Bis-heterocyclic WRN inhibitor, preparation method therefor, and use thereof

Abstract

The present invention relates to a bis-heterocyclic WRN inhibitor represented by formula (I), a preparation method therefor, and a use thereof. The present invention also relates to a pharmaceutical composition comprising the compound as an active ingredient, and a use of the compound or the pharmaceutical composition in the treatment and / or prevention of diseases associated with the biological activity of WRN.

Description

Bis-heterocyclic WRN inhibitor, preparation method and application thereof Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and particularly relates to a biheterocyclic WRN inhibitor, a preparation method thereof, and an application thereof. Background Art

[0002] WRN is a member of the RecQ family of DNA helicases and is involved in DNA damage repair, genomic stability maintenance, and telomere maintenance. Defects in the WRN gene can cause Werner syndrome, a recessive genetic disorder characterized by staged premature aging and increased cancer susceptibility (Nat Rev Cancer 2003, 3, 169-178). In 2019, the Cancer Dependency Map (DepMap), a project aimed at identifying new cancer therapeutic targets through genetic screening, drug sensitivity, and drug prediction models, discovered that WRN is essential for the survival of microsatellite instability (MSI) tumors. The results showed that WRN deficiency causes DNA double-strand breaks (DSBs) in MSI cells, cell cycle arrest, and subsequent apoptosis, while having no effect on microsatellite stable (MSS) cells (Nature 2019, 568, 511-516). Further mechanistic studies have shown that the integrity of the WRN exonuclease domain function has no significant effect on the survival of MSI cells, and the integrity of the WRN unwinding domain function is necessary for the survival of MSI cells (Life Sci Alliance 4). In addition, MSI cells contain large-scale expanded and highly unstable TA-dinucleotide repeat sequences. The expanded TA repeat sequences form non-B-shaped cross-shaped DNA secondary structures, which hinder the extension of the replication fork. WRN can unwind this structure to restore normal DNA replication. When WRN is lost, the cross-shaped structure will be cut by the nuclease MUS81, resulting in chromosomal genome damage and cell death. In MSS cells, the TA repeat sequence does not form a cross-shaped structure, so WRN loss will specifically cause MSI cell death (Nature 2020, 586, 292-298). Therefore, inhibitors targeting the WRN unwinding domain may be an effective measure for the treatment of MSI cancers.

[0003] Currently, no drugs related to WRN target inhibition have been found on the market. Based on the potential medical value of WRN target inhibition in the treatment of microsatellite instability (MSI) tumors, the development of compounds related to WRN target inhibition has very important social significance and medical value. Summary of the Invention

[0004] Problems to be solved by the invention:

[0005] Although several patent applications for WRN inhibitors have been published, new compounds are still under development, given the current lack of marketed WRN-targeting drugs and the potential medical value of WRN inhibition in treating microsatellite instability (MSI) tumors. Through continuous research, the inventors of this application have designed compounds with the structures represented by the general formula of the present invention and discovered that compounds with such structures exhibit excellent WRN inhibitory effects and potential for treating MSI tumor-related diseases.

[0006] Solutions used to solve the problem:

[0007] In order to solve the above problems, the inventors of the present application have attempted to conduct in-depth research and found that the bicyclic structure compound represented by the general formula of the present invention and its derivatives can achieve the desired purpose, resulting in the completion of the present invention.

[0008] The present invention protects the following specific embodiments:

[0009] A compound represented by formula (I), or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, or pharmaceutically acceptable salt thereof:

[0010] wherein Ring A is selected from a 5-membered or 6-membered heteroaryl group; each of the 5-membered or 6-membered heteroaryl groups independently contains 1-3 heteroatoms selected from N, O and S; each of the 5-membered or 6-membered heteroaryl groups is independently unsubstituted or substituted with one or more R1; each R1 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl;

[0011] R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0012] R4, R5, R6, R7, R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl.

[0013] In some specific embodiments, in the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Ring A is selected from a 5-membered heteroaryl group and a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O, and S, and the 6-membered heteroaryl group contains 1-2 N heteroatoms; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R1.

[0014] In some specific embodiments, ring A is selected from a 5-membered heteroaryl group and a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N and O, and the 6-membered heteroaryl group contains 1 N heteroatom; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R1.

[0015] In some specific embodiments, ring A is selected from pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl; and the pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl are each independently unsubstituted or substituted with one or more R1.

[0016] In some specific embodiments, ring A is selected from pyrrolyl, furanyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, and pyridinyl; and the pyrrolyl, furanyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, and pyridinyl are each independently unsubstituted or substituted with one or more R1.

[0017] In some specific embodiments, ring A is selected from pyrrolyl, furanyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, and pyridinyl; and the pyrrolyl, pyrazolyl, and imidazolyl groups are each independently unsubstituted or substituted with one or more R1.

[0018] In some embodiments, Ring A is selected from Among them 1 and 1 is connected to the The two phases are connected. That is, ring A is selected from Among them 1 and 1 is connected to the 2 are connected, so that the A ring and the pyridine ring are fused to form a 9-membered or 10-membered heteroaryl group.

[0019] In some embodiments, Ring A is selected from Among them 1 and 1 is connected to the 2 are connected, so that the A ring and the pyridine ring are fused to form a 9-membered or 10-membered heteroaryl group.

[0020] In some embodiments, each R1 is independently selected from the same or different C 1-6 Alkyl, C 3-6 Cycloalkyl.

[0021] In some embodiments, each R1 is independently selected from the same or different C 1-3 Alkyl, C 4-6 Alkyl, C 3-6 Cycloalkyl.

[0022] In some embodiments, each R1 is independently selected from the same or different C 1-3 Alkyl, C 3-6 Cycloalkyl.

[0023] In some embodiments, each R1 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0024] In some embodiments, each R1 is independently selected from the same or different methyl, ethyl, and cyclopropyl groups.

[0025] In some embodiments, each R1 is independently selected from the same or different C 1-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0026] In some embodiments, each R1 is independently selected from the same or different C 1-3 Alkyl, C 4-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0027] In some embodiments, each R1 is independently selected from the same or different C 1-3 Alkyl, halogen, C 3-6 Cycloalkyl.

[0028] In some embodiments, each R1 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0029] In some embodiments, each R1 is independently selected from the same or different methyl, ethyl, F, Cl, and cyclopropyl.

[0030] In some embodiments, Ring A is selected from Among them 1 and 1 is connected to the The two phases are connected. Selected from

[0031] In some embodiments, Selected from

[0032] In some embodiments, Selected from

[0033] In some embodiments, Selected from

[0034] In some embodiments, Selected from

[0035] The present invention also provides a compound represented by formula (IX), or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, or pharmaceutically acceptable salt thereof:

[0036] wherein Ring A is selected from a 5-membered or 6-membered heteroaromatic ring or a benzene ring, Ring B is selected from a pyridine ring or an N-containing heterocyclic alkenyl group, the N on Ring B and the substituent hydroxyl group are respectively located at the ortho position of the carbon atom connected to the carbonyl group, the 5-membered heteroaromatic ring contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaromatic ring contains 1-3 N atoms; Ring B and Ring A are fused to form a 9-membered or 10-membered heterocyclic ring;

[0037] Ring A is unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substitution; the 5-membered or 6-membered heteroaryl group, the 4-6-membered heterocycloalkyl group each independently contains 1-3 heteroatoms selected from N, O and S;

[0038] R2 and R3 are each independently selected from H, C 1-6 Alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 Cycloalkyl;

[0039] R4, R5, R6, R7, R8, each R 11 Each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2- 6 alkenyl, C 2-6 Alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 Aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O and S;

[0040] R 10 Selected from

[0041] Among them, when Selected from hour, Not selected

[0042] The present invention also provides a compound represented by formula (IX), or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, or pharmaceutically acceptable salt thereof:

[0043] wherein ring A is selected from a 5-membered or 6-membered heteroaromatic ring or a benzene ring, ring B is a pyridine ring, the nitrogen and the substituent hydroxyl group on ring B are respectively located at the ortho position of the carbon atom connected to the carbonyl group, the 5-membered heteroaromatic ring contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaromatic ring contains 1-3 nitrogen atoms;

[0044] Alternatively, ring A is a 5-membered or 6-membered heteroaromatic ring or a benzene ring, ring B is an N-containing heterocycloalkenyl group, the N on ring B and the substituent hydroxyl group are respectively located at the ortho position of the carbon atom connected to the carbonyl group, the 5-membered heteroaromatic ring contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaromatic ring contains 1-3 N atoms;

[0045] Ring A is unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substitution; the 5-membered or 6-membered heteroaryl group, the 4-6-membered heterocycloalkyl group each independently contains 1-3 heteroatoms selected from N, O and S;

[0046] R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0047] R4, R5, R6, R7, R8, each R 11 Each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl;

[0048] R 10 Selected from

[0049] The present invention also provides a compound represented by formula (X), or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, or pharmaceutically acceptable salt thereof:

[0050] wherein Ring A is a 5-membered or 6-membered heteroaryl group, Ring B is a pyridyl group, the N group and the substituent hydroxyl group on Ring B are respectively ortho to the carbon atom connected to the carbonyl group, the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms;

[0051] Or ring A is a 5-membered or 6-membered heteroaryl group, ring B is an N-containing heterocycloalkenyl group, the N on ring B and the substituent hydroxyl group are respectively ortho to the carbon atom attached to the carbonyl group, the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms;

[0052] Ring A is unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl;

[0053] R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0054] R4, R5, R6, R7, R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl.

[0055] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XI), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0056] wherein Ring A is a 5-membered or 6-membered heteroaryl group, wherein the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms; the 5-membered or 6-membered heteroaryl groups are each independently unsubstituted or substituted with one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl;

[0057] R2 and R3 are each independently selected from H, C 1-6 Alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 Cycloalkyl;

[0058] R4, R5, R6, R7, R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2- 6 alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10Aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O and S;

[0059] Among them, when Selected from hour, Not selected

[0060] In some specific embodiments, in the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Ring A is selected from a 5-membered heteroaryl group and a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O, and S, and the 6-membered heteroaryl group contains 1-2 N atoms; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R9.

[0061] In some specific embodiments, ring A is selected from furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl; and the furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, and pyridazinyl are each independently unsubstituted or substituted with one or more R9.

[0062] In some embodiments, Selected from

[0063] In some embodiments, each R9 is independently selected from the same or different C 1-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0064] In some embodiments, each R9 is independently selected from the same or different C 1-3 Alkyl, C 4-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0065] In some embodiments, each R9 is independently selected from the same or different C 1-3 Alkyl, halogen, C 3-6 Cycloalkyl.

[0066] In some embodiments, each R9 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0067] In some embodiments, each R9 is independently selected from the same or different methyl, ethyl, F, Cl, and cyclopropyl.

[0068] In some embodiments, Selected from

[0069] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XI), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0070] wherein Ring A is a 5-membered or 6-membered heteroaryl group, wherein the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms; the 5-membered or 6-membered heteroaryl groups are each independently unsubstituted or substituted with one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl;

[0071] R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0072] R4, R5, R6, R7, R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl.

[0073] In some specific embodiments, in the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Ring A is selected from a 5-membered heteroaryl group and a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O, and S, and the 6-membered heteroaryl group contains 1-2 N atoms; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R9.

[0074] In some specific embodiments, ring A is selected from a 5-membered heteroaryl group and a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 2 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1 N atom; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R9.

[0075] In some specific embodiments, ring A is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl; and the pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl are each independently unsubstituted or substituted with one or more R9.

[0076] In some specific embodiments, ring A is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, and pyridinyl; and the pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, and pyridinyl are each independently unsubstituted or substituted with one or more R9.

[0077] In some embodiments, Selected from

[0078] In some embodiments, each R9 is independently selected from the same or different C 1-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0079] In some embodiments, each R9 is independently selected from the same or different C 1-3 Alkyl, C 4-6 Alkyl, halogen, C 3-6 Cycloalkyl.

[0080] In some embodiments, each R9 is independently selected from the same or different C 1-3 Alkyl, halogen, C 3-6 Cycloalkyl.

[0081] In some embodiments, each R9 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0082] In some embodiments, each R9 is independently selected from the same or different methyl, ethyl, F, Cl, and cyclopropyl.

[0083] In some embodiments, Selected from

[0084] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XIII), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0085] Wherein, R2 and R3 are each independently selected from H, C 1-6 Alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 Cycloalkyl;

[0086] R4, R5, R6, R7, R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2- 6 alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 Aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O and S;

[0087] Each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl; m is 0, 1, 2, 3 or 4.

[0088] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XIII), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0089] Wherein, R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0090] R4, R5, R6, R7, R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl;

[0091] Each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl; m is 0, 1, 2, 3 or 4.

[0092] In some specific embodiments, the compound represented by the above formula (XIII), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, each R9 is independently selected from the same or different cyano, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl.

[0093] In some embodiments, each R9 is independently selected from the same or different cyano, halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl.

[0094] In some embodiments, each R9 is independently selected from the same or different cyano, halogen, C 1-2 Alkyl, C 3-4 Alkyl, C 1-2 Alkoxy, C 3-4 Alkoxy, C 1-2 Fluorinated alkyl, C 3-4 Fluoroalkyl.

[0095] In some embodiments, each R9 is independently selected from the same or different cyano, halogen, C 1-2 Alkyl, C 1-2 Alkoxy, C 1-2 Fluoroalkyl.

[0096] In some embodiments, each R9 is independently selected from the same or different cyano, F, Cl, Br, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl.

[0097] In some embodiments, each R9 is independently selected from the same or different cyano, F, Cl, methyl, methoxy, -CF3, -CHF2.

[0098] In some embodiments, m is 0, 1, 2, or 3.

[0099] In some embodiments, m is 0, 1 or 2.

[0100] In some embodiments, m is 0 or 1.

[0101] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XV), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0102] Wherein, R9 is selected from cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substitution; the 5-membered or 6-membered heteroaryl group, the 4-6-membered heterocycloalkyl group each independently contains 1-3 heteroatoms selected from N, O and S;

[0103] R2 and R3 are each independently selected from H, C 1-6 Alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 Cycloalkyl;

[0104] R4, R5, R6, R7, R8, each R 11 Each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2- 6 alkenyl, C 2-6 Alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic groups each independently contain 1-3 heteroatoms selected from N, O and S.

[0105] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XV), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0106] Wherein, R9 is selected from cyano, amino, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substitution; the 5-membered or 6-membered heteroaryl group, the 4-6-membered heterocycloalkyl group each independently contains 1-3 heteroatoms selected from N, O and S;

[0107] R2 and R3 are each independently selected from H, C 1-6 alkyl;

[0108] R4, R5, R6, R7, R8, each R 11 Each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl.

[0109] In some embodiments, the compound represented by the above formula (XV), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, R9 is selected from C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 3-6 Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substituted; the 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain 1-2 heteroatoms selected from N, O and S.

[0110] In some embodiments, R9 is selected from C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 3-6Cycloalkyl, phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substituted; the 5-membered or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contains 1 heteroatom selected from N, O and S.

[0111] In some embodiments, R9 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, fluoromethyl, fluoroethyl, C 3-4 Fluoroalkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, furyl, thienyl, pyrrolyl, pyridyl, 4-6 membered heterocycloalkyl; the phenyl, pyrrolyl, pyridyl, 4-6 membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 Substituted; the 4-6 membered heterocycloalkyl contains 1 heteroatom selected from N and O.

[0112] In some embodiments, each R 11 Each independently selected from halogen, hydroxy, cyano, amino, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl.

[0113] In some embodiments, each R 11 are each independently selected from halogen, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl, C 1-4 Halogenated alkoxy, C 3-6 Cycloalkyl.

[0114] In some embodiments, each R 11 are each independently selected from halogen, C 1-4 alkyl.

[0115] In some embodiments, each R 11 Each is independently selected from F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, and tert-butyl.

[0116] In some embodiments, each R 11 Each is independently selected from F, methyl.

[0117] In some specific embodiments, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CHF2, -CH2CF3, -CH2CHF2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, oxolanyl, oxhexyl, N-methylcyclobutyl, N-methylcyclohexyl, phenyl, 4-fluorophenyl, furyl, thienyl, and pyridyl.

[0118] In some embodiments, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, oxolanyl, oxhexyl, phenyl, 4-fluorophenyl.

[0119] In some embodiments, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, Phenyl,

[0120] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XX), or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts:

[0121] Wherein, R2 and R3 are each independently selected from H, C 1-6 Alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 Cycloalkyl;

[0122] R4, R5, R6, R7, R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2- 6 alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic groups each independently contain 1-3 heteroatoms selected from N, O and S.

[0123] As a preferred embodiment of the present invention, the present invention further provides a compound represented by formula (XXI), or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, or pharmaceutically acceptable salt thereof:

[0124] Wherein, R4, R5, R6, R7, R8 are each independently selected from H, D, halogen, hydroxyl, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Halogenated alkoxy, C 3-6 Cycloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl; or R4 and R8 are combined with the carbon atom to form C 3-10 Cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic groups each independently contain 1-3 heteroatoms selected from N, O and S.

[0125] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R2 and R3 are each independently selected from H, C 1-4 Alkyl, C 5-6 alkyl.

[0126] In some embodiments, R2 and R3 are each independently selected from H, C 1-2 Alkyl, C 3-4 alkyl.

[0127] In some embodiments, R2 and R3 are each independently selected from H, C 1-2 alkyl.

[0128] In some embodiments, R2 and R3 are each independently selected from H, methyl, and ethyl.

[0129] In some embodiments, R2 and R3 are each independently selected from H.

[0130] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R4 is selected from H, halogen, C 1-6 Alkyl, C 2-6 Alkynyl.

[0131] In some embodiments, R4 is selected from H, halogen, C 1-4 Alkyl, C 5-6 Alkyl, C 2-4 Alkynyl, C 5-6 Alkynyl.

[0132] In some embodiments, R4 is selected from H, halogen, C 1-4 Alkyl, C 2-4 Alkynyl.

[0133] In some embodiments, R4 is selected from H, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, ethynyl, propynyl, butynyl.

[0134] In some embodiments, R4 is selected from Cl, Br, methyl, and ethynyl.

[0135] In some embodiments, R4 is selected from H, Cl, methyl, and ethynyl.

[0136] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R5 is selected from H, halogen, C 1-4 alkyl.

[0137] In some embodiments, R5 is selected from H, F, Cl, Br, methyl, and ethyl.

[0138] In some embodiments, R5 is selected from H and F.

[0139] In some embodiments, R5 is selected from H.

[0140] In some embodiments, R5 is selected from H, D, F, Cl, Br, methyl, and ethyl.

[0141] In some embodiments, R5 is selected from H, D, and F.

[0142] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R6 is selected from H, halogen, C 1-4 alkyl.

[0143] In some embodiments, R6 is selected from H, halogen.

[0144] In some embodiments, R6 is selected from H, F, Cl, and Br.

[0145] In some embodiments, R6 is selected from H, Cl, and F.

[0146] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R7 is selected from H, halogen, C 1-4 Alkyl, C 1-4 Halogenated alkyl.

[0147] In some embodiments, R7 is selected from C 1-4 Halogenated alkyl.

[0148] In some embodiments, R7 is selected from C 1-2 Fluorinated alkyl, C 3-4 Fluoroalkyl.

[0149] In some embodiments, R7 is selected from fluoromethyl and fluoroethyl.

[0150] In some embodiments, R7 is selected from -CF3.

[0151] In some embodiments, the above compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, R8 is selected from H, halogen, C 1-4 alkyl.

[0152] In some embodiments, R8 is selected from H, halogen.

[0153] In some embodiments, R8 is selected from H, F, Cl, and Br.

[0154] In some embodiments, R8 is selected from H, F.

[0155] In some specific embodiments, in the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, R4 and R8 are combined with the attached carbon atoms to form a 5-10 membered heteroaryl group; the 5-10 membered heteroaryl group contains 1-3 heteroatoms selected from N, O and S.

[0156] In some embodiments, R4 and R8 are combined with the attached carbon atom to form a 5-6 membered heteroaryl group; the 5-6 membered heteroaryl group contains 1-3 heteroatoms selected from N, O and S.

[0157] In some embodiments, R4 and R8 are combined with the attached carbon atom to form a 5-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O and S.

[0158] In some embodiments, R4 and R8 are combined with the attached carbon atom to form furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, or isoxazolyl.

[0159] In some embodiments, R4 and R8 are combined with the attached carbon atom to form furanyl or thienyl.

[0160] In some embodiments, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Selected from

[0161] In some embodiments, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Selected from

[0162] In some embodiments, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, Selected from

[0163] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts, has the structure represented by the following formula (XIIIa):

[0164] wherein R9, m, R4, R5, R6, R7, and R8 are as defined in formula (XIII).

[0165] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts, has the structure represented by the following formula (XIIIb):

[0166] wherein R9, m, R4, R5, R6, R7, and R8 are as defined in formula (XIII).

[0167] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts, has the structure represented by the following formula (XIIIc):

[0168] wherein R9, m, R4, R5, R6, R7, and R8 are as defined in formula (XIII).

[0169] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, has the structure represented by the following formula (XVa):

[0170] wherein R9, R4, R5, R6, R7, and R8 are as defined in formula (XV).

[0171] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, has the structure represented by the following formula (XVb):

[0172] wherein R9, R4, R5, R6, R7, and R8 are as defined in formula (XV).

[0173] In a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, or pharmaceutically acceptable salts, has the structure represented by the following formula (XVc):

[0174] wherein R9, R4, R5, R6, R7, and R8 are as defined in formula (XV).

[0175] As a preferred technical solution of the present invention, the above-mentioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts, is selected from the following structures:

[0176] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared by reacting the compounds discovered herein with the specified substituents with relatively nontoxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent.

[0177] The term "prodrug" refers to derivatives of the compound represented by formula (I) with specific substituents discovered in the present invention. They themselves may have weak activity or even no activity, but after administration, they are converted into the compound with specific substituents discovered in the present invention under physiological conditions (for example, by metabolism, solvent decomposition or other means) and produce corresponding biological activity in the body.

[0178] The term "metabolite" refers to a product obtained by in vivo metabolism of a compound of formula (I) having specific substituents discovered by the present invention. Metabolites of a compound can be identified by techniques known in the art, and their activity can be characterized by assays as described herein. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, and the like. Accordingly, the present invention includes metabolites of the compound, including metabolites produced by contacting the compound of the present invention with a mammal for a sufficient period of time.

[0179] The term "deuterated compound" refers to a compound of the present invention that includes at least one deuterium atom, specifically one or more hydrogen atoms in a compound of the present invention that can be replaced or substituted with a deuterium atom. In some embodiments, the compound includes two or more deuterium atoms. In some embodiments, the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Synthetic methods for incorporating isotopes into organic compounds are known in the art.

[0180] Preparation method:

[0181] The present invention also provides a method for preparing the compound. The preparation of the compound described in the general formula (I) of the present invention can be completed by the following illustrative methods and examples, but these methods and examples should not be considered in any way to limit the scope of the present invention. The compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of synthetic methods known in the art and the method described in the present invention. The product obtained by each step of the reaction is obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized according to the literature (such as provided by Scifinder) or purchased.

[0182] Preparation steps of general formula (I):

[0183] 1) Substitution reaction of A and C to obtain intermediate D;

[0184] 2) Removing the protecting group from intermediate D to obtain intermediate E;

[0185] 3) Intermediate E is condensed with the corresponding carboxylic acid F to obtain intermediate G;

[0186] 4) Removing the protecting group from the intermediate G to obtain the compound represented by the general formula (I);

[0187] 5) Alternatively, the intermediate E is directly condensed with the carboxylic acid H to obtain the compound represented by the general formula (I).

[0188] wherein PG is a protecting group, such as -Boc (tert-butyloxycarbonyl), benzyl, PMB, etc., and ring A, R2, R3, R4, R5, R6, R7 and R8 are as defined in formula (I).

[0189] The synthesis methods of other compounds of the general formula in the present invention can refer to the above synthesis methods.

[0190] Pharmaceutical composition

[0191] The present invention also provides a pharmaceutical composition comprising the aforementioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, and optional pharmaceutical excipients. Preferably, the pharmaceutical excipient is a pharmaceutically acceptable carrier, diluent, excipient, or a combination thereof.

[0192] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredient are known or will be apparent to those skilled in the art based on the disclosure of the present invention. As described in REMINGTON'S PHARMACEUTICAL SCIENCES, Martin, EW, ed., Mack Publishing Company, 19th ed. (1995), the method for preparing the pharmaceutical composition includes incorporating appropriate pharmaceutical excipients, carriers, diluents, etc.

[0193] The present invention also provides a WRN inhibitor, which includes the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or pharmaceutical compositions.

[0194] Medical uses:

[0195] The present invention also provides the use of the aforementioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions in the preparation of WRN inhibitors.

[0196] The present invention also provides the use of the aforementioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions in the preparation of drugs for treating and / or preventing tumors or cancers.

[0197] The present invention also provides the use of the aforementioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions in the treatment and / or prevention of tumors or cancers.

[0198] The present invention also provides the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the pharmaceutical composition as described above, for use in treating and / or preventing tumors or cancers.

[0199] The present invention also provides a method for treating and / or preventing tumors or cancers, comprising administering to an individual in need thereof a therapeutically and / or preventatively effective amount of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, hydrates, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or pharmaceutical compositions as described above.

[0200] In some embodiments, the tumor or cancer is associated with WRN biological activity.

[0201] In some embodiments, the tumor or cancer is an MSI tumor or MSI cancer.

[0202] In some embodiments, the tumor or cancer is an MSI tumor or MSI cancer associated with WRN biological activity.

[0203] As used herein, "treat" generally refers to obtaining a desired pharmacological and / or physiological effect. This effect can be prophylactic, in terms of completely or partially preventing a disease or its symptoms; and / or therapeutic, in terms of partially or completely stabilizing or curing a disease and / or causing side effects due to the disease. As used herein, "treat" encompasses any treatment of a disease in a patient, including: (a) preventing the onset of a disease or symptom in a patient who is susceptible to the disease or symptom but has not yet been diagnosed with the disease; (b) suppressing the symptoms of a disease, i.e., arresting its development; or (c) alleviating the symptoms of a disease, i.e., causing the disease or symptom to regress.

