Cgas inhibitor, preparation method therefor and use thereof

Abstract

The present invention relates to a cGAS inhibitor, a preparation method therefor and a use thereof. Specifically, the present invention provides a compound having a novel structure represented by formula (I), and the definition of each group is as described in the description. The compound of the present invention has strong activity as a cGAS inhibitor compound, and has good clinical application prospects for diseases related to abnormal cGAS activity levels and / or expression levels.

Description

CgAS inhibitors, methods of making and uses thereof TECHNICAL FIELD

[0001] The present invention belongs to the field of medicinal chemistry, in particular to a methylene derivative as a cGAS inhibitor, and its preparation method, pharmaceutical composition and use. BACKGROUND

[0002] The cGAS-STING signaling pathway, as an important inflammation regulatory pathway, is involved in a variety of pathological processes such as infection, cell stress and tissue damage. In this signaling pathway, cyclic GMP-AMP synthase (cGAS) activates its catalytic activity by recognizing cytosolic double-stranded DNA (dsDNA) derived from pathogens or cell stress to produce the second messenger 2'3' cyclic GMP-AMP (cGAMP). cGAMP is then recognized by interferon gene stimulatory protein (STING) and causes its activation and oligomerization, further recruiting TBK1 to phosphorylate the transcription factor IRF3, triggering the transcription of inflammatory genes. (Nat. Rev. Immunol. 2021, 21, 548-569.)

[0003] Overactivation of the cGAS-STING pathway is involved in the pathogenesis of viral infection, sepsis, acute pancreatitis, rheumatoid arthritis, systemic lupus erythematosus and Aicardi-Goutieres syndrome, tumors and many other diseases (Nature, 2022, 603(7899), 145-151.; J. Exp. Med., 2016, 213, 2527-2538.; Annu. Rev. Immunol., 2017, 35, 313-336.; Nature, 2018, 563(7729), 131-136.). In a mouse model of Aicardi-Goutieres syndrome, knock-out of cGAS can reverse the increase in interferon levels caused by knock-out of 3'-nucleic acid repair exonuclease 1 (Trex1) (J. Immunol., 2015, 195, 1939-1943.). In addition, knock-out of cGAS can also effectively inhibit the inflammatory response caused by mitochondrial stress (Nature., 2015, 520, 553-557). Since abnormal activation of the cGAS-STING pathway is an important factor in triggering inflammation and autoimmune diseases and many other diseases, the development of new cGAS inhibitors has received widespread attention from academia and industry in recent years.

[0004] However, only a few structural types of inhibitors that specifically bind to cGAS have been reported so far (WO 2017 / 176812 Al; WO 2019 / 153002 Al; CN 115197207 A; WO 2023US15728). Some inhibitors such as Ru.521 only have inhibitory activity against mouse-derived cGAS and weak inhibitory activity against human-derived cGAS (Nat. Commun., 2017, 8, 750). Therefore, it is of great scientific significance and broad clinical transformation prospect to develop novel cGAS inhibitors with novel structure and strong activity. SUMMARY

[0005] An object of the present application is to provide a compound having a structure shown in Formula I.

[0006] Another object of the present application is to provide a novel cGAS inhibitor and its use for preventing and treating diseases related to abnormal levels of cyclic cGAS activity and / or expression.

[0007] Still another object of the present application is to provide a method for synergistically treating diseases related to abnormal levels of cyclic cGAS activity and / or expression.

[0008] In a first aspect of the present application, a compound shown in Formula I, or a stereoisomer, tautomer, deuterated compound, solvate, hydrate, prodrug, metabolite, pharmaceutically acceptable salt thereof, or a combination thereof, is provided,

[0009] in the formula,

[0010] R1is selected from the group consisting of H, C1-C 10 alkyl, C3-C 10 cycloalkyl, 5-12 membered heteroaryl, C6-C 14 aryl, 4-12 membered heterocyclyl, C6-C 10 cycloalkenyl;

[0011] said alkyl, cycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocyclyl are optionally substituted with one or more substituents selected from the group consisting of hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), sulfonate (-S03Na), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester (-COO-Ci-C6alkyl), Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(Ci-C6alkyl), carboxyl substituted Ci-C6alkoxy, ester (-COO-Ci-C6alkyl) substituted Ci-C6alkoxy, -(Ci-C6alkylene)-COO-(Ci-C6alkyl); wherein said alkylene, alkoxy and alkyl are substituted with carboxyl, ester (-COO-Ci-C6alkyl), oxo;

[0012] R2is selected from the group consisting of H, D, halogen, Ci-C 10 alkyl, Ci-C 10 alkoxy, C3-C8cycloalkyl, Ci-C 10 haloalkyl, Ci-C 10 haloalkoxy;

[0013] R3and R4are each independently selected from a compound of Formula II:

[0014] wherein,

[0015] X is selected from the group consisting of N, O or S;

[0016] Y is selected from the group consisting of CH, C or N;

[0017] Ring B is selected from a five-membered heteroaryl and a five-membered heterocyclyl;

[0018] R5is selected from the group consisting of substituted or unsubstituted H, D, halogen, hydroxyl, amino, amine, thiol, cyano, nitro, -C(O)R', Ci-C 10 alkyl, Ci-C 10 haloalkyl, Ci-C 10 alkoxy, Ci-C 10 haloalkoxy, Ci-C 10 alkylthio, C3-C 10 cycloalkyl, 4-11 membered heterocyclyl, C3-C8cycloalkyl-O-, C3-C8cycloalkyl-S-, Ci-C 10 alkyl-(C=O)-O-, Ci-C 10 alkyl-(C=S)-O-;

[0019] R' is selected from the group consisting of H, D, halogen, hydroxyl, amino, amine, thiol, cyano, nitro, acyl, sulfonyl, amido, sulfonamido, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl) substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl), -(C1-C6alkylene)-5-6 membered heteroaryl, -(C1-C6alkylene)-C3-C6cycloalkyl, -(C1-C6alkylene)-4-7 membered heterocyclyl, -(C1-C6alkylene)-phenyl, wherein the alkyl, alkylene, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, phenyl, heteroaryl groups are optionally substituted with a group selected from hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), acyl, sulfonyl, sulfonamido, amine, carboxyl, amido (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl; 10 alkyl, C1-C 10 haloalkyl, C3-C 10 cycloalkyl;

[0020] R6 is selected from the group consisting of H, D, halogen, hydroxyl, amino, amine, thiol, cyano, nitro, acyl, sulfonyl, amido, sulfonamido, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl) substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl), -(C1-C6alkylene)-5-6 membered heteroaryl, -(C1-C6alkylene)-C3-C6cycloalkyl, -(C1-C6alkylene)-4-7 membered heterocyclyl, -(C1-C6alkylene)-phenyl, wherein the alkyl, alkylene, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, phenyl, heteroaryl groups are optionally substituted with a group selected from hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), acyl, sulfonyl, sulfonamido, amine, carboxyl, amido (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl; 10 alkyl, C1-C 10 alkoxy, C3-C 10 cycloalkyl, 4-11 membered heterocyclyl, C6-C10 aryl, 5-11 membered heteroaryl, C3-C8cycloalkyl-O-, C3-C8cycloalkyl-S-;

[0021] Ring A is selected from the group consisting of 8-14 membered bicyclic or tricyclic heteroaryl, 8-14 membered saturated bicyclic or tricyclic heterocyclyl, 8-14 membered partially unsaturated bicyclic or tricyclic heterocyclyl, wherein the bicyclic or tricyclic rings are connected by spiro, bridged or fused;

[0022] The substitution means that one or more hydrogens on the group are replaced with a group selected from hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), acyl, sulfonyl, sulfonamido, amine, carboxyl, amido (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl;

[0023] The heteroaryl or heterocyclyl contains 1-3 heteroatoms each independently selected from N, O and S.

[0024] In another preferred embodiment, R1is selected from the group consisting of C1-C6alkyl, C3-C6cycloalkyl, 5-10 membered heteroaryl, C6-C7cycloalkenyl; 10 aryl, 5-8 membered heterocyclyl, C6-C7cycloalkenyl;

[0025] The alkyl, cycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocyclyl groups are optionally substituted with one or more substituents selected from the group consisting of hydroxy, halogen, cyano, amino, oxo (=0), sulfonate (-SO3Na), carboxyl, amido (-CONH2), ester (-COO-C1-C6alkyl), C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl) substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl); wherein the alkylene, alkoxy and alkyl groups are substituted with carboxyl, ester (-COO-C1-C6alkyl), oxo groups.

[0026] In another preferred embodiment, R1is selected from the group consisting of substituted or unsubstituted methyl, phenyl, naphthyl, cyclohexyl, cyclopentyl, pyridyl, thienyl, imidazolyl, indolyl, thiazole, quinolinyl,

[0027] The substitution means that one or more hydrogens on the group are replaced by a substituent selected from the group consisting of hydroxy, halogen, cyano, amino, oxo (=0), sulfonate (-SO3Na), carboxyl, amido (-CONH2), ester (-COO-C1-C6alkyl), C1-C6alkyl, C1-C6alkoxy, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl) substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl); wherein the alkylene, alkoxy and alkyl groups are substituted with carboxyl, ester (-COO-C1-C6alkyl), oxo groups.

[0028] In another preferred embodiment, R2is selected from the group consisting of H, D, halogen, C1-C4alkyl, C1-C4alkoxy, C3-C6cycloalkyl, C1-C4haloalkyl, C1-C4haloalkoxy.

[0029] In another preferred embodiment, R2is selected from H.

[0030] In another preferred embodiment, R5is selected from the group consisting of H, hydroxyl, amino, amine, thiol, cyano, nitro, -C(O)R', C1-C4alkyl, C1-C4haloalkyl, C1-C4alkoxy, C1-C4haloalkoxy, C1-C4alkylthio, C3-C8cycloalkyl-O-, C3-C8cycloalkyl-S-, C1-C4alkyl-(C=O)-O-, C1-C4alkyl-(C=S)-O-;

[0031] R' is selected from the group consisting of H, C1-C4alkyl, C1-C4haloalkyl, C3-C6cycloalkyl.

[0032] In another preferred embodiment, R5is selected from the group consisting of hydroxyl, trifluoromethyl, methoxy or acetoxy.

[0033] In another preferred embodiment, R6is selected from H, halogen, substituted or unsubstituted C1-C4alkyl, said substitution being with one or more halogen, cyano, hydroxyl, nitro, C1-C4alkoxy; preferably methyl.

[0034] In another preferred embodiment, R3and R4are each independently selected from the structure shown in Formula II-A:

[0035] wherein,

[0036] Ring A, R5and R6are as described in the first aspect of the application.

[0037] In another preferred embodiment, R3and R4are the same or different.

[0038] In another preferred embodiment, R3and R4are the same.

[0039] In another preferred embodiment, Ring A is selected from the group consisting of substituted or unsubstituted C6-C 10 aryl, five-membered heteroaromatic ring fused to a six-membered heteroaromatic ring, benzo five-membered heteroaromatic ring, benzo six-membered heteroaromatic ring, six-membered heteroaromatic ring fused to a six-membered heteroaromatic ring, wherein the six-membered heteroaromatic ring is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, pyrylium; the five-membered heteroaromatic ring is selected from the group consisting of imidazole, thiazole, oxazole, furan, pyrazole, thiophene, pyrrole;

[0040] Preferably, ring A is selected from the group consisting of substituted or unsubstituted: benzimidazole, benzothiazole, benzoxazole, benzofuran, benzopyrazole, benzothiophene, benzopyrrole, pyridoimidazole, pyridopyrazole, purine, quinoline, isoquinoline, indole;

[0041] wherein the substitution means that one or more (e.g., 2, 3, 4, or 5) hydrogens on the group are replaced by a group independently selected from the group consisting of: hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), sulfonate (-SO3Na), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester (-COO-C1-C6alkyl), urea, sulfone, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl-substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl)-substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl), -(C1-C6alkylene)-5-6 membered heteroaryl, -(C1-C6alkylene)-C3-C6cycloalkyl, -(C1-C6alkylene)-4-7 membered heterocyclyl, -(C1-C6alkylene)-phenyl, wherein the alkyl, alkylene, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, phenyl, heteroaryl groups are optionally substituted with a group selected from: hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl;

[0042] More preferably, ring A is selected from the group consisting of:

[0043] In another preferred embodiment, the compound is selected from the group consisting of:

[0044] In another preferred embodiment, the compound is a compound selected from the group consisting of A4-A50.

[0045] In another preferred embodiment, the compound is a compound prepared in the Examples.

[0046] In a second aspect of the present application, there is provided a method for preparing a compound of the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt thereof, characterized in that it comprises the following steps:

[0047] reacting a compound of formula IV with a compound of formula V in an inert solvent to obtain a compound of formula I;

[0048] wherein ring A, ring B, X, Y, R1, R2, R5 and R6 are as described in the first aspect of the present application.

[0049] In another preferred embodiment, the molar ratio of the compound of formula IV to the compound of formula V is 2-4:1; preferably 2-4:1; more preferably 2:1.

[0050] In another preferred embodiment, the inert solvent is selected from the group consisting of water, C1-C6 halogenated hydrocarbon solvents, C1-C6 ketone solvents, C1-C6 ester solvents, C1-C6 alcohol solvents, C2-C6 ether solvents, amide solvents, or a combination thereof; preferably dichloromethane, acetone, ethyl acetate, methanol, ethanol, water, 1,4-dioxane, N,N-dimethylformamide; preferably methanol, ethanol; more preferably ethanol.

[0051] In another preferred embodiment, the reaction temperature is 50-100°C, preferably 75-85°C.

[0052] In another preferred embodiment, the method comprises the following steps:

[0053] reacting a compound of formula IV-A with a compound of formula V-A in an inert solvent to obtain a compound of formula I;

[0054] wherein ring A, R1, R5 and R6 are as described in the first aspect of the present application.

[0055] In a third aspect of the present application, there is provided a pharmaceutical composition comprising a compound of formula I of the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, as an active ingredient; and a pharmaceutically acceptable carrier.

[0056] In another preferred embodiment, in the pharmaceutical composition, the mass ratio of the compound of formula I, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof to the carrier is 0.001-10.

[0057] In a fourth aspect of the present application, there is provided a pharmaceutical composition comprising:

[0058] a compound of Formula I as described in the first aspect of the present application, or a stereoisomer, tautomer, deuterated analog, solvate, hydrate, prodrug, metabolite, pharmaceutically acceptable salt thereof, or a combination thereof, as a first active ingredient;

[0059] a further cGAS inhibitor as a second active ingredient; and

[0060] optionally a pharmaceutically acceptable carrier.

[0061] In another preferred embodiment, the further cGAS inhibitor is a small molecule inhibitor that has inhibitory effect on cGAS enzyme activity.

[0062] In another preferred embodiment, the further cGAS inhibitor is G150, having the structural formula

[0063] In another preferred embodiment, the further cGAS inhibitor inhibits the activity of cGAS by binding to the active site of the catalytic domain of cGAS.

[0064] In another preferred embodiment, the further cGAS inhibitor is selected from the group consisting of WO2023288253A1; CN116209442A; WO2023081441A1; WO2023183275A1; US20230192623A1; US20230192631A1; US11414422B2; WO2022238327A1; WO2022066851A1; WO2022137085A1; WO2021209484A1; WO2021233852A1; WO2021233851A1; WO2021209475A1; WO2020186027A1; WO2019055750A1.

[0065] In another preferred embodiment, the molar ratio of the first active ingredient to the second active ingredient is 0.01-100; preferably 0.1-10, more preferably 1-10.

[0066] In a fifth aspect of the present application, there is provided a use of a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt, or a combination thereof, or a pharmaceutical composition according to the third aspect of the present application, or a pharmaceutical composition according to the fourth aspect of the present application, for the manufacture of (a) a cGAS inhibitor; (b) a medicament for preventing, treating and / or adjuvant treatment of a disease associated with abnormal level of activity and / or expression of cGAS.

[0067] In another preferred embodiment, the disease is selected from the group consisting of an infectious disease, an inflammatory disease, an autoimmune disease, a neurodegenerative disease, a metabolic disease, a rare disease, a tumor, a radiation injury.

