d,J=8.0Hz,1H),7.51(d,J=8.0Hz,1H),6.81 (s,1H),6.78(s,1H),5.36(s,2H),4.53(d,J=8.0Hz,1H),4.23(d,J=8.0Hz,1H), 3.79(t,J=8Hz,2H),3.56-3.42(m,4H),3.26-3.21(m,3H),3.02-2.95(m,3H),2. 82(d,J=8.0Hz,1H),2.63(s,1H),2.56(s,3H),2.39(s,3H),1.19(t,J=8Hz,3H).

[0968] Example 41 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-(methoxymethyl)-piperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0969]

[0970] First step 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-(methoxymethyl)piperazine-1-carboxylic acid tert-butyl ester 41b

[0971] Compound 4a (400 mg, 713.32 μmol) and compound 41a (1.6 g, 7.13 mmol) were dissolved in NMP (0.03 mL) and Ag was added. 2 O (330 mg, 1.43 mmol), deoxygenated three times, reacted at 140 ° C for 2 h. The reaction solution was cooled to room temperature, EtOAc (30 mL) was added to dissolve, and saturated NH4 The reaction mixture was washed with aqueous Cl solution (20 mL), the organic phase was separated, and the crude product was separated by preparative liquid chromatography to give compound 41b (42 mg) as a white solid.

[0972] MS (ESI) m / z 710.2 [M+H] +

[0973] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-(methoxymethyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4-(7H)-yl)acetamide 41c

[0974] Compound 41b (41 mg, 57.73 μmol) was dissolved in DCM (1 mL), TFA (1 mL) was added, and the mixture was reacted at room temperature for 0.5 h. The reaction solution was concentrated under reduced pressure, and DCM (20 mL) and saturated NaHCO were added. 3 Aqueous solution (5 mL). The organic phase was separated and concentrated under reduced pressure to give compound 41c (42 mg) as a yellow solid.

[0975] MS (ESI) m / z 610.3 [M+H] +

[0976] Step 3 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)-3-(methoxymethyl)piperazin-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-chloro-4-(trifluoromethyl)phenyl)acetamide 41d

[0977] Compound 41c (42 mg, 68.85 μmol) and compound 1e (20 mg, 82.62 μmol) were dissolved in DMF (1 mL), and DIPEA (44 mg, 344.24 μmol) and T3P (50% wt in EtOAc, 74 mg, 103.27 μmol) were added. The reaction mixture was reacted at room temperature for 0.5 h. The reaction mixture was concentrated under reduced pressure, and DCM (15 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by column chromatography to give compound 41d (58 mg) as a yellow solid.

[0978] MS (ESI) m / z 836.3 [M+H] +

[0979] Step 4: N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)-3-(methoxymethyl)-piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 41

[0980] Compound 41d (58 mg, 69.36 μmol) was dissolved in TFA (2 mL) and reacted at 50 °C for 3 h. The reaction solution was concentrated under reduced pressure, and DCM (10 mL) and saturated NaHCO 3 The organic phase was separated and concentrated under reduced pressure to give a crude product, which was purified by preparative liquid chromatography to give compound 41 (15.1 mg) as a light yellow solid.

[0981] MS (ESI) m / z 746.2 [M+H] +

[0982] 1 H NMR (400 MHz, DMSO-d 6 )δ10.36(br s,2H),8.54(d,J=4.4Hz,1H),8.05(d,J=8.8Hz,1H),7.97(d,J=1.6Hz,1H),7.72(d,J=8.8Hz,1H),6.83(s,1H),5.37-5.26(m,2H), 4.78-4.73(m,0.5H),4.44-4.37(m,0.5H),4.26(s,1H),4.25(s,1H),3.92(dd,J=17.6,8.8Hz,1H),3.80(t,J=5.2Hz,2H),3.66(dt ,J=11.6,3.6Hz,1H),3.59-3.48(m,2H),3.43-3.37(m,1H),3.32(s,3H),3.17(m,0.5H),3.11-3.05(m,1H),2.93(d,J=11.8Hz,0.5 H),2.88-2.77(m,2H),2.62(d,J=10.8Hz,0.5H),2.52(m,2H),2.44(s,1.5H),2.43(s,1.5H),1.99(m,0.5H),1.20(t,J=7.6Hz,3H)

[0983] Example 42 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1,5,6-trimethyl-2-oxo-1,2-dihydropyridine-3-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0984]

[0985] Compound 42a (190 mg, 0.96 mmol) was dissolved in DMF (5 mL), EDCI (76 mg, 0.40 mmol), HOBt (54 mg, 0.40 mmol) and DIEA (154 mg, 1.20 mmol) were added at room temperature, and then compound 6f was added, and the mixture was reacted at 50 °C for 16 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative liquid chromatography to obtain compound 42 (1.4 mg).