[0204] In the present invention, "subject" refers to a vertebrate. In certain embodiments, the vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In certain embodiments, the mammal refers to a human.

[0205] In the present invention, an "effective amount" refers to an amount that is effective at the necessary dosage and time to achieve the desired therapeutic or preventive effect. The "therapeutically effective amount" of the substance / molecule of the present invention may vary according to factors such as the disease state, age, sex and weight of the individual and the ability of the substance / molecule to elicit the desired response in the individual. A therapeutically effective amount also encompasses an amount in which the therapeutically beneficial effects of the substance / molecule outweigh any toxic or deleterious consequences. A "prophylactically effective amount" refers to an amount that is effective at the necessary dosage and time to achieve the desired preventive effect. Typically, but not necessarily, a prophylactic dose is used in subjects before the onset of disease or in the early stages of the disease, so the prophylactic effective amount will be lower than the therapeutically effective amount. In the case of cancer, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down to a certain extent, preferably stop) cancer cell infiltration into surrounding organs; inhibit (i.e., slow down to a certain extent, preferably stop) tumor metastasis; inhibit tumor growth to a certain extent; and / or alleviate one or more symptoms associated with cancer to a certain extent.

[0206] Definition of terms:

[0207] According to the common practice in this field, The bonds used in the formulae herein to describe the points of attachment of the moiety or substituent to the parent core or structure.

[0208] A dash “-” that does not appear between two letters or symbols is used to indicate the point of attachment of a substituent. For example, C 3-6 Cycloalkyl-(C 1-6 alkyl) r - means (C1-C6 alkyl) r - connected to the rest of the molecule.

[0209] As used herein, the term "substituted" means that any one or more hydrogens on a designated atom or group are replaced with the selection of a designated group, provided that the normal valence of the designated atom is not exceeded.

[0210] In various parts of this specification, substituents of compounds disclosed herein are disclosed in terms of group types or ranges. It is specifically noted that the present invention includes every independent subcombination of the individual members of these group types and ranges. For example, the term "C 1-6 "Alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl disclosed independently, or "C 1-4 Alkyl", or independently disclosed "C 1-3 alkyl".

[0211] The term "alkyl" is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. For example, "C 1-6 "Alkyl" refers to C1, C2, C3, C4, C5 and C6. In addition, for example, "C 1-6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms. The alkyl group may be unsubstituted or substituted so that one or more of its hydrogen atoms are replaced by another chemical group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like. It will be understood by those skilled in the art that C 1-6 Alkyl groups contain a monovalent C 1-6 Alkyl, divalent C 1-6 Alkylene, such as C3-C6 cycloalkyl-(C1-C6 alkyl) r -The C1-C6 alkyl group refers to a C1-C6 alkylene group.

[0212] The term "alkoxy" refers to any of the above alkyl groups (e.g., C 1-6 Alkyl, C 1-4 Alkyl, C 1-3 alkyl, etc.), which is attached to the rest of the molecule through an oxygen atom (—O—).

[0213] The term "C 1-6 Haloalkyl" or "C 1-6"Haloalkoxy" refers to an alkyl or alkoxy group in which one or more (such as 2 or 3) hydrogen atoms are replaced by halogen atoms, such as fluorine, chlorine, or bromine. The alkyl or alkoxy group is defined as above. In some embodiments, the term "haloalkoxy" refers to an alkyl or alkoxy group in which one or more (such as 2 or 3) hydrogen atoms are replaced by halogen atoms, such as fluorine, chlorine, or bromine. 1-6 "Alkyl" is preferably fluorinated, for example, -CF3, -CHF2, -CH2F, -CH2CH2F, -CH2CHF2, -CH2CF3, etc. In some embodiments, the term "halogenated C 1-6 The "alkoxy" is preferably fluorinated, for example, it can be -OCF3, -OCHF2, -OCH2F, -OCH2CH2F, -OCH2CHF2, -OCH2CF3, etc.

[0214] The term "alkenyl" refers to hydrocarbon groups including straight or branched configurations and having one or more carbon-carbon double bonds that may be present at any stable point along the chain. For example, "C 2-6 The term "alkenyl" is intended to include C2, C3, C4, C5 and C6. Examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, and the like.

[0215] The term "alkynyl" refers to hydrocarbon groups including straight or branched configurations and having one or more carbon-carbon triple bonds that may be present at any stable point along the chain. For example, "C 2-6 The term "alkynyl" is intended to include C2, C3, C4, C5 and C6 alkynyl groups. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0216] The term "cycloalkyl" refers to a cyclized alkyl group, including monocyclic, bicyclic or polycyclic ring systems. When the cycloalkyl group is bicyclic or polycyclic, each ring should be a saturated carbocyclic ring or a carbocyclic ring residue. The two rings of the bicyclic or polycyclic cycloalkyl group can be connected in a manner including bridging, fusion or spiro connection. For example, C 3-10 Cycloalkyl refers to C3, C4, C5, C6, C7, C8, C9 and C 10 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wait.

[0217] The term "heterocycloalkyl" refers to a saturated ring system having at least one heteroatom ring member independently selected from B, P, N, S and O, and having 4 to 10 ring members, 4 to 7 ring members or 4 to 6 ring members. The term "heterocycloalkyl" includes monocyclic 4-, 5-, 6- and 7-membered heterocycloalkyls. Heterocycloalkyl can include monocyclic or bicyclic (e.g., having two fused or bridged rings, spiro rings) ring systems. In some embodiments, heterocycloalkyl is a monocyclic group having 1, 2 or 3 heteroatoms independently selected from N, S and O. Heterocycloalkyl can be connected via ring-forming carbon atoms or ring-forming heteroatoms. Examples of heterocycloalkyl groups include azetidinyl, azepanyl, N-morpholinyl, 3-oxa-9-azaspiro[5.5]undecyl, 1-oxa-8-azaspiro[4.5]decyl, piperidinyl, piperazinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tropanediyl.

[0218] The term "heteroaryl" refers to substituted and unsubstituted aromatic groups having at least one heteroatom (O, N, or S) in at least one ring, including aromatic 5-8 membered monocyclic groups, 8-10 membered bicyclic groups, and 10-14 membered tricyclic groups, wherein the heteroatom-containing ring preferably has 1, 2, or 3 heteroatoms selected from O, N, or S. Each heteroatom-containing ring of the heteroaryl group may contain 1 or 2 oxygen or sulfur atoms and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less and each ring has at least one carbon atom. Each ring of a bicyclic or tricyclic heteroaryl group is aromatic.

[0219] The term "heteroatom" shall include oxygen, sulfur and nitrogen.

[0220] The term "halogen" shall include "F, Cl, Br, I".

[0221] Throughout the specification, groups and substituents thereof may be chosen by one skilled in the art to provide stable moieties and compounds and compounds useful as pharmaceutically acceptable compounds and / or intermediate compounds useful in preparing pharmaceutically acceptable compounds.

[0222] Herein, unless otherwise expressly stated, the description “…are independently selected from” used throughout this document may mean that in different groups, the specific options expressed by the same or different symbols do not affect each other, or that in the same group, the specific options expressed by the same or different symbols do not affect each other.

[0223] Effects of the invention:

[0224] The compounds of the present invention demonstrate potent WRN inhibitory activity and are suitable for use as therapeutic and / or preventive agents for diseases associated with this activity. The compounds of the present invention also demonstrate high WRN exposure and potent anti-tumor efficacy, making them useful for the treatment and / or prevention of MSI-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0225] Figure 1 Pharmacodynamics of the human colon cancer cell line SW48 CDX model DETAILED DESCRIPTION

[0226] It should be understood that the terminology used herein is intended to describe specific embodiments and is not intended to be limiting. In addition, although any method, device, and material similar or equivalent to those described herein may be used for implementing or testing the present invention, preferred methods, devices, and materials are now described.

[0227] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker ASCENA-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were calculated based on a 10 - 6 The units are given in ppm.

[0228] Reaction monitoring and MS measurements were performed using a Thermofisher ESQ (ESI) mass spectrometer.

[0229] HPLC determination was performed using a Thermo Fisher U3000 DAD high pressure liquid chromatograph (GL Sciences ODS-HL HP 3 μm 3.0*100 mm column).

[0230] Qingdao Ocean GF254 silica gel plates were used for thin-layer chromatography (TLC). Silica gel plates with a size of 0.15-0.2 mm were used for thin-layer chromatography (TLC). High-performance thin-layer chromatography (HPLC) preparative plates with a size of 0.9-1.0 mm were used for TLC separation and purification. Column chromatography used Qingdao Ocean 200-300 mesh silica gel as the carrier. The developing solvents used were A: dichloromethane and methanol; B: petroleum ether and ethyl acetate. The solvent volume ratio was adjusted according to the polarity of the compound. A Biotage Isera One preparative liquid phase was used for medium-pressure preparative liquid chromatography (HPLC). An Agilent 1290 Infinity II preparative liquid chromatograph was used.

[0231] In the following examples, unless otherwise specified, all reaction raw materials can be purchased from manufacturers such as San Chemical Technology (Shanghai) Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., Jiangsu Aikon Biopharmaceutical R&D Co., Ltd., and Shanghai Bid Pharmaceutical Technology Co., Ltd.

[0232] Abbreviated description

[0233] Preparation of intermediate int-1:

[0234] Step 1: Dissolve int-1a (70 g, 538.05 mmol) in DCM (500 mL), add NBS (100.6 g, 564.95 mmol), then add TSOH.H2O (20.4 g, 107.61 mmol), and stir at room temperature for 3 hours. Filter and concentrate the filtrate under reduced pressure to obtain crude int-1b (126.5 g, 602.87 mmol). MS Calcd: 207.97; MS Found: 206.98 ([MH] - ).

[0235] Step 2: Dissolve int-1b (10 g, 47.85 mmol), int-1c (49.0 g, 263.16 mmol), and potassium carbonate (39.7 g, 287.08 mmol) in MeCN (100 mL) and stir at room temperature for 30 minutes. Filter, neutralize the filtrate with dilute hydrochloric acid, wash with water, extract with DCM, concentrate the organic phase, and filter the residue to obtain int-1d (10.97 g, 34.89 mmol, 72.9%). MS Calcd: 314.18; MS Found: 315.23 ([M+H] + ).

[0236] Step 3: At room temperature, H3PO4 (4.5 g, 46.01 mmol) was added to a solution of int-1e (5 g, 30.67 mmol) and int-1d (10.6 g, 33.74 mmol) in anhydrous ethanol (50 mL). The mixture was heated to 80°C and stirred for two days. Saturated sodium bicarbonate was added to quench the mixture and the pH was adjusted to 6-7. The reaction mixture was concentrated and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate and concentrated by flash column chromatography (DCM = 95%, MeOH = 5%) to give int-1 (2.09 g, 4.89 mmol, 15.9%). MS Calcd: 426.10; MS Found: 427.07 ([M+H] + ).

[0237] Preparation of intermediate int-2:

[0238] Step 1: Int-1 (400 mg, 0.94 mmol) and dioxane (5 mL) were added to a 50 mL eggplant-shaped flask. N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (374 mg, 1.03 mmol) and DIPEA (383 mg, 2.81 mmol) were then added. The mixture was stirred at 85°C for 5 hours. LC-MS confirmed the completion of the reaction. Water (20 mL) was added and the mixture was extracted with EA (20 mL × 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and dried under reduced pressure to afford the crude product. The crude product was purified by column chromatography (PE:EA = 1:2) to afford int-2a (430 mg, 0.65 mmol, 69.3% yield). MS Calcd: 662.89; MS Found: 564.15 ([M+H-100] + ).

[0239] Step 2: To a 50 mL eggplant-shaped flask, int-2a (350 mg, 0.53 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (166 mg, 0.79 mmol), dioxane (3 mL), and H2O (1.5 mL) were added. Pd(dppf)Cl2 (39 mg, 0.053 mmol) and potassium phosphate (365 mg, 1.58 mmol) were then added. Under nitrogen, the mixture was stirred at 80°C overnight. Water (20 mL) was added, and the mixture was extracted with EA (20 mL × 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 1:2) to obtain int-2b (220 mg, 0.33 mmol, 61.8% yield). MS Calcd:666.10; MS Found:610.19([M-56] + ).

[0240] Step 3: Int-2b (220 mg, 0.33 mmol) and DCM (5 mL) were added to a 50 mL eggplant-shaped flask. Trifluoroacetic acid (1 mL) was then added and the mixture was stirred at room temperature overnight. LCMS confirmed the completion of the reaction. Saturated sodium bicarbonate solution (20 mL) was added and the mixture was extracted with EA (20 mL × 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain int-2 (140 mg, 0.25 mmol, 75.7% yield). MS Calcd: 565.18; MS Found: 566.26 ([M+H] + ).

[0241] Preparation of intermediate int-3a:

[0242] Step 1: Int-1e (2.0 g, 12.3 mmol) and int-1a (2.2 g, 15.3 mmol) were added to a 25 mL round-bottom flask, and AcOH (7 mL) was added. The mixture was heated to 80°C and stirred overnight. After cooling to room temperature, the mixture was stirred at 0°C for 1 h, filtered, and the filter cake was washed with a small amount of EtOH (10 mL) and dried. The filtrate was concentrated to dryness, washed with a small amount of EtOH (10 mL), and the solid was dried to obtain the second portion of product. The product was combined to give int-3a (1.63 g, 6.71 mmol, 54.5% yield). MS Calcd: 241.98, 243.98; MS Found: 242.98, 244.98 ([M+H] + ).

[0243] Preparation of intermediate int-4:

[0244] Step 1: Int-3a (1.63 g, 6.71 mmol) and N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (2.67 g, 7.35 mmol) were added to a 25 mL round-bottom flask, and dioxane (7 mL) was added. The mixture was stirred at 80°C for 4 h under nitrogen. The mixture was cooled to room temperature, and a white precipitate was precipitated. The filtrate was filtered and concentrated. After adding EA, a white solid precipitated again. The solid was filtered and washed with EA (10 mL). The solids were combined and the solvent was removed under reduced pressure to obtain int-4a (3.02 g, 6.31 mmol, 94.0% yield). MS Calcd: 476.98, 478.98; MS Found: 477.94, 479.95 ([M+H] + ).

[0245] Step 2: Int-4a (3.0 g, 6.27 mmol) and 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (1.71 g, 8.14 mmol) were added to a 100 mL two-necked flask, and potassium phosphate (3.99 g, 18.80 mmol), Pd(dppf)Cl2 (458 mg, 0.63 mmol), dioxane (40 mL) and purified water (20 mL) were added in sequence. The system showed a brown suspension and was stirred at 80 ° C for 2 h under nitrogen protection. Cool to room temperature, remove excess solvent by vortexing, add DCM (100 mL) and water (100 mL), separate the layers, extract the aqueous phase with DCM (50 mL*3), combine the organic phases, wash with saturated brine (100 mL), remove the solvent under reduced pressure, add EA / PE (2:1) to form a brown precipitate, and column chromatography (DCM:MeOH=10:1) afforded int-4b (1.58 g, 3.28 mmol, 52.3% yield). MS Calcd: 481.11; MS Found: 479.94 ([MH] - ).

[0246] Step 3: Int-4b (1.35 g, 2.80 mmol) was added to a 50 mL flask, followed by NBS (548 mg) and acetonitrile (50 mL). The mixture was heated to 80°C and the reaction continued for 6 h. After cooling, a solid precipitated and was filtered. The filter cake was purified by column chromatography (MeOH:DCM = 1:10) to afford int-4c (0.51 g, 0.91 mmol, 32.5% yield). MS Calcd: 559.02, 561.02; MS Found: 559.8, 562.0 ([M+H] + ).

[0247] Step 4: Int-4c (100 mg, 0.18 mmol), (R)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (180 mg, 0.9 mmol), and silver tetrafluoroborate (35 mg, 0.18 mmol) were added to dry DMSO (2 mL). The mixture was heated to 120°C under nitrogen and stirred for 4 hours. After the reaction was complete, the mixture was cooled to room temperature and saturated aqueous sodium bicarbonate solution was added. The mixture was then extracted with ethyl acetate (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (EA:PE = 1:1) to give int-4d (74 mg, 0.11 mmol, 60%). MS Calcd: 679.25; MS Found: 680.33 ([M+H] + ).

[0248] Step 5: Add int-4d (74 mg, 0.11 mmol) to DCM (5 mL), then add trifluoroacetic acid (1 mL), and stir at room temperature for 4 hours. After the reaction is complete, concentrate under reduced pressure, add saturated aqueous sodium bicarbonate solution to the residue, and then extract with DCM (5 mL x 3). The organic layer is dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure to obtain int-4 (51 mg, 0.09 mmol, 80%). MS Calcd: 579.20; MS Found: 580.31 ([M+H] + ).

[0249] Preparation of intermediate int-5:

[0250] Step 1: Place int-1 (200 mg, 0.47 mmol) in a 100 mL eggplant flask and add 10 mL of 1,4-dioxane solution. Then, add N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (185.9 mg, 0.51 mmol) and DIPEA (0.23 mL, 1.4 mmol) sequentially. Stir at 75°C for 5 hours. The reaction mixture was concentrated and purified by column chromatography (PE:EA = 60%:40%) to obtain int-5a (180 mg, 0.27 mmol, 58% yield). MS Calcd: 661.15; MS Found: 662.30 ([M+H] + ).

[0251] Step 2: Dissolve int-5a (180 mg, 0.27 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (85.9 mg, 0.41 mmol) in 1,4-dioxane (6 mL) and H2O (3 mL), and then add Pd(dppf)Cl2 (20 mg, 0.03 mmol) and potassium phosphate (188 mg, 0.82 mmol) in sequence. Replace nitrogen three times and react at 80°C for 5.0 hours. The reaction solution was concentrated, and 30 mL of ethyl acetate was added to the residue, which was filtered through celite. The filtrate was concentrated and purified by column chromatography (DCM:MeOH=4:1) to give int-5b (153 mg, 0.23 mmol, 85% yield) MS Calcd: 663.28; MS Found: 664.20 ([M+H] + ).

[0252] Step 3: Dissolve int-5b (153 mg, 0.23 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. The reaction solution was directly concentrated, and 20 mL of saturated sodium bicarbonate solution was added to the residue. Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, dried, and concentrated to obtain int-5 (150 mg, 0.27 mmol, 98% yield). MS Calcd: 563.23; MS Found: 564.31 ([M+H] + ).

[0253] Preparation of intermediate int-6:

[0254] Step 1: Weigh int-6a (6.0 g, 28.63 mmol) into a 50 mL single-necked flask, add DCM (30 mL), then triethylamine (7.94 mL, 57.25 mmol). Add chloroacetyl chloride (3.40 mL, 34.35 mmol) in DCM (10 mL) dropwise under an ice bath, then allow to react at room temperature overnight. Concentrate the reaction mixture, and the residue is purified by flash column chromatography (PE:EA = 20:1) to afford int-6b (3.7 g, 12.93 mmol, 45.1% yield). MS Calcd: 284.99; MS Found: 283.97 ([MH] - ).

[0255] Step 2: Int-6b (3.7 g, 12.93 mmol) and acetone (15 mL) were added to a 100 mL eggplant-shaped flask. Potassium iodide (5.4 g, 32.33 mmol) was then added and stirred at 50°C for 3 hours. The solvent was evaporated to dryness under reduced pressure to obtain a crude product, which was purified by column chromatography (PE:EA = 3:1) to obtain int-6c (3.4 g, 9.01 mmol, 69.4% yield). MS Calcd: 376.93; MS Found: 375.97 ([MH] - ).

[0256] Step 3: Int-6c (1.1 g, 2.81 mmol), int-1 (1.0 g, 2.34 mmol), and 1,4-dioxane (15 mL) were added to a 100 mL eggplant flask. DIPEA (0.9 g, 7.02 mmol) was then added and stirred at 90°C for 2 hours. The solvent was evaporated to dryness under reduced pressure to obtain a crude product, which was purified by column chromatography (PE:EA = 1:4) to afford int-6d (940 mg, 1.39 mmol, 59.3% yield). MS Calcd: 676.92.; MS Found: 578.12 ([M+H-100]+ ).

[0257] Step 4: To a 100 mL eggplant-shaped flask, int-6d (940 mg, 1.39 mmol), 1,4-dioxane (10 mL), and H2O (5 mL) were added. Then, 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (438 mg, 2.08 mmol), Pd(dppf)Cl2 (102 mg, 0.14 mmol), and potassium phosphate (960 mg, 4.17 mmol) were added. The mixture was stirred at 100°C under nitrogen for 6 hours. Water (50 mL) was added thereto, and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (DCM:MeOH=30:1) to give int-6e (800 mg, 1.18 mmol, 84.7% yield) MS Calcd: 679.25.; MS Found: 624.21 ([M+H-56] + ).

[0258] Step 5: Int-6e (800 mg, 1.18 mmol) and DCM (5 mL) were added to a 100 mL eggplant flask. TFA (0.5 mL) was then added and the mixture was stirred at room temperature overnight. H2O (20 mL) was added and the pH was adjusted to alkaline with saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane (30 mL × 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure to give int-6 (500 mg, 0.86 mmol, 73.3% yield). MS Calcd: 579.20.; MS Found: 580.22 ([M+H] + ).

[0259] Preparation of intermediate int-8:

[0260] Step 1: Int-3a (4 g, 16.5 mmol), N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (6 g, 16.5 mmol), and DIPEA (6.4 g, 49.5 mmol) were added to DMF (50 mL). The temperature was raised to 80°C and stirred for 2 hours. The mixture was cooled to room temperature and poured into water. Solid precipitated and was filtered. The filter cake was washed with water (50 mL x 2). The solid was collected and purified by slurrying with a mixed solvent of ethyl acetate and petroleum ether (1:3) to obtain int-8a (5.9 g, 12.4 mmol, 75% yield). MS Calcd: 475.03; MS Found: 476.11 ([M+H]+ ).

[0261] Step 2: Int-8a (5.9 g, 12.4 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (2.6 g, 12.4 mmol), potassium phosphate (7.9 g, 37.2 mmol), and 1,1-bis(diphenylphosphino)diborane iron palladium dichloride (906 mg, 1.24 mmol) were added to a mixed solvent of dioxane (40 mL) and water (10 mL). The mixture was purged with nitrogen for 3 minutes, the temperature was raised to 85°C, and the mixture was stirred for 3 hours. The mixture was concentrated under reduced pressure and the residue was poured into water. The solid was collected by filtration and then purified by ethanol slurry to obtain int-8b (4.8 g, 10.0 mmol, 81% yield). MS Calcd: 479.16; MS Found: 480.21 ([M+H] + ).

[0262] Step 3: Int-8b (4.8 g, 10.0 mmol) was added to DMF (50 mL), followed by NBS (1.78 g, 10.0 mmol), the temperature was raised to 60°C, and the mixture was stirred for 3 hours. The reaction system was then poured into water, whereupon a solid precipitated. The solid was collected by filtration and then purified by ethanol slurry to afford int-8c (3.4 g, 6.1 mmol, 61%). MS Calcd: 557.07; MS Found: 558.07 ([M+H] + ).

[0263] Step 4: Int-8c (1 g, 1.8 mmol), (R)-tert-butyl 2-methylpiperazine-1-carboxylate (3.6 g, 18 mmol), and silver tetrafluoroborate (351 mg, 1.8 mmol) were added to dry DMSO (10 mL). The temperature was then raised to 120°C under nitrogen, stirred for 4 hours, cooled to room temperature, and saturated aqueous sodium bicarbonate was added. The mixture was then extracted with ethyl acetate (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography (EA:PE = 1:1) to give int-8d (810 mg, 1.20 mmol, 66%). MS Calcd: 677.29; MS Found: 678.31 ([M+H] + ).

[0264] Step 5: Int-8d (810 mg, 1.20 mmol) was added to DCM (10 mL), followed by trifluoroacetic acid (3 mL). The mixture was stirred at room temperature for 4 hours and concentrated under reduced pressure. The residue was added to saturated aqueous sodium bicarbonate solution and extracted with DCM (10 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain int-8 (483 mg, 0.84 mmol, 70%). MS Calcd: 577.24; MS Found: 578.28 ([M+H]+).

[0265] Preparation of intermediate int-9:

[0266] Step 1: In a 1 L single-necked flask, int-9a (60.00 g, 268.98 mmol) was dissolved in acetonitrile (300 mL). K2CO3 (74.36 g, 537.96 mmol) and (R)-1-N-Boc-2-methylpiperazine (56.56 g, 282.43 mmol) were added sequentially. The resulting reaction mixture was stirred at room temperature overnight. Undissolved inorganic salts were filtered off, and the filtrate was concentrated to dryness. The resulting residue was purified on a flash silica gel column (petroleum ether / ethyl acetate = 5:1) to afford int-9b (64.0 g, 69.5% yield). MS Calcd.: 342.22; MS Found: 343.22 [M+H] + .

[0267] Step 2: Weigh int-9b (91.32 g, 266.68 mmol) into a 1 L single-necked flask and dissolve it in ethanol (150 mL). Then, add 5-bromo-2H-1,2,4-triazole-3-amine (47.81 g, 293.35 mmol) and phosphoric acid (27.44 g, 280.01 mmol) sequentially. Switch the reaction system to nitrogen three times. Stir the reaction at 90°C for 44 hours before cooling to room temperature. Add DIPEA (103.40 g, 800.04 mmol) and Boc2O (29.10 g, 133.34 mmol) to the single-necked flask and react at room temperature for 2 hours. After completion of the reaction, pour the reaction mixture into saturated brine (500 mL) and extract it three times with ethyl acetate (1 L x 3). The combined organic phases are backwashed twice with brine, dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a residue. The resulting residue was purified by flash silica gel column chromatography (dichloromethane / methanol = 17:3) to afford int-9 (50.0 g, yield 38.6%). MS Calcd.: 442.12; MS Found: 443.2 [M+H] +.1HNMR (400MHz, DMSO-d6) δ4.16 (d, J = 23.6Hz, 1H), 3.81-3.69 (m, 1H), 3.58-3.49 (m, 1H), 3.41-3.31 (m, 1H), 3.19-3.09 (m,1H), 3.09-2.89(m,1H), 2.88-2.77(m,1H), 2.66-2.54(m,2H), 1.42(s,9H), 1.29-1.26(m,3H), 1.18(t,J=7.6Hz,3H).