[0068] In another preferred embodiment, the infectious disease is selected from a viral infection or a bacterial infection, preferably an HIV infection, a SARS-CoV-2 infection.

[0069] In another preferred embodiment, the inflammatory disease is selected from the group consisting of non-alcoholic fatty liver, acute or chronic nephritis, chronic obstructive pulmonary disease, myocardial infarction, aortic aneurysm and aortic dissection, heart failure, inflammatory bowel disease, arthritis, allergy, cutaneous dermatosis such as dermatomyositis or cutaneous lupus, interstitial pulmonary fibrosis, Sjogren's syndrome, psoriasis, juvenile inflammatory arthritis, gout, aging, type I interferonopathy, age-related macular degeneration, ischemic stroke, or a combination thereof.

[0070] In another preferred embodiment, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus, ankylosing spondylitis, scleroderma, rheumatoid arthritis, multiple sclerosis, or a combination thereof.

[0071] In another preferred embodiment, the neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, or a combination thereof.

[0072] In another preferred embodiment, the metabolic disease is selected from the group consisting of type I diabetes, obesity, fatty liver degeneration, or a combination thereof.

[0073] In another preferred embodiment, the rare disease is selected from the group consisting of Aicardi-Goutieres syndrome, familial chilblain lupus (FCL), retinal vasculopathy with cerebral leukodystrophy (RVCL), STING-associated vasculopathy with onset in infancy (SAVI), Bloom syndrome, Hutchinson-Gilford progeria syndrome, COPA syndrome, Sjogren syndrome, or a combination thereof.

[0074] In another preferred embodiment, the tumor is selected from the group consisting of head and neck squamous cell carcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, cardiac cancer, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, Kaposi's sarcoma, kidney cancer, pancreatic cancer, penile cancer, testicular germ cell cancer, thymus cancer, lung cancer, ovarian cancer, prostate cancer, or a combination thereof.

[0075] In a sixth aspect of the present application, a method for inhibiting cGAS activity or expression is provided, comprising: administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, or a pharmaceutical composition according to the third aspect of the present application, or a pharmaceutical composition according to the fourth aspect of the present application.

[0076] In a seventh aspect of the present application, a method for preventing, treating and / or adjuvant therapy of a disease associated with abnormal level of cGMP-AMP synthase (cGAS) activity and / or expression is provided, comprising: administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, or a pharmaceutical composition according to the third aspect of the present application, or a pharmaceutical composition according to the fourth aspect of the present application.

[0077] In an eighth aspect of the present application, a potentiator is provided, which is a compound of Formula I, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, for use in potentiating the inhibitory effect of other cGAS inhibitors.

[0078] In another preferred embodiment, the present application provides a method of synergistically inhibiting cGAS activity or expression, comprising: administering to a subject in need thereof a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, and a cGAS inhibitor as a second active ingredient.

[0079] In another preferred embodiment, the cGAS inhibitor as a second active ingredient can be administered simultaneously, prior to, or after the administration of the compound of Formula I.

[0080] In another preferred embodiment, the cGAS inhibitor as a second active ingredient inhibits the activity of cGAS by binding to the active site of the catalytic domain of cGAS.

[0081] In another preferred embodiment, the compound of Formula I enhances the efficiency of DNA delivery in cells or animals by inhibiting the activity or expression of cGAS.

[0082] In another preferred embodiment, the present application provides a method of enhancing the efficiency of DNA delivery in cells or animals, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, or a pharmaceutical composition comprising the same.

[0083] In another preferred embodiment, the present application provides a method of enhancing the efficiency of production of chimeric antigen receptor T cells, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, or a pharmaceutical composition comprising the same.

[0084] In another preferred embodiment, the compound of Formula I enhances the efficiency of production of chimeric antigen receptor T cells by inhibiting the activity of cGAS, thereby improving the survival rate of chimeric antigen receptor T cells after production.

[0085] In another preferred embodiment, the present application provides a method of assisting tumor immunotherapy, comprising: providing to a subject in need thereof a therapeutically effective amount of a compound according to the first aspect of the present application, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the third aspect of the present application.

[0086] It should be understood that, in the scope of the present application, each of the technical features described above and each of the technical features described in detail below (such as the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they will not be listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0087] Figure 1 shows the synergistic inhibitory effect of the compound A46 of the present application and G150 on cells.

[0088] Figure 2 shows the protective effect of the compound A02 of the present application on mouse cerulein-induced pancreatitis, wherein A shows the induction of acute pancreatitis and the administration method; B shows the pancreas to body weight ratio of mice; C shows the serum amylase activity of mice; D shows the HE staining of mouse pancreatic tissue sections, the black arrow is the inflammatory cell infiltration, the green arrow is the tissue necrosis, and the red is the pancreatic edema performance, the scale is 10 μm; E shows the scoring according to the pathological condition of D. DETAILED DESCRIPTION

[0089] The inventors have made extensive and in-depth research and for the first time provided a cGAS inhibitor compound with a completely new structure. The compound of the present application has excellent cGAS inhibitory activity at the enzymatic and cellular levels. The compound of the present application can also be used as a synergist to improve the therapeutic effect of cGAS inhibitors. Based on this, the inventors have completed the present application.

[0090] TERMS

[0091] In the present application, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0092] The term "alkyl" refers to a straight-chain or branched alkane group or a cyclic monovalent saturated hydrocarbon group (including a hydrocarbon group connected to other moieties through a carbon in the ring or non-ring), preferably a straight-chain or branched alkane group, containing 1-20 carbon atoms, such as 1-18 carbon atoms, especially 1-18 carbon atoms, preferably containing 1-10 carbon atoms (C1-C10), more preferably containing 1-6 carbon atoms (C1-C6). Typical "alkyl" groups include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl, isobutyl, pentyl, isopentyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and the like. In the present application, alkyl also includes substituted alkyl. "Substituted alkyl" refers to one or more positions in the alkyl group being substituted, especially 1-4 substituents, which can be substituted at any position.

[0093] In this document, the chemical structure written from left to right (such as (CR Le R Lf)-O) unless otherwise defined, includes the left form as well, i.e. (CR Le R Lf )-O can be (CR Le R Lf )-O or O-(CR Le R Lf ).

[0094] As used herein, the term "alkylene" (also referred to herein as "alkylene chain"), especially those "alkylene" groups which are located internally or non-terminally in a molecule or group (including as a substituent group (e.g. C3-C 10 alkylene") means a linking group for attaching two different atoms or a group formed by removing one hydrogen atom from an "alkyl or substituted alkyl" group which is not at the atom where the "alkyl or substituted alkyl" group is unsaturated, such as methylene or methylidene (-CH2-), ethylene propylene isopropylene butylene (e.g. ), pentylene (e.g. ), hexylene (e.g. ), heptylene (e.g. ), and the like.

[0095] As used herein, the term "alkylidene" (also referred to herein as "terminal alkylidene"), especially those "alkylidene" groups which are terminal substituents (e.g. in R), means a group formed by removing one hydrogen atom from an "alkyl or substituted alkyl" group which is at the atom where the "alkyl or substituted alkyl" group is unsaturated, which is attached to the rest of the molecule by a double bond, such as methylene (also referred to as terminal methylene or methylidene, ), ethylene (e.g. ), propylene (e.g. ), isopropylene (e.g. ), butylene, pentylene, hexylene, heptylene and the like. In addition, the term also includes an alkylidene (e.g. C1-C18 alkylidene) in which one methylene group is replaced by a cycloalkylidene (e.g. C3-C20 cycloalkylidene), such as "C1-C18 alkylidene C3-C20 cycloalkylidene" or "C3-C20 cycloalkylidene C1-C18 alkylidene". In the present application, alkylidene also includes substituted alkylidene groups, the substituents of which can be halo (e.g. -CHF- or -CF2-), hydroxy, cyano, nitro and the like.

[0096] The term "C1-C6alkyleneC3-C6cycloalkylene" or "C3-C6cycloalkyleneC1-C6alkylene" has the same meaning and refers to a cycloalkylalkyl or alkylcycloalkyl radical, which has been deprived of two hydrogen atoms, such as etc. Preferably C1-C4alkyleneC3-C6cycloalkylene. In the present application, "C1-C6alkyleneC3-C6cycloalkylene" or "C3-C6cycloalkyleneC1-C6alkylene" also includes substituted "C1-C6alkyleneC3-C6cycloalkylene" or "C3-C6cycloalkyleneC1-C6alkylene", the substituents can be halo, hydroxy, cyano, nitro, etc. The term "C1-C6alkylene-4- to 6-membered heterocyclylene" or "4- to 6-membered heterocyclylene-C1-C6alkylene" has a similar meaning, preferably "C1-C4alkylene-4- to 6-membered heterocyclylene" or "4- to 6-membered heterocyclylene-C1-C4alkylene".

[0097] In the present application, the term "alkenyl" denotes a straight-chain or branched hydrocarbon group containing one or more double bonds and usually having a length of from 2 to 20 carbon atoms. Alkenyl is preferably C2-C6alkenyl, more preferably C2-C4alkenyl. Alkenyl includes, but is not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl and the like. In the present application, alkenyl includes substituted alkenyl. In the present application, alkenyl also includes substituted alkenyl, the substituents can be halo, hydroxy, cyano, nitro, etc.

[0098] The term "alkynyl" denotes a straight-chain or branched hydrocarbon group containing one or more triple bonds and usually having a length of from 2 to 20 carbon atoms. Alkynyl is preferably C2-C6alkynyl, more preferably C2-C4alkynyl. Alkynyl includes, but is not limited to, ethynyl, propynyl or the like. In the present application, alkynyl also includes substituted alkynyl, the substituents can be halo, hydroxy, cyano, nitro, etc.

[0099] In the present application, the term "cycloalkyl" refers to a cyclic hydrocarbon compound group which is fully saturated or partially unsaturated (preferably fully saturated), including 1-4 rings, each ring containing 3-8 carbon atoms. The term "C3-C 20 " refers to a cycloalkyl group comprising 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. Cycloalkyl is preferably C3-C 14 cycloalkyl, more preferably C3-C 10 cycloalkyl, more preferably C3-C6monocyclic cycloalkyl, C7-C 10Bicyclic or tricyclic cycloalkyl. "Substituted cycloalkyl" refers to a cycloalkyl group in which one or more positions are substituted, particularly 1-4 substituents, which can be substituted at any position. In this invention, "cycloalkyl" includes substituted cycloalkyl groups, and typical substitutions include, but are not limited to, one or more of the following groups: such as hydrogen, deuterium, halogen (e.g., monohalogen substituents or polyhalogen substituents, the latter such as trifluoromethyl or alkyl containing Cl3), nitrile, nitro, oxygen (e.g., =O), trifluoromethyl, trifluoromethoxy, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocyclic, aromatic, OR a SR a S(=O)R e S(=O)2R e P(=O)2R e S(=O)2OR e P(=O)2OR e NR b R c NR b S(=O)2R e NR b P(=O)2R e S(=O)2NR b R c P(=O)2NR b R c C(=O)OR d C(=O)R a C(=O)NR b R c OC(=O)R a OC (=O)NR b R c NR b C(=O)OR e NR d C(=O)NR b R c NR d S(=O)2NR b R c NR d P(=O)2NR b R c NR b C(=O)R a , or NR b P(=O)2R e R appears here a It can independently represent hydrogen, deuterium, alkyl, cycloalkyl, alkenyl, cycloalkenyl, ynyl, heterocyclic, aryl, or heteroaryl, R b R c and R dmay independently represent hydrogen, deuterium, alkyl, cycloalkyl, heterocycle, or aryl, or R b and R c together with the N atom can form a heterocycle; R e may independently represent hydrogen, deuterium, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocyclyl, aryl, or heteroaryl. The typical substituents described above can be optionally substituted. Typical substitutions also include spirocyclic, pyrimidinocyclic, or fused cyclic substituents, especially spirocycloalkyl, spirocycloalkenyl, spiroheterocycle (excluding heteroaromatic rings), pyrimidinocycloalkyl, pyrimidinocycloalkenyl, pyrimidinoheterocycle (excluding heteroaromatic rings), fused cycloalkyl, fused cycloalkenyl, fused heterocyclyl, or fused aryl rings, which can be optionally substituted. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl, and the like.

[0100] The term "C3-C20cycloalkylene" refers to a group formed by removing two hydrogen atoms from a cycloalkyl group, such as:

[0101] and the like.

[0102] In the present invention, the term "heterocyclyl" refers to a cyclic group that is fully saturated or partially unsaturated (including but not limited to, e.g., 3-7 membered monocyclic, 4-7 membered monocyclic, 6-11 membered bicyclic, or 8-16 membered tricyclic or polycyclic systems) in which at least one heteroatom is present in at least one carbon atom-containing ring. The term "4-20 membered heterocyclyl" refers to a heterocyclyl group comprising 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ring atoms. "Heterocyclyl" has the same meaning as "saturated or unsaturated heterocyclyl". "Heterocyclyl" is preferably 4-14 membered heterocyclyl (including but not limited to, e.g., 4-6 membered monocyclic, 7-10 membered bicyclic, or 8-14 membered tricyclic or polycyclic systems), more preferably 4-12 membered heterocyclyl, more preferably 4-10 membered heterocyclyl, such as 4-6 membered monocyclic heterocyclyl, 7-11 membered bicyclic or tricyclic heterocyclyl, more preferably 4-8 membered heterocyclyl, more preferably 4-6 membered heterocyclyl. The heterocyclic ring containing heteroatoms of each heterocyclyl group can bear 1, 2, 3, or 4 heteroatoms each independently selected from a nitrogen atom, an oxygen atom, or a sulfur atom, wherein the nitrogen atom or the sulfur atom can be oxidized and the nitrogen atom can also be quaternized. The heterocyclyl group can be attached to the residue of any heteroatom or carbon atom of the ring or ring system, preferably to the N or C atom of the ring or ring system. Typical monocyclic heterocycles include, but are not limited to, azetidinyl, pyrrolidinyl, oxetanyl, pyrazolinyl, imidazolinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl, hexahydroazepinyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxanyl, and tetrahydro-l,l-dioxothiopyran, and the like. Polycyclic heterocyclyl groups include spiro, fused, and bridged ring heterocyclyl groups; wherein the spiro, fused, and bridged ring heterocyclyl groups are optionally connected to other groups by a single bond or further annulated to other cycloalkyl, heterocyclyl, aryl, and heteroaryl groups through any two or more atoms of the rings; the heterocyclyl group can be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxyl, thiol, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylthio, oxo, carboxyl, and carboxylate.

[0103] The term "4-20 membered heterocyclyl ene" refers to a group formed by removing two hydrogen atoms from a heterocyclyl group, such as:

[0104] and the like.

[0105] In the present invention, the term "aryl" refers to an aromatic cyclic hydrocarbon group having 1-5 rings, especially monocyclic and bicyclic groups. Within this, "C6-C 14 "aryl" refers to an aromatic cyclic hydrocarbon group containing 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring carbon atoms. Preferably, the aryl group is C6-C 10 "aryl". Aryl includes phenyl, biphenyl, or naphthyl. Where there are two or more aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group can be linked by single bonds (as in biphenyl), or fused (as in naphthalene, anthracene, etc.). "Substituted aryl" refers to an aryl group in which one or more positions are substituted, especially 1-3 substituents, which can be substituted at any position.

[0106] The term "heteroaryl" refers to an aromatic cyclic hydrocarbon group containing 1-4 heteroatoms, selected from oxygen, nitrogen, and sulfur. Within this, "5-14 membered heteroaryl" refers to a heteroaromatic system containing 1-4 heteroatoms, 5-14 ring atoms. Preferably, the heteroaryl group is a 5- to 10-membered ring, more preferably 5- or 6-membered, such as pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl, and tetrazolyl. The "heteroaryl" group can be substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halo, amino, nitro, hydroxy, mercapto, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylthio, oxo, carboxy, and carboxylate.

[0107] In the present invention, the term "alkoxy" refers to a straight or branched chain alkoxy group, including alkyl-O-, alkyl-O-alkyl, wherein "C1-C 18 "alkoxy" refers to a straight or branched chain alkoxy group having 1 to 18 carbon atoms, including C1-C 18 alkyl-O-, -C1-C6alkyl-O-C1-C6alkyl, including without limitation methoxy, ethoxy, propoxy, isopropoxy, and butoxy, and the like. Preferably, the alkoxy group is C1-C8alkoxy, more preferably C1-C6alkoxy.