[0986] MS (ESI) m / z 745.2 [M+H] + .

[0987] 1 H NMR (400 MHz, CDCl 3 )δ8.99(s,1H),8.50(d,J=8.6Hz,1H),7.63(s,1H),7.52(d,J=8.6Hz,1H) ,7.52(s,1H),6.96(s,1H),5.35(m,2H),5.08(s,2H),4.36(d,J=2.7Hz,2 H),3.91(t,J=5.4Hz,2H),3.65(m,2H),3.51(s,3H),3.16(m,2H),2.73(m ,4H),2.34(s,3H),2.22(t,J=7.6Hz,2H),2.07(s,3H),1.35-1.31(m,3H)

[0988] Example 43 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0989]

[0990] Step 1 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 43c

[0991] Compound 43b (80 mg, 0.29 mmol) was dissolved in 1,4-dioxane (4 mL) and water (0.5 mL), and compound 43a (190 mg, 0.24 mmol), potassium phosphate (153 mg, 0.72 mmol) and Pd(dppf)Cl were added. 2 (22 mg, 0.03 mmol). The reaction mixture was replaced with nitrogen three times and then reacted at 100 °C for 3 hours. After the reaction solution was cooled, ethyl acetate (30 mL) was added to dilute the reaction solution, and then washed three times with saturated brine. The organic phase was concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by column chromatography (DCM / MeOH=10:1) to obtain compound 43c (90 mg).

[0992] MS (ESI) m / z 861.5 [M+H] + .

[0993] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(6-methyl-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 43

[0994] Compound 43c (90 mg, 0.10 mmol) was dissolved in TFA (5 mL) and reacted at 50°C for 3 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC to obtain compound 43 (38.2 mg) as a white solid.

[0995] MS (ESI) m / z 771.3 [M+H] + .

[0996] 1 H NMR (400 MHz, MeOD-d 4)δ8.55(s,1H),8.16(d,J=8.5Hz,1H),7.82(s,1H),7.64-7.59(m,2H),5.42(s,2H),4.72(d,J=11.9Hz,2H),4.23(s,2H),4.11(d,J=13.0Hz, 1H),3.74(br.,2H),3.47(d,J=13.9Hz,1H),3.17-3.06(m,4H),3.04-2 .95(m,3H),2.86(s,3H),2.80(m,1H),2.52(s,3H),1.36-1.31(m,3H).

[0997] Example 44 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[0998]

[0999] Step 1 2-(6-(4-(5-(benzyloxy)-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-5-ethyl-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide 44b

[1000] Compound 44a (50 mg, 0.2537 mmol) and compound 43a (139.77 mg, 0.2114 mmol) were dissolved in 1,4-dioxane (2 mL) and water (0.4 mL), and Pd(dppf)Cl was added. 2 .CH 2 Cl 2 (17.31 mg, 0.0212 mmol) and sodium carbonate (67.23 mg, 0.6343 mmol), replaced with nitrogen three times, heated to 100 ° C for 18 h, cooled to room temperature, diluted with water (5 mL), extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was separated by column chromatography (DCM / MeOH=10:1) to give compound 44b (42 mg) as a light yellow solid.

[1001] MS (ESI) m / z 861.3 [M+H] + .

[1002] Step 2 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(5-methyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide 44

[1003] Compound 44b (42 mg, 0.0488 mmol) was dissolved in trifluoroacetic acid (1 mL), heated to 50°C for 3 h, and the reaction solution was concentrated under reduced pressure. The crude product was separated by preparative HPLC to obtain compound 44 (15.7 mg) as a light yellow solid.

[1004] MS (ESI) m / z 771.2 [M+H] + .

[1005] 1 H NMR (400 MHz, DMSO-d 6 )δ10.40(s,1H),8.59(s,1H),8.15(s,1H),8.06(d,J=8.0Hz,1H),7.98(d,J =8.0Hz,1H),7.72(dd,J=8.0Hz,1H),7.48(s,1H),5.35(s,2H),4.53(d,J=8 Hz,1H),3.60(s,2H),3.54-3.46(m,4H),3.05-2.96(m,4H),2.92(d,J=8.0H z,2H),2.83(d,J=12Hz,3H),2.48(s,3H),2.45(s,3H),1.20(t,J=8Hz,3H).