[0268] Preparation of intermediate int-10:

[0269] Step 1: 2-Iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (234 mg, 0.68 mmol) and int-9 (300 mg, 0.68 mmol) were weighed into a 25 mL single-necked flask. 1,4-Dioxane (5 mL) and DIEA (0.34 mL, 2.04 mmol) were added and reacted at 80°C for 2 hours. The mixture was concentrated under reduced pressure and the residue was purified by column chromatography (DCM:MeOH = 20:1) to obtain int-10a (389 mg, 0.56 mmol, 82.8% Yield). MS Calcd: 655.17, 657.17; MS Found: 556.12, 558.09 ([M+H-100] + ).

[0270] Step 2: Int-10a (389 mg, 0.56 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (177.4 mg, 0.84 mmol), potassium phosphate (388.9 mg, 1.69 mmol), and Pd(dppf)Cl2 (82.4 mg, 0.11 mmol) were weighed into a 50 mL single-necked flask. Dioxane (5 mL) and water (2 mL) were added. After the addition, the atmosphere was purged with nitrogen three times and the reaction was continued at 90°C for 2 hours. The reaction mixture was mixed and purified by column chromatography (DCM:MeOH = 20:1) to obtain int-10b (312 mg, 0.43 mmol, 75.6% yield). MS Calcd: 659.30; MS Found: 604.24 ([M+H-56] + ).

[0271] Step 3: Weigh int-10b (312 mg, 0.43 mmol) into a 50 mL single-necked flask, add DCM (5 mL) and trifluoroacetic acid (2 mL), and stir at room temperature for 1 hour. The reaction solution was partially concentrated to remove the solvent, and then saturated sodium bicarbonate solution was added. The mixture was extracted with ethyl acetate. The organic phase was dehydrated and concentrated. The residue was purified by column chromatography (DCM:MeOH = 10:1) to obtain int-10 (211 mg, 0.36 mmol, 84.2% Yield). MS Calcd: 559.25; MS Found: 560.20 ([M+H] + ).

[0272] Preparation of intermediate int-11:

[0273] Step 1: int-1 (14 g, 32.8 mmol) was added to a 250 mL three-necked flask containing dioxane (150 mL) and water (50 mL), followed by the addition of 3,6-dihydro-2H-pyran-4-boronic acid pyranoside (10.3 g, 49.1 mmol), K2CO3 (9.1 g, 65.6 mmol), and Pd(dppf)Cl2 (1.2 g, 1.64 mmol). The atmosphere was replaced three times with nitrogen, heated to 100°C for 14 hours, cooled to room temperature, and 300 mL of water was added. The mixture was extracted three times with 300 mL of dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried. The residue was purified by column chromatography (DCM / MeOH = 100 / 0 to 95 / 5) to obtain a crude product. The crude product was slurried with methyl tert-butyl ether to obtain pure int-11 (9.2 g, 65% yield). MS Calcd.: 430.2; MS Found: 431.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ13.00(s,br,1H),6.81(br,1H),4.28-4.27(m,2H),3.93-3.90(m,2H),3.83-3.80(m,2H),3.37-3. 33(m,2H),2.90-2.89(m,2H),2.79-2.73(m,2H),2.64-2.60(m,2H),2.51-2.50(m,2H),1.42(s,9H),1.19(t,J=7.2Hz,3H).

[0274] Intermediate int-12:

[0275] Step 1: To a 50 mL eggplant-shaped flask, int-11 (205.2 mg, 0.48 mmol), dioxane (10 mL), 75-d (200 mg, 0.55 mmol), and DIPEA (0.25 mL, 1.50 mmol) were added. The mixture was reacted at 80°C for 2 hours under nitrogen. The reaction solution was concentrated, and the residue was purified by flash column chromatography (DCM:MeOH = 20:1) to obtain int-12a (292 mg, 0.43 mmol, 89.6%). MS Calcd: 683.22; MS Found: 684.36 ([M+H] + ).

[0276] Step 2: Int-12a (290 mg, 0.42 mmol) and DCM (10 mL) were added to a 50 mL eggplant flask. TFA (3 mL) was added and the mixture was stirred at room temperature for 2 hours. Saturated sodium bicarbonate solution was added to the reaction mixture and the pH was adjusted to 8. The mixture was extracted with DCM (3*50 mL). The organic phases were combined to obtain int-12 (221 mg, 0.38 mmol, 89.3%). MS Calcd: 583.17; MS Found: 584.2 ([M+H] + ).

[0277] Preparation of intermediate int-13:

[0278] Step 1: Int-3a (700 mg, 2.88 mmol) and int-6c (1087.0 mg, 2.88 mmol) were added to a 100 mL round-bottom flask. Dioxane (20 mL) and DIPEA (1.43 mL, 8.64 mmol) were added sequentially. The mixture was stirred at 80°C under nitrogen for 3 hours. The reaction mixture was cooled to room temperature, and a solid precipitated. The solid was filtered, washed with EA, and the combined filtrates were concentrated and purified by flash column chromatography (DCM:MeOH = 5:1) to afford int-13a (603 mg, 1.22 mmol, 42.5% yield).

[0279] Step 2: Dissolve int-13a (600 mg, 1.22 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (383.8 mg, 1.83 mmol) in 1,4-dioxane (8 mL) and H2O (4 mL). Add Pd(dppf)Cl2 (89 mg, 0.09 mmol) and potassium phosphate (860 mg, 3.65 mmol) sequentially. Replace nitrogen atmosphere three times and react at 80°C for 5.0 hours. Concentrate the reaction mixture, add 30 mL of ethyl acetate to the residue, filter through celite, concentrate the filtrate, and purify by column chromatography (DCM:MeOH = 4:1) to afford int-13b (200 mg, 0.4 mmol, 33% yield). MS Calcd:495.13; MS Found:496.10([M+H] + ).

[0280] Step 3: Int-13b (450 mg, 0.91 mmol) and N-bromosuccinimide (192.7 mg, 1.09 mmol) were placed in a 100 mL eggplant flask, 10 mL of acetonitrile was added, and the temperature was raised to 60°C for 1 hour. The reaction solution was poured into 20 mL of water and extracted with EA (3×20 mL). The organic phases were combined, dried, concentrated, and purified by column chromatography (DCM:MeOH=6:1) to give int-13c (300 mg, 0.52 mmol, 57% yield). MS Calcd: 575.04; MS Found: 576.20 ([M+H] + ).

[0281] Step 4: Int-13c (250 mg, 0.43 mmol) and (R)-1-N-Boc-2-methylpiperazine (871 mg, 4.35 mmol) and AgBF4 (101.3 mg, 0.52 mmol) were added to dry DMSO (3 mL), replaced with nitrogen for 1 minute, and then the temperature was raised to 120°C for 18 hours. The reaction system was poured into water (20 mL), then extracted with EA (20 mL x 3), the organic phases were combined, dried and concentrated, and the residue was purified by Prep-TLC (DCM:MeOH=20:1) to give int-13d (127 mg, 0.18 mmol, 42% yield). MS Calcd: 693.27; MS Found: 694.10 ([M+H] + ).

[0282] Step 5: Dissolve int-13d (127 mg, 0.18 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. The reaction solution was directly concentrated, and 20 mL of saturated sodium bicarbonate solution was added to the residue. Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, dried, and concentrated to obtain crude int-13 (90 mg, 0.15 mmol, 82% yield). MS Calcd: 593.21; MS Found: 594.27 ([M+H] + ).

[0283] Preparation of intermediate 72-e:

[0284] Step 1: Int-3a (2.0 g, 8.23 ​​mmol), N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (3.0 g, 8.23 ​​mmol), and DIPEA (3.2 g, 24.68 mmol) were added to 60 mL of dioxane solution and stirred at 80°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 72-a (3.6 g, 7.56 mmol, 91.9% yield). MS Calcd: 475.03; MS Found: 476.05 ([M+H] + ).

[0285] Step 2: 72-a (3.6 g, 7.56 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (1.58 g, 7.5 mmol), Pd(dppf)Cl2·CH2Cl2 (1.07 g, 1.32 mmol), and potassium phosphate (4.77 g, 22.5 mmol) were added sequentially to a 40 mL dioxane / water (4:1) mixture. The mixture was evacuated and replaced with nitrogen three times. The temperature was raised to 80°C and stirred for 2 h. The reaction solution was completely concentrated, and the residue was isolated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 72-b (1.5 g, 3.2 mmol, 41.6% yield). MS Calcd: 479.16; MS Found: 480.19 ([M+H] + ).

[0286] Step 3: Dissolve 72-b (1.34 g, 2.8 mmol) in DMF (10 mL), then slowly add NBS (0.55 g, 3.07 mmol), heat to 60°C, and stir for 3 h. Add water to the reaction solution, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (EA) to obtain 72-c (1.3 g, 2.33 mmol, 83.3% yield). MS Calcd: 557.07; MS Found: 558.15 ([M+H] + ).

[0287] Step 4: 72-c (1.3 g, 2.33 mmol), (R)-1-N-Boc-2-methylpiperazine (5.02 g, 25.08 mmol), and silver tetrafluoroborate (0.58 g, 3.01 mmol) were added sequentially to a DMSO (10 mL) solution. The mixture was evacuated and replaced with nitrogen three times. The temperature was raised to 120°C and stirred overnight. Water was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography to obtain 72-d (1.2 g, 1.77 mmol, 75.9% yield). MS Calcd: 677.29; MS Found: 678.32 ([M+H] + ).

[0288] Step 5: Add 72-d (1.2 g, 1.77 mmol) to a hydrochloric acid-dioxane solution (20 mL) and stir at room temperature overnight. The reaction solution was completely concentrated, and a saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product 72-e (0.6 g, 1.03 mmol, 58.8% yield). MS Calcd: 577.24; MS Found: 578.06 ([M+H] + ).

[0289] Preparation of intermediate 75-d:

[0290] Step 1: To a 250 mL eggplant flask, add 75-a (10 g, 55.83 mmol) and DCM (50 mL), followed by BF3OEt2 (7.9 g, 55.83 mmol) and NCS (8.2 g, 61.42 mmol). Stir and react at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by flash column chromatography (PE:EA = 95:5) to obtain 75-b (8.1 g, 37.92 mmol, 67.9%). MS Calcd: 213.00; MS Found: 212.12 ([MH]- ).

[0291] Step 2: To a 250 mL eggplant flask, add 75-b (8.1 g, 37.92 mmol) and DCM (50 mL), followed by triethylamine (10.51 mL, 75.84 mmol). Slowly add a solution of chloroacetyl chloride (3.04 mL, 37.92 mmol) in DCM (10 mL) under an ice bath. Allow to react overnight at room temperature. The reaction mixture was concentrated, and the residue was purified by flash column chromatography (PE:EA = 92:8) to afford 75-c (4.5 g, 15.52 mmol, 40.9%). MS Calcd: 288.97; MS Found: 288.18 ([M+H] - ).

[0292] Step 3: To a 250 mL eggplant-shaped flask, 75-c (4.5 g, 15.52 mmol), potassium iodide (5.15 g, 31.04 mol), and acetone (30 mL) were added and reacted at 50°C for 3 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by flash column chromatography (PE:DCM = 70:30) to give 75-d (2.23 g, 5.85 mmol, 37.7%). MS Calcd: 380.90; MS Found: 379.93 ([MH] - ).

[0293] Preparation of intermediate 76-e:

[0294] Step 1: Int-3a (600 mg, 2.47 mmol) and 75-d (986.9 mg, 2.71 mmol) were added to a 100 mL round-bottom flask. Dioxane (30 mL) and DIPEA (1.23 mL, 7.40 mmol) were added and stirred at 80°C under nitrogen for 6 h. The reaction mixture was concentrated and the residue was purified by flash column chromatography (PE:EA = 1:5) to afford 76-a (1.04 g, 2.09 mmol, 84.8%). MS Calcd: 494.97, 496.97; MS Found: 495.99, 497.94 ([M+H] + ).

[0295] Step 2: Add 76-a (1.0 g, 2.01 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (0.6 g, 3.02 mmol), Pd(dppf)Cl2 (0.1 g, 0.20 mmol), and potassium phosphate (1.4 g, 6.04 mmol) to a mixed solvent of water (5 mL) and dioxane (20 mL). Raise the temperature to 85°C and stir for 2 hours. The reaction mixture was concentrated, and the residue was purified by flash column chromatography (PE:EA = 1:5) to afford 76-b (508 mg, 1.02 mmol, 50.5%). MS Calcd: 499.10; MS Found: 500.15 ([M+H] + ).

[0296] Step 3: Dissolve 76-b (0.45 g, 0.9 mmol) in DMF (5 mL), then slowly add NBS (0.18 g, 1.0 mmol), heat to 60°C, and stir for 3 h. Add water to the reaction solution, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (EA% 100) to obtain 76-c (0.45 g, 0.78 mmol, 86.3% yield). MS Calcd: 577.01; MS Found: 578.07 ([M+H] + ).

[0297] Step 4: 76-c (0.45 g, 0.78 mmol), (R)-1-N-Boc-2-methylpiperazine (1.56 g, 7.78 mmol), and silver tetrafluoroborate (0.18 g, 0.93 mmol) were added sequentially to a DMSO (4 mL) solution. The mixture was evacuated and replaced with nitrogen three times. The temperature was raised to 120°C and stirred overnight. Water was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA% 100) to obtain 76-d (0.48 g, 0.69 mmol, 88.4% yield). MS Calcd: 697.24; MS Found: 643.0 ([M-56+H] + ).

[0298] Step 5: Add 76-d (0.48 g, 0.69 mmol) to a hydrochloric acid-dioxane solution (20 mL) and stir at room temperature overnight. The reaction solution was completely concentrated, and a saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 76-e (0.3 g, 0.5 mmol, 73.0% yield). MS Calcd: 597.19; MS Found: 598.16 ([M+H] +).

[0299] Preparation of intermediate 114-g:

[0300] Step 1: Dissolve 114-a (1.52 g, 9.92 mmol), benzenesulfonyl chloride (2.63 g, 14.88 mmol), and tetrabutylammonium bromide (0.32 g, 0.99 mmol) in a toluene / H2O (10:1) mixture (50 mL). Slowly add sodium hydroxide (3.97 g, 99.22 mmol) at room temperature and stir at room temperature for 1 h. Aqueous solution was added to the reaction mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (EA:PE = 1:6) to obtain 114-b (2.3 g, 7.84 mmol, 79.3% yield). MS Calcd: 293.07; MS Found: 294.2 ([M+H] + ).

[0301] Step 2: 114-b (1.22 g, 4.16 mmol), BPO (100 mg, 0.42 mmol), and NBS (1.11 g, 6.24 mmol) were added sequentially to a carbon tetrachloride solution (50 mL) and stirred at 80°C overnight. The reaction solution was completely concentrated, and water was added to the residue. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA:PE = 1:5) to obtain 114-c (0.8 g, 2.15 mmol, 51.7% yield). MS Calcd: 370.98; MS Found: 372.0 ([M+H] + ).

[0302] Step 3: 114-c (1.95 g, 5.24 mmol) and N-(p-toluenesulfonyl)glycine methyl ester (1.25 g, 5.14 mmol) were added sequentially to an acetonitrile solution (10 mL), followed by K2CO3 (1.808 g, 13.10 mmol). The mixture was stirred at room temperature for 1 h. Saturated ammonium chloride solution was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (EA:PE = 1:2) to afford 114-d (0.68 g, 1.27 mmol, 25.2% yield).

[0303] Step 4: Dissolve 114-d (0.68 g, 1.27 mmol) in THF (5 mL), evacuate the solution and replace the atmosphere with nitrogen three times. Slowly add LiHMDS (3.82 mL, 3.82 mmol) dropwise at -78°C. Continue stirring at this temperature for 1 h. Add saturated ammonium chloride to the reaction solution to terminate the reaction. Extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (EA:PE = 1:2) to obtain 114-e (0.2 g, 0.6 mmol, 47.3% yield). MS Calcd: 332.05; MS Found: 332.97 ([M+H] + ).

[0304] Step 5: Dissolve 114-e (0.18 g, 0.54 mmol) and benzyl bromide (111 mg, 0.65 mmol) in THF (5 mL). Then add cesium carbonate (0.44 g, 1.35 mmol) and heat to 70°C with stirring for 2 h. Saturated ammonium chloride was added to the reaction solution to terminate the reaction. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA:PE = 1:2) to obtain 114-f (0.21 g, 0.5 mmol, 91.7% yield). MS Calcd: 422.09; MS Found: 422.93 ([M+H] + ).

[0305] Step 6: Dissolve 114-f (0.18 g, 0.43 mmol) in 2 mL of acetonitrile, then add potassium methoxide (60 mg, 0.85 mmol) and stir at room temperature for 4 h. The reaction solution was completely concentrated to obtain 114-g (120 mg, 0.43 mmol, 99.8% yield). MS Calcd: 282.10; MS Found: 283.05 ([M+H] + )

[0306] Preparation of intermediate 127-b:

[0307] Step 1: Int-9 (3000 mg, 6.80 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (3570.7 mg, 17.00 mmol), Pd(dppf)Cl2·CH2Cl2 (1113.1 mg, 1.36 mmol), and K3PO4 (4329.7 mg, 20.39 mmol) were added sequentially to dioxane / water (4:1, 40 mL). The atmosphere was purged with nitrogen three times and stirred at 80°C for 24 h. The reaction mixture was concentrated completely, and the residue was isolated by silica gel column chromatography (DCM:MeOH = 20:1) to afford 12-a (0.7 g, 1.57 mmol, 23.2% yield). MS Calcd: 444.25; MS Found: 445.27 ([M+H] + ).

[0308] Step 2: Place 126-c (1376 mg, 3.37 mmol) in a 100 mL eggplant flask and add 20 mL of 1,4-dioxane solution. Then, add 12-a (1500 mg, 3.37 mmol) and DIPEA (1.67 mL, 10.12 mmol) sequentially. Stir at 80°C for 5 hours. TLC monitors the reaction completion. The reaction solution is directly concentrated and purified by column chromatography (PE:EA = 2:3) to obtain 127-a (1050 mg, 1.45 mmol, 42% yield). MS Calcd: 725.20; MS Found: 726.21 ([M+H] + ).

[0309] Step 3: Dissolve 127-a (1050 mg, 1.45 mmol) in 4 mL of dichloromethane solution, add 1 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. The reaction solution was directly concentrated, and 20 mL of saturated sodium bicarbonate solution was added to the residue. Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, dried, and concentrated to obtain 127-b (670 mg, 1.07 mmol, 74% yield). MS Calcd: 625.18; MS Found: 625.20 ([M+H] + ).

[0310] Example 1 Preparation of Compound 129

[0311] Step 1: Compound 114-g (102 mg, 0.36 mmol), DIPEA (116 mg, 0.9 mmol), PyBOP (188 mg, 0.36 mmol), and 76-e (180 mg, 0.36 mmol) were added to DMF (4 mL) and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain compound 129-a (210 mg, 0.24 mmol, 80.9% yield). MS Calcd: 861.28; MS Found: 862.32 ([M+H] + ).

[0312] Step 2: Dissolve 129-a (210 mg, 0.24 mmol) in 3 mL of DCM and slowly add boron trichloride (0.97 mmol, 0.97 mL) dropwise at -78°C. After completion of the addition, slowly warm the mixture to room temperature and stir overnight. The reaction was terminated by adding a small amount of methanol, concentrated under reduced pressure, and the residue purified by prep-HPLC to afford the title compound: 129 (38 mg, 0.05 mmol, 20.2% yield). MS Calcd: 771.23; MS Found: 772.23 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.37 (s, 1H), 8.04 (d, J = 12.8 Hz, 1H), 7.93 (d, J = 7.2 Hz, 1H), 7.44 (d, J = 3.2 Hz, 1H), 6.82-6.80 (m, 1H), 6.69 (d, J = 3.2 Hz, 1H), 5.30 (s, 2H), 4.85 (brs, 0.5H), 4.55 (brs, 0.5H), 4.45 (b rs,0.5H),4.24-4.17(m,2.5H),3.87(s,3H),3.79–3.69(m,3H),3.56-3.50(m,2H),3.27-3.11(m,1.5H), 2.93–2.81(m,1.5H),2.72–2.67(m,1H),2.53-2.51(m,2H),1.43(t,J=7.2Hz,3H),1.20(t,J=7.2Hz,3H).

[0313] Example 2 Preparation of Compound 130

[0314] Step 1: The operation steps were the same as those for the synthesis of 129-a. 114-g (100 mg, 0.35 mmol) was replaced with 72-e (185 mg, 0.32 mmol) to give 130-a (180 mg, 0.21 mmol, 66.7% yield). MS Calcd: 841.33; MS Found: 842.33 ([M+H] + ).

[0315] Step 2: The operation steps were the same as those for the synthesis of compound 129, except that 129-a was replaced with 130-a (180 mg, 0.21 mmol) to obtain compound 130 (35 mg, 0.05 mmol, 21.8% yield). MS Calcd: 751.29; MS Found: 752.31 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.38 (s, 1H), 7.73 (d, J = 12.8 Hz, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.45 (d, J = 3.2 Hz, 1H), 6.82 (t, J = 2.0 Hz, 1H), 6.69 (d, J = 3.2 Hz, 1H), 5.28 (s, 2H), 4.86 (brs, 0.5H), 4.60 (brs, 0.5H), 4.45 (d, J = 13.2 Hz ,0.5H),4.25-4.20(m,2.5H),3.87(s,3H),3.81–3.70(m,3H),3.56-3.49(m,1.5H),3.28-3.12(m,1.5H),2.96–2 .80(m,1.5H),2.71-2.64(m,1H),2.53-2.49(m,2.5H),2.32(s,3H),1.43(t,J=6.4Hz,3H),1.21(t,J=7.2Hz,3H).

[0316] Example 3 Preparation of Compound 18

[0317] Step 1: Dissolve 18-a (24 g, 171.4 mmol) in CCl4 (200 mL), add NBS (36.6 g, 205.71 mmol) and AIBN (1.4 g, 8.57 mmol), replace the atmosphere with nitrogen three times, and stir in a 50°C oil bath overnight. Add 100 mL of DCM to the reaction mixture, then wash twice with 100 mL of water. The organic phases are combined, dried, and concentrated. The crude product is purified by flash column chromatography (PE:EA = 10:1) to afford 18-b (25 g, 114.16 mmol, 66.6% yield).

[0318] Step 2: Dissolve 18-b (25 g, 114.16 mmol), methyl tosylglycine (27.7 g, 114.16 mmol), and potassium carbonate (31.5 g, 228.31 mmol) in acetonitrile (200 mL) and stir at 30°C overnight. The reaction mixture was directly filtered, the filtrate was concentrated, and purified by column chromatography (PE:EA = 2:1) to give 18-c (36 g, 94.39 mmol, 82% yield). MS Calcd: 381.08; MS Found: 382.08 ([M+H] + ).

[0319] Step 3: Dissolve 18-c (36 g, 94.39 mmol) in THF (400 mL) and replace the atmosphere with nitrogen three times under vacuum. Slowly add LiHMDS (400 mL) at -78°C. After completion, continue stirring at this temperature for 2 h. Saturated ammonium chloride solution is slowly added to the reaction solution to terminate the reaction. The pH is adjusted to 7 with 5N HCl solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (PE:EA=1:1) to obtain 18-d (12.5 g, 64.7 mmol, 61% yield). MS Calcd: 193.03; MS Found: 194.09 ([M+H] + ).

[0320] Step 4: Dissolve 18-d (10 g, 51.76 mmol) in 100 mL of methanol, then add dropwise a solution of NaOH (6.2 g, 155.28 mmol) in 25 mL of water. Heat to 60°C and stir overnight. After cooling, a solid precipitated. Filter and obtain 18-e (6.5 g, 36.29 mmol, 70% yield). After drying, use directly in the next step. MS Calcd: 179.02; MS Found: 180.03 ([M+H] + ).

[0321] Step 5: 18-e (60.3 mg, 0.34 mmol), DIPEA (0.14 mL, 0.78 mmol), and HATU (127.8 mg, 0.34 mmol) were added sequentially to a DMF (3 mL) solution and stirred at room temperature for 2 h. Then, intermediate int-4 (150 mg, 0.26 mmol) was added and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was purified by prep-HPLC to obtain the title compound: 18 (8 mg, 0.01 mmol, 4% yield). MS Calcd: 740.21; MS Found: 741.25 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.42 (s, 1H), 8.11 (s, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.96 (d, J = 2.0 Hz, 1H), 7.72 (dd, J = 8.8, 2.0 Hz, 1H), 7.09 (s, 1H), 6.84 (s, 1H), 5.32 (s, 2H), 4.85 (brs, 0. 5H),4.46(brs,0.5H),4.26(q,J=2.8Hz,2H),3.99(brs,0.5H),3.82–3.67(m,3.5H),3.26–3 .15(m,2H),2.97–2.79(m,2H),2.74–2.60(m,4H),1.45-1.39(m,3H),1.22(m,J=7.2Hz,3H).