[0108] In the present invention, the term "cycloalkyloxy" refers to a cycloalkyl-O- group, wherein "C3-C 20 "cycloalkyloxy" refers to a C3-C 20 cycloalkyl-O- group, wherein C3-C 20 cycloalkyl is as defined above.

[0109] In the present application, the term "heterocyclyloxy" means heterocyclyl-O-, wherein "4-20 membered heterocyclyloxy" means 4-20 membered heterocyclyl-O-, wherein 4-20 membered heterocyclyl is as defined above.

[0110] In the present application, the term "C1-C 18 alkyleneoxy" means a group "-(C1-C6alkylene)-O-". 18 alkoxy" means a group "-O-C1-C6alkyl".

[0111] In the present application, the term "halogen" or "halo" means chloro, bromo, fluoro, iodo.

[0112] In the present application, the term "halo" means substituted with halogen.

[0113] In the present application, the term "deutero" means substituted with deuterium.

[0114] In the present application, the term "hydroxy" means a group bearing the structure OH.

[0115] In the present application, the term "nitro" means a group bearing the structure NO2.

[0116] In the present application, the term "cyano" means a group bearing the structure CN.

[0117] In the present application, the term "ester" means a group bearing the structure -COOR, wherein R represents hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heterocycle or substituted heterocycle. Preferably, the ester is -COO-C1-C6alkyl.

[0118] The term "amine" means a group bearing the structure -NR'R", wherein R' and R" can independently represent hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or R' and R" together with the nitrogen atom to which they are both attached form a substituted or unsubstituted heterocyclyl, as defined above. In one embodiment, R' or R" is each independently selected from the group consisting of H, deuterium, C1-C6alkyl, haloC1-C6alkyl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, or R' and R" together with the nitrogen atom to which they are both attached form a 4 to 7 membered heterocyclyl (preferably a saturated 4 to 7 membered heterocyclyl containing only one nitrogen heteroatom as a ring atom). In one embodiment, at least one of R' and R" is other than H. R' and R" can be the same or different in a dialkylamine moiety. Preferably, the amine is NH2, NHC1-C6alkyl, N(C1-C6alkyl)2; more preferably NHC1-C6alkyl, N(C1-C6alkyl)2.

[0119] The term "amido" refers to a group having the structure -CONR'R" wherein R' and R" can independently represent hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, or R' and R" together with the nitrogen atom to which they are both attached form a substituted or unsubstituted heterocyclyl group, as defined above. In one embodiment, R' or R" are each independently selected from the group consisting of H, deuterium, C1-C6 alkyl, halo C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or R' and R" together with the nitrogen atom to which they are both attached form a 4 to 7 membered heterocyclyl group (preferably a saturated 4 to 7 membered heterocyclyl group containing only one nitrogen heteroatom as a ring atom). R' and R" in the dialkylamine fragment can be the same or different. The amido group is preferably -CONH2, -CONH(C1-C6 alkyl), -CONH(C3-C6 cycloalkyl).

[0120] The term "sulfone" refers to a group having the structure -SO2R' wherein R' can independently represent hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heterocyclyl, as defined above. The sulfone group is preferably -SO2C1-C6 alkyl.

[0121] The term "ureido" refers to a group having the structure -NRCONR'R" wherein R, R' and R" can independently represent hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, heterocyclyl or substituted heterocyclyl, as defined above. R, R' and R" in the dialkylamine fragment can be the same or different. The ureido group is preferably -NHCONH2, -NHCONH(C1-C6 alkyl).

[0122] The term "heterocyclylalkyl" refers to a group having the structure -RR' wherein R can independently represent alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl; and R' represents heterocyclyl or substituted heterocyclyl.

[0123] In the present application, the term "substituted" means that one or more hydrogen atoms on a particular group are replaced with a particular substituent. The particular substituents are those described in the foregoing, or those substituents appearing in the various embodiments. Unless otherwise specified, a substituted group can have at each substitutable position one substituent selected from the indicated group, which itself can be substituted or unsubstituted. It will be understood by those skilled in the art that combinations of substituents that are not chemically feasible or that are inherently unstable are not intended within the scope of the application. The substituents include, but are not limited to, halogen, hydroxyl, cyano, carboxyl (-COOH), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, 3- to 12-membered heterocyclyl, aryl, heteroaryl, C1-C8 aldehyde, C2-C10 acyl, C2-C10 ester, amine, C1-C6 alkoxy, C1-C10 sulfonyl, and C1-C6 urea, and the like.

[0124] The term "sulfonyl" refers to a group with the structure -S(O)2R, where R can independently represent alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted cycloalkenyl, aryl or substituted aryl; R' represents heterocycle or substituted heterocycle. As defined above. The sulfonyl group is preferably -S(O)2C1-C6 alkyl.

[0125] Unless otherwise indicated, it is assumed that any heteroatom of unsatisfied valence has sufficient hydrogen atoms to satisfy its valence.

[0126] When a substituent is a non-terminal substituent, it is an "alkyl" group, for example, an "alkyl" group corresponds to an "alkylene" group, a "cycloalkyl" group corresponds to a "cycloalkylene" group, a "heterocyclyl" group corresponds to a "heterocyclylene" group, an "alkoxy" group corresponds to an "alkyleneoxy" group, and the like.

[0127] In the present application, multiple means 2, 3, 4, 5.

[0128] It is understood that the definition of any substituent or variable for a molecule at a particular location is independent of the definition of any substituent or variable for the same molecule at another location. It is readily understood by those skilled in the art that the substituents or substitution patterns of the compounds of the present application can be selected by those skilled in the art through the use of known techniques and methods described in the present application to arrive at a chemically stable and synthetically feasible compound.

[0129] Active ingredient

[0130] As used herein, "a compound of the present application" refers to a compound of Formula I, and also includes a compound of Formula I, or a pharmaceutically acceptable salt, stereoisomer, tautomer, deuterated isotope, crystal, hydrate, solvate, prodrug, or combination thereof.

[0131] Salts of the compounds of the present application are also within the scope of the application. Unless otherwise specified, the compounds of the present application are understood to include salts thereof. The term "salt," as used herein, refers to an acid or base salt formed by reaction with inorganic or organic acids and bases. In addition, when the compounds of the present application contain a basic moiety, such as, but not limited to, a pyridine or imidazole, and an acidic moiety, such as, but not limited to, a carboxylic acid, zwitterions ("inner salts") can be formed and are included within the scope of the term "salt." Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, e.g., in isolation and purification steps. Salts of the compounds of the present application can be formed by conventional methods, for example, by reacting a compound of the present application with an amount of acid or base, in a medium which is either neat or contains an inert solvent, and isolating the salt thus formed, or by co-precipitation with a salt under conditions which allow the salt to form, or by freeze-drying a solution of the salt in aqueous solution.

[0132] Compounds of the present application containing a basic moiety, such as, but not limited to, an amine or pyridine or imidazole ring, can form salts with organic or inorganic acids. Exemplary acids which can form salts include acetic acid (e.g., with acetic acid or a trihaloacetic acid, such as trifluoroacetic acid), adipic acid, alginic acid, ascorbic acid, aspartic acid, benzoic acid, benzenesulfonic acid, bisulfic acid, boric acid, butyric acid, citric acid, camphoric acid, camphorsulfonic acid, cyclopentanepropionic acid, digluconic acid, dodecylsulfic acid, ethanesulfonic acid, fumaric acid, glucoheptanoic acid, glycerophosphoric acid, hemisulfic acid, heptanoic acid, hexanoic acid, hydrochloric acid, hydrobromic acid, hydroiodic acid, isethionic acid (e.g., 2-hydroxyethanesulfonic acid), lactic acid, maleic acid, methanesulfonic acid, naphthalenesulfonic acids (e.g., 2-naphthalenesulfonic acid), nicotinic acid, nitric acid, oxalic acid, pectinic acid, persulfic acid, phenylpropionic acid (e.g., 3-phenylpropionic acid), phosphoric acid, picric acid, pivalic acid, propionic acid, salicylic acid, succinic acid, sulfuric acid (e.g., with sulfuric acid), sulfonic acids, tartaric acid, thiocyanic acid, toluenesulfonic acid (e.g., para-toluenesulfonic acid), dodecanoic acid, and the like.

[0133] Certain compounds of the present application can contain acidic moieties, including, but not limited to, carboxylic acids, which can form salts with a variety of organic and inorganic bases. Typical bases which form salts include ammonium salts, alkali metal salts such as sodium, lithium, potassium salts, alkaline earth metal salts such as calcium, magnesium salts, and salts formed with organic bases (e.g. organic amines) such as benzathines, bicyclohexylamines, hydrabamines (formed with N,N-bis(dehydroabietyl)ethylenediamine), N-methyl-D-glucamines, N-methyl-D-glucamides, t-butyl amines, and salts with amino acids such as arginine, lysine, and the like. Basic nitrogen-containing groups can be quaternized with, for example, alkyl halides (e.g. methyl, ethyl, propyl, and butyl chlorides, bromides and iodides), dialkyl sulfates (e.g. dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g. decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), aralkyl halides (e.g. benzyl and phenyl bromides), and others.

[0134] Prodrugs and solvates of the compounds of the present application are also within the scope of the application. The term "prodrug" as used herein refers to a compound which, upon administration, is metabolized or chemically converted in the body to form a compound, salt, or solvate of the present application. The compounds of the present application include solvates, such as hydrates.

[0135] The compounds, salts or solvates of the present application can exist in tautomeric forms (e.g. amide and imine ether). All such tautomers are intended to be encompassed by the present application.

[0136] When the compounds of Formula (I) and pharmaceutically acceptable salts thereof exist as solvates or polymorphs, the present application includes any possible solvates and polymorphs. The type of solvent forming the solvate is not particularly limited as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone and the like can be used.

[0137] Stereoisomers (for example, those caused by the presence of an asymmetric carbon atom in the various substituents) of all compounds, including their enantiomeric and diastereomeric forms, are intended to be encompassed within the scope of the present application. The individual stereoisomers of the compounds of the present application can be isolated by physical methods such as, for example, fractional crystallization or by formation of a mixture with another enantiomer or a diastereomeric form, which can be separated by chromatography or other techniques. The chiral centers of the present application have the S or R configuration as defined by the International Union of Pure and Applied Chemistry (IUPAC) Recommendations 1974. The racemic forms can be resolved by physical methods such as, for example, fractional crystallization or by formation of diastereomeric salts with an optically active acid, which can be separated by chromatography or other techniques. The individual optical isomers can be obtained by the application of the appropriate methods such as those described by Schmidt, U. (1986) Fortsch. Chem. Org. Naturst. 35: 433-597 to the racemates. The present application includes all possible diastereomers and mixtures thereof, substantially pure resolved enantiomers, all possible geometric isomers and pharmaceutically acceptable salts thereof.

[0138] When tautomers exist for the compounds of formula (I), the present application includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof, unless specifically stated otherwise.

[0139] Since the compounds of formula (I) are intended for use as pharmaceuticals, it is preferred to use a certain degree of purity, for example, at least 60% pure, more suitably at least 75% pure, and especially suitably at least 98% pure (% by weight).

[0140] The compounds of the present application, obtained by preparation, isolation and purification, are intended to be in a weight amount equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 99% ("very pure") as described in the text. Such "very pure" compounds of the present application are also intended to be part of the present application.

[0141] All configurational isomers of the compounds of the present application are intended to be within the scope of the present application, whether in mixtures, pure or very pure form. The definition of the compounds of the present application includes both cis (Z) and trans (E) olefinic isomers, as well as cis and trans isomers of carbocyclic and heterocyclic rings.

[0142] Throughout the specification, groups and substituents can be chosen to provide stable moieties and compounds.

[0143] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this application, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75thEd. th Definitions of specific functional groups are also described in more detail below. Additionally, general principles of organic chemistry, and specific functional groups and reactivity are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, the entire contents of which are incorporated herein by reference.

[0144] Certain compounds of the application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-isomers, R- and S- enantiomers, diastereomers, (D)- isomers, (L)-isomers, racemic mixtures, and other mixtures thereof. Additionally, asymmetric carbon atoms can be designated as substituents, such as alkyl groups. All isomers and mixtures thereof are encompassed by the present application.

[0145] According to the present application, mixtures of isomers can have a variety of ratios of isomers. For example, mixtures of only two isomers can have the following compositions: 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0, with all ratios of isomers being within the scope of the present application. Similar ratios, as would be understood by those of ordinary skill in the art, and ratios for more complex mixtures of isomers are also within the scope of the present application.

[0146] The present application also includes isotopically-labelled compounds, which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36Cl. The compounds of the present application, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates thereof, wherein the isotopes or other isotopic atoms of the compounds described above are within the scope of the present application. Certain isotopically-labeled compounds of the present application, for example, 3 H and 14 C are useful in drug and substrate tissue distribution assays. Tritiated, i.e., 3 H, and carbon-14, i.e., 14 C, isotopes are particularly preferred for their ease of preparation and detectability. Carbon- 14 is particularly preferred. Further, substitution with heavier isotopes such as deuterium, i.e., 2 H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements. The isotopically labeled compounds of this application can generally be prepared by carrying out the procedures disclosed in the Schemes and Examples described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0147] If it is desired to design a synthesis of a particular enantiomer of a compound of the present application, it can be prepared by asymmetric synthesis, or derivatized with a chiral auxiliary, separating the resulting diastereomeric mixture, and then removing the chiral auxiliary to yield the pure enantiomer. Alternatively, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, it can be derivatized with an appropriate optically active acid or base to form a diastereomeric salt which is then separated by conventional means, such as fractional crystallization or chromatography, and the pure enantiomer is then recovered by removing the chiral auxiliary.

[0148] As used herein, the compounds of the present application can be expanded to include any number of substituents or functional groups. In general, the term "substituted" whether preceded by an "optionally" or not, is intended to refer to the replacement of hydrogen radicals in the general formula of the compounds of the present application with the indicated structural moieties. When a number of moieties are substituted in a particular structure, each position can be occupied by the same or different moiety. As used herein, the term "substituted" includes all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic organic moieties. In the present application, for example, the heteroatom nitrogen can have a hydrogen substituent or any permissible organic moiety as described above to complete its valence. In no way is the present application intended to be limited to any specific allowed substituents of organic compounds. The present application contemplates that the combination of substituents and variables in a stable compound is well-tolerated, for example, in the treatment of infectious or proliferative diseases. The term "stable" is intended to refer to compounds having a sufficient longevity in time to be detectable for a sufficient period of time to maintain the integrity of the compound structure, and preferably, to be effective for a sufficient period of time. As used herein, this term is intended to encompass the above purposes.

[0149] Metabolites of compounds of the present application, as well as prodrugs of the structures of the compounds of the present application and pharmaceutically acceptable salts thereof, that can be converted in vivo to the compounds of the present application, are also within the scope of the claims.

[0150] Preparation methods

[0151] The preparation of the compounds of formula (I) of the present application is described in more detail below, but these specific methods do not limit the present application in any way. The compounds of the present application can also be prepared conveniently by optionally combining various synthetic methods described in this specification or known in the art, which can be easily performed by those skilled in the art to which the present application pertains.

[0152] Typically, the preparation of the compounds of the present application follows the process flow as shown below, wherein the starting materials and reagents used, if not otherwise specified, can be purchased through commercial channels.

[0153] Preferably, the compounds of the present application are prepared by the following methods:

[0154] In an inert solvent, the compound of formula IV is reacted with the compound of formula V to obtain the compound of formula I;

[0155] wherein ring A, ring B, X, Y, R1, R2, R5and R6are as described in the first aspect of the present application.

[0156] In a specific embodiment, the molar ratio of the compound of formula IV to the compound of formula V is 2-4:1; preferably 2-4:1; more preferably 2:1.

[0157] In a specific embodiment, the reaction solvent of the present application can be selected according to the reaction convention, in particular, the inert solvent is selected from the group consisting of water, C1-C6halogenated hydrocarbon solvents, C1-C6ketone solvents, C1-C6ester solvents, C1-C6alcohol solvents, C2-C6ether solvents, amide solvents, or combinations thereof; preferably dichloromethane, acetone, ethyl acetate, methanol, ethanol, water, 1,4-dioxane, N,N-dimethylformamide; preferably methanol, ethanol; more preferably ethanol.