[1006] Example 45 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(5-(methyl-d 3 )-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[1007]

[1008] The synthesis of intermediate 45a refers to intermediate 43b, except that methyl p-toluenesulfonate is replaced by D 3- p-Toluenesulfonylmethyl ester and replace 4,5,6,7-tetrahydrothieno[2,3-c]pyridine hydrochloride (43b-1) with 4,5,6,7-tetrahydrothieno[3,2-c]pyridine hydrochloride (44a-1); MS (ESI) m / z 283.2 [M+H] + .

[1009] Synthesis of Example 45 Referring to Example 44, intermediate 44a was replaced by 5-(methyl-d 3 )-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine (45a).

[1010] MS (ESI) m / z 774.3 [M+H] + .

[1011] 1 H NMR (400 MHz, CDCl 3 )δ9.06(s,1H),8.59(s,1H),8.45(d,J=8.7Hz,1H),7.62(d,J=2.0Hz,1H),7.52(d,J=8.8Hz,1H),7.49(s,1H),5.67(d,J=13.2Hz,1H),5.1 6(s,2H),4.78(s,1H),4.01(s,2H),3.80(m,2H),3.49(m,2H),3.27(s,2H),3.16(s,4H),2.81(m,2H),2.57(s,3H),1.33(t,J=7.5Hz,3H).

[1012] Example 46 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(5-ethyl-4,5,6,7-tetrahydrothieno[3,2-c]pyridin-2-yl))-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[1013]

[1014] The synthesis of intermediate 46a refers to intermediate 43b; methyl p-toluenesulfonate is replaced by ethyl p-toluenesulfonate and 4,5,6,7-tetrahydrothieno[2,3-c]pyridine hydrochloride (43b-1) is replaced by 4,5,6,7-tetrahydrothieno[3,2-c]pyridine hydrochloride (44a-1); MS (ESI) m / z 294.2 [M+H] + .

[1015] Synthesis of Example 46 Referring to Example 44, intermediate 44a was replaced with 5-ethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4,5,6,7-tetrahydrothieno[3,2-c]pyridine (46a).

[1016] MS (ESI) m / z 785.3 [M+H] + .

[1017] 1 H NMR (400 MHz, CDCl 3 )δ9.07(s,1H),8.60(s,1H),8.48(d,J=8.63Hz,1H),8.20(br.,1H),7.63(s,1H),7.57-7.50(m,2H),5.66(br.,1H),5.15( s,2H),4.81(m,1H),3.94-3.75(m,4H),3.26-3.01(m,8H),2.92(m,2H),2.84-2.78(m,2H),2.58(s,3H),1.39-1.28(m,6H).

[1018] Example 47 N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(4-(5-hydroxy-6-methylpyrimidine-4-carbonyl)piperazin-1-yl)-2-(6-(methyl-d 3 )-4,5,6,7-tetrahydrothieno[2,3-c]pyridin-2-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide

[1019]

[1020] The synthesis of intermediate 47a refers to intermediate 43b, except that methyl p-toluenesulfonate is replaced by p-D 3 -p-Toluenesulfonyl methyl ester; MS (ESI) m / z 283.2 [M+H] + .

[1021] The synthesis of Example 47 was carried out with reference to Example 44; Intermediate 44a was replaced with 6-(methyl-d 3 )-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-4,5,6,7-tetrahydrothieno[2,3-c]pyridine (47a).

[1022] MS (ESI) m / z 774.3 [M+H] + .

[1023] 1H NMR (400 MHz, DMSO-d 6 )δ10.40(s,1H),10.24(s,1H),8.59(s,1H),8.06(d,J=8.0Hz,1H),7.98(d ,J=8.0Hz,1H),7.73(dd,J=8.0Hz,1H),7.61(s,1H),5.35(s,2H),4.53(d,J =8Hz,1H),3.51(d,J=8Hz,3H),3.35~3.42(m,4H),3.32~3.30(m,4H),3.01 (s,2H),2.84(d,J=8Hz,1H),2.65(s,1H),2.45(s,3H),1.20(t,J=8Hz,3H).

[1024] NSC617145 was purchased from a commercial company, manufacturer: Bidex Pharmaceuticals, catalog number: BD628938

[1025] The synthesis of control compound 1 refers to patent WO2022249060A1.