[0322] Example 4 Preparation of Compound 20

[0323] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was replaced with 72-e (150 mg, 0.26 mmol) to obtain compound 20 (25 mg, 0.03 mmol, 13% yield). MS Calcd: 738.25; MS Found: 739.25 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.45 (s, 1H), 8.11 (s, 1H), 7.76 (d, J = 12.8 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.10 (s, 1H), 6.82 (s, 1H), 5.28 (s, 2H), 4.85 (brs, 0.5H), 4.45 (d, J = 12.8 Hz, 0.5H), 4.26 ( m,J=3.2Hz,2H),3.93(brs,0.5H),3.82–3.66(m,5.5H),3.56-3.47(m,2H),3.27–3.11(m,1.5H) ,2.99–2.82(m,1.5H),2.74–2.62(m,1H),2.34(s,3H),1.45-1.38(m,3H),1.22(t,J=7.2Hz,3H).

[0324] Example 5 Preparation of Compound 24

[0325] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (58 mg, 0.33 mmol) was used to replace int-4 with 76-e (150 mg, 0.26 mmol) to obtain compound 24 (17 mg, 0.02 mmol, 9% yield). MS Calcd: 758.20; MS Found: 759.25 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.44 (s, 1H), 8.12–8.08 (m, 2H), 7.98 (d, J = 8.0 Hz, 1H), 7.10 (s, 1H), 6.82 (s, 1H), 5.34 (s, 2H), 4.85 (brs, 0.5H), 4.46 (d, J = 13.2 Hz, 0.5H), 4.25 (d, J=2.8Hz,2H),3.98(brs,0.5H),3.83–3.67(m,3.5H),3.54-3.49(m,2H),3.27–3.06(m,2 .5H),2.97–2.81(m,2.5H),2.74–2.63(m,1H),1.45-1.39(m,3H),1.22(t,J=7.2Hz,3H).

[0326] Example 6 Preparation of Compound 17

[0327] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was replaced with int-2 (150 mg, 0.26 mmol) to obtain compound 17 (20 mg, 0.03 mmol, 9% yield). MS Calcd: 726.19; MS Found: 727.20 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.43 (s, 1H), 8.10 (d, J = 2.0 Hz, 1H), 8.04 (d, J = 8.8 Hz, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.71 (dd, J = 8.8, 2.0 Hz, 1H), 7.09 (d, J = 2.0 Hz, 1H), 6.84–6.82 (m,1H),5.31(s,2H),4.58(brs,1H),4.25(q,J=2.8Hz,2H),3.82-3.79(m,3H),3.54-3. 47(m,3H),3.28(brs,1H),3.02-2.96(m,4H),2.82–2.63(m,2H),1.20(t,J=7.2Hz,3H).

[0328] Example 7 Preparation of Compound 19

[0329] Step 1: Int-1 (2.2 g, 5.15 mmol), N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (2.0 g, 5.67 mmol), and DIPEA (2.0 g, 15.45 mmol) were added to 60 mL of dioxane solution and stirred at 80°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (EA:PE = 1:1) to obtain 19-a (1.44 g, 2.18 mmol, 42.3% yield). MS Calcd: 659.15; MS Found: 560.21 ([M-100+H] + ).

[0330] Step 2: 19-a (1.44 g, 2.18 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (687 mg, 3.27 mmol), Pd(dppf)Cl2·CH2Cl2 (357 mg, 0.44 mmol), and potassium phosphate (1.39 g, 6.54 mmol) were added sequentially to a 20 mL dioxane / water (4:1) mixture. The mixture was evacuated and replaced with nitrogen three times. The temperature was raised to 80°C and stirred for 2 h. The reaction solution was completely concentrated, and the residue was isolated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 19-b (1.4 g, 2.11 mmol, 96.8% yield). MS Calcd: 663.28; MS Found: 664.32 ([M+H] + ).

[0331] Step 3: Add 19-b (1.4 g, 2.11 mmol) to 20 mL of hydrochloric acid-dioxane solution and stir at room temperature overnight. The reaction solution was completely concentrated, and saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude product 19-c (0.73 g, 1.30 mmol, 61.4% yield). MS Calcd: 563.23; MS Found: 564.21 ([M+H] + ).

[0332] Step 4: The operation steps were the same as those for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was replaced with 19-c (150 mg, 0.26 mmol) to obtain compound 19 (18 mg, 0.03 mmol, 9% yield). MS Calcd: 724.24; MS Found: 725.20 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.39 (s, 1H), 8.08 (s, 1H), 7.76 (d, J = 12.8 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 7.07 (s, 1H), 6.82 (s, 1H), 5.29 (s, 2H), 4.58 (brs, 1H), 4.25 (s, 2H), 3.84-3.78 (m, 4H), 3.54-3.48 (m, 3H), 3.28 (brs, 1H), 3.03-2.96 (m, 3H), 2.81–2.65 (m, 2H), 2.34 (s, 3H), 1.20 (t, J = 7.2 Hz, 3H).

[0333] Example 8 Preparation of Compound 23

[0334] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (58 mg, 0.33 mmol) was replaced with int-4 by int-12 (150 mg, 0.26 mmol) to obtain compound 23 (18 mg, 0.02 mmol, 94% yield). MS Calcd: 744.14; MS Found: 745.22 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.43 (s, 1H), 8.10–8.05 (m, 2H), 7.97 (d, J = 7.6 Hz, 1H), 7.09 (s, 1H), 6.82 (t, J = 2.4 Hz, 1H), 5.34 (s, 2H), 4.59 (brs, 1H), 4.25 (q, J = 2.8 Hz, 2H), 3.80 (t, J = 5.6 Hz, 3H), 3.49 (t, J = 12.0 Hz, 2H), 3.28 (brs, 1H), 3.03–2.95 (m, 3H), 2.86–2.65 (m, 2H), 2.53–2.51 (m, 2H), 1.19 (t, J = 7.2 Hz, 3H).

[0335] Example 9 Preparation of Compound 89

[0336] Step 1: The operation steps were the same as those for the synthesis of 129-a. 114-g (96.8 mg, 0.34 mmol) was replaced with int-8 (180 mg, 0.31 mmol) to give 89-a (210 mg, 0.25 mmol, 80.0% yield). MS Calcd: 841.33; MS Found: 842.38 ([M+H] + ).

[0337] Step 2: The operation steps were the same as those for the synthesis of compound 129, except that 129-a was replaced with 89-a (210 mg, 0.25 mmol). The residue was purified by prep-TLC to obtain compound 89 (25 mg, 0.03 mmol, 13.3% yield). MS Calcd: 751.29; MS Found: 752.29 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.38 (s, 1H), 7.60–7.53 (m, 2H), 7.45 (d, J = 2.8 Hz, 1H), 6.83–6.81 (m, 1H), 6.69 (d, J = 2.8 Hz, 1H), 5.25 (s, 2H), 4.86 (brs, 0.5H), 4.60 (brs, 0.5H), 4.45 (d, J = 12.8 Hz, 0.5H), 4.26–4.24 (m, 2.5H), 3. 88(s,3H),3.80(t,J=5.6Hz,2H),3.74-3.69(m,1.5H),3.56–3.51(m,1.5H),3.28–3.14(m,1.5H),2.93-2.81(m, 1.5H),2.71-2.67(m,1H),2.53-2.51(m,2H),2.22(d,J=2.4Hz,3H),1.44(t,J=2.8Hz,3H),1.21(t,J=7.2Hz,3H).

[0338] Example 10 Preparation of Compound 48

[0339] Step 1: Dissolve 48-a (100 g, 714 mmol) in CCl4 (1000 mL), add NBS (152.6 g, 857.14 mmol) and AIBN (5.9 g, 35.71 mmol), and stir overnight at 50°C under nitrogen. Concentrate the reaction mixture, and the residue is purified by flash column chromatography (PE:EA = 20:1) to obtain 48-b (150 g, 684.93 mmol, 95% yield).

[0340] Step 2: Dissolve 48-b (70 g, 319.63 mmol), p-toluenesulfonylglycine methyl ester (93.3 g, 383.56 mmol), and K2CO3 (97.2 g, 703.20 mmol) in acetonitrile (500 mL) and stir overnight at 30°C. The reaction mixture was filtered and concentrated, and the residue was purified by flash column chromatography (PE:EA = 6:1) to afford 48-c (109 g, 285.79 mmol, 89% yield).

[0341] Step 3: Dissolve 48-c (4.0 g, 10.49 mmol) in THF (40 mL) and slowly add LiHMDS (31.46 mL, 31.46 mmol) dropwise at room temperature. Stir at room temperature for 1 h. Add saturated ammonium chloride to the reaction solution to terminate the reaction. Extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (EA:PE = 4:1) to obtain 48-d (0.6 g, 3.11 mmol, 29.6% yield). MS Calcd: 193.04; MS Found: 194.1 ([M+H] + ).

[0342] Step 4: 48-d (0.12 g, 0.62 mmol) and sodium hydroxide (75 mg, 1.86 mmol) were added sequentially to a 4 mL MeOH / H2O (4:1) mixture, heated to 80°C, and stirred overnight. The reaction solution was completely concentrated, and 5N HCl solution was added to the residue until a solid precipitated. 48-e (80 mg, 0.45 mmol, 71.9% yield) was obtained by filtration. MS Calcd: 179.02; MS Found: 180.02 ([M+H] + ).

[0343] Step 5: The operation steps were the same as those for the synthesis of compound 18, except that int-4 was replaced by int-8 (100 mg, 0.17 mmol), and 18-e was replaced by 48-e (40 mg, 0.23 mmol). The residue was purified by prep-TLC to obtain compound 48 (21 mg, 0.02 mmol, 14.4% yield). MS Calcd: 738.25; MS Found: 739.28 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.43 (s, 1H), 8.07 (t, J = 2.8 Hz, 1H), 7.61-7.54 (m, 2H), 7.21 (s, 1H), 6.84–6.82 (m, 1H), 5.26 (s, 2H), 4.85 (d, J = 7.2 Hz, 0.5H), 4.45 (d, J = 12.8 Hz, 0.5H), 4.26 (q,J=2.8Hz,2H),3.88–3.67(m,3.5H),3.55–3.46(m,1.5H),3.25–3.10(m,1.5H),2.94-2.81 (m,1.5H),2.71-2.60(m,4H),2.23(d,J=2.4Hz,3H),1.44–1.37(m,3H),1.21(t,J=7.2Hz,3H).

[0344] Example 11 Preparation of Compound 36

[0345] Step 1: Dissolve 36-a (5.0 g, 35.69 mmol) in a DCM / MeOH (1:1) mixture (70 mL). Slowly add trimethylsilylated diazomethane (21.41 mL, 42.82 mmol) at 0°C and stir overnight at room temperature. Add aqueous solution to the reaction mixture, extract with dichloromethane, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography (EA:PE = 1:5) to obtain 36-b (2.2 g, 14.27 mmol, 40.0% yield). MS Calcd: 154.07; MS Found: 155.1 ([M+H] + ).

[0346] Step 2: 36-b (1.8 g, 11.67 mmol), BPO (283 mg, 1.17 mmol), and NBS (3.12 g, 17.51 ​​mmol) were added sequentially to a carbon tetrachloride solution (30 mL) and stirred at 80°C overnight. The reaction solution was completely concentrated, and water was added to the residue. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA:PE = 1:5) to obtain 36-c (1.05 g, 4.5 mmol, 38.6% yield). MS Calcd: 231.98; MS Found: 233.0 ([M+H] + ).

[0347] Step 3: 36-c (2.28 g, 9.78 mmol) and N-(p-toluenesulfonyl)glycine methyl ester (2.62 g, 10.76 mmol) were added sequentially to an acetonitrile solution (30 mL), followed by potassium carbonate (3.38 g, 24.45 mmol). The mixture was stirred at room temperature overnight. Saturated ammonium chloride solution was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (EA:PE = 1:2) to obtain 36-d (3.5 g, 8.85 mmol, 90.8% yield). MS Calcd: 395.12; MS Found: 396.09 ([M+H] + ).

[0348] Step 4: Dissolve 36-d (1.12 g, 2.83 mmol) in THF (10 mL), evacuate the solution and replace the atmosphere with nitrogen three times. Slowly add LiHMDS (8.5 mL, 8.5 mmol) dropwise at -78°C. Continue stirring at this temperature for 1 h. Add saturated ammonium chloride to the reaction solution to terminate the reaction. Extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography (EA:PE = 1:2) to obtain 36-e (0.21 g, 1.01 mmol, 35.8% yield). MS Calcd: 207.06; MS Found: 208.14 ([M+H] + ).

[0349] Step 5: 36-e (0.21 g, 1.01 mmol) and sodium hydroxide (122 mg, 3.04 mmol) were added sequentially to a 4 mL MeOH / H2O (4:1) mixture, heated to 80°C, and stirred overnight. The reaction solution was completely concentrated, and 5N HCl solution was added to the residue until a solid precipitated. 36-f (120 mg, 0.62 mmol, 61.3% yield) was obtained by filtration. MS Calcd: 193.05; MS Found: 194.11 ([M+H] + ).

[0350] Step 6: The operation steps were the same as those for the synthesis of compound 18, except that int-4 was replaced by 72-e (100 mg, 0.17 mmol), and 18-e was replaced by 36-f (40 mg, 0.21 mmol). The residue was purified by prep-TLC to obtain compound 36 (16 mg, 0.02 mmol, 12.3% yield). MS Calcd: 752.38; MS Found: 753.27 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.54 (s, 1H), 8.28 (s, 1H), 7.78 (d, J = 12.8 Hz, 1H), 7.66 (d, J = 8.4 Hz, 1H), 6.83 (s, 1H), 5.30 (s, 2H), 4.88 (brs, 0.5H), 4.48 (d, J = 12.8 Hz, 0.5H), 4.33-4.25 (m, 2.5H), 4.12 (s, 3 H),3.94–3.90(m,0.5H),3.82-3.70(m,3.5H),3.56-3.47(m,1.5H),3.29-3.14(m,1.5H),2.95-2.90( m,1.5H),2.72-2.63(m,1H),2.53-2.51(m,2H),2.35(s,3H),1.46–1.41(m,3H),1.22(t,J=7.2Hz,3H).

[0351] Example 12 Preparation of Compound 38

[0352] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that int-4 was replaced by 76-e (120 mg, 0.20 mmol), and 18-e was replaced by 36-f (46 mg, 0.24 mmol). The residue was purified by prep-TLC to obtain compound 38 (16 mg, 0.02 mmol, 9.7% yield). MS Calcd: 772.23; MS Found: 773.22 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.56 (s, 1H), 8.28 (s, 1H), 8.08 (d, J = 12.8 Hz, 1H), 7.93 (d, J = 7.2 Hz, 1H), 6.82 (s, 1H), 5.31 (s, 2H), 4.86 (brs, 0.5H), 4.46 (d, J = 13.2 Hz, 0.5H), 4.29–4.24(m,2.5H),4.12(s,3H),3.89–3.68(m,4.5H),3.56-3.50(m,1H),3.28–3.11 (m,1.5H),2.94–2.81(m,1.5H),2.70(m,3H),1.45-1.40(m,3H),1.21(t,J=7.2Hz,3H).

[0353] Example 13 Preparation of Compound 34

[0354] Step 1: 36-f (58 mg, 0.30 mmol), DIPEA (97 mg, 0.75 mmol), and HATU (114 mg, 0.3 mmol) were added sequentially to a DMF (3 mL) solution, stirred at room temperature for 2 h, and then int-10 (140 mg, 0.25 mmol) was added, and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was purified by prep-TLC to give the title compound: (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-1H-pyrazolo[3,4-c]pyridine-5-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (34, 15 mg, 0.02 mmol, 7.6% yield). MS Calcd: 734.29; MS Found: 735.29 ([M+H] + ).

[0355] Preparation of sodium salt of compound 34: Compound 34 (380 mg, 0.52 mmol) was added to tert-butanol (6 mL) to form a milky white suspension. NaOH solution (5.7 mL, 0.1 M) was then added dropwise. After stirring at room temperature for 10 minutes, the mixture was directly lyophilized to obtain the corresponding sodium salt. 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 7.95 (s, 1H), 7.71 (s, 1H), 7.65–7.58 (m, 2H), 7.50 (dd, J = 8.8, 2.4 Hz, 1H), 6.82 (s, 1H), 5.21 (s, 2H), 4.77 (brs, 0.5H), 4.35 (d, J = 12.4 Hz, 0.5H), 4.24–4.22 (m, 2H), 4.06–2.39 (m, 16H), 2.30 (s, 3H), 1.39–1.16 (m, 6H).

[0356] Example 14 Preparation of Compound 58

[0357] Step 1: Dissolve 58-a (9.5 g, 62.83 mmol) in CCl4 (100 mL), add NBS (13.4 g, 75.40 mmol) and AIBN (0.5 g, 0.05 mmol), replace the atmosphere with nitrogen three times, and stir overnight in an oil bath at 50°C. Add 100 mL of DCM to the reaction solution, wash twice with 100 mL of water, combine the organic phases, dry and concentrate to obtain a crude product. To the crude product, add diethyl phosphite (34.5 g, 251.32 mmol) and DIPEA (32.5 g, 251.32 mmol), stir overnight at room temperature, filter, concentrate the organic phase, and obtain 58-b (5.5 g, 23.90 mmol, 38.0% yield). MS Calcd:228.97;MS Found:230.01([M+H] + ).

[0358] Step 2: Dissolve 58-b (5.3 g, 23.03 mmol), methyl tosylglycine (5.6 g, 23.03 mmol), and potassium carbonate (6.4 g, 46.07 mmol) in acetonitrile (60 mL) and stir at 30°C overnight. The reaction mixture was directly filtered, the filtrate was concentrated, and purified by column chromatography (PE:EA=1:1) to give 58-c (6.75 g, 17.20 mmol, 74.7% yield). MS Calcd: 392.10; MS Found: 393.10 ([M+H] + ).

[0359] Step 3: Dissolve 58-c (6.75 g, 17.20 mmol) in THF (80 mL) and replace the atmosphere with nitrogen three times under vacuum. Slowly add LiHMDS (80 mL) at -78°C. Continue stirring at this temperature for 2 h. Saturated ammonium chloride solution is slowly added to the reaction mixture to terminate the reaction. The pH is adjusted to 7 with 5N HCl solution, and the mixture is extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (PE:EA=1:1) to obtain 58-d (2.7 g, 13.22 mmol, 77.4% yield). MS Calcd: 204.05; MS Found: 205.00 ([M+H] + ).

[0360] Step 4: Dissolve 58-d (2.7 g, 13.22 mmol) in 40 mL of methanol, then add dropwise a solution of NaOH (1.6 g, 39.67 mmol) in 10 mL of water. Heat to 60°C and stir overnight. After cooling, a solid precipitated. Filter to obtain crude 58-e (1.4 g, 7.36 mmol, 55.7% yield), which was used directly in the next step. MS Calcd: 190.04; MS Found: 191.03 ([M+H] + ).

[0361] Step 5: 58-e (80 mg, 0.25 mmol), DIPEA (108 mg, 0.84 mmol), and HATU (95 mg, 0.25 mmol) were added sequentially to a DMF (2 mL) solution and stirred at room temperature for 2 h. Then, 76-e (100 mg, 0.17 mmol) was added and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was purified by C-18 reverse phase column to obtain the title compound: 58 (15 mg, 0.02 mmol, 11.3% yield). MS Calcd: 769.22; MS Found: 770.25 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.00 (s, 1H), 8.68 (s, 1H), 8.33 (s, 1H), 8.10 (d, J = 12.8 Hz, 1H), 8.00 (d, J = 7.2 Hz, 1H), 7.95 (s, 1H), 6.82 (t, J = 2.0 Hz, 1H), 5.37 (s, 2H), 4.9 0(brs,0.5H),4.50(brs,0.5H),4.25(q,J=2.8Hz,2H),3.81-3.70(m,3.5H),3.58-3 .53(m,1H),3.28–3.14(m,1H),2.96-2.83(m,3.5H),2.74(s,3H),1.24–1.16(m,6H).

[0362] Example 15 Preparation of Compound 91

[0363] Step 1: The operation procedure was the same as that for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was used to replace int-4 with int-13 (150 mg, 0.26 mmol). The residue was purified by prep-TLC to obtain compound 91 (15 mg, 0.02 mmol, 7% yield). MS Calcd: 754.22; MS Found: 755.01 ([M+H]+ ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.43 (s, 1H), 8.11 (s, 1H), 7.95 (s, 1H), 7.75 (s, 1H), 7.09 (s, 1H), 6.83 (t, J = 2.0 Hz, 1H), 5.27 (s, 2H), 4.85 (brs, 0.5H), 4.51–4.44 (m, 0.5H), 4.26 (d, J = 2.8 Hz, 2H), 3.96 (brs ,0.5H),3.81(t,J=5.6Hz,2H),3.74-3.67(m,1H),3.59–3.48(m,2H),3.27–3.10(m,1.5H),2.99–2.82 (m,2H),2.72–2.62(m,1H),2.53-2.51(m,2H),2.35(s,3H),1.45-1.39(m,3H),1.22(t,J=7.2Hz,3H).

[0364] Example 16 Preparation of Compound 26

[0365] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was replaced with int-4 by int-8 (150 mg, 0.26 mmol). The residue was purified by prep-TLC to give compound 26 (13 mg, 0.02 mmol, 6.4% yield). MS Calcd: 738.25; MS Found: 739.01 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.45 (d, J = 7.2 Hz, 1H), 8.13 (s, 1H), 7.63-7.57 (m, 2H), 7.11 (s, 1H), 6.84 (s, 1H), 5.28 (s, 2H), 4.86 (brs, 0.5H), 4.47 (d, J = 12.8 Hz, 0.5H), 4.28 (d, J = 3.6 Hz, 2H), 3.98 (brs, 0. 5H),3.83(t,J=5.6Hz,2H),3.76–3.69(m,1H),3.56–3.50(m,2.5H),3.29–3.16(m,1.5H),3.00-2.92( m,1H),2.86–2.64(m,1.5H),2.53-2.51(m,2H),2.26(s,3H),1.46-1.40(m,3H),1.23(t,J=7.2Hz,3H).

[0366] Example 17 Preparation of Compound 22

[0367] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (60.3 mg, 0.34 mmol) was replaced with int-4 by int-10 (150 mg, 0.26 mmol). The residue was purified by prep-TLC to give compound 22 (18 mg, 0.02 mmol, 8% yield). MS Calcd: 720.26; MS Found: 721.21 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.46 (s, 1H), 8.13 (s, 1H), 7.72 (d, J = 8.4 Hz, 1H), 7.63 (d, J = 2.0 Hz, 1H), 7.55 (dd, J = 8.4, 2.0 Hz, 1H), 7.11 (s, 1H), 6.86–6.84 (m, 1H), 5.26 (s, 2H), 4.86 (brs, 0.5H), 4.46 (d, J = 12.8 Hz, 0.5H), 4.28 (q, J = 2 .8Hz,2H),3.96(brs,0.5H),3.82(t,J=5.6Hz,2H),3.75-3.68(m,2H),3.55-3.47(m,1.5H),3.28–3.11(m,1.5H),2. 99–2.91(m,1H),2.84(d,J=10.4Hz,0.5H),2.73–2.63(m,1H),2.36(s,3H),1.46-1.39(m,3H),1.22(t,J=7.2Hz,3H).

[0368] Example 18 Preparation of Compound 32

[0369] Step 1: 36-f (56 mg, 0.30 mmol), DIPEA (94 mg, 0.72 mmol), and HATU (110 mg, 0.29 mmol) were added sequentially to a DMF (3 mL) solution, stirred at room temperature for 2 h, and then int-4 (140 mg, 0.24 mmol) was added, and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and the organic phase was concentrated under reduced pressure. The residue was purified by prep-TLC to obtain the title compound: (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(4-(4-hydroxy-1-methyl-1H-pyrazolo[3,4-c]pyridine-5-carbonyl)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (32,13 mg, 0.02 mmol, 7.1% yield). MS Calcd: 754.24; MS Found: 755.41 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.56 (s, 1H), 8.27 (s, 1H), 8.02 (d, J = 8.4 Hz, 1H), 7.93 (d, J = 2.0 Hz, 1H), 7.70 (dd, J = 8.8, 2.0 Hz, 1H), 6.83 (t, J = 2.0 Hz, 1H), 5.30 (s, 2H), 4.85 (brs, 0.5H), 4.44 (d, J = 12.8 Hz, 0.5H), 4.25-4 .20(m,2.5H),4.11(s,3H),3.80(t,J=5.6Hz,2H),3.75-3.71(m,0.5H),3.54-3.48(m,1.5H),3.27–3.12(m,1 .5H),2.92–2.81(m,1.5H),2.71-2.62(m,3.5H),2.53-2.51(m,1H),1.45–1.38(m,3H),1.22(t,J=7.2Hz,3H).

[0370] Preparation of compound 32 sodium salt: Compound 32 (755 mg, 0.02 mmol) was added to tert-butanol (15.0 mL) to form a milky white suspension. NaOH solution (11.0 mL, 0.1 M) was then added dropwise and stirred at room temperature for 10 minutes to dissolve. The solution was directly lyophilized to obtain the corresponding sodium salt. 1H NMR (400MHz, DMSO-d6) δ8.06(d,J=8.4Hz,1H),7.92(d,J=3.6Hz,1H),7.76( s,1H),7.62(s,1H),7.42(d,J=8.8Hz,1H),6.84(s,1H),5.02(s,2H),4.86( d,J=7.2Hz,0.5H),4.45(d,J=12.4Hz,0.5H),4.27–4.17(m,3H),3.95(s,3H) ),3.82–2.45(m,15H),1.40(dd,J=12.0,6.8Hz,3H),1.22(t,J=7.2Hz,3H).

[0371] Example 19 Preparation of Compound 92

[0372] Step 1: The same operation steps were used for the synthesis of 129-a. 114-g (94 mg, 0.33 mmol) was used to replace 76-e with int-13 (180 mg, 0.30 mmol) to obtain 92-a (150 mg, 0.17 mmol, 57.7% yield). MS Calcd: 857.30; MS Found: 858.32 ([M+H] + ).