[0158] In a specific embodiment, the reaction temperature is 50-100°C, preferably 75-85°C, for example 75°C, 80°C or 85°C.

[0159] In a specific embodiment, the method comprises the steps of:

[0160] In an inert solvent, the compound of formula IV-A is reacted with the compound of formula V-A to obtain the compound of formula I;

[0161] wherein ring A, R1, R5and R6are as described in the first aspect of the application.

[0162] Pharmaceutical composition and administration

[0163] The pharmaceutical composition of the present application is used for preventing and / or treating infectious diseases, inflammatory diseases, autoimmune diseases, neurodegenerative diseases, metabolic diseases, rare diseases, tumors, radiation damage.

[0164] The compounds of general formula (I) can be used in combination with other drugs known to be of use in the treatment or amelioration of the same conditions. When administered in combination, the previously known drugs can be administered at the same time as the compound of formula I or sequentially. When the compound of formula I is administered concurrently with the other drug(s), a pharmaceutical composition containing both the known drug(s) and the compound of formula I can be used. Pharmaceutical combinations also include administration of the compound of formula I and the other known drug(s) within overlapping time periods. When the compound of formula I is used in combination with other drugs, the dosage of the compound of formula I or the known drug(s) can be lower than that required when used alone.

[0165] In particular, the compounds of formula I of the present application can be used as potentiators to enhance the therapeutic effect of existing cGAS inhibitors (e.g. G150).

[0166] Drugs or active ingredients that can be used in combination with the compounds of general formula (I) include, but are not limited to, G150, G140, RU.521, etc. The dosage forms of the pharmaceutical composition of the present application include, but are not limited to, injections, tablets, capsules, aerosols, suppositories, films, dripping pills, external liniments, controlled release or sustained release or nano-preparations.

[0167] The pharmaceutical composition of the present application comprises a safe and effective amount of the compound of the present application or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. The "safe and effective amount" refers to the amount of the compound sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1-2000 mg of the compound of the present application per dose, more preferably 10-1000 mg of the compound of the present application per dose. Preferably, the "dose" is a capsule or a tablet.

[0168] "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid filler substances or gel materials, which are suitable for human use and which are of sufficient purity and sufficiently low toxicity. By "compatible" it is meant that the components of the composition are capable of being commingled with the compounds of the application, with each other, and with other ingredients, and that the resulting mixture is stable enough to be manufactured, used, and / or stored. Examples of suitable pharmaceutically acceptable carriers are celluloses and their derivatives (e.g., sodium carboxymethylcellulose, ethylcellulose sodium, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween 80®, Span 80®, etc.), wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like. ) emulsifiers, wetting agents, coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

[0169] The mode of administration of the compounds or pharmaceutical compositions of the present application is not narrowly critical and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0170] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with such other ingredients as binders, (a) fillers or extenders, e.g., starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, e.g., glycerol; (d) disintegrating agents, e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, e.g., paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) absorbents, e.g., kaolin and bentonite clay; and (i) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms can also comprise buffering agents.

[0171] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings and shells known in the art. They can contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0172] Tablets containing the compounds or pharmaceutical compositions of the application can be obtained by compression or molding, optionally, with one or more accessory ingredients or adjuvants. The active compounds can be compressed into tablets from a free-flowing form such as powder or granules. The tables can be prepared from a free-flowing form of the compound or composition of matter using suitable binders, lubricants, inert diluents, surface active or dispersing agents. Molded tablets can be made directly from the powdered compound or composition of matter using a suitable polymeric binder such as povidone, hydroxypropylmethyl cellulose, sucrose, acacia, corn starch or gelatin, in a manner known per se. Alternatively, capsules can be used, made of gelatin or other suitable absorbent material, with the compound or composition of matter being mixed with suitable inert fillers such as lactose, binders such as starch or polyvinylpyrrolidone, lubricants such as stearic acid or talc or disintegrating agents such as potato starch or sodium starch glycolate in a suitable machine. Preferably, each tablet contains about 0.05 mg to 5 g of active compound, each capsule contains about 0.05 mg to 5 g of active compound. For oral administration, the compositions can take the form of, for example, tablets or capsules. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients are, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as potato starch, or

[0173] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms can include inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and the like, as well as mixtures thereof.

[0174] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0175] Suspensions, in addition to the active compounds, can contain suspending agents, as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, and agar-agar, as well as mixtures thereof.

[0176] Compositions for parenteral injection can contain physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyol, glycols, oils, and the like, as well as combinations thereof.

[0177] Dosage forms for topical administration of a compound of this application include ointments, powders, sprays, and inhalers. The active component is admixed with a carrier, which can be a solid, a liquid, or a gas, and is applied in a manner compatible with the formulation to be administered. For example, the carrier can be a transdermal patch, an ointment, or a cream. Transdermal patches have the added advantage of providing controlled delivery of a compound of the application at a predetermined level that can be adjusted as necessary. Dosage forms for topical administration of a compound of this application include ointments, powders, sprays, and inhalers. The active component is admixed with a carrier, which can be a solid, a liquid, or a gas, and is applied in a manner compatible with the formulation to be administered. For example, the carrier can be a transdermal patch, an ointment, or a cream. Transdermal patches have the added advantage of providing controlled delivery of a compound of the application at a predetermined level that can be adjusted as necessary.

[0178] The therapeutic method of the present application can be used alone or in combination with other therapeutic methods or therapeutic drugs.

[0179] When the pharmaceutical composition is used, a safe and effective amount of the compound of the present application is applied to a mammal (e.g., human) in need of treatment, wherein the dosage is a pharmaceutically effective dosage, and the daily dosage is usually 1-2000 mg, preferably 50-1000 mg, for a 60 kg body weight. Of course, the specific dosage should also consider health status, gender, diet, administration time, administration route, excretion rate, drug combination, etc., which are within the skill of a skilled physician.

[0180] Compared with the prior art, the main advantages of the present application include:

[0181] 1. The present application provides a class of cGAS inhibitor compounds with a completely new structure.

[0182] 2. The compounds of the present application have excellent inhibitory effect on cGAS at the enzyme and cellular levels.

[0183] 3. The compounds of the present application can also be used as synergists to improve the therapeutic effect of cGAS inhibitors.

[0184] The present application will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not to limit the scope of the present application. The experimental methods in the following examples, if not specified, are usually carried out according to the conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight.

[0185] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. In addition, any method and material similar or equivalent to those described herein can be used in the present application. The preferred methods and materials described herein are only used as examples.

[0186] In the present application, 1 H NMR spectra were measured by Bruker-600 MHz, 500 MHz or 400 MHz nuclear magnetic resonance spectrometer, and LRMS spectra were measured on Finnigan LTQ linear ion trap mass spectrometer. The reactions were tracked by TLC, HPLC preparation and purity determination were carried out on Dalian Eliet P3140A / P type instrument, and all reactions were carried out under air atmosphere unless otherwise specified.

[0187] The abbreviations used in this invention and their Chinese meanings.

[0188] CDI: N,N'-carbonyldiimidazole;

[0189] THF: Tetrahydrofuran;

[0190] DCM: Dichloromethane;

[0191] EA: Ethyl acetate;

[0192] TFA: Trifluoroacetic acid;

[0193] TEA: Triethylamine;

[0194] DMF: N,N-dimethylformamide.

[0195] Example 1

[0196] Synthesis of intermediate 4,5-dichloro-1,3-dihydro-2H-benzo[d]imidazol-2-one (2)

[0197] 3,4-Dichlorophenyl-1,2-diamine (240 mg, 1.36 mmol) and pyridine (219 μL, 2.71 mmol) were dissolved in THF (3.5 mL), and CDI (440 mg, 2.71 mmol) dissolved in DCM (3.5 mL) was slowly added dropwise. The reaction was carried out at room temperature for 15 h. After the reaction was completed, 220 mg of a pale yellow solid was obtained by filtration, with a yield of 79.9%. 1 H NMR (500MHz, DMSO-d6) δ11.39(s,1H),11.05(s,1H),7.17(d,J=8.4,1H),6.91(d,J=8.3Hz,1H).

[0198] Synthesis of intermediate 2-bromo-6,7-dichloro-1H-benzo[d]imidazole (3)

[0199] Intermediate 2 (680 mg, 3.35 mmol) and phosphorus oxybromide (8.07 mg, 28.13 mmol) were added to a reaction flask, and the mixture was heated to 110 °C and reacted for 18 h. After the reaction was complete, saturated sodium bicarbonate was slowly added dropwise under ice bath conditions to adjust the pH to 6-7. The mixture was then filtered to obtain 720 mg of a white solid, with a yield of 80.1%. 1 H NMR (500MHz, DMSO-d6) δ13.85(brs,1H),7.50(d,J=8.7Hz,1H),7.43(d,J=8.6Hz,1H).

[0200] Synthesis of intermediate 6,7-dichloro-2-hydrazino-1H-benzimidazole (4)

[0201] To intermediate 3 (720 mg, 2.71 mmol) was added hydrazine hydrate (2 mL, 40.61 mmol) and warmed to 100 °C for 12 h. After completion of the reaction, it was cooled to room temperature and filtered to get white solid 450 mg, yield 76.6%. 1 H NMR (600 MHz, DMSO-d6) δ 11.41 (s, 1H), 8.27 (s, 1H), 7.07 (d, J = 8.2 Hz, 1H), 6.99 (d, J = 8.2 Hz, 1H), 4.59 (s, 2H).

[0202] Synthesis of intermediate 1 -(6,7-dichloro- 1 H-benzo[d]imidazol-2-yl)-3-methyl- 1 H-pyrazol-5-ol (5)

[0203] To intermediate 4 (200 mg, 0.92 mmol) and ethyl acetoacetate (116 μL, 0.92 mmol) was added glacial acetic acid (1.5 mL) and warmed to 100 °C for 3 h. After completion of the reaction, it was cooled to room temperature and white solid was precipitated. The solid was filtered and recrystallized using ethanol to get white solid 200 mg, yield 76.7%. 1 H NMR (400 MHz, DMSO-d6) δ 11.96 (s, 1H), 7.50 (d, J = 8.5 Hz, 1H), 7.36 (d, J = 8.5 Hz, 1H), 5.30 (s, 1H), 2.22 (s, 3H).

[0204] Synthesis of 4,4'-(phenylmethylene)bis( 1 -(4,5-dichloro- 1 H-benzo[d]imidazol-2-yl)-3-methyl- 1 H-pyrazol-5-ol) (A00)

[0205] To intermediate 5 (80 mg, 0.28 mmol) and benzaldehyde (14.4 μL, 0.14 mmol) was added ethanol and warmed to 80 °C for 12 h. After completion of the reaction, the solid was filtered and recrystallized using ethanol to get white solid 78 mg, yield 84.4%. ESI-MS: 652.7 [M+H] + 1 H NMR (500 MHz, DMSO-d6) δ 12.79 (s, 2H), 12.22 (br s, 2H), 7.49 (d, J = 8.5 Hz, 2H), 7.41 - 7.28 (m, 6H), 7.24 (t, J = 6.6 Hz, 1H), 5.13 (s, 1H), 2.14 (s, 6H).

[0206] Example 2

[0207] ​Synthesis of intermediate 1 -(1 H-benzo[d]imidazol-2-yl)-3-methyl- 1 H-pyrazol-5-ol (7)

[0208] Refer to the synthesis method of intermediate 5, using 2-hydrazinyl-1 H-benzo[d]imidazole as the starting material, a white solid was obtained. 1 H NMR (400 MHz, DMSO-d6) δ 7.61 - 7.45 (m, 2H), 7.17 (dd, J = 6.0, 3.2 Hz, 2H), 5.24 (s, 1 H), 2.20 (s, 3H).

[0209] Synthesis of 4,4'-((3-hydroxyphenyl)methylene)bis(1 -(1 H-benzo[d]imidazol-2-yl)-3-methyl- 1 H-pyrazol-5-ol) (A01 )

[0210] Refer to the synthesis method of A00, using intermediate 7 and m-hydroxybenzaldehyde as the starting material, a white solid was obtained in 82.1 % yield. ESI-MS: 530.8 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.22 (br s, 1 H), 7.57 - 7.48 (m, 4H), 7.26 - 7.14 (m, 4H), 7.07 (t, J = 7.9 Hz, 1 H), 6.79 - 6.72 (m, 2H), 6.62 - 6.51 (m, 1 H), 4.88 (s, 1 H), 2.22 (s, 6H).

[0211] Example 3

[0212] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(1 -(1 H-benzo[d]imidazol-2-yl)-3-methyl- 1 H-pyrazol-5-ol) (A02)

[0213] Refer to the synthesis method of A00, using intermediate 7 and pyridine-2-carboxaldehyde as the starting material, a white solid was obtained in 85.3% yield. ESI-MS: 516.1 [M-H]; 1 H NMR (400 MHz, DMSO-d6) δ 8.77 (d, J = 5.7 Hz, 1 H), 8.49 (t, J = 8.0 Hz, 1 H), 8.11 (d, J = 8.2 Hz, 1 H), 7.87 (t, J = 6.7 Hz, 1 H), 7.69 - 7.57 (m, 4H), 7.46 - 7.33 (m, 4H), 5.51 (s, 1 H), 2.41 (s, 6H).

[0214] Example 4

[0215] Synthesis of 4,4'-(thiophen-3-ylmethylidene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A03)

[0216] Refer to the synthesis method of A00, with intermediate 7 and 3-thiophene carboxaldehyde as starting materials, white solid was obtained, yield 82.1%. ESI-MS: 521.8 [M-H]; 1 H NMR (400 MHz, DMSO-d6) δ 7.52 (dd, J = 6.0, 3.2 Hz, 4H), 7.44 (dd, J = 5.0, 3.0 Hz, 1H), 7.24 - 7.15 (m, 5H), 7.07 (d, J = 5.0 Hz, 1H), 4.95 (s, 1H), 2.23 (s, 6H).

[0217] Example 5

[0218] Synthesis of 4,4'-(4-fluorophenyl)methylidene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A04)

[0219] Refer to the synthesis method of A00, with intermediate 7 and p-fluorobenzaldehyde as starting materials, white solid was obtained, yield 90.6%. ESI-MS: 534.8 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.58 - 7.49 (m, 4H), 7.39 (dd, J = 8.6, 5.6 Hz, 2H), 7.25 - 7.17 (m, 4H), 7.12 (t, J = 8.9 Hz, 2H), 4.96 (s, 1H), 2.23 (s, 6H).

[0220] Example 6

[0221] Synthesis of 4,4'-((lH-imidazol-4-yl)methylidene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A05)

[0222] Refer to the synthesis method of compound 1, with intermediate 7 and 4-imidazole carboxaldehyde as starting materials, light yellow solid was obtained, yield 78.8%. 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1H), 7.52 - 7.41 (m, 4H), 7.17 (s, 1H), 7.12 (dt, J = 6.1, 3.6 Hz, 4H), 4.87 (s, 1H), 2.21 (s, 6H).

[0223] Example 7

[0224] Synthesis of 4,4'-((2-hydroxyphenyl)methylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (Compound 8)

[0225] Refer to the synthesis method of A00, using intermediate 7 and 2-hydroxybenzaldehyde as starting materials, white solid was obtained in 82.5% yield. ESI-MS: 533.9 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.59 - 7.45 (m, 5H), 7.19 (dt, J = 6.1, 3.6 Hz, 4H), 7.01 (t, J = 7.6 Hz, 1H), 6.80 - 6.67 (m, 2H), 5.16 (s, 1H), 2.18 (s, 6H).

[0226] Example 8

[0227] Synthesis of 4,4'-(pyridin-3-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A07)

[0228] Refer to the synthesis method of A00, using intermediate 7 and 3-pyridinecarboxaldehyde as starting materials, pink solid was obtained in 89.6% yield. ESI-MS: 518.0 [M+H] + ; HRMS (ESI) calculated for C 28 H 24 N9O2 + ([M+H] + ), 518.2047; found, 518.2044. 1 H NMR (500 MHz, DMSO-d6) δ 8.60 (d, J = 2.4 Hz, 1H), 8.42 (dd, J = 4.7, 1.7 Hz, 1H), 7.81 (dt, J = 8.0, 2.2 Hz, 1H), 7.59 - 7.48 (m, 4H), 7.39 - 7.32 (m, 1H), 7.21 (dt, J = 6.0, 3.4 Hz, 4H), 5.01 (s, 1H), 2.26 (s, 6H). 13 C NMR (126 MHz, DMSO) δ 159.60, 151.56, 149.35, 147.32, 143.95, 138.76, 135.85, 135.35, 123.67, 122.75, 114.62, 102.44, 31.83, 12.93.