[1026]

[1027] Test Example 1: Test of Compounds on WRN (517-1238) Enzyme Activity

[1028] This experiment studies the inhibitory effect of compounds on WRN (517-1238) enzyme activity by detecting the effects of compounds on WRN (517-1238) enzyme activity in vitro. NSC617145 is the internal control compound in this project.

[1029] Experimental reagents and consumables

[1030] Reagents factory Part Number WRN(517-1238) ICE S2212T-H56HZ dsDNA Genscript custom made ATP Ark Pharm Ak-54737

[1031] Consumables supplier Part Number 96Well Plates Nunc 249944 384PP Plate LABCYTE PP-0200 384-well plate Corning 4514

[1032]

[1033] Compounds were serially diluted using DMSO in the 384PP Plate compound dilution plate, and 0.15 μL of compound was transferred to a 384-reaction microplate (Corning 4514) using an Echo, ensuring that the final DMSO content was 1% (duplicate). 5 μL of WRN (517-1238) enzyme solution was added to each well of the 384-reaction microplate and incubated at 25°C for 10 minutes. Wells containing 1% DMSO and enzyme were used as high controls, and wells containing the same amount of DMSO and buffer were used as low controls. 5 μL of dsDNA solution and 5 μL of ATP solution were added to each well and incubated at 25°C for 20 minutes (final concentrations: 1 nM WRN, 5 nM dsDNA, 50 μM ATP). Fluorescence signal values ​​were read on a BMG (CLARIO Star Plusacu) microplate reader (Ex: 620 nm, Em: 685 nm). The percentage of inhibition of compound-treated wells was standardized between high control and low control %inhibition = ((signal value high control -Signal value 化合物处理 ) / (signal value high control -Signal value low control )*100). Then the four-parameter IC was fitted by XLfit 5.5.0 50 Curve and analyze.

[1034] Table 1 Inhibition results of test compounds on WRN(517-1238)_FI

[1035]

[1036]

[1037]

[1038] As can be seen from the above table, the compounds of the present invention have a good inhibitory effect on WRN(517-1238)_FI.

[1039] Test Example 2: Test of the inhibitory activity of compounds on SW48 cell proliferation

[1040] Complete medium (L-15 + 10% FBS + 1% P / S) was prepared to revive SW48 cells (ATCC), cells in the logarithmic growth phase were collected by centrifugation for two generations and counted, the cells were resuspended to an appropriate concentration, the cell suspension was inoculated in a 96-well plate, 100 μL of cell suspension was added to each well, the seeding density was 5000 cells / well, the 96-well plate was placed in a carbon dioxide incubator (0% carbon dioxide) overnight, 50 μL of the upper culture medium was discarded, and then 40 μL of fresh culture medium was added. The test compound was prepared into a storage solution with DMSO, and DMSO was used to gradually dilute the solution 4 times with 2.5mM as the highest concentration to obtain 9 concentration gradients; the test compound was diluted 25 times with culture medium, and 10μL was added to a 96-well cell plate containing 90μL cells, and culture medium without cells (containing 0.4% DMSO) was added to the Min control well, and 10μL DMSO-cell culture medium mixture (DMSO final concentration was 0.4%) was added to the Max control well, and incubated for 120h at 37℃, 0%, and a carbon dioxide incubator with a relative humidity of more than 90%. 50μL / well CellTiter Glo was added to terminate the reaction, incubated at room temperature in the dark for 10min, and detected in Envision after gentle shaking. Read the fluorescence value RLU of each well, and calculate the cell inhibition rate according to the formula cell growth inhibition rate % = (1-As / Ac) × 100. Where As: RLU sample (cells + CTG + test compound) - RLU Min (culture solution without cells), Ac: RLU normal growth cell control (cells + CTG + DMSO) - RLU Min (culture solution without cells). Input the inhibition rate Inh% (Y) corresponding to each concentration (X) in EXCEL, and use Graphpad Prism 8 software according to the built-in four-parameter fitting formula Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)*HillSlope) to calculate the half inhibitory concentration IC of each compound 50 value.

[1041] Table 2 Inhibition results of test compounds on SW48 cell proliferation

[1042]

[1043]

[1044] The experimental results show that compared with the control compound 1, the compound of the present invention has a good inhibitory effect on the proliferation of SW48 cells.