[0373] Step 2: The operation procedure was the same as that for the synthesis of compound 129, except that 129-a was replaced with 92-a (150 mg, 0.17 mmol). The residue was purified by prep-TLC to obtain compound 92 (18 mg, 0.02 mmol, 11.5% yield). MS Calcd: 767.26; MS Found: 768.26 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.40 (s, 1H), 7.93 (s, 1H), 7.74 (s, 1H), 7.46 (d, J = 3.2 Hz, 1H), 6.84-6.81 (m, 1H), 6.70 (d, J = 3.2 Hz, 1H), 5.27 (s, 2H), 4.87 (brs, 0.5H), 4.66 (brs, 0.5H), 4.47–4.44 (m, 0.5H), 4.30–4.25 (m, 2.5 H),3.89(s,3H),3.81(t,J=5.6Hz,2H),3.73-3.67(m,1.5H),3.57–3.52(m,1.5H),3.28-3.15(m,1.5H),2.98–2 .83(m,1.5H),2.72–2.68(m,1H),2.53-2.51(m,2H),2.34(s,3H),1.44(d,J=5.6Hz,3H),1.22(t,J=7.2Hz,3H).

[0374] Example 20 Preparation of Compound 90

[0375] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that int-4 was replaced by int-13 (140 mg, 0.24 mmol), and 18-e was replaced by 36-f (55 mg, 0.28 mmol). The residue was purified by prep-TLC to obtain compound 90 (16 mg, 0.02 mmol, 7.5% yield). MS Calcd: 768.25; MS Found: 769.08 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.60 (s, 1H), 8.29 (s, 1H), 7.92 (s, 1H), 7.74 (s, 1H), 6.83-6.81 (m, 1H), 5.27 (s, 2H), 4.86 (brs, 0.5H), 4.46 (d, J = 12.8 Hz, 0.5H), 4.25 (d, J = 3.2 Hz, 2H), 4.13 (s,3H),3.88–3.79(m,3H),3.76–3.72(m,0.5H),3.56–3.50(m,2H),3.29–3.14(m,1.5H),2.9 7-2.82(m,1.5H),2.74-2.64(m,3.5H),2.34(s,3H),1.45–1.40(m,3H),1.22(t,J=7.2Hz,3H).

[0376] Example 21 Preparation of Compound 54

[0377] Step 1: The operation steps were the same as those for the synthesis of compound 58. 58-e (50 mg, 0.26 mmol) was replaced with int-4 (100 mg, 0.17 mmol) to give compound 54 (18.2 mg, 0.02 mmol, 13.6% yield). MS Calcd: 751.22; MS Found: 752.21 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.90 (s, 1H), 8.67 (s, 1H), 8.36 (s, 1H), 8.02 (d, J = 8.8 Hz, 1H), 7.94 (d, J = 2.0 Hz, 1H), 7.71 (dd, J = 8.8, 2.0 Hz, 1H), 7.58 (s, 1H), 6.83 (t, J = 2.0 Hz, 1H), 5.32 (s, 2H), 4.92 (brs, 0.5H ),4.50(brs,0.5H),4.25(d,J=3.2Hz,2H),3.81-3.67(m,3.5H),3.61-3.51(m,1.5H),3.27–3.12(m,1.5 H),2.97–2.82(m,1.5H),2.73–2.67(m,1H),2.59-2.49(m,3H),1.55-1.46(m,3H),1.23(t,J=7.2Hz,3H).

[0378] Example 22 Preparation of Compound 50

[0379] Step 1: The operation steps were the same as those for the synthesis of compound 58. 58-e (51 mg, 0.27 mmol) was replaced with int-10 (100 mg, 0.17 mmol) to obtain compound 50 (16.3 mg, 0.02 mmol, 11.7% yield). MS Calcd: 731.28; MS Found: 732.33 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.89 (s, 1H), 8.63 (s, 1H), 8.36 (s, 1H), 7.67–7.60 (m, 3H), 7.52 (d, J = 8.4 Hz, 1H), 6.83 (t, J = 2.0 Hz, 1H), 5.24 (s, 2H), 4.91 (brs, 0.5H), 4.49 (d, J = 10.8 Hz, 0.5H), 4.24 (d,J=4.0Hz,2H),3.81–3.67(m,4.5H),3.59-3.49(m,1.5H),3.26–3.12(m,1.5H),2.97–2.82(m,1 .5H),2.72–2.66(m,1H),2.32(s,3H),2.55-2.48(m,2H),1.51–1.45(m,3H),1.22(t,J=7.2Hz,3H).

[0380] Example 23 Preparation of Compound 2

[0381] Step 1: Dissolve 114-f (1.5 g, 3.55 mmol) in EtOH (15 mL), then add 20% sodium ethoxide solution (1.57 g, 4.62 mmol) and stir overnight at room temperature. Add aqueous solution to the reaction mixture, extract with dichloromethane, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 2-a (0.8 g, 2.83 mmol, 79.8% yield). MS Calcd: 282.10; MS Found: 283.14 ([M+H] + ).

[0382] Step 2: Dissolve 2-a (0.8 g, 2.83 mmol) in DMF (10 mL). Add sodium hydroxide (170 mg, 4.25 mmol) at room temperature and continue stirring for 10 min. Then, add iodoethane (0.53 g, 3.40 mmol) and stir at room temperature for 3 h. Add water to the residue, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 2-b (0.7 g, 2.26 mmol, 79.6% yield). MS Calcd: 310.13; MS Found: 311.20 ([M+H] + ).

[0383] Step 3: 2-b (0.15 g, 0.48 mmol) and sodium hydroxide (77 mg, 1.93 mmol) were added sequentially to a 4 mL MeOH / H2O (4:1) mixture, heated to 60°C, and stirred overnight. The reaction solution was completely concentrated, and 5N HCl solution was added to the residue. The mixture was concentrated under reduced pressure to afford product 2-c (130 mg, 0.44 mmol, 90.8% yield). MS Calcd: 296.12; MS Found: 297.13 ([M+H] + ).

[0384] Step 4: The operation steps were the same as those for the synthesis of 129-a, except that 114-g was replaced by 2-c (105 mg, 0.25 mmol) and 76-e was replaced by int-4 (120 mg, 0.21 mmol) to give 2-d (150 mg, 0.17 mmol, 84.5% yield). MS Calcd: 857.30; MS Found: 858.33 ([M+H] + ).

[0385] Step 5: The same procedure as compound 129 was used, except that 129-a was replaced with 2-d (150 mg, 0.17 mmol). The residue was purified by prep-TLC to obtain compound 2 (18 mg, 0.02 mmol, 10.2% yield). MS Calcd: 767.26; MS Found: 768.26 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.42 (s, 1H), 8.03 (d, J = 8.8 Hz, 1H), 7.94 (d, J = 2.0 Hz, 1H), 7.71 (dd, J = 8.8, 2.0 Hz, 1H), 7.53 (d, J = 3.2 Hz, 1H), 6.84–6.82 (m, 1H), 6.70 (d, J = 3.2 Hz, 1H), 5.30 (s, 2H), 4.86 (brs, 0.5H), 4.65 (brs, 0.5H), 4.45(d,J=13.2Hz,0.5H),4.32–4.23(m,5H),3.80(t,J=5.6Hz,2H),3.73–3.70(m,1H),3.57–3.51(m,1.5H),3.28–3 .13(m,1.5H),2.95–2.82(m,1.5H),2.74–2.67(m,1H),2.53-2.51(m,2H),1.45-1.38(m,6H),1.23(t,J=7.2Hz,3H).

[0386] Example 24 Preparation of Compound 33

[0387] Step 1: In a 100 mL single-necked bottle, int-1 (1000 mg, 2.34 mmol), 2-iodo-N-(2-methyl-4-trifluoromethylphenyl)acetamide (880 mg, 2.57 mmol), DIPEA (907 mg, 7.02 mmol), and DMF (5 mL) were added sequentially and stirred at room temperature overnight. The reaction mixture was added with 100 mL of DCM and washed with 30 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, and the crude product was purified by flash column chromatography (MeOH:DCM = 0:100% to 10%:90% in 30 min) to afford 33-a (1200 mg, 1.87 mmol, 79% yield). MS Calcd: 641.16; MS Found: 640.05 ([MH] - ).

[0388] Step 2: In a 30 mL microwave tube, reactant 33-a (900 mg, 1.40 mmol), 2-(3,6-dihydropyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (441 mg, 2.10 mmol), Pd(dppf)Cl2 (102 mg, 0.14 mmol), anhydrous potassium phosphate (967 mg, 4.20 mmol), 1,4-dioxane (10 mL), and water (2 mL) were added. The atmosphere was purged with nitrogen three times and stirred in a microwave at 90°C for 2 hr. The reaction mixture was filtered through Celite, and the filter cake was washed with 3 x 10 mL of methanol. The filtrates were combined and evaporated to dryness under reduced pressure. The crude product was purified by flash column chromatography (MeOH:DCM = 0:100% to 10%:90% in 30 min) to give 33-b (820 mg, 1.27 mmol, 90% yield), MS Calcd: 645.29; MS Found: 644.23 ([MH] - ).

[0389] Step 3: 33-b (500 mg, 0.77 mmol), dioxane hydrochloride solution (4 M, 5 mL), and DCM (10 mL) were added to a 100 mL single-necked bottle and stirred at room temperature for 2 hours. The solution became turbid. The reaction mixture was evaporated to dryness under reduced pressure to obtain crude product 33-c (380 mg, 0.7 mmol, 89% yield). MS Calcd: 545.24; MS Found: 546.21 ([M+H] + ).

[0390] Step 4: The operation steps were the same as those for the synthesis of compound 18, except that int-4 was replaced by 33-c (120 mg, 0.22 mmol), and 18-e was replaced by 36-f (51 mg, 0.26 mmol). The residue was purified by prep-TLC to obtain compound 33 (12 mg, 0.02 mmol, 7.2% yield). MS Calcd: 720.27; MS Found: 721.33 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.55 (s, 1H), 8.27 (s, 1H), 7.69 (d, J = 8.8 Hz, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.53 (dd, J = 8.8, 2.0 Hz, 1H), 6.83 (s, 1H), 5.24 (s, 2H), 4.53 (brs, 2H) ,4.26(d,J=2.8Hz,2H),4.11(s,3H),3.80(t,J=5.6Hz,2H),3.54-3.47(m,2H),3.22-2 .97(m,4H),2.74-2.68(m,2H),2.53-2.51(m,2H),2.33(s,3H),1.21(t,J=7.2Hz,3H).

[0391] Example 25 Preparation of Compound 120

[0392] Step 1: 120-a (2.5 g, 8.71 mmol), Pd(Ph3P)2Cl2 (0.61 g, 0.87 mmol), and cuprous iodide (166 mg, 0.87 mmol) were added sequentially to a 20 mL TEA / THF (1:1) mixture. Trimethylethynylsilane (1.28 g, 13.07 mmol) was then slowly added dropwise. The mixture was stirred at room temperature overnight. Water was added to the reaction mixture, extracted with ethyl acetate, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (PE) to afford 120-b (1.9 g, 7.38 mmol, 84.8% yield). MS Calcd: 257.08; MS Found: 258.3 ([M+H] + ).

[0393] Step 2: Dissolve 120-b (1.7 g, 6.61 mmol) in MeOH (20 mL), then add potassium carbonate (1.83 g, 13.21 mmol) and stir at room temperature overnight. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EA:PE = 1:10) to obtain 120-c (1.2 g, 6.48 mmol, 98.1% yield). MS Calcd: 185.05; MS Found: 186.1 ([M+H] + ).

[0394] Step 3: Dissolve 120-c (1.2 g, 6.48 mmol) and triethylamine (2.87 g, 28.36 mmol) in DCM (20 mL). Slowly add chloroacetyl chloride (1.92 g, 17.02 mmol) at 0°C, slowly warm to room temperature, and stir overnight. Add water to the reaction mixture, extract with DCM, wash with saturated brine, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (EA:PE = 1:10) to obtain 120-d (0.72 g, 2.75 mmol, 42.6% yield). MS Calcd: 261.02; MS Found: 262.0 ([M+H] + ).

[0395] Step 4: Dissolve 120-d (0.6 g, 2.29 mmol) in acetone (10 mL), then add potassium iodide (0.76 g, 4.59 mmol) and stir at 60°C for 3 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA:PE = 1:10) to obtain 120-e (0.72 g, 2.04 mmol, 88.9% yield). MS Calcd: 352.95; MS Found: 353.9 ([M+H] + ).

[0396] Step 5: 120-e (1.24 g, 3.51 mmol), 12-a (2.6 g, 3.51 mmol), and DIPEA (1.36 g, 10.53 mmol) were added to a dioxane solution (20 mL) and stirred at 60°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE:EA=5:1) to obtain 120-f (0.35 g, 0.52 mmol, 14.9% yield). MS Calcd: 669.29; MS Found: 614.5 ([M-56+H] + ).

[0397] Step 6: Add 120-f (0.35 g, 0.52 mmol) to a hydrochloric acid-dioxane solution (4 mL) and stir at room temperature overnight. The reaction solution was completely concentrated, and a saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 120-g (0.25 g, 0.44 mmol, 84.0% yield). MS Calcd: 569.24; MS Found: 570.26 ([M+H] + ).

[0398] Step 7: The operation steps were the same as those for the synthesis of compound 18. 18-e (45 mg, 0.25 mmol) was replaced with 120-g (0.11 g, 0.19 mmol) to obtain compound 120 (13 mg, 0.02 mmol, 8.4% yield). MS Calcd: 730.25; MS Found: 731.47 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.37 (d, J = 11.2 Hz, 1H), 8.08–8.02 (m, 2H), 7.87 (d, J = 2.0 Hz, 1H), 7.75 (dd, J = 8.8, 2.0 Hz, 1H), 7.07 (t, J = 2.8 Hz, 1H), 6.84 (t, J = 2.0 Hz, 1H), 5.32 (s, 2H), 4.85 (d, J = 6.0 Hz, 0.5H), 4.69 (s, 1H), 4.45 ( d,J=12.8Hz,0.5H),4.25(d,J=2.8Hz,2H),3.94(brs,0.5H),3.80(t,J=5.6Hz,2H),3.75-3.48(m,3.5H),3.26–3. 09(m,1.5H),2.97–2.81(m,1.5H),2.71-2.67(m,1H),2.53–2.51(m,2H),1.44-1.37(m,3H),1.23(t,J=7.2Hz,3H).

[0399] Example 26 Preparation of Compound 121

[0400] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 48-e (59 mg, 0.33 mmol) and int-4 was replaced by int-13 (150 mg, 0.25 mmol) to give compound 121 (4.3 mg, 0.01 mmol, 2.2% yield). MS Calcd: 754.22; MS Found: 755.25 ([M+H]+ ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.35 (s, 1H), 8.01 (s, 1H), 7.84 (s, 1H), 7.70 (s, 1H), 7.16 (d, J = 2.8 Hz, 1H), 6.81 (s, 1H), 5.23 (s, 2H), 4.81 (s, 0.5H), 4.40 (d, J = 12.8 Hz, 0.5H), 4.23 (d, J = 3.6 Hz, 2H), 4.07-3.99 (m, 0.5H), 3 .80-3.74(m,3H),3.66(t,J=10.0Hz,1H),3.51–3.44(m,1.5H),3.32–3.20(m,0.5H),3.12-3.07(m,1H),2.91 –2.80(m,1.5H),2.70-2.60(m,1H),2.53-2.51(m,2H),2.29(s,3H),1.42-1.33(m,3H),1.17(t,J=7.2Hz,3H).

[0401] Example 27 Preparation of Compound 52

[0402] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (50 mg, 0.26 mmol) and int-4 was replaced by int-8 (100 mg, 0.17 mmol) to obtain compound 52 (11.3 mg, 0.01 mmol, 8.2% yield). MS Calcd: 749.27; MS Found: 750.25 ([M+H] + ).

[0403] Example 28 Preparation of Compound 118

[0404] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (48 mg, 0.25 mmol) and int-4 was replaced by int-13 (100 mg, 0.17 mmol) to obtain compound 118 (13.5 mg, 0.02 mmol, 9.8% yield). MS Calcd: 765.24; MS Found: 766.24 ([M+H] + ).

[0405] Example 29 Preparation of Compound 53

[0406] Step 1: 58-e (50 mg, 0.27 mmol), DIPEA (0.15 mL, 0.88 mmol), and HATU (100 mg, 0.27 mmol) were added to a DMF (2 mL) solution in sequence and stirred at room temperature for 2 h. Then, int-2 (100 mg, 0.18 mmol) was added and stirred at room temperature overnight. The reaction was monitored by LCMS. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was purified by medium pressure preparative purification to give the product as a light yellow solid 53 (42.4 mg, 0.05 mmol, 29.7% yield). MS Calcd: 737.21; MS Found: 738.24 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.99 (s, 1H), 8.67 (s, 1H), 8.34 (s, 1H), 8.08 (d, J = 8.8 Hz, 1H), 7.98 (d, J = 2.0 Hz, 1H), 7.74 (dd, J = 8.8, 2.0 Hz, 1H), 7.66 (s, 1H), 6.85 (s, 1H), 5 .36(s,2H),4.58(brs,2H),4.27(d,J=2.8Hz,2H),3.82(t,J=5.6Hz,2H),3.59-3.55(m ,3H),3.04-3.00(m,3H),2.83–2.68(m,3H),2.53–2.51(m,2H),1.23(t,J=7.2Hz,3H).

[0407] Example 30 Preparation of Compound 56

[0408] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (49 mg, 0.26 mmol) and int-4 was replaced by 72-e (100 mg, 0.17 mmol) to obtain compound 56 (20.3 mg, 0.03 mmol, 14.7% yield). MS Calcd: 749.27; MS Found: 750.34 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.10 (dd, J = 4.0, 1.6 Hz, 1H), 8.93 (d, J = 4.0 Hz, 1H), 8.70 (d, J = 8.4 Hz, 1H), 7.83–7.76 (m, 2H), 7.65 (d, J = 8.4 Hz, 1H), 6.82 (s, 1H), 5.30 (s, 2H), 4.91 (s, 0.5H), 4.62–4.50 (m, 1H), 4.27–4.24 (m, 2.5H), 3 .82–3.74(m,3.5H),3.62-3.54(m,1H),3.33(t,J=12.4Hz,0.5H),3.18-3.14(m,1H),2.98-2.85(m,1.5H),2.75–2. 68(m,1H),2.59(d,J=11.2Hz,0.5H),2.53-2.51(m,2H),2.35(s,3H),1.49(d,J=6.8Hz,3H),1.23(t,J=7.2Hz,3H).

[0409] Example 31 Preparation of Compound 57

[0410] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (49 mg, 0.26 mmol) and int-4 was replaced by int-12 (100 mg, 0.17 mmol) to obtain compound 57 (15.5 mg, 0.02 mmol, 11.3% yield). MS Calcd: 755.20; MS Found: 756.31 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.09 (dd, J = 4.0, 1.6 Hz, 1H), 8.90 (s, 1H), 8.70 (d, J = 8.4 Hz, 1H), 8.07 (d, J = 12.4 Hz, 1H), 7.99 (d, J = 7.6 Hz, 1H), 7.82 (dd, J = 8.4, 4.0 Hz, 1H), 6.82 (s, 1H), 5.35 ( s,2H),4.56(brs,2H),4.24(q,J=2.8Hz,2H),3.80(t,J=5.6Hz,2H),3.59-3.53(m,2H),3.36-3 .16(m,2H),3.00(q,J=7.2Hz,2H),2.83-2.73(m,2H),2.53-2.51(m,2H),1.20(t,J=7.2Hz,3H).

[0411] Example 32 Preparation of Compound 60

[0412] Step 1: Dissolve 60-a (24 g, 158.73 mmol) in CCl4 (200 mL), add NBS (42.4 g, 238.10 mmol) and AIBN (1.3 g, 7.94 mmol), replace the atmosphere with nitrogen three times, and stir overnight in a 50°C oil bath. Add 100 mL of DCM to the reaction solution, then wash twice with 100 mL of water. Combine the organic phases, dry and concentrate to obtain the crude product, which is then purified by flash column chromatography (PE:EA=2:1) ​​to obtain 60-b (15.7 g, 68.23 mmol, 68.23% yield). MS Calcd: 230.97; MS Found: 231.83 ([M+H] + ).

[0413] Step 2: Dissolve 60-b (13.5 g, 58.67 mmol), methyl tosylglycine (14.3 g, 58.67 mmol), and potassium carbonate (16.2 g, 117.34 mmol) in acetonitrile (150 mL) and stir at 30°C overnight. The reaction mixture was directly filtered, the filtrate was concentrated, and purified by column chromatography (PE:EA=1:1) to give 60-c (18.2 g, 46.38 mmol, 79.1% yield). MS Calcd: 392.10; MS Found: 393.19 ([M+H] + ).

[0414] Step 3: Dissolve 60-c (18 g, 45.87 mmol) in THF (180 mL) and replace the atmosphere with nitrogen three times. Slowly add LiHMDS (184 mL) at -78°C. Continue stirring at this temperature for 2 h. Saturated ammonium chloride solution is slowly added to the reaction solution to terminate the reaction. Adjust the pH to 7 with 5N HCl solution, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by silica gel column chromatography (PE:EA=1:1) to obtain 60-d (3.3 g, 16.16 mmol, 35.2% yield). MS Calcd: 204.05; MS Found: 205.13 ([M+H] + ).

[0415] Step 4: Dissolve 60-d (3.3 g, 16.16 mmol) in 40 mL of methanol, then dropwise add NaOH (1.6 g, 39.67 mmol) dissolved in 10 mL of water. Heat to 60°C and stir overnight. After cooling, a solid precipitates. Slowly add water to dissolve the solid. Adjust the pH to 6 with 1N hydrochloric acid and filter to obtain crude product 60-e (2.2 g, 11.5 mmol, 71.6% yield), which is used directly in the next step. MS Calcd: 190.04; MS Found: 191.21 ([M+H] + ).

[0416] Step 5: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 60-e (51 mg, 0.27 mmol) and int-4 was replaced by int-10 (100 mg, 0.17 mmol) to obtain compound 60 (2.6 mg, 0.01 mmol, 2.05% yield). MS Calcd: 731.28; MS Found: 732.35 ([M+H] + ).

[0417] Example 33 Preparation of Compound 21

[0418] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (64 mg, 0.36 mmol) was used to replace int-4 with 33-c (150 mg, 0.26 mmol) to obtain compound 21 (48 mg, 0.06 mmol, 23% yield). MS Calcd: 706.25; MS Found: 707.20 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.49 (s, 1H), 8.13 (d, J = 2.0 Hz, 1H), 7.70 (d, J = 8.4 Hz, 1H), 7.62 (d, J = 2.0 Hz, 1H), 7.55–7.52 (m, 1H), 7.12 (d, J = 2.0 Hz, 1H), 6.84 (s, 1H), 5.24 (s, 2H), 4.26 (d, J = 2.8 Hz,2H),3.83-3.75(m,3H),3.68-3.65(m,1H),3.53-3.47(m,2H),3.27(brs,1H),3.03-2.97(m,3 H),2.83-2.78(m,1H),2.69-2.64(m,1H),2.53-2.51(m,2H),2.34(s,3H),1.20(t,J=7.2Hz,3H).

[0419] Example 34 Preparation of Compound 25

[0420] Step 1: The operation steps were the same as those for the synthesis of compound 18. 18-e (62 mg, 0.35 mmol) was replaced with int-4 by int-5 (150 mg, 0.26 mmol) to obtain compound 25 (28 mg, 0.04 mmol, 13% yield). MS Calcd: 724.24; MS Found: 725.20 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.47 (s, 1H), 8.10 (d, J = 2.0 Hz, 1H), 7.60-7.52 (m, 2H), 7.10 (d, J = 2.0 Hz, 1H), 6.83 (s, 1H), 5.24 (s, 2H), 4.58 (s, 1H), 4.26 (d, J = 2.8 Hz, 2H), 3. 82-3.76(m,3H),3.52-3.47(m,2H),3.27(brs,1H),3.03-2.97(m,3H),2.85-2.81(m,1 H),2.69-2.64(m,1H),2.53-2.51(m,2H),2.22(d,J=2.3Hz,3H),1.19(t,J=7.2Hz,3H).

[0421] Example 35 Preparation of Compound 31

[0422] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (50 mg, 0.25 mmol) and int-4 was replaced by int-2 (120 mg, 0.21 mmol) to obtain compound 31 (8 mg, 0.01 mmol, 4.8% yield). MS Calcd: 740.22; MS Found: 741.23 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.58 (s, 1H), 8.28 (s, 1H), 8.03 (d, J = 8.8 Hz, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.72 (dd, J = 8.8, 2.0 Hz, 1H), 6.83 (s, 1H), 5.31 (s, 2H), 4.55-4.25 (m, 3H), 4.13 (s, 3H), 3.80 (t, J = 5.6 Hz, 2H), 3.52 (dd, J = 13.2, 9.4 Hz, 2H), 3.25-2.97 (m, 4H), 2.75-2.68 (m, 2H), 2.53-2.51 (m, 2H), 1.20 (t, J = 7.2 Hz, 3H).

[0423] Example 36 Preparation of Compound 186

[0424] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 60-e (48 mg, 0.25 mmol) and int-4 was replaced by int-13 (100 mg, 0.17 mmol) to obtain compound 186 (10 mg, 0.01 mmol, 7.6% yield). MS Calcd: 765.24; MS Found: 766.35 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.14 (s, 1H), 8.92 (d, J = 4.8 Hz, 1H), 8.58 (d, J = 8.4 Hz, 1H), 7.93 (s, 1H), 7.78–7.674 (m, 2H), 6.82 (s, 1H), 5.26 (s, 2H), 4.87 (brs, 0.5H), 4.48 (d, J = 12.8 Hz, 0.5H ),4.25(s,2H),3.82-3.70(m,3.5H),3.57–3.47(m,1.5H),3.31–3.13(m,2H),2.96–2.84(m,2H ),2.74-2.61(m,1H),2.53-2.51(m,2H),2.34(s,3H),1.47-1.37(m,3H),1.21(t,J=7.2Hz,3H).