[0229] Example 9

[0230] Synthesis of 4,4'-((1H-indol-5-yl)methylene)bis(1-(1H-benzo[d]imidazol-2-yl)-3-methyl- 1H-pyrazol-5-ol) (A08)

[0231] Refer to the synthesis method of A00, yellow solid was obtained with intermediate 7 and 5-indolecarboxaldehyde as starting materials in 72.4% yield. ESI-MS: 554.1 [M-H] - ; HRMS (ESI) calculated for C 31 H 25 N9NaO2 + ([M+Na] + ), 578.2023; found, 578.2022; 1 H NMR (500 MHz, DMSO-d6) δ 10.98 (s, 1H), 7.64 - 7.41 (m, 5H), 7.35 - 7.26 (m, 2H), 7.24 - 7.05 (m, 5H), 6.35 (s, 1H), 5.06 (s, 1H), 2.20 (s, 6H).

[0232] Example 10

[0233] Synthesis of 4,4'-(cyclohexylmethylene)bis(1-(1H-benzo[d]imidazol-2-yl)-3-methyl-1H- pyrazol-5-ol) (A09)

[0234] Refer to the synthesis method of A00, white solid was obtained with intermediate 7 and cyclohexanecarboxaldehyde as starting materials in 90.3% yield. ESI-MS: 523.2 [M+H] + ; HRMS (ESI) calculated for C 29 H 31 N8O2 + ([M+H] + ), 523.2564; found, 523.2560; 1 H NMR (500 MHz, DMSO-d6) δ 7.60 - 7.47 (m, 4H), 7.23 - 7.14 (m, 4H), 3.15 (d, J = 11.0 Hz, 1H), 2.41 - 2.33 (m, 1H), 2.28 (s, 6H), 1.74 - 1.56 (m, 6H), 1.19 - 1.13 (m, 2H), 0.91 - 0.82 (m, 2H). 13C NMR (126 MHz, DMSO) δ 159.12, 151.27, 144.02, 135.83, 122.53, 114.66, 103.78, 35.58, 32.10, 26.54, 26.39, 25.91, 12.74.

[0235] Example 11

[0236] Synthesis of 4,4'-(thiazol-4-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A10)

[0237] According to the synthetic method of A00, using intermediate 7 and thiazole-4-carboxaldehyde as starting materials, white solid was obtained in 77.2% yield. ESI-MS: 524.0 [M+H] + ; HRMS (ESI) calculated for C 26 H 22 N9O2S + ([M+H] + ), 524.1612; found, 524.1615; 1 H NMR (500 MHz, DMSO-d6) δ 9.04 (d, J = 2.0 Hz, 1H), 7.57 - 7.49 (m, 4H), 7.44 - 7.38 (m, 1H), 7.24 - 7.14 (m, 4H), 5.11 (s, 1H), 2.22 (s, 6H). 13 CNMR (126 MHz, DMSO) δ 159.39, 157.20, 153.48, 150.74, 143.40, 135.32, 122.00, 114.72, 114.20, 102.52, 30.90, 11.99.

[0238] Example 12

[0239] Synthesis of 4,4'-((l-methyl-lH-imidazol-4-yl)methylene)bis(l-(lH-benzo[d]imidazol-2- yl)-3-methyl-lH-pyrazol-5-ol) (Al l)

[0240] According to the synthetic method of A00, using intermediate 7 and l-methyl-lH-imidazole-4- carboxaldehyde as starting materials, yellow solid was obtained in 88.4% yield. ESI-MS: 521.3 [M+H] + ; HRMS (ESI) calculated for C 27 H 25 N10 O2 + ([M+H] + ),521.2156;found,521.2155; 1 H NMR (400 MHz, DMSO-d6) δ 7.50 (dd, J = 6.0, 3.2 Hz, 4H), 7.42 (s, 1H), 7.24 (s, 1H), 7.20 - 7.09 (m, 4H), 5.37 (s, 1H), 3.72 (s, 3H), 2.16 (s, 6H). 13 C NMR (126 MHz, DMSO) δ 158.00, 150.37, 148.57, 144.55, 135.97, 122.75, 121.53, 120.14, 114.12, 97.21, 33.45, 26.86, 12.42.

[0241] Example 13

[0242] Synthesis of 4,4'-((6-methylpyridin-2-yl)methylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A12)

[0243] Refer to the synthesis method of A00, using intermediate 7 and 6-methyl-2-pyridinecarboxaldehyde as starting materials, white solid was obtained in 90.5% yield. ESI-MS: 532.1 [M+H] + ; HRMS (ESI) calculated for C 29 H 26 N9O2 + ([M+H] + ),532.2204; found, 532.2211; 1 H NMR (400 MHz, DMSO-d6) δ 7.67 (t, J = 7.7 Hz, 1H), 7.51 (dd, J = 5.9, 3.2 Hz, 4H), 7.38 (d, J = 7.9 Hz, 1H), 7.18 (dd, J = 5.9, 3.2 Hz, 4H), 7.13 (d, J = 7.6 Hz, 1H), 5.07 (s, 1H), 2.47 (s, 3H), 2.19 (s, 6H). 13 C NMR (126 MHz, DMSO) δ 160.55, 158.96, 156.41, 150.84, 143.58, 137.33, 135.42, 121.89, 120.96, 119.32, 114.14, 101.97, 36.65, 23.70, 12.28.

[0244] Example 14

[0245] Synthesis of 2-(bis(l-(lH-benzo[d]imidazol-2-yl)-5-hydroxy-3-methyl-lH-pyrazol-4- yl)methyl)benzoic acid (A13)

[0246] Refer to the synthesis method of A00, with intermediate 7 and o-carboxybenzaldehyde as starting materials, white solid was obtained in 77.1% yield. ESI-MS: 561.0 [M+H] + ; HRMS (ESI) calculated for C 30 H 24 N8NaO4 + [M+Na] + , 583.1813; found, 583.1821; 1 H NMR (500 MHz, DMSO-d6) δ 7.66 - 7.60 (m, 2H), 7.54 - 7.48 (m, 4H), 7.45 (td, J = 7.6, 1.6 Hz, 1H), 7.30 (td, J = 7.5, 1.3 Hz, 1H), 7.20 - 7.14 (m, 4H), 5.90 (s, 1H), 2.11 (s, 6H). 13 C NMR (126 MHz, DMSO) δ 170.33, 160.77, 151.22, 144.00, 141.78, 135.83, 132.35, 130.87, 129.75, 129.68, 126.58, 122.44, 114.91, 103.95, 30.89, 12.45.

[0247] Example 15

[0248] Synthesis of 4,4'-(ethane-l,l-diyl)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl-lH- pyrazol-5-ol) (A14)

[0249] Refer to the synthesis method of A00, with intermediate 7 and acetaldehyde as starting materials, white solid was obtained in 73.6% yield. ESI-MS: 455.0 [M+H] + ; HRMS (ESI) calculated for C 24 H 23 N8O2 + [M+H] + , 455.1938; found, 455.1947; 1H NMR (500 MHz, DMSO-d6) δ 7.57 - 7.49 (m, 4H), 7.23 - 7.13 (m, 4H), 3.78 (q, J = 7.4 Hz, 1H), 2.35 (s, 6H), 1.61 (d, J = 7.5 Hz, 3H). 13 C NMR (126 MHz, DMSO) δ 159.83, 149.59, 143.53, 135.88, 121.78, 114.24, 105.66, 22.83, 17.84, 11.94.

[0250] Example 16

[0251] Synthesis of 4,4'-(quinoline-8-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A16)

[0252] Referring to the synthesis method of A00, intermediate 7 and cyclopentanecarboxaldehyde were used as starting materials to obtain a white solid with a yield of 85.1%. ESI-MS: 509.1 [M+H] + ; HRMS (ESI) calculated for C 28 H 29 N8O2 + ([M+H] + ), 509.2408; found, 509.2412; 1 H NMR (500 MHz, DMSO-d6) δ 7.56 - 7.48 (m, 4H), 7.18 (dd, J = 6.0, 3.2 Hz, 4H), 3.24 (d, J = 11.6 Hz, 1H), 3.12 - 3.04 (m, 1H), 2.32 (s, 6H), 1.70 - 1.59 (m, 4H), 1.55 - 1.48 (m, 2H), 1.18 - 1.10 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 159.93, 150.71, 144.08, 136.35, 122.46, 114.82, 105.30, 34.94, 32.10, 25.32, 12.66.

[0253] Example 17

[0254] Synthesis of 4,4'-(quinoline-8-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A16)

[0255] A00 as a yellow solid in 73.1% yield. ESI-MS: 568.0 [M+H] + ; HRMS (ESI) calculated for C 32 H 26 N9O2 + ([M+H] + ), 568.2204; found, 508.2202; 1 H NMR (500 MHz, DMSO-d6) δ 8.98 (dd, J = 4.2, 1.8 Hz, 1H), 8.35 (dd, J = 8.3, 1.8 Hz, 1H), 8.13 (d, J = 7.3 Hz, 1H), 7.84 (dd, J = 8.2, 1.4 Hz, 1H), 7.60 - 7.47 (m, 6H), 7.19 (dd, J = 6.0, 3.2 Hz, 4H), 6.31 (s, 1H), 2.26 (s, 6H).

[0256] Example 18

[0257] Synthesis of 4,4'-((3-methoxyphenyl)methylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A17)

[0258] A00 as a white solid in 82.2% yield. ESI-MS: 546.8 [M+H] + ; HRMS (ESI) calculated for C 30 H 26 N8NaO3 + ([M+Na] + ), 569.2020; found, 569.2029; 1 H NMR (500 MHz, DMSO-d6) δ 7.56 - 7.49 (m, 4H), 7.24 - 7.16 (m, 5H), 6.96 - 6.91 (m, 2H), 6.77 (dd, J = 8.1, 2.5 Hz, 1H), 4.91 (s, 1H), 3.70 (s, 3H), 2.22 (s, 6H).

[0259] Example 19

[0260] Synthesis of 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(lH- benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A18)

[0261] Referring to the synthesis method of A00, using intermediate 7 and m- trifluoromethylbenzaldehyde as starting materials, white solid was obtained in 82.7% yield. ESI-MS: 584.9 [M+H] + ; HRMS (ESI) calculated for C 30 H 24 F3N8O2 + ([M+H] + ), 585.1969; found, 585.1974; 1 H NMR (500 MHz, DMSO-d6) δ 7.72 - 7.66 (m, 2H), 7.59 - 7.49 (m, 6H), 7.25 - 7.15 (m, 4H), 5.06 (s, 1H), 2.24 (s, 6H).

[0262] Example 20

[0263] Synthesis of 4,4'-(bicyclo[2.2.1]hept-5-en-2-ylmethylene)bis(l-(lH- benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A19)

[0264] Referring to the synthesis method of A00, using intermediate 7 and 5- norbornene-2-carboxaldehyde as starting materials, white solid was obtained in 80.2% yield. ESI-MS: 533.1 [M+H] + ; HRMS (ESI) calculated for C 30 H 29 N8O2 + ([M+H] + ), 533.2408; found, 533.2404.

[0265] Example 21

[0266] Synthesis of intermediate l-(benzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol (9)

[0267] The synthesis of intermediate 9 refers to the synthesis method of compound 5, using 2-hydrazinylbenzo[d]thiazole (8) as starting material, white solid was obtained in 73.6% yield. 1H NMR (400 MHz, DMSO-d6) δ 12.81 (s, 1H), 8.05 (d, J = 7.9 Hz, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.53 - 7.45 (m, 1H), 7.40 - 7.33 (m, 1H), 5.28 (s, 1H), 2.24 (s, 3H).

[0268] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(3-methyl-l-(4-methylbenzo[d]thiazol-2-yl)-lH-pyrazol-5-ol) (A21)

[0269] The synthesis was performed according to the procedure for A20, using intermediate 9 and pyridine-2-carbaldehyde as starting materials to give a white solid in 86.1% yield. HRMS (ESI) calculated for C 28 H 22 N7O2S2 + ([M+H] + ), 552.1271; found 552.1275; 1 H NMR (400 MHz, DMSO-d6) δ 12.59 (s, 2H), 8.54 (d, J = 4.8 Hz, 1H), 8.04 (d, J = 7.9 Hz, 2H), 7.84 (d, J = 8.0 Hz, 2H), 7.81 - 7.73 (m, 1H), 7.53 - 7.45 (m, 2H), 7.44 - 7.33 (m, 3H), 7.32 - 7.26 (m, 1H), 5.21 (s, 1H), 2.09 (s, 6H).

[0270] Example 22

[0271] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(3-methyl-l-(4-methylbenzo[d]thiazol-2-yl)-lH-pyrazol-5-ol) (A21)

[0272] The synthesis was performed according to the procedure for A20, using 2-hydrazinyl-4-methylbenzo[d]thiazole (10) as starting material to give a white solid in 70.4% yield. HRMS (ESI) calculated for C 30 H 26 N7O2S2 + ([M+H] + ), 580.1584; found 580.1580; 1H NMR (400 MHz, DMSO-d6) δ 12.20 (s, 2H), 8.55 (d, J = 4.8 Hz, 1H), 7.90 - 7.72 (m, 3H), 7.48 - 7.37 (m, 1H), 7.35 - 7.27 (m, 3H), 7.24 (t, J = 7.6 Hz, 2H), 5.23 (s, 1H), 2.65 (s, 6H), 2.13 (s, 6H).

[0273] Example 23

[0274] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(6-chloro-lH-benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A23)

[0275] Refer to the synthesis method of A00, starting from 6-chloro-2-hydrazinyl-lH- benzo[d]imidazole (14) to react to give white solid, yield 66.3%. HRMS (ESI) calculated for C 28 H 20 F4N9O2 + ([M+H] + ), 586.1268; found 586.1264; 1 H NMR (400 MHz, DMSO-d6) δ 12.64 (br s, 2H), 8.54 (d, J = 4.9 Hz, 1H), 7.86 - 7.74 (m, 1H), 7.55 - 7.43 (m, 1H), 7.37 - 7.23 (m, 3H), 7.17 (dt, J = 11.7, 7.9 Hz, 2H), 5.21 (s, 1H), 2.16 (s, 6H).

[0276] Example 24

[0277] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(6-chloro-lH-benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A23)

[0278] Refer to the synthesis method of A00, starting from 6-chloro-2-hydrazinyl-lH- benzo[d]imidazole (14) to react to give white solid, yield 66.3%. HRMS (ESI) calculated for C 28 H 22 Cl2N9O2 + ([M+H] + ), 586.1268; found 586.1264;1 H NMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 4.9 Hz, 1H), 7.78 (t, J = 7.8 Hz, 1H), 7.64 - 7.41 (m, 5H), 7.28 (dd, J = 7.5, 4.9 Hz, 1H), 7.18 (dd, J = 8.5, 2.1 Hz, 2H), 5.17 (s, 1H), 2.16 (s, 6H).

[0279] Example 25

[0280] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A25)

[0281] Refer to the synthesis method of A00, using 2-hydrazinyl-lH-benzo[d]imidazole and ethyl trifluoroacetylacetate as starting materials, white solid was obtained in 57.7% yield. HRMS (ESI) calculated for C 28 H 18 F6N9O2 + ([M+H] + ), 626.1482; found 626.1475; 1 H NMR (400 MHz, DMSO-d6) δ 8.55 - 8.48 (m, 1H), 7.79 (td, J = 7.7, 1.9 Hz, 1H), 7.60 (dd, J = 6.0, 3.2 Hz, 4H), 7.45 (d, J = 8.0 Hz, 1H), 7.32 (dt, J = 6.0, 3.6 Hz, 4H), 7.26 (dd, J = 7.5, 4.9 Hz, 1H), 5.41 (s, 1H).