[1045] Test Example 3: Thermodynamic Solubility Test of Compounds

[1046] Simulated artificial gastrointestinal fluid (FaSSGF (pH = 1.6), FaSSIF (pH = 6.5)) is a commercial reagent purchased from Biorelevant or other commonly used commercial companies. Accurately weigh 1 part / pH of about 2 mg of the compound, add appropriate volumes of simulated artificial gastrointestinal fluid FaSSGF (pH = 1.6) and FaSSIF (pH = 6.5) to obtain a solution with a concentration of 2 mg / mL. After ultrasonication for 10 minutes, fix it on a shaker and shake it overnight at room temperature (at least 8 hours). After shaking, ultrasonicate for 10 minutes and centrifuge at 13000rpm for 15 minutes. Pipette 0.1mL of supernatant into a new tube, shake and rinse for 5 minutes, and then discard the liquid. Transfer 0.5mL of supernatant to a new tube again, centrifuge at 13000rpm for 15 minutes, take the supernatant (if necessary, add water to dilute) and analyze it by LC-UV (if UV cannot be detected, use LC-MS detection). The sample concentration is quantified using a fitted calibration line (3 to 5 points).

[1047] Table 3 Thermodynamic solubility test results of test compounds

[1048]

[1049] The experimental results show that compared with the control compound 1, the compound of the present invention has better solubility in FaSSGF (pH=1.6) solvent.

[1050] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound as represented by formula (I), its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, in, X is selected from CH or N; and when X is CH, the CH may be substituted by R5 or R6 (i.e., X is C-R5 or C-R6); m, n, e and q are each independently selected from: 0, 1, 2 or 3; R1 is selected from the following group: C 3-6 Cycloalkyl, C 4-6 Alkynyl, C 4-10 Cycloalkenyl, C6-C 10 aryl, 5-10 membered heteroaryl, saturated or partially unsaturated 5-10 membered heterocyclic group; wherein said R1 may be optionally replaced by 0, 1, 2 or 3 R 1a , R 1b , R 1c , R 1d , R 1e , R 1f , R 1g , R i or R h replace; R2 is selected from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl; R3 and R4 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Alkoxy, C 6-10 Alkylamino, C 1-4 R3 and R4 can form a 4-6 membered carbocyclic ring or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic ring or heterocyclic ring formed above can be independently substituted by 0, 1, 2 or 3 R7; R5 and R6 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, -SO2CF3, -SF5, C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R5 and R6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heterocyclic ring contains heteroatoms selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, which are optionally substituted by 0, 1, 2 or 3 R8, respectively and independently; R7 and R8 are each independently selected from the group consisting of hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, or two R7 or R8 together form =O; Cy A Select from the following group: Among them, the Cy A Can be optionally replaced by one or more R 12 Substituents are substituted; d is selected from 0, 1, 2 or 3; each R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, or two R 12 Together they form =O; B is selected from O or S; Cy B R3 and R4 are each independently selected from the following groups: H, D, hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 6-10 Alkylamino, C 1-4 R3 and R4 can form a 4-6 membered carbocyclic or heterocyclic ring with the carbon atoms to which they are attached, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently and optionally replaced by 0, 1, 2 or 3 R 42 replace; R 42 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, or two R 42 Together form = O; Wherein, the heteroaryl or heterocyclic group includes 1, 2, 3, 4 or 5 heteroatoms selected from the following group as ring atoms: N, O, S or Se; the carbocyclic ring, heterocyclic ring, heterocyclic group may be a monocyclic ring, a fused ring, a spirocyclic ring or a bridged ring structure, which may be saturated or partially unsaturated, but not aromatic; R 1a and R 1b Each independently selected from the group consisting of H, D, oxo, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1b and R 1c Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino; R 1e and R 1f Each independently selected from the group consisting of H, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 alkyl; Each R 1g Each independently selected from the group consisting of H, D, hydroxyl, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy; R i C 3-6 Cycloalkoxy; Each R h Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