[0425] Example 37 Preparation of Compound 66

[0426] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 60-e (48 mg, 0.25 mmol) and int-4 was replaced by 76-e (100 mg, 0.17 mmol) to obtain compound 66 (10 mg, 0.01 mmol, 7.6% yield). MS Calcd: 769.22; MS Found: 770.35 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 9.06-9.04 (m, 1H), 8.83 (d, J = 4.4 Hz, 1H), 8.49 (d, J = 8.4 Hz, 1H), 7.69-7.64 (m, 2H), 7.56 (d, J = 8.4 Hz, 1H), 6.73 (s, 1H), 5.19 (s, 2H), 4.78 (brs, 0.5H), 4.39 (d, J = 12.8 Hz, 0.5H), 4.16 (d,J=3.2Hz,2.5H),3.73–3.59(m,3H),3.49–3.37(m,1.5H),3.22-3.15(m,1H),3.07-3.00(m,1H),2.85- 2.75(m,2H),2.65–2.51(m,1H),2.44-2.41(m,2H),2.25(s,3H),1.38-1.28(m,3H),1.12(t,J=7.2Hz,3H).

[0427] Example 38 Preparation of Compound 159

[0428] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 48-e (55.9 mg, 0.31 mmol) and int-4 was replaced by 127-b (150 mg, 0.24 mmol) to obtain compound 159 (35 mg, 0.04 mmol, 18% yield). MS Calcd: 786.16; MS Found: 787.68 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.50 (d, J = 4.8 Hz, 1H), 8.11 (d, J = 2.0 Hz, 1H), 8.07 (d, J = 2.0 Hz, 1H), 7.90 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 2.0 Hz, 1H), 7.21 (s, 1H), 6.85 (s, 1H), 5.28 (s, 2H), 4.85 (s, 0.5H), 4.45 (d, J = 12.8 Hz ,0.5H),4.26(d,J=2.8Hz,2H),3.88–3.68(m,4H),3.55–3.47(m,2H),3.27-3.10(m,1.5H),2.94–2.81(m,1.5H) ,2.72–2.62(m,1H),2.53–2.51(m,2H),1.44(d,J=6.8Hz,1.5H),1.41(d,J=6.8Hz,1.5H),1.22(t,J=7.2Hz,3H).

[0429] Example 39 Preparation of Compound 158

[0430] Step 1: 126-a (9500 mg, 39.58 mmol) was placed in a 250 mL eggplant flask. 50 mL of dichloromethane and triethylamine (16.46 mL, 118.75 mmol) were added. Chloroacetyl chloride (4.79 mL, 59.37 mmol) was then weighed and dissolved in 10 mL of dichloromethane. The mixture was slowly added to the reaction system under an ice bath. After complete addition, the mixture was allowed to return to room temperature and stirred for 12 hours. The reaction solution was concentrated, and 50 mL of saturated ammonium chloride solution was added. The mixture was extracted with DCM (3 × 50 mL). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE:EA = 20%) to afford 126-b (10 g, 31.6 mmol, 55% yield). MS Calcd: 314.93; MS Found: 315.98 ([M+H] + ).

[0431] Step 2: 126-b (10 g, 31.6 mmol) was placed in a 100 mL eggplant-shaped flask. 20 mL of acetone solution was added, followed by potassium iodide (10.5 g, 63.2 mmol). The mixture was then heated to 50°C and stirred for 3 hours. 20 mL of saturated sodium thiosulfate solution was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE:EA = 5:1) to give 126-c (7.2 g, 17.65 mmol, 56% yield). MS Calcd: 406.86; MS Found: 407.98 ([M+H] + ).

[0432] Step 3: 126-c (1895 mg, 4.65 mmol) was placed in a 100 mL eggplant flask and 20 mL of 1,4-dioxane solution was added. Int-11 (2000 mg, 4.65 mmol) and DIPEA (2.31 mL, 13.48 mmol) were then added sequentially. The mixture was stirred at 80°C for 5 hours. The reaction mixture was directly concentrated and purified by column chromatography (PE:EA = 2:3) to obtain 126-d (2500 mg, 3.52 mmol, 75% yield). MS Calcd: 709.18; MS Found: 710.20 ([M+H] + ).

[0433] Step 4: Dissolve 126-d (2500 mg, 3.52 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. The reaction solution was directly concentrated, and 20 mL of saturated sodium bicarbonate solution was added to the residue. Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, dried, and concentrated to obtain 126-e (1950 mg, 3.19 mmol, 90% yield). MS Calcd: 609.18; MS Found: 610.20 ([M+H] + ).

[0434] Step 5: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 48-e (57.2 mg, 0.32 mmol) and int-4 was replaced by 126-e (150 mg, 0.24 mmol) to obtain compound 158 (21 mg, 0.03 mmol, 10% yield). MS Calcd: 772.16; MS Found: 773.05 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.50 (s, 1H), 8.12 (d, J = 2.0 Hz, 1H), 8.08 (d, J = 2.0 Hz, 1H), 7.91 (d, J = 8.8 Hz, 1H), 7.76 (dd, J = 8.8, 2.0 Hz, 1H), 7.21 (d, J = 2.0 Hz, 1H), 6.86 (s, 1H), 5.29 (s, 2H ),4.60(brs,1H),4.26(q,J=2.8Hz,2H),3.82-3.77(m,3H),3.54–3.48(m,2H),3.26(brs,1H) ,3.03-2.97(m,3H),2.83(brs,1H),2.65(brs,1H),2.53–2.51(m,2H),1.21(t,J=7.2Hz,3H).

[0435] Example 40 Preparation of Compound 166

[0436] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (68.5 mg, 0.36 mmol) and int-4 was replaced by 127-b (150 mg, 0.26 mmol) to obtain compound 166 (38 mg, 0.05 mmol, 18% yield). MS Calcd: 797.17; MS Found: 798.17 ([M+H] + ). 1 H NMR (400MHz, DMSO-d6) δ9.10(dd,J=4.0,1.6Hz,1H),8.92(d,J=5.2Hz,1H),8.71(d,J=8.8Hz,1H),8.09(d,J=2.0Hz,1 H),7.92(d,J=8.8Hz,1H),7.82(dd,J=8.8,4.0Hz,1H),7.77(dd,J=8.8,2.0Hz,1H),6.86(s,1H),5.30(s,2H),4.91(b rs,0.5H),4.58–4.49(m,1H),4.27-4.21(m,3H),3.82–3.73(m,3H),3.62–3.59(m,0.5H),3.35–3.29(m,0.5H),3.19- 3.13(m,1H),2.98–2.85(m,1.5H),2.75–2.68(m,1H),2.53–2.51(m,2H),1.49(d,J=6.8Hz,3H),1.25(t,J=7.2Hz,3H).

[0437] Example 41 Preparation of Compound 62

[0438] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced with 60-e (48 mg, 0.25 mmol) to give compound 62 (5.6 mg, 0.01 mmol, 2.8% yield). MS Calcd: 751.22; MS Found: 752.35 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.14 (s, 1H), 8.91 (d, J = 4.8 Hz, 1H), 8.58 (d, J = 8.4 Hz, 1H), 8.02 (d, J = 8.4 Hz, 1H), 7.94 (s, 1H), 7.78–7.75 (m, 1H), 7.71 (d, J = 8.8 Hz, 1H), 6.83 (s, 1H), 5.29 (s, 2H), 4.86 (brs, 0.5H), 4.47 (d, J = 12.8 Hz, 0. 5H),4.25(s,3H),3.83–3.69(m,3H),3.56-3.47(m,1.5H),3.30-3.24(m,1H),3.12(t,J=7.2Hz,1H),2.92–2.84(m,1 .5H),2.74–2.68(m,1H),2.53–2.51(m,2H),1.46(d,J=6.8Hz,1.5H),1.38(d,J=6.8Hz,1.5H),1.20(t,J=7.2Hz,3H).

[0439] Example 42 Preparation of Compound 65

[0440] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 60-e (48 mg, 0.25 mmol) and int-4 was replaced by int-12 (100 mg, 0.17 mmol) to obtain compound 65 (13 mg, 0.02 mmol, 10% yield). MS Calcd: 755.20; MS Found: 756.35 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.15 (dd, J = 4.0, 1.6 Hz, 1H), 8.93 (s, 1H), 8.60 (dd, J = 8.4, 1.6 Hz, 1H), 8.09 (d, J = 12.8 Hz, 1H), 8.00 (d, J = 7.2 Hz, 1H), 7.78 (dd, J = 8.4, 4.0 Hz, 1H), 6.83 (s, 1H), 5.35 (s, 2H), 4 .63(d,J=12.6Hz,1H),4.28-4.24(m,3H),3.84–3.79(m,3H),3.54-3.46(m,1H),3.32-3.27(m,1H),3. 07-2.95(m,3H),2.88-2.74(m,1H),2.63(d,J=10.8Hz,1H),2.53-2.51(m,2H),1.19(t,J=7.2Hz,3H).

[0441] Example 43 Preparation of Compound 39

[0442] Step 1: The operation steps were the same as those for the synthesis of 129-a, except that 114-g was replaced by 2-c (104 mg, 0.35 mmol) and 76-e was replaced by int-12 (120 mg, 0.21 mmol) to give 39-a (90 mg, 0.1 mmol, 50.8% yield). MS Calcd: 861.28; MS Found: 862.28 ([M+H] + ).

[0443] Step 2: The operation steps were the same as those for the synthesis of compound 129, except that 129-a was replaced with 39-a (80 mg, 0.09 mmol). The residue was purified by prep-TLC to obtain compound 39 (18 mg, 0.02 mmol, 24.2% yield). MS Calcd: 771.23; MS Found: 772.31 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6 heavy water exchange) δ8.43 (s, 1H), 8.07 (d, J = 12.8 Hz, 1H), 7.95 (d, J = 7.2 Hz, 1H), 7.54 (d, J = 3.2 Hz, 1H), 6.82 (dq, J = 3.2, 1.6 Hz, 1H), 6.70 (d, J = 3.2 Hz, 1H), 5.32 (s, 2H), 4.66 ( d,J=11.2Hz,2H),4.33–4.23(m,4H),3.80(t,J=5.6Hz,2H),3.57-3.50(m,2H),3.17–2.96( m,4H),2.76–2.69(m,2H),2.53–2.51(m,2H),1.40(t,J=7.2Hz,3H),1.20(t,J=7.2Hz,3H).

[0444] Example 44 Preparation of Compound 270

[0445] Step 1: Dissolve 270-a (9.5 g, 41.47 mmol), methyl tosylglycine (11.1 g, 45.61 mmol), and potassium carbonate (11.5 g, 82.93 mmol) in DMF (100 mL) and stir at 30°C overnight. Filter the reaction mixture, dilute with 200 mL of ethyl acetate, wash four times with saturated brine, dry over anhydrous sodium sulfate, and concentrate the organic phase. Purify by column chromatography (PE:EA = 3:1) to afford 270-b (13 g, 33.21 mmol, 80.1% yield). MS Calcd: 391.11; MS Found: 392.01 ([M+H] + ).

[0446] Step 2: Dissolve 270-b (13 g, 33.21 mmol) in THF (150 mL) and replace the atmosphere with nitrogen three times under vacuum. Slowly add LiHMDS (133 mL, 1 mol / L) at -78°C. Continue stirring at this temperature for 2 h. Saturated ammonium chloride solution is slowly added to the reaction mixture to terminate the reaction. The pH is adjusted to 7 with 5N HCl solution, and the mixture is extracted with ethyl acetate. The mixture is washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue is purified by silica gel column chromatography (DCM:MeOH=25:1) to obtain 270-c (3.3 g, 15.26 mmol, 45.9% yield). MS Calcd: 203.06; MS Found: 204.00 ([M+H] + ).

[0447] Step 3: Dissolve 270-c (3.1 g, 15.26 mmol) in 25 mL of methanol, then add dropwise a solution of NaOH (2.4 g, 61.02 mmol) in 25 mL of water. Heat to 50°C and stir overnight. After cooling, a solid precipitated. The solid was filtered, washed with a small amount of methanol and water, and dried to give 270-d (2.5 g, 13.21 mmol, 86.6% yield), which was used directly in the next step. MS Calcd: 189.04; MS Found: 190.00 ([M+H] + ).

[0448] Step 4: The operation procedure was the same as that for the synthesis of compound 18, except that int-4 (100 mg, 0.17 mmol) was replaced with 270-d (49 mg, 0.26 mmol). The residue was purified by prep-TLC to give compound 270 (15.6 mg, 0.02 mmol, 11.1% yield). MS Calcd: 750.23; MS Found: 751.25 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.80 (s, 1H), 8.28 (d, J = 8.0 Hz, 1H), 8.11-8.03 (m, 2H), 7.93 (d, J = 2.0 Hz, 1H), 7.82–7.69 (m, 3H), 6.83 (s, 1H), 5.31 (s, 2H), 4.89–4.76 (m, 1H), 4.50–4.40 (m, 1H) ,4.25–4.23(m,2H),3.81-3.73(m,3H),3.61-3.54(m,1.5H),3.33-3.14(m,1.5H),2.95–2.84( m,1.5H),2.73-2.69(m,1H),2.59–2.51(m,2.5H),1.48(d,J=6.8Hz,3H),1.22(t,J=7.2Hz,3H).

[0449] Example 45 Preparation of Compound 398

[0450] Step 1: Dissolve 398-a (8.5 g, 47.9 mmol) in MeOH (25 mL) and add a solution of ICl (11.56 g, 71.19 mmol) in dichloromethane (25 mL) dropwise at 0°C. After addition, stir at room temperature for 2 hours. Concentrate the reaction mixture, add 30 mL of dichloromethane, and wash with saturated sodium thiosulfate solution, sodium bicarbonate solution, and brine in sequence. The organic phase is dried, concentrated, and purified by column chromatography (PE:EA = 50:1) to give 398-b (10.5 g, 34.4 mmol, 72% yield). MS Calcd: 304.93; MS Found: 305.98 ([M+H] + ).

[0451] Step 2: Dissolve 398-b (10 g, 32.7 mmol), methylboronic acid (2.9 g, 49.18 mmol), potassium phosphate (20.9 g, 4.92 mmol), and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.4 g, 3.28 mmol) in 1,4-dioxane (80 mL) and react at 80°C under nitrogen overnight. The reaction solution was filtered, the filtrate was concentrated, and purified by column chromatography (PE:EA=9:1) to give 398-c (3.8 g, 19.68 mmol, 60% yield). MS Calcd: 193.05; MS Found: 194.08 ([M+H] + ).

[0452] Step 3: 398-c (3.8 g, 19.68 mmol) was placed in a 250 mL eggplant flask, and 50 mL of dichloromethane solution and triethylamine (6.74 mL, 48.68 mmol) were added. Chloroacetyl chloride (2.36 mL, 29.52 mmol) was then weighed and dissolved in 10 mL of dichloromethane. The mixture was slowly added to the reaction system under an ice bath. After the addition was complete, the mixture was allowed to react at room temperature. Stir for 12 hours. The reaction solution was concentrated, and 50 mL of saturated ammonium chloride solution was added. The mixture was extracted with DCM (3 × 50 mL). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE:EA = 20%) to afford 398-d (3800 mg, 14.09 mmol, 71% yield). MS Calcd: 269.02; MS Found: 270.08 ([M+H] + ).

[0453] Step 4: 398-d (3800 mg, 14.09 mmol) was placed in a 100 mL eggplant flask. 20 mL of acetone solution was added, followed by potassium iodide (4680 mg, 28.19 mmol). The mixture was then heated to 50°C and stirred for 3 hours. 20 mL of saturated sodium thiosulfate solution was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried, concentrated, and purified by column chromatography (PE:EA = 5:1) to give 398-e (3.3 g, 9.14 mmol, 64% yield). MS Calcd: 360.96; MS Found: 361.98 ([M+H] + ).

[0454] Step 5: 398-e (2.5 g, 6.92 mmol) was placed in a 100 mL eggplant flask and 20 mL of 1,4-dioxane solution was added. Int-9 (3055.2 mg, 6.92 mmol) and DIPEA (3.63 mL, 20.77 mmol) were then added sequentially. The mixture was stirred at 80°C for 5 hours. The reaction mixture was directly concentrated and purified by column chromatography (PE:EA = 2:3) to obtain 398-f (3.3 g, 4.89 mmol, 77.0%). MS Calcd: 673.16; MS Found: 674.20 ([M+H] + ).

[0455] Step 6: 398-f (3.3 g, 4.89 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (1541.9 mg, 7.34 mmol), Pd(dppf)Cl2 (354 mg, 0.49 mmol), and potassium phosphate (3375 mg, 6.04 mmol) were added to a solution of 1,4-dioxane (30 mL) and H2O (6 mL). The atmosphere was replaced with N2 three times, then heated to 90°C and stirred for 4 hours. The reaction mixture was spin-dried, the residue was dissolved, filtered, and the filtrate was concentrated and purified by column chromatography (DCM:MeOH = 50:1) to afford 398-g (2950 mg, 4.35 mmol, 89% yield). MS Calcd:677.29; MS Found:678.20([M+H] + ).

[0456] Step 7: Dissolve 398-g (2950 mg, 4.35 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. The reaction solution was directly concentrated, and 20 mL of saturated sodium bicarbonate solution was added to the residue. Extraction was performed with dichloromethane (20 mL × 3). The organic phases were combined, dried, and concentrated to obtain 398-h (2050 mg, 3.55 mmol, 81% yield). MS Calcd: 577.24; MS Found: 578.20 ([M+H] + ).

[0457] Step 8: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (87.0 mg, 0.45 mmol) and int-4 was replaced by 398-h (200 mg, 0.35 mmol) to obtain compound 398 (25 mg, 0.03 mmol, 9.8% yield). MS Calcd: 752.28; MS Found: 753.20 ([M+H] + ).

[0458] Example 46 Preparation of Compound 163

[0459] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 48-e (80.6 mg, 0.45 mmol) and int-4 was replaced by 398-h (200 mg, 0.35 mmol) to obtain compound 163 (30 mg, 0.04 mmol, 11.7% yield). MS Calcd: 738.26; MS Found: 739.38 ([M+H] + ).

[0460] Example 47 Preparation of Compound 168

[0461] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 58-e (85.6 mg, 0.45 mmol) and int-4 was replaced by 398-h (200 mg, 0.35 mmol) to obtain compound 168 (35 mg, 0.05 mmol, 13% yield). MS Calcd: 749.37; MS Found: 750.38 ([M+H] + ).

[0462] Example 48 Preparation of Compound 194

[0463] Step 1: Dissolve 194-a (9.5 g, 51.30 mmol) in ethanol (150 mL), add ethylhydrazine dihydrochloride (7.5 g, 56.43 mmol), and stir overnight in a 70°C oil bath. After concentrating the ethanol, dissolve the residue in ethyl acetate. The organic phase is washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product is purified by flash column chromatography (PE:EA=2:1) ​​to obtain 194-b (7.83 g, 42.97 mmol, 83.8% yield). MS Calcd: 182.11; MS Found: 183.11 ([M+H] + ).

[0464] Step 2: 194-b (6.8 g, 37.32 mmol) was dissolved in CCl4 (80 mL), and NBS (9.9 g, 55.98 mmol) and AIBN (0.3 g, 1.87 mmol) were added. The atmosphere was replaced with nitrogen three times, and the mixture was stirred in an oil bath at 90°C for 1 hour. 100 mL of DCM was added to the reaction solution, and the mixture was washed twice with 100 mL of water. The organic phases were combined, dried, and concentrated to give the crude product. The crude product was then purified by flash column chromatography (PE:EA=1:1) to give 194-c (4.0 g, 15.32 mmol, 41.0% yield). MS Calcd: 260.02; MS Found: 260.94 ([M+H] + ).

[0465] Step 3: 194-c (4.0 g, 15.32 mmol), tosylglycine methyl ester (4.1 g, 16.85 mmol), and potassium carbonate (4.2 g, 30.64 mmol) were dissolved in acetonitrile (60 mL) and stirred at 30°C overnight. The reaction solution was directly filtered, the filtrate was concentrated, and purified by column chromatography (PE:EA = 1:1) to give 194-d (5.5 g, 12.99 mmol, 84.8% yield) MS Calcd: 423.15; MS Found: 423.98 ([M+H] + ).

[0466] Step 4: 194-d (5.5 g, 12.99 mmol) was dissolved in THF (55 mL) and the atmosphere was replaced with nitrogen three times under vacuum. LiHMDS (52 mL, 1 mmol / L) was slowly added dropwise at -78°C and stirred for 2 h at the same temperature. Saturated ammonium chloride solution was slowly added to the reaction solution to terminate the reaction. The pH was adjusted to 7 with 5N HCl solution, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 25:1) to obtain 194-e (190 mg, 0.86 mmol, 6.6% yield). MS Calcd: 221.08; MS Found: 202.02 ([M+H] + ).

[0467] Step 5: Dissolve 194-e (190 mg, 0.86 mmol) in 4 mL of methanol, then add dropwise a solution of NaOH (187 mg, 3.44 mmol) in 4 mL of water. Heat to 50°C and stir overnight. Concentrate the methanol under reduced pressure, adjust the pH to acidic with 1N hydrochloric acid, and a solid precipitates. Filter to obtain crude 194-f (100 mg, 0.48 mmol, 56.2% yield). MS Calcd: 207.06; MS Found: 208.06 ([M+H] + ).

[0468] Step 6: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced with 194-f (86 mg, 0.41 mmol) to give compound 194 (23 mg, 0.03 mmol, 8.8% yield). MS Calcd: 768.25; MS Found: 769.35 ([M+H] + ).

[0469] Example 49 Preparation of Compound 136

[0470] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (48 mg, 0.25 mmol) and int-4 was replaced by 127-b (130 mg, 0.21 mmol) to obtain compound 136 (10 mg, 0.012 mmol, 6.0% yield). MS Calcd: 800.18; MS Found: 801.27 ([M+H] + ).

[0471] Example 50 Preparation of Compound 35

[0472] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (54 mg, 0.28 mmol) and int-4 was replaced by 19-c (130 mg, 0.23 mmol) to obtain compound 35 (11 mg, 0.015 mmol, 6.5% yield). MS Calcd: 738.26; MS Found: 739.33 ([M+H] + ).

[0473] Example 51 Preparation of Compound 37

[0474] Step 1: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (100 mg, 0.51 mmol) and int-4 was replaced by int-12 (150 mg, 0.26 mmol) to obtain compound 37 (12 mg, 0.02 mmol, 6.2% yield). MS Calcd: 758.21; MS Found: 759.23 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.61 (s, 1H), 8.29 (s, 1H), 8.08 (d, J = 12.4 Hz, 1H), 7.99 (d, J = 7.2 Hz, 1H), 6.83–6.81 (m, 1H), 5.35 (s, 2H), 5.53 (s, 2H), 4.25 (q, J = 2.8 Hz, 2H), 4.14 (s, 3H), 3.80 (t, J = 5.6 Hz, 2H), 3.56–3.49 (m, 2H), 3.33–2.96 (m, 4H), 2.76–2.69 (m, 2H), 2.53–2.49 (m, 2H), 1.19 (t, J = 7.2 Hz, 3H).

[0475] Example 52 Preparation of Compound 139

[0476] Step 1: 139-a (4.0 g, 25.45 mmol), BPO (0.62 g, 2.54 mmol), and NBS (5.43 g, 30.53 mmol) were added sequentially to a carbon tetrachloride solution (50 mL) and stirred at 80°C overnight. The reaction solution was completely concentrated, and water was added to the residue. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA:PE = 1:5) to obtain 139-b (5.8 g, 24.57 mmol, 96.5% yield). MS Calcd: 234.93; MS Found: 236.0 ([M+H] + ).

[0477] Step 2: 139-b (5.56 g, 23.55 mmol) and N-(p-toluenesulfonyl)glycine methyl ester (4.73 g, 19.43 mmol) were added sequentially to an acetonitrile solution (50 mL), followed by potassium carbonate (6.1 g, 44.16 mmol). The mixture was stirred at room temperature overnight. Saturated ammonium chloride solution was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (EA:PE = 5:1) to obtain 139-c (4.6 g, 11.55 mmol, 49.1% yield). MS Calcd: 398.06; MS Found: 399.12 ([M+H] + ).

[0478] Step 3: Dissolve 139-c (2.0 g, 5.02 mmol) in DMSO (20 mL), then add potassium carbonate (4.16 g, 30.12 mmol). Heat to 60°C and continue stirring overnight. Saturated ammonium chloride was added to the reaction solution to terminate the reaction, and the mixture was acidified with 5N hydrochloric acid. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain 139-d (0.18 g, 0.86 mmol, 17.1% yield). MS Calcd: 210.01; MS Found: 211.11 ([M+H] + ).

[0479] Step 4: Dissolve 139-d (0.18 g, 0.86 mmol) and sodium hydroxide (137 mg, 3.43 mmol) in a mixture of MeOH / H2O (4:1) (10 mL), heat to 60°C, and stir overnight. The reaction solution was concentrated, and 5N dilute hydrochloric acid was added to the residue to precipitate a solid, which was filtered to obtain 139-e (0.11 g, 0.56 mmol, 65.5% yield). MS Calcd: 195.99; MS Found: 197.09 ([M+H] + ).

[0480] Step 5: 139-e (47 mg, 0.24 mmol), DIPEA (84 mg, 0.64 mmol), and HATU (92 mg, 0.24 mmol) were added sequentially to a DMF (2 mL) solution and stirred at room temperature for 2 h. Then, int-10 (90 mg, 0.16 mmol) was added and stirred at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain a crude product. The crude product was further purified by prep-HPLC to obtain the title compound 139 (8 mg, 0.01 mmol, 6.7% yield). MS Calcd: 737.24; MS Found: 738.28 ([M+H] + ).