[0282] Example 26

[0283] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A25)

[0284] Refer to the synthesis method of A00, using 6-fluoro-2-hydrazinyl-lH-benzo[d]imidazole (17) as starting material, white solid was obtained in 76.4% yield. HRMS (ESI) calculated for C 28 H 22 F2N9O2 + ([M+H] +), 554.1859; found 554.1845; 1 H NMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 4.9 Hz, 1H), 7.82 - 7.74 (m, 1H), 7.49 (dd, J = 8.7, 5.1 Hz, 3H), 7.34 - 7.25 (m, 3H), 7.01 (td, J = 9.3, 2.6 Hz, 2H), 5.16 (s, 1H), 2.17 (s, 6H).

[0285] Example 27

[0286] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(4-fluoro-lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A27)

[0287] Referring to the synthetic method of A00, using 2-hydrazinyl-lH-imidazo[4,5- b]pyridine as the starting material, white solid was obtained in 61.6% yield. HRMS (ESI) calculated for C 26 H 22 N 11 O2 + ([M+H] + ), 520.1952; found 520.1958; 1 H NMR (400 MHz, DMSO-d6) δ 8.52 (d, J = 4.9 Hz, 1H), 7.82 - 7.74 (m, 1H), 7.49 (dd, J = 8.7, 5.1 Hz, 3H), 7.34 - 7.25 (m, 3H), 7.01 (td, J = 9.3, 2.6 Hz, 2H), 5.16 (s, 1H), 2.17 (s, 6H).

[0288] Example 28

[0289] Synthesis of 4,4'-(pyridin-2-ylmethylene)bis(l-(4-fluoro-lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A27)

[0290] Referring to the synthetic method of A00, using 4-fluoro-2-hydrazinyl-lH-benzo[d]imidazole as the starting material, white solid was obtained in 70.6% yield.

[0291] Example 29

[0292] 4,4'-(phenylmethylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A28)

[0293] Refer to the synthesis method of A00, using intermediate 7 and benzaldehyde as starting materials, white solid was obtained in 80.3% yield. ESI-MS: 517.0 [M+H] + ; 1 H NMR (500 MHz, DMSO-d6) δ 7.57 - 7.49 (m, 4H), 7.38 (d, J = 7.7 Hz, 2H), 7.30 (t, J = 7.6 Hz, 2H), 7.24 - 7.17 (m, 5H), 4.97 (s, 1H), 2.22 (s, 6H).

[0294] Example 30

[0295] Synthesis of 4,4'-((2-fluorophenyl)methylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A29)

[0296] Refer to the synthesis method of A00, using intermediate 7 and o-fluorobenzaldehyde as starting materials, white solid was obtained in 68.9% yield. 1 H NMR (500 MHz, DMSO-d6) δ 7.67 (s, 1H), 7.52 (dd, J = 6.0, 3.2 Hz, 4H), 7.26 (d, J = 7.3 Hz, 1H), 7.23 - 7.17 (m, 4H), 7.17 - 7.10 (m, 2H), 5.11 (s, 1H), 2.20 (s, 6H).

[0297] Example 31

[0298] Synthesis of 4,4'-((2-chlorophenyl)methylene)bis(l-(lH-benzo[d]imidazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A30)

[0299] Refer to the synthesis method of A00, using intermediate 7 and o-chlorobenzaldehyde as starting materials, white solid was obtained in 75.1% yield. 1 H NMR (500 MHz, DMSO-d6) δ 7.79 - 7.71 (m, 1H), 7.55 - 7.49 (m, 4H), 7.43 (dd, J = 7.8, 1.5 Hz, 1H), 7.34 - 7.28 (m, 1H), 7.28 - 7.23 (m, 1H), 7.22 - 7.15 (m, 4H), 5.17 (s, 1H), 2.15 (s, 6H).

[0300] Example 32

[0301] Synthesis of methyl 2-(bis(1-(1H-benzo[d]imidazol-2-yl)-5-hydroxy-3-methyl-1H- pyrazol-4-yl)methyl)benzoate (A31)

[0302] According to the synthetic method of A00, using intermediate 7 and methyl 2- formylbenzoate as starting materials, white solid was obtained in 68.1% yield. 1 H NMR (600 MHz, DMSO-d6) d 7.63 - 7.57 (m, 2H), 7.54 - 7.47 (m, 5H), 7.34 (t, J = 7.5 Hz, 1H), 7.18 (dt, J = 6.1, 3.6 Hz, 4H), 5.72 (s, 1H), 3.67 (s, 3H), 2.11 (s, 6H).

[0303] Example 33

[0304] Synthesis of 4,4'-((3-fluoropyridin-2-yl)methylene)bis(1-(1H-benzo[d]imidazol-2-yl)-3- methyl-1H-pyrazol-5-ol) (A32)

[0305] According to the synthetic method of A00, using intermediate 7 and 3-fluoropyridine-2- carboxaldehyde as starting materials, white solid was obtained in 65.9% yield. ESI-MS: 536.0 [M+H] + ; 1 H NMR (500 MHz, Methanol-d4) d 8.37 - 8.32 (m, 1H), 7.64 (ddd, J = 10.1, 8.2, 1.4 Hz, 1H), 7.51 (dd, J = 5.9, 3.2 Hz, 4H), 7.36 (dt, J = 8.4, 4.3 Hz, 1H), 7.19 (dt, J = 6.0, 3.6 Hz, 4H), 5.32 (s, 1H), 2.17 (s, 6H).

[0306] Example 34

[0307] Synthesis of 4,4'-((5-fluoropyridin-2-yl)methylene)bis(1-(1H-benzo[d]imidazol-2-yl)-3- methyl-1H-pyrazol-5-ol) (A33)

[0308] According to the synthetic method of A00, using intermediate 7 and 5-fluoropyridine-2- carboxaldehyde as starting materials, white solid was obtained in 78.2% yield. HRMS (ESI) calculated for C28 H 23 FN9O2 + ([M+H] + ), 536.1953; found 536.1957; 1 H NMR (500 MHz, DMSO-d6) δ 8.47 (d, J = 3.0 Hz, 1H), 7.66 (td, J = 8.8, 3.0 Hz, 1H), 7.57 (dd, J = 8.9, 4.5 Hz, 1H), 7.52 (dd, J = 5.9, 3.2 Hz, 4H), 7.20 (dt, J = 5.9, 3.7 Hz, 4H), 5.08 (s, 1H), 2.22 (s, 6H).

[0309] Example 35

[0310] Synthesis of 4,4'-((6-fluoropyridin-2-yl)methylene)bis(l-(lH-benzo[d]imidazol-2- yl)-3-methyl-lH-pyrazol-5-ol) (A34)

[0311] Following the procedure for the synthesis of A00, using intermediate 7 and 6-fluoropyridine-2-carboxaldehyde as starting materials, afforded white solid in 77.9% yield. HRMS (ESI) calculated for C 28 H 23 FN9O2 + ([M+H] + ), 536.1953; found 536.1956; 1 H NMR (500 MHz, Methanol-d4) δ 7.92 (q, J = 8.1 Hz, 1H), 7.53 (dt, J = 5.9, 3.7 Hz, 4H), 7.43 (dd, J = 7.6, 2.5 Hz, 1H), 7.21 (dt, J = 6.0, 3.6 Hz, 4H), 6.99 (dd, J = 8.2, 2.7 Hz, 1H), 4.99 (s, 1H), 2.23 (s, 6H).

[0312] Example 36

[0313] Synthesis of 4,4'-((6,7-dihydro-5H-pyrrolo[l,2-a]imidazol-2-yl)methylene)bis(l-(lH- benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A35)

[0314] The synthesis method of A00 was referred to, with intermediate 7 and 6,7-dihydro-5H- pyrrolo [1,2-a] imidazole-2-carbaldehyde as starting materials to obtain a light yellow solid with a yield of 70.5%. HRMS (ESI) calculated for C 29 H 27 N 10 O2 + ([M+H] + ), 547.2313; found 537.2314; 1 H NMR (500 MHz, Methanol-d4) δ 7.47 (dd, J = 5.9, 3.2 Hz, 4H), 7.16 (s, 1H), 7.15 - 7.07 (m, 4H), 4.83 (s, 1H), 4.03 (t, J = 7.2 Hz, 2H), 2.95 (t, J = 7.5 Hz, 2H), 2.60 - 2.53 (m, 2H), 2.21 (s, 6H).

[0315] Example 37

[0316] Synthesis of methyl 2-(bis(1-(4,5-difluoro-1H-benzo[d]imidazol-2-yl)-5-hydroxy-3- methyl-1H-pyrazol-4-yl)methyl)benzoate (A36)

[0317] The synthesis method of A00 was referred to, with intermediate 13 and methyl 2- formylbenzoate as starting materials to obtain a white solid with a yield of 83.2%. 1 H NMR (500 MHz, DMSO-d6) δ 12.67 (s, 2H), 12.40 (br s, 2H), 7.63 (d, J = 7.5 Hz, 1H), 7.50 (d, J = 4.9 Hz, 2H), 7.41 - 7.34 (m, 1H), 7.30 (dd, J = 8.8, 3.8 Hz, 2H), 7.22 - 7.13 (m, 2H), 5.75 (s, 1H), 3.62 (s, 3H), 2.04 (s, 6H).

[0318] Example 38

[0319] Synthesis of 2-(bis(1-(4,5-difluoro-1H-benzo[d]imidazol-2-yl)-5-hydroxy-3-methyl- 1H-pyrazol-4-yl)methyl)benzoic acid (A37)

[0320] The synthesis method of A00 was referred to, with intermediate 13 and 2-formylbenzoic acid as starting materials to obtain a white solid with a yield of 49.5%. ESI-MS: [M-H] - ;1 H NMR (500 MHz, DMSO-d6) δ 12.90 (br s, 1H), 12.67 (s, 2H), 12.32 (br s, 2H), 7.66 (d, J = 7.6 Hz, 1H), 7.50 (s, 1H), 7.46 (t, J = 7.6 Hz, 1H), 7.34 (s, 1H), 7.31 - 7.20 (m, 2H), 7.20 - 7.09 (m, 2H), 5.93 (s, 1H), 2.02 (s, 6H).

[0321] Example 39

[0322] Synthesis of 2-(bis(l-(benzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-lH-pyrazol-4-yl)methyl)benzoic acid (A38)

[0323] Refer to the synthetic method of A20, using intermediate 9 and 2-formylbenzoic acid as starting materials, to give a light yellow solid in 47.5% yield. ESI-MS: 592.3 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.89 (br s, 1H), 12.51 (s, 2H), 8.03 (d, J = 7.9 Hz, 2H), 7.83 (d, J = 8.1 Hz, 2H), 7.68 (d, J = 7.6 Hz, 1H), 7.48 (t, J = 7.6 Hz, 3H), 7.41 (d, J = 7.9 Hz, 1H), 7.38 - 7.32 (m, 3H), 5.95 (s, 1H), 1.99 (s, 6H).

[0324] Example 40

[0325] Synthesis of 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(6,7-difluoro-lH- benzo[d]imidazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A39)

[0326] Refer to the synthetic method of A00, using intermediate 13 and m-trifluoromethylbenzaldehyde as starting materials, to give a white solid in 87.7% yield. ESI-MS: 657.6 [M+H] - ; 1H NMR (500 MHz, DMSO-d6) δ 12.68 (br s, 4H), 7.67 (d, J = 6.0 Hz, 2H), 7.61 (d, J = 7.7 Hz, 1H), 7.57 (t, J = 7.9 Hz, 1H), 7.31 (dd, J = 8.9, 3.8 Hz, 2H), 7.22 - 7.14 (m, 2H), 5.22 (s, 1H), 2.16 (s, 6H).

[0327] Example 41

[0328] Synthesis of 4,4'-(cyclohexylmethylene)bis(l-(6,7-difluoro-lH-benzo[d]imidazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A40)

[0329] Refer to the synthesis method of A00, using intermediate 13 and cyclohexanecarboxaldehyde as starting materials, white solid was obtained in 84.2% yield. ESI-MS: 593.3 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.64 (s, 2H), 12.16 (s, 2H), 7.31 (s, 2H), 7.18 (d, J = 10.4 Hz, 2H), 3.17 (s, 1H), 2.46 - 2.29 (m, 6H), 1.76 - 1.56 (m, 5H), 1.25 - 1.12 (m, 3H), 0.86 (d, J = 11.5 Hz, 2H).

[0330] Example 42

[0331] Synthesis of 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(4-fluorobenzo[d]thiazol-2-yl)-3- methyl-lH-pyrazol-5-ol) (A41)

[0332] Refer to the synthesis method of A20, using 4-fluoro-2-hydrazinylbenzo[d]thiazole as starting material, white solid was obtained in 54.4% yield. ESI-MS: 652.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.72 (s, 2H), 7.87 (dd, J = 6.9, 1.9 Hz, 2H), 7.64 (d, J = 7.9 Hz, 3H), 7.59 (d, J = 7.5 Hz, 1H), 7.38 - 7.32 (m, 4H), 5.26 (s, 1H), 2.13 (s, 6H).

[0333] Example 43

[0334] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H]

[0335] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.73 (s, 2H), 8.04 (d, J = 8.0 Hz, 2H), 7.84 (d, J = 8.1 Hz, 2H), 7.69 - 7.54 (m, 4H), 7.49 (t, J = 7.7 Hz, 2H), 7.36 (t, J = 7.6 Hz, 2H), 5.25 (s, 1H), 2.05 (s, 6H).

[0336] Example 44

[0337] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H]

[0338] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.73 (s, 2H), 8.04 (d, J = 8.0 Hz, 2H), 7.84 (d, J = 8.1 Hz, 2H), 7.69 - 7.54 (m, 4H), 7.49 (t, J = 7.7 Hz, 2H), 7.36 (t, J = 7.6 Hz, 2H), 5.25 (s, 1H), 2.05 (s, 6H).

[0339] Example 44

[0340] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H]

[0341] 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(benzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A42) was synthesized according to the similar procedure as A20, using intermediate 9 and m-trifluoromethylbenzaldehyde as starting materials in yield of 67.3% as a white solid. ESI-MS: 617.1 [M-H] - ; 1H NMR (500 MHz, DMSO-d6) δ 12.70 (s, 2H), 7.86 (dd, J = 8.4, 2.4 Hz, 2H), 7.81 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.44-7.42 (m, 2H), 5.38 (s, 1H), 2.01 (s, 6H).

[0342] Example 46

[0343] Synthesis of 4,4'-((3-(trifluoromethyl)phenyl)methylene)bis(l-(4,6-difluoro- benzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A45)

[0344] Referring to the synthesis method of A20, intermediate 24 and m-trifluoromethylbenzoic acid were used as starting materials to obtain a white solid with a yield of 70.6%. ESI-MS: 688.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.70 (s, 2H), 7.86 (dd, J = 8.4, 2.4 Hz, 2H), 7.81 (d, J = 8.0 Hz, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.46-7.42 (m, 2H), 5.38 (s, 1H), 2.01 (s, 6H).

[0345] Example 47

[0346] Synthesis of 4,4'-((4-(trifluoromethyl)phenyl)methylene)bis(l-(4,6-difluoro- benzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A46)

[0347] Referring to the synthesis method of A20, intermediate 24 and p-trifluoromethylbenzaldehyde were used as starting materials to obtain a white solid with a yield of 75.33%. ESI-MS: 688.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.76 (s, 2H), 7.88-7.83 (m, 2H), 7.69 (d, J = 8.1 Hz, 2H), 7.51 (d, J = 8.1 Hz, 2H), 7.44 (t, J = 9.6 Hz, 2H), 5.23 (s, 1H), 2.12 (s, 6H).

[0348] Example 48

[0349] Synthesis of 4,4'-((2-(trifluoromethyl)phenyl)methylene)bis(l-(4- fluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A49)

[0350] Refer to the synthesis method of A20, using intermediate 9 and o-trifluoromethylbenzaldehyde as starting materials, white powdery solid was obtained, yield 68.8%. ESI-MS: 617.0 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.68 (s, 2H), 8.04 (dd, J = 8.1, 1.2 Hz, 2H), 7.84 (t, J = 7.4 Hz, 3H), 7.77-7.74 (m, 1H), 7.67 (t, J = 7.6 Hz, 1H), 7.52-7.47 (m, 3H), 7.38-7.34 (m, 2H), 5.39 (s, 1H), 2.01 (s, 6H).