2. The compound according to claim 1, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: R1 is selected from the following group: (1)C 3-6 Cycloalkyl, the C 3-6 The cycloalkyl group may be optionally substituted with 0, 1, 2 or 3 R 1a or R 1b Replacement; R 1a and R 1b Each independently selected from the group consisting of H, D, oxo, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1b and R 1c Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; (2)C 4-6 Alkynyl, C 4-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino; (3) R is selected from CH, CD, CR 1e or N; Y, Z, W, U are independently selected from CR 1e R 1f NR 1e or oxygen atom, R 1e and R 1f Each independently selected from the group consisting of H, deuterium, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl; R 1e and R 1f Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; (4) R is selected from CH, CD, CR 1e or N; K, F, G are independently selected from CR 1e R 1f NR 1e or oxygen atom, R 1e and R 1f Each independently selected from the group consisting of H, deuterium, halogen, hydroxyl, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 3-6 Halogenated cycloalkyl, Methylamino-substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl; R 1e and R 1f Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; (5) (6) a selenium-containing aromatic heterocycle, wherein the selenium-containing aromatic heterocycle is independently optionally substituted with 0, 1, 2 or 3 R 1g Replacement; R 1g Each independently selected from the group consisting of H, D, hydroxyl, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy; (7) R i Selected from C 3-6 Cycloalkoxy or halogen substituted C 3-6 Cycloalkoxy, and Cy C Selected from the following group: phenyl, 5-12 membered heteroaryl, C 4-10 Cycloalkenyl, C 4-10 heterocyclic group; f is selected from 0, 1, 2 or 3; R h Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy; (8) Cy D is selected from the following group: benzo five-membered heterocycle, benzo five-membered aromatic heterocycle, benzo six-membered heterocycle, benzo six-membered aromatic heterocycle, pyrido five-membered heterocycle, pyrido five-membered aromatic heterocycle, pyrimido five-membered heterocycle, pyrimido five-membered aromatic heterocycle, pyrido six-membered heterocycle, pyrido six-membered aromatic heterocycle, pyrimido six-membered heterocycle, pyrimido six-membered aromatic heterocycle; the heteroatoms of the above heterocycles and aromatic heterocycles are selected from O, N and S; p and t are each independently selected from 0, 1, 2, 3 or 4; R m and R n Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy; R m and R n Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; R m Can be 0, 1, 2, 3 or 4 R m1 Replacement, R n Can be 0, 1, 2, 3 or 4 R n1 Replacement; R m1 and R n1 Each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, C 3-6 Cycloalkoxy; R2 is selected from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl; Cy A Select from the following group: R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, cyano substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl, C 1-2 Haloalkoxy substituted C 1-4 Alkyl; B is selected from O or S; d is selected from 0, 1, 2 or 3; in represents a covalent bond to the parent nucleus, and represents a covalent bond connected to the carbonyl group; -* represents a covalent bond connected; Cy B is selected from the group consisting of 6-9 membered heteroaryl; R3 and R4 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 6-10 Alkylamino, C 1-4 R3 and R4 can form a 4-6 membered carbocyclic ring or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic ring or heterocyclic ring formed above can be independently substituted by 0, 1, 2 or 3 R7; R7 are each independently selected from the following group: hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy; X is selected from CH or N; and when X is CH, the CH may be substituted by R5 or R6 (i.e., X is C-R5 or C-R6); R5 and R6 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, -SO2CF3, -SF5, C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R5 and R6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heterocyclic ring contains heteroatoms selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, which are optionally substituted by 0, 1, 2 or 3 R8, respectively and independently; R8 are each independently selected from the following group: hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

3. The compound according to claim 2, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: R1 is preferably selected from the following group:

4. The compound according to claim 2, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy A Preferably from the following group: Among them, R 12 and d are as defined in claim 1.