[0481] Example 53 Preparation of Compound 140

[0482] Step 1: The operation procedure was the same as that for the synthesis of compound 139. 139-e (53 mg, 0.27 mmol) was replaced with int-4 (120 mg, 0.21 mmol) to obtain compound 140 (7 mg, 0.01 mmol, 4.3% yield). MS Calcd: 757.18; MS Found: 758.12 ([M+H] + ).

[0483] Example 54 Preparation of Compound 400

[0484] Step 1: To a 100 mL eggplant-shaped flask, 400-a (3.0 g, 11.32 mmol) (raw material 400-a was prepared according to CN111278283A), tert-butyl carbamate (1.99 g, 16.98 mmol), and 1,4-dioxane (20 mL) were added. Xantphos (1.4 g, 2.45 mmol), Pd2(dba)3 (1.6 g, 1.75 mmol), and Cs2CO3 (17.1 g, 52.53 mmol) were then added. The mixture was heated to 85°C under nitrogen for 5 hours. The mixture was cooled to room temperature, filtered, and concentrated under reduced pressure to afford the crude product. Flash column chromatography (PE:EA = 20:1) afforded 400-b (2.56 g, 8.52 mmol, 75.3% yield). MS Calcd: 301.09; MS Found: 300.12 ([MH] - ).

[0485] Step 2: Add 400-b (2.56 g, 8.52 mmol) and methanolic hydrochloric acid (10 mL) to a 100 mL eggplant-shaped flask and react at room temperature for 2 h. Add sodium hydroxide to the reaction solution to adjust the pH to neutral, filter, and wash the organic phase with 2 x 30 mL of water, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to obtain crude product 400-c (1.54 g, 7.65 mmol, 89.9% yield). MS Calcd: 201.04; MS Found: 200.09 ([MH] - ).

[0486] Step 3: To a 100 mL eggplant-shaped flask, 400-c (1.54 g, 7.65 mmol), dichloromethane (15 mL), and triethylamine (1.55 g, 15.3 mmol) were added dropwise. A dichloromethane solution (5 mL) of chloroacetyl chloride (1.03 g, 9.18 mmol) was added dropwise under an ice bath. The mixture was then allowed to react overnight at room temperature. The reaction mixture was evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by flash column chromatography (PE:EA = 5:1) to obtain 400-d (1.45 g, 5.25 mmol, 68.6% yield). MS Calcd: 277.01; MS Found: 276.05 ([MH] - ).

[0487] Step 4: To a 100 mL eggplant-shaped flask, 400-d (1.45 g, 5.25 mmol), potassium iodide (2.18 g, 13.12 mmol), and acetone (20 mL) were added and reacted at 50°C for 4 hours. The reaction solution was evaporated to dryness under reduced pressure to obtain a crude product, which was purified by flash column chromatography (PE:EA = 5:1) to afford 400-e (1.54 g, 4.19 mmol, 79.8% yield). MS Calcd: 368.95; MS Found: 367.92 ([MH] - ).

[0488] Step 5: 400-e (1.54 g, 4.19 mmol), 12-a (1.86 g, 4.19 mmol), and DIPEA (1.62 g, 12.57 mmol) were added to a dioxane solution (20 mL) and stirred at 60°C for 4 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (PE:EA=5:1) to obtain 400-f (2.17 g, 3.16 mmol, 75.6% yield). MS Calcd: 685.28; MS Found: 630.32 ([M-56+H] + ).

[0489] Step 6: Add 400-f (2.17 g, 3.16 mmol) to a hydrochloric acid-dioxane solution (4N, 10 mL) and stir at room temperature overnight. The reaction solution was concentrated, and a saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude product 400-g (1.71 g, 2.92 mmol, 92.5% yield). MS Calcd: 585.23; MS Found: 586.28 ([M+H] + ).

[0490] Step 7: The operation steps were the same as those for the synthesis of compound 18, except that 18-e was replaced by 36-f (40 mg, 0.21 mmol), and int-4 was replaced by 400-g (100 mg, 0.17 mmol). The residue was purified by prep-TLC to obtain compound 400 (13 mg, 0.018 mmol, 10.6% yield). MS Calcd: 760.27; MS Found: 761.31 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.64 (s, 1H), 8.30–8.29 (m, 2H), 8.03 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.13 (s, 1H), 6.81 (s, 1H), 5.36 (s, 2H), 4.87 (brs, 0.5H), 4.47 (d, J = 13.2 Hz, 0.5H), 4.32 (brs, 0.5H) ,4.23(d,J=2.8Hz,2H),4.14(s,3H),3.95(d,J=10.4Hz,0.5H),3.81–3.72(m,3H),3.60-3.52(m,2H),3. 29–3.16(m,1.5H),2.92–2.84(m,1.5H),2.74–2.68(m,1H),1.44(d,J=6.8Hz,3H),1.22(t,J=7.2Hz,3H).

[0491] Example 55 Preparation of Compound 401

[0492] Step 1: To a 25 mL eggplant flask, 58-e (24.4 mg, 0.13 mmol) and DMF (2 mL) were added. HATU (48.7 mg, 0.13 mmol) and DIPEA (33.1 mg, 0.26 mmol) were then added and allowed to react at room temperature for 10 minutes. 400-g (50 mg, 0.09 mmol) was then added and allowed to react overnight at room temperature. The mixture was diluted with DCM (30 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude product. The crude product was purified by prep-HPLC to afford the title compound 401 (33 mg, 0.04 mmol, 50.1% yield). MS Calcd: 757.26; MS Found: 758.3 ([M+H] + ). 1H NMR (400 MHz, DMSO-d6, heavy water exchange) δ9.06 (s, 1H), 8.88 (s, 1H), 8.70 (d, J = 8.4 Hz, 1H), 8.27–8.25 (m, 1H), 8.03–8.00 (m, 1H), 7.81–7.77 (m, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.13 (t, J = 2.0 Hz, 1H), 6.81 (s, 1H), 5.35 (s, 2H), 4.90 (br s,0.5H),4.50(d,J=12.4Hz,1H),4.23(d,J=2.8Hz,2H),4.10(s,0.5H),3.81–3.50(m,5.5H),3.35-3.28(m,0 .5H),3.17-3.12(m,1H),2.96-2.86(m,2H),2.76-2.57(m,2H),1.48(d,J=6.8Hz,3H),1.23(t,J=7.2Hz,3H).

[0493] Example 56 Preparation of Compound 402

[0494] Step 1: The operation procedure was the same as that for the synthesis of compound 401. 400-g (50 mg, 0.09 mmol) was replaced with 48-e (22.9 mg, 0.13 mmol) to obtain compound 402 (25 mg, 0.03 mmol, 39.2% yield). MS Calcd: 746.24; MS Found: 747.3 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ8.49 (d, J = 5.2 Hz, 1H), 8.27 (d, J = 2.4 Hz, 1H), 8.11 (d, J = 2.4 Hz, 1H), 8.02 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.21 (s, 1H), 7.13–7.12 (m, 1H), 6.81 (s, 1H), 5.35 (s ,2H),4.85(brs,0.5H),4.45(d,J=12.8Hz,0.5H),4.29-4.22(m,2.5H),3.87–3.72(m,4.5H),3.53 -3.46(m,1.5H),3.27–3.12(m,1.5H),2.92–2.63(m,3H),1.45-1.37(m,3H),1.21(t,J=7.2Hz,3H).

[0495] Example 57 Preparation of Compound 403

[0496] Step 1: To a 50 mL eggplant-shaped flask, 400-e (85.7 mg, 0.23 mmol), int-1 (100 mg, 0.23 mmol), DIPEA (90 mg, 0.70 mmol), and dioxane (15 mL) were added. The mixture was then reacted at 80°C overnight. The reaction mixture was filtered, and the filtrate was diluted with 30 mL of ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (EA:PE = 100%) to obtain 403-a (110 mg, 0.16 mmol, 70.5% yield); MS Calcd: 671.27; MS Found: 672.3 ([M+H] + ).

[0497] Step 2: To a 50 mL eggplant-shaped flask, 403-a (110 mg, 0.16 mmol) and HCl / 1,4-dioxane solution (50 mL) were added. Stir at room temperature for 1 hour. The reaction solution was concentrated, and the residue was dissolved in EA (50 mL). Saturated sodium bicarbonate solution was then added to adjust the pH to approximately pH 7. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford 403-b (73 mg, 0.13 mmol, 78.0% yield). MS Calcd: 571.22; MS Found: 572.3 ([M+H] + ).

[0498] Step 3: The operation steps were the same as those for the synthesis of compound 401, except that 400-g was replaced by 403-b (73 mg, 0.13 mmol) and 58-e was replaced by 36-f (37.0 mg, 0.19 mmol) to obtain compound 403 (16 mg, 0.02 mmol, 22.5% yield). MS Calcd: 760.27; MS Found: 761.4 ([M+H] + ). 1H NMR (400MHz, DMSO-d6) δ12.43(s,1H),11.09(s,1H),8.67(s,1H),8.37(d,J=2.4Hz,1H ),8.33(s,1H),8.08(d,J=8.4Hz,1H),7.63(d,J=8.4Hz,1H),7.17–7.15(m,1H),6.84–6 .82(m,1H),5.39(s,2H),4.59(s,2H),4.26(d,J=2.8Hz,2H),4.28(s,3H),3.82(t,J=5 .6Hz,3H),3.61–3.55(m,3H),3.06-3.02(m,3H),2.79(brs,3H),1.23(t,J=7.2Hz,3H).

[0499] Example 58 Preparation of Compound 404

[0500] Step 1: To a 25 mL eggplant flask, 48-e (23.5 mg, 0.13 mmol) and DMF (2 mL) were added. HATU (49.9 mg, 0.13 mmol) and DIPEA (33.9 mg, 0.26 mmol) were then added and allowed to react at room temperature for 10 minutes. 403-b (50 mg, 0.09 mmol) was then added and allowed to react at room temperature overnight. The mixture was diluted with DCM (30 mL). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford the crude product. The crude product was purified by prep-HPLC to afford the title compound 404 (4 mg, 0.005 mmol, 5.7% yield). MS Calcd: 732.23; MS Found: 733.3 ([M+H] + ). 1 H NMR (400MHz, DMSO-d6) δ11.10(s,1H),8.46(s,1H),8.37(d,J=2.4Hz,1H),8.12(s,1H),8. 07(d,J=8.4Hz,1H),7.62(d,J=8.4Hz,1H),7.23(s,1H),7.15(t,J=2.0Hz,1H),6.83(t,J= 2.4Hz,1H),5.38(s,2H),4.63(s,1H),4.28-4.24(m,3H),3.85-3.80(m,3H),3.55(t,J=11 .2Hz,2H),3.37–3.27(m,2H),3.03–3.00(m,3H),2.85–2.68(m,3H),1.22(t,J=7.2Hz,3H).

[0501] Example 59 Preparation of Compound 405

[0502] Step 1: The operation procedure was the same as that for the synthesis of compound 401. 58-e (25.0 mg, 0.13 mmol) was replaced by 403-b (50 mg, 0.09 mmol) to obtain compound 405 (19 mg, 0.02 mmol, 27.1% yield). MS Calcd: 743.24; MS Found: 744.3 ([M+H] + ). 1 H NMR(400MHz, DMSO-d6)δ12.50(s,1H),11.08(s,1H),9.12(dd,J=4.0,1.6Hz,1H),8.93(s,1H),8.73–8.70(m, 1H),8.37(d,J=2.0Hz,1H),8.07(d,J=8.4Hz,1H),7.83(dd,J=8.4,4.0Hz,1H),7.62(d,J=8.4Hz,1H),7.15(t, J=2.0Hz,1H),6.83(t,J=2.0Hz,1H),5.39(s,2H),4.67(d,J=12.4Hz,2H),4.25(d,J=2.8Hz,2H),3.81(t,J=5 .6Hz,2H),3.64-3.58(m,2H),3.44-3.25(m,4H),3.04(q,J=7.2Hz,2H),2.82(brs,2H),1.24(t,J=7.2Hz,3H).

[0503] Example 60 Preparation of Compound 442

[0504] Step 1: Sodium deuterated formate (2.01 g, 29.14 mmol), 442-a (2.0 g, 7.29 mmol), Pd2(dba)3 (0.67 g, 0.73 mmol), and tri-tert-butylphosphine (0.15 g, 0.73 mmol) were added sequentially to 20 mL of dimethyl sulfoxide solution. The solution was evacuated and replaced with nitrogen three times. The temperature was raised to 80°C and stirred overnight. The solution was diluted with 70 mL of ethyl acetate. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The organic phase was completely concentrated and the residue was purified by silica gel column chromatography (PE:EA = 6:1) to obtain 442-b (1.6 g, 6.51 mmol, 89.3% yield). MS Calcd: 196.01; MS Found: 195.15 ([MH] - ).

[0505] Step 2: 442-b (1.6 g, 6.51 mmol) and triethylamine (0.67 g, 0.73 mmol) were added sequentially to a 20 mL dichloromethane solution. The atmosphere was replaced with nitrogen under vacuum. Chloroacetyl chloride (1.1 g, 9.77 mmol) was slowly added to the reaction solution under ice-cooling. The temperature was raised to room temperature and stirred overnight. The reaction was quenched by adding 50 mL of water and extracted with dichloromethane. The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate. The organic phase was completely concentrated and the residue was purified by silica gel column chromatography (PE:EA = 15:1) to obtain 442-c (1.5 g, 5.49 mmol, 84.4% yield). MS Calcd: 271.98; MS Found: 271.10 ([MH] - ).

[0506] Step 3: 442-c (1.5 g, 5.49 mmol) and potassium iodide (1.37 g, 8.24 mmol) were added sequentially to a 20 mL acetone solution, heated to 50°C, and stirred overnight. The mixture was filtered, and the solid was washed with dichloromethane. The organic phases were combined and concentrated completely. The residue was separated and purified by silica gel column chromatography (PE:EA = 10:1) to obtain 442-d (1.6 g, 4.39 mmol, 79.9% yield). MS Calcd: 363.92; MS Found: 363.12 ([MH] - ).

[0507] Step 4: Compound 442-d (854 mg, 1.19 mmol), compound 12-a (700 mg, 1.92 mmol), and compound DIPEA (744 mg, 5.76 mmol) were added sequentially to a dioxane solution (20 mL). The mixture was heated to 80°C and stirred for 4 hours. The reaction solution was completely concentrated, and the residue was purified by silica gel column chromatography (PE:EA = 1:1) to afford compound 442-e (1.01 g, 1.47 mmol, 76.5% yield). MS Calcd: 680.26; MS Found: 681.25 ([M+H] + ).

[0508] Step 5: 442-e (1.01 g, 1.47 mmol) was added to a DCM solution (10 mL), and TFA (2 mL) was added, followed by stirring at room temperature for 3 hr. The reaction solution was completely concentrated, and saturated sodium bicarbonate solution was added to the residue. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to afford crude product 442-f (0.7 g, 1.20 mmol, 82.1% yield). MS Calcd: 580.20; MS Found: 581.35 ([M+H] + ).

[0509] Step 6: 36-f (43 mg, 0.22 mmol) and 1-chloro-N,N,2-trimethylpropenamine (82 mg, 0.62 mmol) were added sequentially to a dichloromethane solution (3 mL). The mixture was stirred at room temperature for 2 hours. 442-f (150 mg, 0.26 mmol) and DIPEA (100 mg, 0.77 mmol) were then added sequentially. Stirring was continued at room temperature overnight. Water was added to the reaction solution, extracted with DCM, washed with saturated brine, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (DCM:MeOH = 20:1) to obtain a crude product. The crude product was further purified by prep-HPLC to obtain the title compound 442 (10.07 mg, 0.01 mmol, 4.7% yield). MS Calcd: 755.24; MS Found: 756.35 ([M+H] + ). 1 H NMR (400 MHz, DMSO-d6, heavy water exchange) δ 8.63 (d, J = 4.0 Hz, 1H), 8.30 (s, 1H), 7.95 (d, J = 2.0 Hz, 1H), 7.72 (d, J = 2.0 Hz, 1H), 6.84 (tt, J = 3.2, 1.6 Hz, 1H), 5.32 (d, J = 3.2 Hz, 2H), 4.87 (brs, 1H), 4.47 (d, J = 13.2 Hz, 1H), 4.33-4.24 (m,3H),4.14(s,3H),3.96(d,J=9.2Hz,0.5H),3.81(t,J=5.6Hz,2H),3.74-3.69(m,1H),3.56–3.51(m,2H) ,3.29–3.14(m,1.5H),2.93–2.83(m,1.5H),2.72–2.56(m,1.5H),1.46–1.41(m,3H),1.22(t,J=7.2Hz,3H).

[0510] Example 61 In vitro enzymatic activity evaluation:

[0511] By measuring IC 50 The inhibitory activity of the compounds against human WRN helicase was evaluated by PCR.

[0512] Main reagents and consumables:

[0513] Experimental Protocol: Two single-stranded DNA oligos were annealed to form dsDNA, which served as the substrate for the WRN (aa 500-946) enzymatic reaction. Reaction buffer composition and concentrations: 5 mM NaCl, 25 mM Tris base (pH 8.0), 0.01% Tween 20, 2 mM MgCl2, 1 mM TCEP, 0.02% BSA. All reactions were performed in a microplate shaker set at 25°C and 300 rpm. The experiment used a 5 μL reaction system. At the beginning of the experiment, 2 μL WRN (aa500-946) solution (final concentration 1.2 nM) and 1 μL compound solution (maximum final concentration of compound 10 μM, 3× gradient dilution, 10 concentration points, final DMSO concentration 1%) were added to the compound group in a 384-well plate. 2 μL WRN (aa500-946) solution (final concentration 1.2 nM) and 1 μL reaction buffer (final DMSO concentration 1%) were added to the maximum signal group. 2 μL reaction buffer and 1 μL reaction buffer (final DMSO concentration 1%) were added to the minimum signal group. Each group was incubated for 30 min, and then 2 μL substrate solution (containing dsDNA with a final concentration of 0.1 nM and 10 μM ATP) was added to react for 60 min. Subsequently, 5 μL ADP Glo Reagent was added and incubated for 40 min. Finally, 10 μL Kinase Detection Reagent was added and incubated for 30 min. After incubation, the Tecan Luminescence was read using Spark. Inhibition rate (%) = {1-[(luminescence signal of compound group - minimum signal group) / (maximum signal group - minimum signal group)]}*100%. Data were analyzed using GraphPad Prism 8 software, using the "log (inhibitor) vs. response - variable slope (four parameters)" method to fit the data and obtain the compound IC. 50 value.

[0514] Table 1. Half maximal inhibitory concentration (IC) of the compounds of the present invention on WRN 50

[0515] Conclusion: The results show that the IC of the compounds of the present invention for WRN 50 The value is below 50 nM, indicating that the compound of the present invention has good inhibitory activity against WRN.

[0516] Example 62 Cell activity and selectivity test experiment:

[0517] The cell activity and selectivity of the inhibitors were evaluated by testing the proliferation inhibitory activity of WRN non-covalent inhibitors on the MSI-H cell line SW48 and the MSS cell line SW620.

[0518] Main reagents and consumables:

[0519] When SW48 and SW620 cells were cultured to a growth density of approximately 80%, the cells were digested with trypsin and counted. The cells were diluted to 1*10^4 cells / mL with DMEM (the corresponding culture medium for SW48 cells) and DMEM-F12 (the corresponding culture medium for SW620 cells). Then, 100 μL of the diluted cell suspension was added to a 96-well plate to ensure that the number of cells in each well was 1000. The plates were then placed in a carbon dioxide incubator (37°C, 5% CO2) and cultured overnight until the cells adhered. Test compounds were dissolved in DMSO and diluted to 5 mM. Using an automated microplate pipetting system, the compounds were serially diluted eight times, starting at 5 mM and ending at 0.0008 mM (a total of nine concentration points), in 96-well V-bottom plates. These serially diluted compounds were then diluted 500-fold with the corresponding culture medium of SW48 and SW620 cells, to a final DMSO concentration of 0.2% and a compound concentration range of 10 μM to 0.0015 μM. The corresponding culture medium containing only 0.2% DMSO served as a blank control. The culture medium in the SW48 and SW620 cell culture plates was thoroughly aspirated. Subsequently, 100 μL of culture medium containing the corresponding compound concentration was added to the wells of the compound control group, while culture medium containing only 0.2% DMSO was added to the wells of the DMSO control group. The blank, compound, and DMSO control groups were cultured in a CO2 incubator for 4 days. After 4 days, remove the culture plate from the incubator and allow it to cool to room temperature. Once it returns to room temperature, add 50 μL of the prepared CellTiter-Glo solution to each well. The plate is then placed in a microplate shaker at 25°C and 800 rpm for 10 minutes. After incubation, pipette 100 μL of the reaction solution from each well into a black 96-well plate and read the luminescence using a microplate reader.

[0520] The inhibition percentage (%) of each concentration of compound was calculated based on the signals of the minimum fluorescence signal group (blank group) and the maximum fluorescence signal group (DMSO control group) contained in each test plate. The minimum fluorescence signal group contained no cells and only SW48 and SW620 cell culture medium (0.2% DMSO), while the maximum fluorescence signal group contained no compound and only SW48 and SW620 cells (0.2% DMSO). The inhibition rate of each compound concentration was calculated as follows: inhibition rate % = {1-[(luminescence signal of compound group - minimum fluorescence signal group) / (maximum luminescence signal group - minimum luminescence signal group)]} * 100%, and GI was obtained using GraphPad Prism software. 50 Values ​​and nonlinear regression curve fitting, compound cell selectivity = SW620 GI 50 / SW48 GI 50 , GI 50 The selectivity values ​​are shown in the table below.

[0521] Reference Examples 1 and 2 (HRO761) were prepared with reference to Examples 58 and 42 in WO2022249060A1, and the structures are as follows:

[0522] Table 2 Cell proliferation inhibitory activity and selectivity of compounds SW48 and SW620 of the present invention Note: NT means not tested.

[0523] Conclusion: The results showed that the compounds of the present invention had strong proliferation inhibition activity against MSI-H cells SW48, but had very weak proliferation inhibition against MSS cells SW620, indicating that the compounds of the present invention had good selectivity.

[0524] Example 63 Pharmacokinetic Experiment:

[0525] Experimental materials, methods and results analysis:

[0526] The experimental animals were healthy adult BALB / c female mice (provided by Beijing Huafukang Biotechnology Co., Ltd.);

[0527] Mouse medication and sample collection:

[0528] BALB / c female mice were given a single oral administration (10 mg / kg, solvent: 5% DMSO + 5% solutol + 90% saline) (DMSO, Chengdu Kelong Chemical Co., Ltd.; Solutol, BASF, 42966-1KG; Saline, Sichuan Meida Kangjiale Pharmaceutical Co., Ltd., 100 mL_0.9 g). 60 μL of whole blood was collected from the fundus venous plexus of the mice at different time points at 0.25, 0.5, 1, 2, 4, 6, and 8 h after administration, and the blood was centrifuged at 4000 rpm for 10 min to obtain plasma.

[0529] Sample analysis:

[0530] 10 μL of mouse plasma sample was added to 190 μL of acetonitrile solution containing internal standard to precipitate proteins. The samples were vortexed for 10 minutes and then centrifuged at 4000 rpm for 10 minutes. The supernatant was collected and transferred to a 96-well plate for analysis using LC-MS / MS (mass spectrometer, TQ4500, liquid chromatograph, Shimadzu 30AD).

[0531] The LC-MS / MS method was used to determine the drug concentration in the plasma of mice at different times after oral administration of the example compound, and the relevant pharmacokinetic parameters were calculated to study the pharmacokinetic behavior of the compound in mice and evaluate its pharmacokinetic characteristics.

[0532] Table 3. Pharmacokinetic parameters of the compounds of the present invention in mice

[0533] Conclusion: The results showed that the compounds of the present invention had higher exposure in mice.

[0534] Example 64 In vivo efficacy: evaluation of antitumor efficacy in a human colon cancer cell line SW48 transplanted tumor model

[0535] Experimental Materials:

[0536] Human colon cancer cell line SW48 was purchased from Nanjing Kebai Biotechnology Co., Ltd. (CBP60018) and cultured in DMEM medium (Gibco) containing 10% fetal bovine serum (Gibco).

[0537] The sodium salt of Control Example 2 (HRO761), the sodium salt of Compound 32, and the sodium salt of Compound 34 were prepared into clear solutions with a concentration of 6 mg / mL using sterile water and then administered.

[0538] Experimental methods:

[0539] SW48 cells in the logarithmic growth phase were digested and collected, and after counting, the cell density was adjusted to 3×10 cells using a 1:1 ratio of serum-free medium and Matrigel (Corning). 7Twenty-four Balb / c nude mice (18 ± 2 g, Beijing Huafukang Biotechnology Co., Ltd.) were subcutaneously inoculated with 100 μL of SW48 cell suspension. On the 10th day after inoculation, the average tumor volume reached 129 mm 3 Animals were randomly divided into four groups (N=6): a vehicle control group and groups receiving the sodium salt of Control Example 2, the sodium salt of Compound 32, and the sodium salt of Compound 34. The sodium salt of Control Example 2, the sodium salt of Compound 32, and the sodium salt of Compound 34 were all administered orally at 60 mg / kg once daily. The vehicle control group was given an equal volume of sterile water. After 16 days of continuous administration, the animals were observed after drug withdrawal.

[0540] After the animals were inoculated with SW48 cells, the tumor volume was measured 2 to 3 times a week. The tumor volume was calculated as length × width. 2 ×0.5 calculation.

[0541] The tumor inhibition results of each group are shown in the attached figure:

[0542] As shown in Figure 1, compound 32, compound 34 and control example 2 were administered for 16 days, significantly reducing tumor volume (P < 0.0001) and causing tumor regression. After 25 days of observation (D41), 60 mg / kg of compound 32 and compound 34 continued to cause tumor regression, and were superior to control example 2. After 41 days of observation (D57), 60 mg / kg of control example 2 showed a significant rebound, with an average tumor volume of 776 mm 3 The tumor volumes of compound 32 and compound 34 at 60 mg / kg were significantly lower than those of control 2, with an average tumor volume of <200 mm 3 .