[0351] Example 49

[0352] Synthesis of 4,4'-((4-(trifluoromethyl)phenyl)methylene)bis(l-(4- fluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A49)

[0353] Refer to the synthesis method of A20, using intermediate 9 and p-trifluoromethylbenzaldehyde as starting materials, white powdery solid was obtained, yield 74.8%. ESI-MS: 617.0 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.74 (s, 2H), 8.04 (d, J = 7.9 Hz, 2H), 7.84 (d, J = 8.1 Hz, 2H), 7.69 (d, J = 8.1 Hz, 2H), 7.53 (s, 1H), 7.51 (s, 1H), 7.51-7.47 (m, 2H), 7.36 (t, J = 7.7 Hz, 2H), 5.22 (s, 1H), 2.12 (s, 6H).

[0354] Example 50

[0355] Synthesis of 4,4'-((4-(trifluoromethyl)phenyl)methylene)bis(l-(4- fluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A49)

[0356] Refer to the synthesis method of A20, with intermediate 22 and m-trifluoromethylbenzaldehyde as starting materials, to obtain white solid, yield 35.0%. ESI-MS: 652.8 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.69 (s, 2H), 7.87 (dd, J = 7.1, 2.0 Hz, 2H), 7.83 (d, J = 7.9 Hz, 1H), 7.78-7.74 (m, 1H), 7.67 (t, J = 7.7 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.39-7.31 (m, 4H), 5.39 (s, 1H), 2.02 (s, 6H).

[0357] Example 51

[0358] Synthesis of methyl 2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)benzoate (A52) and 2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5- hydroxy-3-methyl-lH-pyrazol-4-yl)methyl)benzoic acid (A50)

[0359] Refer to the synthesis method of A20, with intermediate 22 and methyl 2-formylbenzoate as starting materials, to obtain A52, light yellow solid, yield 30.1%. ESI-MS: 642.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.69 (s, 2H), 7.87 (dd, J = 7.1, 2.0 Hz, 2H), 7.83 (d, J = 7.9 Hz, 1H), 7.78-7.74 (m, 1H), 7.67 (t, J = 7.7 Hz, 1H), 7.51 (t, J = 7.6 Hz, 1H), 7.39-7.31 (m, 4H), 5.39 (s, 1H), 2.02 (s, 6H).

[0360] Dissolve A52 (30 mg, 0.047 mmol) in methanol (1 mL), add sodium hydroxide aqueous solution (7.5 mg in 250 μL H2O, 0.19 mmol), and warm to 70 °C for 1 h. After the reaction is completed, filter out the solid to obtain A50, light yellow solid 27 mg, yield 92.0%. ESI-MS: 628.8 [M-H] - ; 1H NMR (500 MHz, DMSO-d6) δ 12.90 (s, 1H), 12.51 (s, 2H), 7.87 (dd, J = 7.0, 2.0 Hz, 2H), 7.72 - 7.67 (m, 1H), 7.51 - 7.46 (m, 1H), 7.41 - 7.31 (m, 6H), 5.97 (s, 1H), 2.01 (s, 6H).

[0361] Example 52

[0362] Synthesis of 4,4'-((3-hydroxyphenyl)methylene)bis(l-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl- lH-pyrazol-5-ol) (A51)

[0363] Refer to the synthesis method of A20, using intermediate 22 and m-hydroxybenzaldehyde as starting materials, white solid was obtained in 64.5% yield. ESI-MS: 600.8 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.58 (s, 2H), 9.29 (s, 1H), 7.88 (dd, J = 7.0, 2.0 Hz, 2H), 7.38 - 7.32 (m, 4H), 7.12 (t, J = 7.8 Hz, 1H), 6.74 - 6.66 (m, 2H), 6.63 (dd, J = 7.9, 2.4 Hz, 1H), 5.06 (s, 1H), 2.11 (s, 6H).

[0364] Example 53

[0365] Synthesis of 3-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-lH-pyrazol-4-yl)methyl)benzamide (A53)

[0366] Refer to the synthesis method of A20, using intermediate 22 and 3-formylaminomethyl formamide as starting materials, white solid was obtained in 93.4% yield. ESI-MS: 627.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.65 (s, 2H), 7.98 (s, 1H), 7.88 (dd, J = 7.1, 1.9 Hz, 2H), 7.80 (s, 1H), 7.76 (d, J = 7.5 Hz, 1H), 7.45 - 7.40 (m, 2H), 7.37 - 7.32 (m, 5H), 5.20 (s, 1H), 2.10 (s, 6H).

[0367] Example 54

[0368] 4,4'-((2-(1H-1,2,4-triazol-1-yl)phenyl)methylene)bis(1-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-5-ol) (A54)

[0369] Refer to the synthesis method of A20, using intermediate 22 and 2-(1H-1,2,4-triazol-1-yl)benzaldehyde as starting materials, white solid was obtained in 53.4% yield. ESI-MS: 651.8 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.53 (s, 2H), 8.64 (s, 1H), 8.03 (s, 1H), 7.86 (dd, J = 7.1, 1.9 Hz, 2H), 7.57-7.52 (m, 2H), 7.49 (m, 1H), 7.43 (dd, J = 7.5, 1.6 Hz, 1H), 7.37-7.32 (m, 4H), 5.42 (s, 1H), 2.05 (s, 6H).

[0370] Example 55

[0371] Synthesis of tert-butyl (2-(bis(1-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-4-yl)methyl)phenyl)carbamate (A55) and 4,4'-((2-aminophenyl)methylene)bis(1-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-5-ol) (A57)

[0372] Refer to the synthesis method of A20, using intermediate 22 and tert-butyl (2-formylphenyl)carbamate as starting materials, A55 was obtained as white solid in 63.9% yield. ESI-MS: 699.8 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.45 (s, 2H), 8.50 (s, 1H), 7.87 (dd, J = 7.0, 2.1 Hz, 2H), 7.36-7.31 (m, 4H), 7.29-7.23 (m, 2H), 7.18-7.15 (m, 2H), 5.47 (s, 1H), 2.02 (s, 6H), 1.21 (s, 9H).

[0373] To A55 (40 mg, 0.057 mmol) was added trifluoroacetic acid (1 mL) and reacted at room temperature for 3 h. After the reaction was completed, it was rotary evaporated to give a yellow solid 40 mg, which was A57 as trifluoroacetate salt form in 98.1% yield. ESI-MS: 713.5 [M-H]- ; 1 H NMR (500 MHz, DMSO-d6) δ 12.66 (br s, 1H), 10.53 (s, 1H), 7.92 (dd, J = 7.9, 1.1 Hz, 1H), 7.40 - 7.27 (m, 3H), 7.24 - 7.20 (m, 1H), 7.07 - 6.94 (m, 5H), 6.91 - 6.84 (m, 1H), 5.31 (s, 1H), 4.51 (d, J = 13.3 Hz, 1H), 4.06 (d, J = 13.2 Hz, 1H), 2.18 (s, 3H), 1.99 (s, 3H).

[0374] Example 56

[0375] Synthesis of tert-butyl (3-(bis(1-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-1H- pyrazol-4-yl)methyl)phenyl)carbamate (A56)

[0376] Refer to the synthetic method of A20, using intermediate 22 and tert-butyl (3-formylphenyl)carbamate as starting materials, white solid was obtained in 95.9% yield. ESI-MS: 699.9 [M-H] - ; 1 H NMR (500 MHz, DMSO-d6) δ 12.66 (br s, 1H), 10.53 (s, 1H), 7.92 (dd, J = 7.9, 1.1 Hz, 1H), 7.40 - 7.27 (m, 3H), 7.24 - 7.20 (m, 1H), 7.07 - 6.94 (m, 5H), 6.91 - 6.84 (m, 1H), 5.31 (s, 1H), 4.51 (d, J = 13.3 Hz, 1H), 4.06 (d, J = 13.2 Hz, 1H), 2.18 (s, 3H), 1.99 (s, 3H).

[0377] Example 57

[0378] Synthesis of 4,4'-((3''-aminophenyl)methylene)bis(1-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl- 1H-pyrazol-5-ol) (A57)

[0379] Refer to the synthetic method of A57, using A56 as starting material, white solid was obtained in 95.3% yield as trifluoroacetate salt. ESI-MS: 713.6 [M-H] - ; 1H NMR (500 MHz, DMSO-d6) δ 12.41 (s, 2H), 7.88 (dd, J = 7.2, 1.9 Hz, 2H), 7.45 - 7.26 (m, 5H), 7.19 - 6.96 (m, 3H), 5.15 (s, 1H), 2.14 (s, 6H).

[0380] Example 58

[0381] Synthesis of 2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-lH- pyrazol-4-yl)methyl)benzenesulfonic acid sodium salt (A59)

[0382] According to the synthetic method of A20, using intermediate 22 and 2-formylbenzenesulfonic acid sodium salt as starting materials, a yellow solid was obtained in 93.6% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.11 (s, 2H), 7.90 (dd, J = 7.8, 1.4 Hz, 1H), 7.86 - 7.82 (m, 3H), 7.35 - 7.29 (m, 5H), 7.24 - 7.20 (m, 1H), 6.23 (s, 1H), 1.97 (s, 6H).

[0383] Example 59

[0384] Synthesis of methyl 3-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)benzoate (A60) and 3-(bis(l-(4-fluorobenzo[d]thiazol-2- yl)-5-hydroxy-3-methyl-lH-pyrazol-4-yl)methyl)benzoic acid (A61)

[0385] According to the synthetic method of A20, using intermediate 22 and methyl 3- formylbenzoate as starting materials, a white solid was obtained, which was A60 in 58.0% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.71 (s, 2H), 7.94 - 7.83 (m, 4H), 7.63 - 7.57 (m, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.39 - 7.32 (m, 4H), 5.24 (s, 1H), 3.82 (s, 3H), 2.12 (s, 6H).

[0386] According to the synthetic method of A50, using A60 as starting material, a white solid was obtained in 95.3% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.94 (br s, 1H), 12.70 (s, 2H), 7.99 - 7.71 (m, 4H), 7.56 (d, J = 7.8 Hz, 1H), 7.47 (t, J = 7.7 Hz, 1H), 7.42 - 7.20 (m, 4H), 5.22 (s, 1H), 2.12 (s, 6H).

[0387] Example 60

[0388] Synthesis of 3-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-lH- pyrazol-4-yl)methyl)benzonitrile (A62)

[0389] According to the synthetic method of A20, using intermediate 22 and m-cyanobenzaldehyde as starting materials, white solid was obtained in 59.6% yield. 1 H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 2H), 7.89 - 7.85 (m, 2H), 7.37 - 7.30 (m, 5H), 7.23 (t, J = 7.8 Hz, 1H), 6.93 (dd, J = 11.4, 7.8 Hz, 2H), 5.34 (s, 1H), 4.81 (s, 2H), 3.61 (s, 3H), 2.04 (s, 6H).

[0390] Example 61

[0391] Synthesis of methyl 2-(2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)phenoxy)acetate (A63) and 2-(2-(bis(l-(4-fluorobenzo[d]thiazol- 2-yl)-5-hydroxy-3-methyl-lH-pyrazol-4-yl)methyl)phenoxy)acetic acid (A64)

[0392] According to the synthetic method of A20, using intermediate 22 and methyl 2-(2- formylphenoxy)acetate as starting materials, white powdery solid, A63, was obtained in 87.9% yield. 1 H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 2H), 7.89 - 7.85 (m, 2H), 7.37 - 7.30 (m, 5H), 7.23 (t, J = 7.8 Hz, 1H), 6.93 (dd, J = 11.4, 7.8 Hz, 2H), 5.34 (s, 1H), 4.81 (s, 2H), 3.61 (s, 3H), 2.04 (s, 6H).

[0393] The synthesis was performed according to the procedure described for A50, using A63 as starting material to give a white solid in 96.7% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.42 (br s, 2H), 7.92-7.83 (m, 2H), 7.40-7.30 (m, 5H), 7.23 (t, J = 7.9 Hz, 1H), 6.97-6.89 (m, 2H), 5.37 (s, 1H), 4.70 (s, 2H), 2.05 (s, 6H).

[0394] Example 62

[0395] Synthesis of intermediate 2-(1H-tetrazol-5-yl)benzaldehyde (25)

[0396] A solution of sec-butyllithium (5.53 mL, 7.18 mmol, 1.3 M in n-Hexane) was added dropwise to a suspension of 5-phenyl-1H-tetrazole (500 mg, 3.42 mmol) and dry tetrahydrofuran (10 mL) at -78 °C under nitrogen protection. After 0.5 h, dry DMF (2 mL) was added dropwise and the reaction was allowed to proceed for 0.5 h. After the reaction was completed, 2N hydrochloric acid (5 mL) was added to quench the reaction and the reaction was allowed to warm to room temperature. The reaction was extracted with 3 x 25 mL ethyl acetate and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated and recrystallized from ethyl acetate-n-hexane to give a white solid 122 mg in 20.5% yield.1H NMR (400 MHz, CDCl3) δ 10.14 (s, 1H), 8.86 (d, J = 7.8 Hz, 1H), 8.10 (dd, J = 7.6, 1.6 Hz, 1H), 7.92 (t, J = 1.6 Hz, 1H), 7.85 (t, J = 1.4 Hz, 1H).

[0397] Synthesis of 4,4'-((2-(1H-tetrazol-5-yl)phenyl)methylene)bis(1-(4- fluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-5-ol) (A65)

[0398] The synthesis was performed according to the procedure described for A20, using intermediate 22 and 25 as starting materials to give a white solid in 89.2% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.54 (s, 2H), 7.86 (dd, J = 6.8, 2.1 Hz, 2H), 7.60-7.46 (m, 4H), 7.38-7.31 (m, 4H), 5.69 (s, 1H), 2.03 (s, 6H).

[0399] Example 63

[0400] 4,4'-(naphthalen-1-ylmethylene)bis(1-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-5-ol) (A66)

[0401] Refer to the synthesis method of A20, using intermediate 22 and 1-naphthaldehyde as starting materials, white solid was obtained with a yield of 81.3%. 1 H NMR (500 MHz, DMSO-d6) δ 12.64 (s, 2H), 8.00 - 7.96 (m, 1H), 7.89 (dd, J = 7.1, 1.8 Hz, 3H), 7.87 - 7.84 (m, 1H), 7.54 - 7.51 (m, 2H), 7.51 - 7.47 (m, 2H), 7.39 - 7.32 (m, 4H), 5.70 (s, 1H), 1.92 (s, 6H).

[0402] Example 64

[0403] Synthesis of 2,2'-((2-(trifluoromethyl)phenyl)methylene)bis(5-methoxy-3-methyl-1H-pyrazole-4,1-diyl))bis(4-fluorobenzo[d]thiazole) (A67)

[0404] A49 (100 mg, 0.153 mmol), potassium carbonate (169 mg, 1.22 mmol) and iodomethane (38 μL, 0.611 mmol) were added into DMF (2 mL), and warmed to 90 °C for 2 h. After the reaction was completed, ethyl acetate was extracted, saturated brine was washed, and the organic phase was collected and concentrated, and column chromatography was separated (petroleum ether: ethyl acetate = 2:1) to obtain white solid 45 mg with a yield of 43.2%. ESI-MS: 683.1 [M+H] + ; 1 H NMR (600 MHz, Chloroform-d) δ 7.78 (d, J = 7.9 Hz, 1H), 7.72 - 7.67 (m, 1H), 7.58 (dd, J = 8.0, 1.0 Hz, 2H), 7.53 - 7.49 (m, 1H), 7.40 (t, J = 7.7 Hz, 1H), 7.28 - 7.27 (m, 1H), 7.26 - 7.24 (m, 1H), 7.17 - 7.13 (m, 2H), 5.60 (s, 1H), 3.69 (s, 6H), 2.15 (s, 6H).