5. The compound according to claim 1, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy A Select from the following group: in represents a covalent bond to the parent nucleus, and represents a covalent bond to the carbonyl group; R1 is selected from the following group: (1)C 3-6 Cycloalkyl, C 3-6 The cycloalkyl group is optionally substituted with 0, 1, 2 or 3 R 1a and R 1b Replacement; R 1a and R 1b Each independently selected from the group consisting of H, D, oxo, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1a and R 1b Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; (2)C 4-6 Alkynyl, C 2-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino; (3) Cycloalkenyl, wherein the cycloalkenyl is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl is unsubstituted or substituted with 1, 2, 3 or 4, preferably 1 or 2 R 33 and R 34 Replacement; R 33 and R 34 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 33 and R 34 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 35 Replacement; R 35 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-4 Alkoxy, C 1-4 Haloalkoxy; (4) heterocyclyl, wherein the heterocyclyl 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 is unbridged or bridged and the bridge is 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or substituted with 1, 2, 3 or 4 R 36 or R 37 Replacement; R 36 and R 37 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 36 and R 37 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 38 Replacement; R 38 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy; (5) phenyl, heteroaryl; wherein the heteroaryl is a 5- or 6-membered fully unsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O, S and Se, 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, wherein the phenyl and heteroaryl are unsubstituted or replaced by 1, 2 or 3 independently selected from R 39 and R 40 Substituents, where R 39 and R 40 are independently selected from H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 39 and R 40 Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-6 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 41 Replacement; R 41 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy; Cy B R3 and R4 are each independently selected from the following groups: H, D, hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Deuterated alkyl, C 6-10 Alkylamino, C 1-4 R3 and R4 can form a 4-6 membered carbocyclic ring or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic ring or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 42 Replacement; R 42 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy; R2 is selected from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl; R3 is selected from the group consisting of halogen, cyano, C 3-6 Alkoxy, C 1-4 Haloalkyl; X is selected from CH or N; and when X is CH, the CH may be substituted by R5 or R6 (i.e., X is C-R5 or C-R6); R5 and R6 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, -SO2CF3, -SF5, C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R5 and R6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heterocyclic ring contains heteroatoms selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, which are optionally substituted by 0, 1, 2 or 3 R8, respectively and independently; R8 are each independently selected from the following group: hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

6. The compound according to claim 5, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy A Preferably from the following group:

7. The compound according to claim 5, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy B Preferably from the following group: Among them, R 44 , R 45 and R 46 Each independently selected from H, D, halogen, CN, OH, C 1-6 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkylamino, C 2-4 Alkenyl, C 1-4 Halogenated alkenyl, C 2-4 Alkynyl; R as described above 45 and R 46 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 47 Replacement; R 47 Independently selected from the group consisting of H, D, halogen, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated cycloalkyl, C 1-4 Halogenated alkoxy.

8. The compound according to claim 5, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: R1 is preferably selected from the following group:

9. The compound according to claim 1, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy B Select from the following group: wherein h is selected from 0, 1, 2 or 3; R 44 , R 45 and R 46 Each independently selected from H, D, halogen, CN, OH, C 1-6 Alkyl, C 1-4 Deuterated alkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy, C 1-6 Alkylamino, C 2-4 Alkenyl, C 1-4 Halogenated alkenyl, C 2-4 Alkynyl; R as described above 45 and R 46 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are connected, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above can be independently optionally replaced by 0, 1, 2 or 3 R 47 Replacement; R 47 Independently selected from the group consisting of H, D, halogen, hydroxyl, cyano, C 1-4 Alkyl, C 1-4 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated cycloalkyl, C 1-4 Haloalkoxy; R1 is selected from the following group: C 3-6 Cycloalkyl, C 3-6 Cycloalkyl, C 4-6 Alkynyl, cycloalkenyl, heterocyclyl, phenyl, heteroaryl; Among them, the C 3-6 The cycloalkyl group is optionally substituted with 0, 1, 2 or 3 R 1a and R 1b Replacement; R 1a and R 1b Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 1a and R 1b Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; The C 4-6 The alkynyl groups are each independently optionally substituted with 0, 1, 2 or 3 R 1c and R 1d Replacement; R 1c and R 1d Each independently selected from the following group: H, D, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, sulfone, C 1-6 Alkylamino; The cycloalkenyl group is a partially unsaturated monocyclic ring containing 5 or 6 ring carbon atoms, and the cycloalkenyl group is unsubstituted or substituted by 1, 2, 3 or 4, preferably 1 or 2 R 33 and R 34 Replacement; R 33 and R 34 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 3-6 Alkynyl, C 3-6 Alkenyl, C 3-6 Halogenated alkenyl, C 1-6 Alkylamino; R 33 and R 34 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 35 Replacement; R 35 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 Cycloalkyl, C 3-6 Cycloalkoxy, C 1-4 Alkoxy, C 1-4 Haloalkoxy; The heterocyclyl is a 5- or 6-membered fully saturated or partially unsaturated group comprising ring carbon atoms and 1 or 2 ring heteroatoms independently selected from N, O and S, and wherein the heterocyclyl is unbridged or bridged and the bridge is 1 or 2 carbon atoms, wherein the heterocyclyl is unsubstituted or substituted by 1, 2, 3 or 4 R 36 or R 37 Replacement; R 36 and R 37 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 36 and R 37 Together with the carbon atoms to which they are attached, they can form a 3-5 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-5 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 38 Replacement; R 38 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy; The heteroaryl group is a 5- or 6-membered fully unsaturated monocyclic group comprising ring carbon atoms and 1, 2, 3 or 4 ring heteroatoms independently selected from N, O, S and Se, 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, wherein the phenyl and heteroaryl groups are unsubstituted or replaced by 1, 2 or 3 groups independently selected from R 39 and R 40 Substituents, where R 39 and R 40 are independently selected from H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkoxy, C 2-6 Halogenated alkenyl, C 1-6 Alkylamino; R 39 and R 40 Together with the carbon atoms to which they are attached, they can form a 3-6 membered carbocyclic or heterocyclic ring, wherein the heteroatoms contained in the heterocyclic ring are selected from O, N and S; the formed 3-6 membered carbocyclic or heterocyclic ring can be substituted by 1, 2, 3 or 4 R 41 Replacement; R 41 Each independently selected from the group consisting of H, D, halogen, cyano, hydroxyl, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 3-6 Cycloalkoxy, C 1-4 Haloalkoxy; Cy A Select from the following group: R 12 Each independently selected from the group consisting of hydroxyl, halogen, cyano, C 1-4 Alkyl, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, cyano substituted C 1-4 Alkyl, methoxy substituted C 1-4 Alkyl, C 1-2 Haloalkoxy substituted C 1-4 Alkyl; B is selected from O or S; in represents a covalent bond to the parent nucleus, and represents a covalent bond to the carbonyl group; R2 is selected from the following group: C 1-4 Alkyl, C 1-4 Deuterated alkyl or C 1-4 Haloalkyl; R3 is selected from the group consisting of halogen, cyano, C 3-6 Alkoxy, C 1-4 Haloalkyl; X is selected from CH or N; and when X is CH, the CH may be substituted by R5 or R6 (i.e., X is C-R5 or C-R6); R5 and R6 are each independently selected from the group consisting of H, D, hydroxy, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, -SO2CF3, -SF5, C 3-6 Cycloalkyl, C 6-10 Alkylamino, C 1-4 Alkenyl, C 2-6 Alkynyl, C 1-4 Halogenated alkenyl, C 2-6 Alkynyl; R5 and R6 can form a 4-6 membered carbocyclic or heterocyclic ring together with the carbon atoms to which they are attached, wherein the heterocyclic ring contains heteroatoms selected from O, N and S; the 4-6 membered carbocyclic or heterocyclic ring formed above contains 0, 1, 2 or 3 unsaturated double bonds, which are optionally substituted by 0, 1, 2 or 3 R8, respectively and independently; R8 are each independently selected from the following group: hydroxyl, halogen, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl, C 1-4 Haloalkyl, C 1-4 Halogenated alkoxy.