[0543] Conclusion: The above experimental results show that compounds 32 and 34 have good anti-tumor effects in the human colon cancer cell line SW48 CDX model; the degree and speed of tumor rebound after discontinuation of 60 mg / kg compound 32 and compound 34 are significantly lower than those in control example 2, indicating that compounds 32 and compound 34 have better anti-tumor efficacy than clinical control example 2.

[0544] Industrial applicability

[0545] The compounds of the present invention have excellent WRN inhibitory activity and can be used as drugs for treating or preventing diseases associated with this activity.

[0546] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A compound of formula (I), or a stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: Among them, Ring A is selected from 5- or 6-membered heteroaryl; each of the 5- or 6-membered heteroaryl independently contains 1-3 heteroatoms selected from N, O, and S; each of the 5- or 6-membered heteroaryl is independently unsubstituted or substituted by one or more R1; each R1 is independently selected from the same or different cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl; R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, and R8 are each independently selected from H, halogen, hydroxyl, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl.

2. The compound according to claim 1, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, characterized in that: Ring A is selected from 5-membered heteroaryl and 6-membered heteroaryl; the 5-membered heteroaryl contains 1-2 heteroatoms selected from N, O and S, and the 6-membered heteroaryl contains 1-2 N heteroatoms; the 5-membered heteroaryl and 6-membered heteroaryl are each independently unsubstituted or substituted by one or more R1; Preferably, ring A is selected from 5-membered heteroaryl and 6-membered heteroaryl; the 5-membered heteroaryl contains 1-2 heteroatoms selected from N and O, and the 6-membered heteroaryl contains 1 N heteroatom; the 5-membered heteroaryl and 6-membered heteroaryl are each independently unsubstituted or substituted by one or more R1; Preferably, ring A is selected from pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl; the pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl are each independently unsubstituted or substituted by one or more R1; Preferably, ring A is selected from pyrrolyl, furyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridyl; the pyrrolyl, furyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridyl are each independently unsubstituted or substituted by one or more R1; Preferably, ring A is selected from pyrrolyl, furyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, pyridyl; the pyrrolyl, pyrazolyl, imidazolyl are each independently unsubstituted or substituted by one or more R1; Preferably, ring A is selected from Among them, 1 and is connected to phase 1, 2 and is connected to 2, so that ring A and the pyridine ring are fused to form a 9-membered or 10-membered heteroaryl; Preferably, ring A is selected from wherein 1 and is connected to phase 1, and 2 and is connected to 2, so that ring A and the pyridine ring are fused to form a 9-membered or 10-membered heteroaryl; Each R1 is independently selected from the same or different C 1-6 alkyl group, C 3-6 cycloalkyl group; Preferably, each R1 is independently selected from the same or different C 1-3 alkyl, C 4-6 alkyl, C 3-6 cycloalkyl; Preferably, each R1 is independently selected from the same or different C 1-3 alkyl, C 3-6 cycloalkyl; Preferably, each R1 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; Preferably, each R1 is independently selected from the same or different methyl, ethyl, cyclopropyl; Each R1 is independently selected from the same or different C 1-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R1 is independently selected from the same or different C 1-3 alkyl, C 4-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R1 is independently selected from the same or different C 1-3 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R1 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; Preferably, each R1 is independently selected from the same or different methyl, ethyl, F, Cl, cyclopropyl; Most preferably, Selected from Or Selected from Preferably, Selected from 3. A compound of formula (IX), or a stereoisomer, tautomer, polymorph, co-crystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: Among them, Ring A is selected from 5-membered or 6-membered heteroaromatic rings, benzene rings, ring B is selected from pyridine rings, N-containing heteroalkenyl groups, the N and the substituent hydroxyl group on ring B are respectively in the ortho position of the carbon atom connected to the carbonyl group, the 5-membered heteroaromatic ring contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaromatic ring contains 1-3 N atoms; ring B and ring A are fused to form a 9-membered or 10-membered bicyclic ring; Ring A is unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; said phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 ; said 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain 1-3 heteroatoms selected from N, O and S; R2 and R3 are each independently selected from H, C 1-6 alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 cycloalkyl; R4, R5, R6, R7, R8, each R 11 is independently selected from H, D, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2- 6-enyl, C 2-6 alkynyl; or R4 and R8 combine with the connected carbon atom to form C 3-10 cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O and S; R 10 selected from Among them, when Selected from When not selected from 4. A compound of formula (IX), or a stereoisomer, tautomer, polymorph, co-crystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: Among them, Ring A is selected from a 5- or 6-membered heteroaryl ring, a benzene ring, ring B is a pyridine ring, the N and the substituent hydroxyl group on ring B are respectively adjacent to the carbon atom connected to the carbonyl group, the 5-membered heteroaryl ring contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl ring contains 1-3 N atoms; Or ring A is a 5- or 6-membered heteroaryl ring, a benzene ring, ring B is a nitrogen-containing heterocyclic alkenyl group, the N and the substituent hydroxyl group on ring B are respectively adjacent to the carbon atom connected to the carbonyl group, the 5-membered heteroaryl ring contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl ring contains 1-3 N atoms; Ring A is unsubstituted or substituted with one or more R9s; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; said phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted with one or more R 11 ; said 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain 1-3 heteroatoms selected from N, O, and S; R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, R8, each R 11 is independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl; R 10 Selected from 5. A compound represented by formula (X), or a stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: Among them, Ring A is a 5- or 6-membered heteroaryl group, ring B is a pyridyl group, the N and the substituent hydroxyl group on ring B are respectively adjacent to the carbon atom connected to the carbonyl group, the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms; Or ring A is a 5- or 6-membered heteroaryl group, ring B is a nitrogen-containing heterocyclic alkenyl group, the N and the substituent hydroxyl group on ring B are respectively adjacent to the carbon atom connected to the carbonyl group, the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms; Ring A is unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl; R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, and R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl.

6. The compound according to claim 3, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: having the structure shown in formula (XI): Among them, Ring A is a 5- or 6-membered heteroaryl group, the 5-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S, and the 6-membered heteroaryl group contains 1-3 N atoms; the 5- or 6-membered heteroaryl group is independently unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano group, amino group, halogen, C 1-6 alkyl group, C 1-6 alkoxy group, C 1-6 haloalkyl group, C 1-6 haloalkoxy group, C 3-6 cycloalkyl group; R2 and R3 are each independently selected from H, C 1-6 alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 cycloalkyl; R4, R5, R6, R7, and R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2- 6 alkynyl; or R4 and R8 together with the connected carbon atom form a C 3-10 cycloalkyl, 5- to 10-membered heteroaryl, 5- to 10-membered heterocyclic group, or C 6-10 aryl; the 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O, and S; Among them, when Selected from When Not selected from 7. The compound according to claim 6, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt: Ring A is selected from a 5-membered heteroaryl group, a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-2 N atoms; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R9; Preferably, ring A is selected from furyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl; the furyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl are each independently unsubstituted or substituted by one or more R9; Preferably, Selected from Each R9 is independently selected from the same or different C 1-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different C 1-3 alkyl, C 4-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different C 1-3 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; Preferably, each R9 is independently selected from the same or different methyl, ethyl, F, Cl, cyclopropyl; Most preferably, Selected from 8. The compound according to claim 5, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof: having the structure shown in formula (XI): Among them, Ring A is a 5- or 6-membered heteroaryl group, the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-3 N atoms; the 5- or 6-membered heteroaryl group is independently unsubstituted or substituted by one or more R9; each R9 is independently selected from the same or different cyano group, amino group, halogen, C 1-6 alkyl group, C 1-6 alkoxy group, C 1-6 haloalkyl group, C 1-6 haloalkoxy group, C 3-6 cycloalkyl group; R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, and R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl.

9. The compound according to claim 8, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt: Ring A is selected from a 5-membered heteroaryl group, a 6-membered heteroaryl group; the 5-membered heteroaryl group contains 2-3 heteroatoms selected from N, O and S, and the 6-membered heteroaryl group contains 1-2 N atoms; the 5-membered heteroaryl group and the 6-membered heteroaryl group are each independently unsubstituted or substituted by one or more R9; Preferably, ring A is selected from 5-membered heteroaryl and 6-membered heteroaryl; the 5-membered heteroaryl contains 2 heteroatoms selected from N, O, and S, and the 6-membered heteroaryl contains 1 N atom; the 5-membered heteroaryl and 6-membered heteroaryl are each independently unsubstituted or substituted with one or more R9; Preferably, ring A is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl; the pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl are each independently unsubstituted or substituted with one or more R9; Preferably, ring A is selected from pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl; the pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, pyridyl are each independently unsubstituted or substituted with one or more R9; Preferably, Selected from Each R9 is independently selected from the same or different C 1-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different C 1-3 alkyl, C 4-6 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different C 1-3 alkyl, halogen, C 3-6 cycloalkyl; Preferably, each R9 is independently selected from the same or different methyl, ethyl, n-propyl, isopropyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl; Preferably, each R9 is independently selected from the same or different methyl, ethyl, F, Cl, cyclopropyl; Most preferably, Selected from 10. The compound according to claim 3, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, has the structure shown in formula (XIII): Among them, R2 and R3 are each independently selected from H, C 1-6 alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 cycloalkyl; R4, R5, R6, R7, and R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2- 6 alkynyl; or R4 and R8 combine with the connected carbon atom to form C 3-10 cycloalkyl, 5-10 membered heteroaryl, 5-10 membered heterocyclic group or C 6-10 aryl; the 5-10 membered heteroaryl and 5-10 membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O, and S; Each R9 is independently selected from the same or different cyano group, amino group, halogen, C 1-6 alkyl group, C 1-6 alkoxy group, C 1-6 haloalkyl group, C 1-6 haloalkoxy group, C 3-6 cycloalkyl group; m is 0, 1, 2, 3 or 4.

11. The compound according to claim 4, or its stereoisomer, tautomer, polymorph, co-crystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, has the structure shown in formula (XIII): Among them, R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, and R8 are each independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl; Each R9 is independently selected from the same or different cyano group, amino group, halogen, C 1-6 alkyl group, C 1-6 alkoxy group, C 1-6 haloalkyl group, C 1-6 haloalkoxy group, C 3-6 cycloalkyl group; m is 0, 1, 2, 3 or 4.

12. The compound according to claim 10 or 11, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, characterized in that: Each R9 is independently selected from the same or different cyano groups, halogens, C 1-6 alkyl groups, C 1-6 alkoxy groups, C 1-6 haloalkyl groups; Preferably, each R9 is independently selected from the same or different cyano groups, halogens, C 1-4 alkyl groups, C 1-4 alkoxy groups, C 1-4 haloalkyl groups; Preferably, each R9 is independently selected from the same or different cyano groups, halogens, C 1-2 alkyl groups, C 3-4 alkyl groups, C 1-2 alkoxy groups, C 3-4 alkoxy groups, C 1-2 fluoroalkyl groups, C 3-4 fluoroalkyl groups; Preferably, each R9 is independently selected from the same or different cyano groups, halogens, C 1-2 alkyl groups, C 1-2 alkoxy groups, C 1-2 fluoroalkyl groups; Preferably, each R9 is independently selected from the same or different cyano, F, Cl, Br, methyl, ethyl, methoxy, ethoxy, fluoromethyl, fluoroethyl; Preferably, each R9 is independently selected from the same or different cyano, F, Cl, methyl, methoxy, -CF3, -CHF2; m is 0, 1, 2, or 3; Preferably, m is 0, 1, or 2; Preferably, m is 0 or 1.

13. The compound according to claim 3, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, has the structure shown in formula (XV): Among them, R9 is selected from the group consisting of cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4- to 6-membered heterocycloalkyl; said phenyl, 5- or 6-membered heteroaryl, 4- to 6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 substituents; said 5- or 6-membered heteroaryl, 4- to 6-membered heterocycloalkyl each independently contain 1-3 heteroatoms selected from N, O, and S; R2 and R3 are each independently selected from H, C 1-6 alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 cycloalkyl; R4, R5, R6, R7, R8, and each R 11 is independently selected from H, D, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2- 6-enyl, C 2-6 alkynyl; or R4 and R8 together with the connected carbon atom form a C 3-10 cycloalkyl, 5- to 10-membered heteroaryl, 5- to 10-membered heterocyclic group or C 6-10 aryl; the 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclic group each independently contain 1 to 3 heteroatoms selected from N, O, and S.

14. The compound according to claim 4, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, has the structure shown in formula (XV): Among them, R9 is selected from cyano, amino, halogen, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by 1 or more R 11 substituents; the 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain 1-3 heteroatoms selected from N, O, and S; R2 and R3 are each independently selected from H, C 1-6 alkyl; R4, R5, R6, R7, R8, each R 11 is independently selected from H, halogen, hydroxy, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl.

15. The compound according to claim 13 or 14, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, characterized in that: R9 is selected from C 1-6 alkyl, C 1-6 haloalkyl, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 substituents; the 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain 1-2 heteroatoms selected from N, O, and S; Preferably, R9 is selected from C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl; the phenyl, 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 substituents; the 5- or 6-membered heteroaryl, 4-6-membered heterocycloalkyl each independently contain one heteroatom selected from N, O, and S; Preferably, R9 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, fluoromethyl, fluoroethyl, C 3-4 fluoroalkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl, furyl, thienyl, pyrrolyl, pyridyl, 4- to 6-membered heterocycloalkyl; the phenyl, pyrrolyl, pyridyl, 4- to 6-membered heterocycloalkyl are each independently unsubstituted or substituted by one or more R 11 substituents; the 4- to 6-membered heterocycloalkyl contains one heteroatom selected from N and O; Each R 11 is independently selected from halogen, hydroxy, cyano, amino, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl, C 1-4 haloalkoxy, C 3-6 cycloalkyl; Preferably, each R 11 is independently selected from halogen, C 1-4 alkyl, C 1-4 alkoxy, C 1-4 haloalkyl, C 1-4 haloalkoxy, C 3-6 cycloalkyl; Preferably, each R 11 is independently selected from halogen, C 1-4 alkyl; Preferably, each R 11 is independently selected from F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; Preferably, each R 11 is independently selected from F, methyl; Preferably, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CHF2, -CH2CF3, -CH2CHF2, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, oxolanyl, oxanyl, N-methylcyclobutyl, N-methylcyclohexyl, phenyl, 4-fluorophenyl, furyl, thienyl, pyridyl; Preferably, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, oxolanyl, oxanyl, phenyl, 4-fluorophenyl; Most preferably, R9 is selected from methyl, ethyl, isopropyl, tert-butyl, -CF3, -CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl, phenyl 16. The compound according to claim 3, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, has the structure shown in formula (XX): Among them, R2 and R3 are each independently selected from H, C 1-6 alkyl; or R2 and R3 together with the carbon atom to which they are attached form C 3-5 cycloalkyl; R4, R5, R6, R7, and R8 are each independently selected from H, D, halogen, hydroxy, cyano, amino, C 1- 6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2- 6 alkynyl; or R4 and R8 together with the attached carbon atom form a C 3-10 cycloalkyl, 5- to 10-membered heteroaryl, 5- to 10-membered heterocyclic group, or C 6-10 aryl; the 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O, and S.

17. The compound according to claim 3, or its stereoisomer, tautomer, polymorph, co-crystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, has the structure shown in formula (XXI): Among them, R4, R5, R6, R7, and R8 are each independently selected from H, D, halogen, hydroxyl, cyano, amino, C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 2-6 alkenyl, C 2-6 alkynyl; or R4 and R8 combine with the connected carbon atom to form C 3-10 cycloalkyl, 5- to 10-membered heteroaryl, 5- to 10-membered heterocyclic group, or C 6-10 aryl; the 5- to 10-membered heteroaryl and 5- to 10-membered heterocyclic group each independently contain 1-3 heteroatoms selected from N, O, and S.

18. The compound according to any one of claims 1-16, or its stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, characterized in that: R2 and R3 are each independently selected from H, C 1-4 alkyl, C 5-6 alkyl; Preferably, R2 and R3 are each independently selected from H, C 1-2 alkyl, C 3-4 alkyl; Preferably, R2 and R3 are each independently selected from H, C 1-2 alkyl; Preferably, R2 and R3 are each independently selected from H, methyl, ethyl; Most preferably, R2 and R3 are each independently selected from H.

19. The compound according to any one of claims 1-18, or its stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, characterized in that: R4 is selected from H, halogen, C 1-6 alkyl, C 2-6 alkynyl; Preferably, R4 is selected from H, halogen, C 1-4 alkyl, C 5-6 alkyl, C 2-4 alkynyl, C 5-6 alkynyl; Preferably, R4 is selected from H, halogen, C 1-4 alkyl, C 2-4 alkynyl; Preferably, R4 is selected from H, F, Cl, Br, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, ethynyl, propynyl, butynyl; Preferably, R4 is selected from Cl, Br, methyl, ethynyl; Most preferably, R4 is selected from H, Cl, methyl, ethynyl; R5 is selected from H, halogen, C 1-4 alkyl; Preferably, R5 is selected from H, F, Cl, Br, methyl, ethyl; Preferably, R5 is selected from H, F; Preferably, R5 is selected from H, D, F, Cl, Br, methyl, ethyl; Preferably, R5 is selected from H, D, F; Most preferably, R5 is selected from H; R6 is selected from H, halogen, C 1-4 alkyl; Preferably, R6 is selected from H, halogen; Preferably, R6 is selected from H, F, Cl, Br; Most preferably, R6 is selected from H, Cl, F; R7 is selected from H, halogen, C 1-4 alkyl, C 1-4 haloalkyl; Preferably, R7 is selected from C 1-4 haloalkyl; Preferably, R7 is selected from C 1-2 fluoroalkyl, C 3-4 fluoroalkyl; Preferably, R7 is selected from fluoromethyl, fluoroethyl; Most preferably, R7 is selected from -CF3; R8 is selected from H, halogen, C 1-4 alkyl; Preferably, R8 is selected from H, halogen; Preferably, R8 is selected from H, F, Cl, Br; Most preferably, R8 is selected from H, F; Alternatively, R4 and R8 together with the connected carbon atom form a 5- to 10-membered heteroaryl group; the 5- to 10-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S; Preferably, R4 and R8 together with the connected carbon atom form a 5- to 6-membered heteroaryl group; the 5- to 6-membered heteroaryl group contains 1-3 heteroatoms selected from N, O, and S; Preferably, R4 and R8 together with the connected carbon atom form a 5-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O, and S; Preferably, R4 and R8 together with the connected carbon atom form furyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl; Preferably, R4 and R8 together with the connected carbon atom form furyl, thienyl; Preferably, Selected from Preferably, Selected from Preferably, Selected from 20. A compound according to any one of claims 1-12, 18, 19, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, characterized in that, Selected from the structures shown below:

21. A compound according to any one of claims 1-9, 13-15, 18, 19, or a stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, characterized in that, Selected from the structures shown below:

22. A bicyclic compound, or a stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, characterized in that: Selected from the following specific compounds: Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9, Compound 10, Compound 11, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, Compound 17, Compound 18, Compound 19, Compound 20, Compound 21, Compound 22, Compound 23, Compound 24, Compound 25, Compound 26, Compound 27, Compound 28, Compound 29, Compound 30, Compound 31, Compound 32, Compound 33, Compound 34, Compound 35, Compound 36, Compound 37, Compound 38, Compound 39, Compound 40, Compound 41, Compound 42, Compound 43, Compound 44, Compound 45, Compound 46, Compound 47, Compound 48, Compound 49, Compound 50, Compound 51, Compound 52, Compound 53, Compound 54, Compound 55, Compound 56, Compound 57, Compound 58, Compound 59, Compound 60, Compound 61, Compound 62, Compound 63, Compound 64, Compound 65, Compound 66, Compound 67, Compound 68, Compound 69, Compound 70, Compound 71, Compound 72, Compound 73, Compound 74, Compound 75, Compound 76, Compound 77, Compound 78, Compound 79, Compound 80, Compound 81, Compound 82, Compound 83, Compound 84, Compound 85, Compound 86, Compound 87, Compound 88, Compound 89, Compound 90, Compound 91, Compound 92, Compound 93, Compound 94, Compound 95, Compound 96, Compound 97, Compound 98, Compound 99, Compound 100, Compound 101, Compound 102, Compound 103, Compound 104, Compound 105, Compound 106, Compound 107, Compound 108, Compound 109, Compound 110, Compound 111, Compound 112, Compound 113, Compound 114, Compound 115, Compound 116, Compound 117, Compound 118, Compound 119, Compound 120, Compound 121, Compound 122, Compound 123, Compound 124, Compound 125, Compound 126, Compound 127, Compound 128, Compound 129, Compound 130, Compound 131, Compound 132, Compound 133, Compound 134, Compound 135, Compound 136, Compound 137, Compound 138, Compound 139, Compound 140, Compound 141, Compound 142, Compound 143, Compound 144, Compound 145, Compound 146, Compound 147, Compound 148, Compound 149, Compound 150, Compound 151, Compound 152, Compound 153, Compound 154, Compound 155, Compound 156, Compound 157, Compound 158Compound 159, Compound 160, Compound 161, Compound 162, Compound 163, Compound 164, Compound 165, Compound 166, Compound 167, Compound 168, Compound 169, Compound 170, Compound 171, Compound 172, Compound 173, Compound 174, Compound 175, Compound 176, Compound 177, Compound 178, Compound 179, Compound 180, Compound 181, Compound 182, Compound 183, Compound 184, Compound 185, Compound 186, Compound 187, Compound 188, Compound 189, Compound 190, Compound 191, Compound 192, Compound 193, Compound 194, Compound 195, Compound 196, Compound 197, Compound 198, Compound 199, Compound 200, Compound 201, Compound 202, Compound 203, Compound 204, Compound 205, Compound 206, Compound 207, Compound 208, Compound 209, Compound 210, Compound 211, Compound 212, Compound 213, Compound 214, Compound 215, Compound 216, Compound 217, Compound 218, Compound 219, Compound 220, Compound 221, Compound 222, Compound 223, Compound 224, Compound 225, Compound 226, Compound 227, Compound 228, Compound 229, Compound 230, Compound 231, Compound 232, Compound 233, Compound 234, Compound 235, Compound 236, Compound 237, Compound 238, Compound 239, Compound 240, Compound 241, Compound 242, Compound 243, Compound 244, Compound 245, Compound 246, Compound 247, Compound 248, Compound 249, Compound 250, Compound 251, Compound 252, Compound 253, Compound 254, Compound 255, Compound 256, Compound 257, Compound 258, Compound 259, Compound 260, Compound 261, Compound 262, Compound 263, Compound 264, Compound 265, Compound 266, Compound 267, Compound 268, Compound 269, Compound 270, Compound 271, Compound 272, Compound 273, Compound 274, Compound 275, Compound 276, Compound 277, Compound 278, Compound 279, Compound 280, Compound 281, Compound 282, Compound 283, Compound 284, Compound 285, Compound 286, Compound 287, Compound 288, Compound 289, Compound 290, Compound 291, Compound 292, Compound 293, Compound 294, Compound 295, Compound 296, Compound 297, Compound 298, Compound 299, Compound 300, Compound 301Compound 302, Compound 303, Compound 304, Compound 305, Compound 306, Compound 307, Compound 308, Compound 309, Compound 310, Compound 311, Compound 312, Compound 313, Compound 314, Compound 315, Compound 316, Compound 317, Compound 318, Compound 319, Compound 320, Compound 321, Compound 322, Compound 323, Compound 324, Compound 325, Compound 326, Compound 327, Compound 328, Compound 329, Compound 330, Compound 331, Compound 332, Compound 333, Compound 334, Compound 335, Compound 336, Compound 337, Compound 338, Compound 339, Compound 340, Compound 341, Compound 342, Compound 343, Compound 344, Compound 345, Compound 346, Compound 347, Compound 348, Compound 349, Compound 350, Compound 351, Compound 352, Compound 353, Compound 354, Compound 355, Compound 356, Compound 357, Compound 358, Compound 359, Compound 360, Compound 361, Compound 362, Compound 363, Compound 364, Compound 365, Compound 366, Compound 367, Compound 368, Compound 369, Compound 370, Compound 371, Compound 372, Compound 373, Compound 374, Compound 375, Compound 376, Compound 377, Compound 378, Compound 379, Compound 380, Compound 381, Compound 382, Compound 383, Compound 384, Compound 385, Compound 386, Compound 387, Compound 388, Compound 389, Compound 390, Compound 391, Compound 392, Compound 393, Compound 394, Compound 395, Compound 396, Compound 397, Compound 398, Compound 399, Compound 400, Compound 401, Compound 402, Compound 403, Compound 404, Compound 405, Compound 406, Compound 407, Compound 408, Compound 409, Compound 410, Compound 411, Compound 412, Compound 413, Compound 414, Compound 415, Compound 416, Compound 417, Compound 418, Compound 419, Compound 420, Compound 421, Compound 422, Compound 423, Compound 424, Compound 425, Compound 426, Compound 427, Compound 428, Compound 429, Compound 430, Compound 431, Compound 432, Compound 433, Compound 434, Compound 435, Compound 436, Compound 437, Compound 438, Compound 439, Compound 440, Compound 441, Compound 442, Compound 443。 23. A pharmaceutical composition comprising the compound according to any one of claims 1-22, or its stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, and optionally a pharmaceutical excipient.

24. Use of the compound according to any one of claims 1-22, or its stereoisomer, tautomer, polymorph, eutectic, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 23 in the preparation of a WRN inhibitor.

25. Use of a compound according to any one of claims 1-22, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 23, in the preparation of a medicament for treating and / or preventing tumors or cancers.

26. A method for treating and / or preventing tumors or cancers, which comprises administering to an individual in need a therapeutically and / or prophylactically effective amount of a compound according to any one of claims 1-22, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 23.

27. A compound according to any one of claims 1-22, or a stereoisomer, tautomer, polymorph, cocrystal, hydrate, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 23, for use in treating and / or preventing tumors or cancers.

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