10. The compound according to claim 9, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: R1 is preferably selected from the following group:

11. The compound according to claim 9, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: Cy A Select from the following group:

12. The compound according to claim 1, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, characterized in that: The compound is selected from:

13. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate; and a pharmaceutically acceptable carrier.

14. Use of the compound according to any one of claims 1 to 12, its stereoisomer, its optical isomer, its pharmaceutically acceptable salt, its crystal form, its isotopic derivative, its prodrug, its metabolite, its solvate or its hydrate, or the pharmaceutical composition according to claim 13 in the preparation of a drug for treating a disease mediated by WRN helicase; Preferably, the disease is a tumor; more preferably, the disease is a microsatellite unstable tumor; further preferably, the tumor is a tumor characterized by microsatellite instability-high (MSI-H), or a tumor with defective mismatch repair (dMMR) or "dMMR signature"; in, The microsatellite unstable tumors include, but are not limited to, uterine body tumors, endometrial cancer, colon adenocarcinoma, gastric adenocarcinoma, rectal adenocarcinoma, adrenal cortical carcinoma, uterine carcinosarcoma, cervical squamous cell carcinoma, endocervical adenocarcinoma, esophageal cancer, breast cancer, renal clear cell carcinoma, and ovarian serous cystic carcinoma; Among them, tumors with "defective mismatch repair" (dMMR) or "dMMR characteristics" include but are not limited to lung cancer, breast cancer, kidney cancer, colorectal cancer, ovarian cancer, prostate cancer, upper aerodigestive tract cancer, gastric cancer, endometrial cancer, liver cancer, pancreatic cancer, hematopoietic and lymphoid tissue cancer, skin cancer, thyroid cancer, pleural cancer, autonomic nervous system tumors, soft tissue tumors, rhabdoid sarcoma, melanoma and other tumors.

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