[0405] Example 65

[0406] Synthesis of 2-(bis(1-(4-fluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-4-yl)methyl)pyrrolidine-1-carboxylic acid (A68)

[0407] A49 (50 mg, 0.076 mmol) and TEA (43 μL, 0.306 mmol) were added into DCM (2 mL), acetyl chloride (14 μL, 0.191 mmol) was added dropwise slowly at 0 °C, and the reaction was allowed to warm to room temperature for 12 h after the addition was completed. After the reaction was completed, column separation (petroleum ether: ethyl acetate = 5:1) gave 18 mg of white solid, with a yield of 32.0%. ESI-MS: 738.7 [M+H] + ; 1 H NMR (500 MHz, Methanol-d4) δ 7.81 (d, J = 8.0 Hz, 1H), 7.74 (d, J = 8.1 Hz, 2H), 7.67 (d, J = 4.2 Hz, 2H), 7.60 - 7.51 (m, 1H), 7.37 (td, J = 8.1, 4.5 Hz, 2H), 7.23 (dd, J = 10.7, 8.1 Hz, 2H), 5.59 (s, 1H), 2.17 (s, 6H), 2.08 (s, 6H).

[0408] Example 66

[0409] Synthesis of 2-(bis(1-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-4-yl)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (A70)

[0410] Referring to the synthesis method of A20, intermediate 22 and tert-butyl 2-formylpyrrolidine-1-carboxylate were used as starting materials to obtain peach red solid with a yield of 61.3%. 1 H NMR (500 MHz, DMSO-d6) δ 12.56 (s, 1H), 12.40 (s, 1H), 7.91 - 7.82 (m, 2H), 7.41 - 7.28 (m, 4H), 4.80 (s, 1H), 3.62 - 3.50 (m, 1H), 2.43 (s, 2H), 2.33 (s, 1H), 2.24 (s, 3H), 1.87 (d, J = 24.2 Hz, 3H), 1.76 (s, 1H), 1.45 - 1.30 (m, 2H), 1.15 (s, 9H).

[0411] Example 67

[0412] Synthesis of tert-butyl 3-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3- methyl-lH-pyrazol-4-yl)methyl)pyrrolidine-l-carboxylate (A71)

[0413] Refer to the synthesis method of A20, using intermediate 22 and tert-butyl 3- formylpyrrolidine-l-carboxylate as starting materials, a white solid was obtained in 80.5% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.56 (s, 1H), 12.51 (s, 1H), 7.88 (dt, J = 7.3, 2.0 Hz, 2H), 7.38 - 7.32 (m, 4H), 3.50 (s, 2H), 3.41 - 3.37 (m, 2H), 3.30 - 3.18 (m, 2H), 2.91 - 2.84 (m, 1H), 2.48 - 2.39 (m, 6H), 1.56 - 1.50 (m, 1H), 1.38 (d, J = 14.3 Hz, 9H).

[0414] Example 68

[0415] Synthesis of 4,4'-(pyrrolidin-3-ylmethylene)bis(l-(4-fluorobenzo[d]thiazol-2-yl)-3- methyl-lH-pyrazol-5-ol) hydrochloride (A72)

[0416] Refer to the synthesis method of A57, using A71 and hydrochloric acid dioxane solution (4 M) as starting materials, an orange-white solid 200 mg was obtained, which was A72 in the form of hydrochloride, in 98.9% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.66 (s, 2H), 9.08 (s, 1H), 8.96 (s, 1H), 7.88 (dd, J = 7.3, 1.7 Hz, 2H), 7.41 - 7.31 (m, 4H), 3.67 - 3.59 (m, 2H), 3.32 - 3.26 (m, 2H), 3.19 - 3.14 (m, 1H), 2.84 - 2.79 (m, 1H), 2.43 (d, J = 3.0 Hz, 6H), 2.06 - 2.00 (m, 1H), 1.60 - 1.54 (m, 1H).

[0417] Example 69

[0418] Synthesis of ethyl 2-(3-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-lH- pyrazol-4-yl)methyl)pyrrolidin-l-yl)-2-oxoacetate (A73)

[0419] A solution of TEA (40 μL, 0.353 mmol) in DCM (1 mL) was added to a solution of A72 (178 mg, 0.294 mmol) in DCM (2 mL) under nitrogen protection, and a solution of ethyl 2-chloro-2-oxoacetate (164 μL, 1.180 mmol) in DCM (2 mL) was added dropwise slowly. After the addition was completed, the reaction was allowed to react at room temperature for 1 h. After the reaction was completed, the product was obtained by washing with ethyl acetate / petroleum ether to obtain a white solid 201 mg, with a yield of 101%. 1 H NMR (500 MHz, DMSO-d6) δ 12.56 (d, J = 21.9 Hz, 2H), 7.94 - 7.82 (m, 2H), 7.45 - 7.24 (m, 4H), 4.72 - 4.55 (m, 2H), 3.33 - 3.22 (m, 1H), 3.05 (s, 1H), 2.97 - 2.88 (m, 1H), 2.48 - 2.39 (m, 6H), 2.06 (d, J = 10.2 Hz, 3H), 1.95 - 1.87 (m, 1H), 1.70 - 1.59 (m, 1H), 1.53 - 1.45 (m, 1H), 1.30 - 1.20 (m, 1H), 0.88 - 0.80 (m, 1H).

[0420] Example 70

[0421] Synthesis of 4,4'-(pyrrolidin-2-ylmethylene)bis(l-(4-fluorobenzo[d]thiazol-2-yl)-3- methyl-lH-pyrazol-5-ol) hydrochloride (A74)

[0422] Referring to the synthesis method of A72, A70 and a solution of hydrogen chloride in dioxane (4 M) were used as raw materials to obtain an orange solid 100 mg, which was A74 in the form of a hydrochloride salt, with a yield of 98.9%. 1 H NMR (500 MHz, DMSO-d6) δ 12.96 (s, 2H), 9.18 (s, 1H), 8.93 (s, 1H), 7.94 - 7.85 (m, 2H), 7.43 - 7.31 (m, 4H), 4.65 - 4.59 (m, 1H), 4.02 (d, J = 10.8 Hz, 1H), 3.31 - 3.19 (m, 2H), 2.45 (d, J = 15.6 Hz, 6H), 2.14 - 2.07 (m, 1H), 2.01 - 1.91 (m, 2H), 1.65 - 1.57 (m, 1H).

[0423] Example 71

[0424] Synthesis of 2-(2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)pyrrolidin-l-yl)-2-oxoethyl acetate (A75)

[0425] Referring to the synthesis method of A73, using TEA, A74 and ethyl 2-chloro-2-oxoacetate as raw materials, 77 mg of white solid was obtained, which was A75, with a yield of 85.6%.

[0426] Example 72

[0427] Synthesis of l-(2-(bis(l-(4-fluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)pyrrolidin-l-yl)-2-hydroxyethan-l-one (A76)

[0428] A solution of NaOH (9.41 mg, 0.235 mmol) in H2O (0.5 mL) was added to a solution of A75 (40 mg, 0.059 mmol) in MeOH (2 mL) and reacted at 70 °C for 1 h. After the reaction was completed, it was rotary evaporated, dissolved in water and suction filtered, and the filtrate was adjusted to pH 1-2 and suction filtered to obtain 26 mg of light red solid with a yield of 68.3%. 1 H NMR (500 MHz, DMSO-d6) δ 12.66 (s, 1H), 12.24 (s, 1H), 7.92 - 7.84 (m, 2H), 7.39 - 7.31 (m, 4H), 5.22 (s, 0.5H), 4.89 (s, 0.5H), 4.00 (s, 0.5H), 3.97 (s, 0.5H), 3.83 (s, 0.5H), 3.80 (s, 0.5H), 3.61 (s, 0.5H), 3.59 (s, 0.5H), 3.47 (s, 1H), 3.44 (s, 1H), 2.40 (s, 1H), 2.30 (s, 2H), 2.25 (d, J = 9.7 Hz, 3H), 1.88 (d, J = 7.5 Hz, 4H).

[0429] Example 73

[0430] Synthesis of 4,4'-((5-bromo-2-fluorophenyl)methylene)bis(l-(4,6-difluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A77)

[0431] Intermediate 24 (50 mg, 0.187 mmol) and 5-bromo-2-fluorobenzaldehyde (22.8 mg, 0.112 mmol) were added into n-butanol and heated to 100 °C for 12 h. After the reaction was completed, the solid was filtered and recrystallized from ethanol to give 62 mg of white solid, yield 86.2%. 1 H NMR (500 MHz, DMSO-d6) δ 12.81 (s, 2H), 7.87-7.84 (m, 2H), 7.52 (d, J = 5.8 Hz, 2H), 7.46-7.42 (m, 2H), 7.20 (t, J = 9.3 Hz, 1H), 5.21 (s, 1H), 2.14 (s, 6H).

[0432] Example 74

[0433] Synthesis of 4,4'-((5-chloro-2-fluorophenyl)methylene)bis(l-(4,6-difluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A78)

[0434] Following the procedure for the synthesis of A77, using intermediate 24 and 5-chloro-2-fluorobenzaldehyde as starting materials, 58 mg of off-white solid was obtained, which was A78, yield 85.9%. 1 H NMR (500 MHz, DMSO-d6) δ 12.81 (s, 2H), 7.87-7.84 (m, 2H), 7.52 (d, J = 5.8 Hz, 2H), 7.46-7.42 (m, 2H), 7.20 (t, J = 9.3 Hz, 1H), 5.21 (s, 1H), 2.14 (s, 6H).

[0435] Example 75

[0436] Synthesis of 4,4'-((2-fluoro-5-(trifluoromethyl)phenyl)methylene)bis(l-(4,6-difluorobenzo[d]thiazol-2-yl)-3-methyl-lH-pyrazol-5-ol) (A79)

[0437] Following the procedure for the synthesis of A78, using intermediate 24 and 2-fluoro-5- trifluoromethylbenzaldehyde as starting materials, 58 mg of off-white solid was obtained, which was A79, yield 61.9%. 1 H NMR (500 MHz, DMSO-d6) δ 12.85 (s, 2H), 7.85 (dd, J = 8.4, 2.5 Hz, 2H), 7.80 (d, J = 6.9 Hz, 1H), 7.78-7.75 (m, 1H), 7.48-7.41 (m, 3H), 5.30 (s, 1H), 2.16 (s, 6H).

[0438] Example 76

[0439] Synthesis of methyl 2-(bis(1-(4,6-difluorobenzo[d]thiazol-2-yl)-5-hydroxy-3- methyl-1H-pyrazol-4-yl)methyl)benzoate (A80)

[0440] Referring to the synthesis method of A52, intermediate 24 and methyl 2-formylbenzoate were used as starting materials to obtain 97 mg of white solid A80, with a yield of 76.2%. 1 H NMR (500 MHz, DMSO-d6) δ 12.62 (s, 2H), 7.87-7.84 (m, 2H), 7.65 (dd, J = 7.9, 1.6 Hz, 1H), 7.54-7.50 (m, 1H), 7.46-7.42 (m, 2H), 7.41-7.37 (m, 2H), 5.75 (s, 1H), 3.59 (s, 3H), 2.01 (s, 6H).

[0441] Example 77

[0442] Synthesis of 2-(bis(1-(4,6-difluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl-1H- pyrazol-4-yl)methyl)benzoic acid (A81)

[0443] Referring to the synthesis method of A51, A80 was used as the starting material to obtain 42 mg of orange-yellow solid A81, with a yield of 95.3%. 1 H NMR (500 MHz, DMSO-d6) δ 12.55 (s, 2H), 7.87-7.84 (m, 2H), 7.70 (dd, J = 7.5, 1.5 Hz, 1H), 7.51-7.47 (m, 1H), 7.45-7.41 (m, 2H), 7.40-7.36 (m, 2H), 5.96 (s, 1H), 2.00 (s, 6H).

[0444] Example 78

[0445] Synthesis of 4,4'-((2-fluoro-6-(trifluoromethyl)phenyl)methylene)bis(1-(4,6-difluorobenzo[d]thiazol-2-yl)-3-methyl-1H-pyrazol-5-ol) (A82)

[0446] Sodium acetate (15.3 mg, 0.187 mmol) was added to a solution of intermediate 24 (50.0 mg, 0.187 mmol) and 2-trifluoromethyl-5-fluorobenzaldehyde (30.70 mg, 0.094 mmol) in acetic acid and the reaction was heated to 100 °C for 1 h. After completion of the reaction, the reaction mixture was filtered and washed with a small amount of DCM to give 33 mg of a white solid in 44.0% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.71 (s, 2H), 7.85 (ddd, J = 8.4, 2.5, 1.0 Hz, 2H), 7.63 (dd, J = 8.0, 1.4 Hz, 1H), 7.59 - 7.54 (m, 1H), 7.49 (ddd, J = 9.9, 8.4, 1.3 Hz, 1H), 7.44 (ddd, J = 11.0, 9.6, 2.5 Hz, 2H), 5.35 (s, 1H), 2.01 (s, 6H).

[0447] Example 79

[0448] Synthesis of 2-(3-(bis(l-(4,6-difluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)phenyl)acetic acid (A83)

[0449] Following the procedure for the synthesis of A37, intermediate 24 and 2-(3- formylphenyl)acetic acid were used as starting materials to give 55 mg of a yellowish solid, A81, in 87.3% yield. 1 H NMR (500 MHz, DMSO-d6) δ 12.62 (s, 2H), 7.86 (ddd, J = 8.2, 2.5, 1.0 Hz, 2H), 7.44 (ddd, J = 10.9, 9.5, 2.5 Hz, 2H), 7.27 (t, J = 7.6 Hz, 1H), 7.18 (d, J = 1.9 Hz, 1H), 7.16 - 7.11 (m, 2H), 5.13 (s, 1H), 3.55 (s, 2H), 2.08 (s, 6H).

[0450] Example 80

[0451] Synthesis of 3-(bis(l-(4,6-difluorobenzo[d]thiazol-2-yl)-5-hydroxy-3-methyl- lH-pyrazol-4-yl)methyl)benzoic acid (A84)

[0452] Following the procedure for the synthesis of A82, intermediate 24, 3- formylbenzoic acid and sodium acetate were used as starting materials to give 47 mg of a yellowish solid, A84, in 75.4% yield. 1H NMR (500 MHz, DMSO-d6) δ 12.93 (s, 1H), 12.71 (s, 2H), 7.87 - 7.81 (m, 4H), 7.55 (d, J = 8.4 Hz, 1H), 7.48 - 7.42 (m, 3H), 5.22 (s, 1H), 2.10 (s, 6H).

[0453] Example 81: Human cGAS enzyme level inhibition activity of compounds of the application

[0454] 1. Experimental methods

[0455] (1) Protein purification

[0456] The nucleic acid sequence (synthesized by GenScript) of human full-length cGAS protein (h-cGAS FL )(amino acids 1-522, Uniprot ID: Q8N884) was cloned into pET-15b vector with N-terminal 6xHis-SUMO2. The constructed plasmid was then transformed into Rosetta 2 (DE3) cells (Sigma-Aldrich) and cultured in 2xYT medium at 37 °C, when the OD 600 reached 0.8-1.0, the culture temperature was reduced to 17 °C, and protein expression was induced by adding 0.4 mM IPTG. After 8-10 hours of induction, the cells were collected by centrifugation at 5000 rpm for 30 min. The cells were resuspended with lysis buffer (20 mM HEPES, 400 mM NaCl, pH 7.5) and lysed in a high-pressure disruptor. After centrifugation of the cell lysate at 22000 rpm for 30 min, the supernatant was collected and loaded onto a manually packed Ni-NTA affinity column (GE Healthcare), which was then washed with wash buffer (20 mM HEPES, 1 M NaCl, 50 mM imidazole, pH 7.5) and eluted with elution buffer (20 mM HEPES, 400 mM NaCl, 300 mM imidazole, pH 7.5). To remove the 6xHis-SUMO2 tag, SENP2 protease was added to the eluted protein and incubated at 4 °C for 4 h. The untagged cGAS was further purified by a Heparin column (GE Healthcare), followed by gel filtration using size exclusion chromatography (HiLoad 16 / 600 Superdex 200 pg, GE Healthcare). The gel filtration buffer for h-cGAS FL was 20 mM HEPES, 250 mM KCl, 1 mM TCEP, pH 7.5.

[0457] (2) Enzyme activity test by pyrophosphate coupling method

[0458] The compound (positive group is DMSO) was incubated with h-cGAS FL After co-incubation for 20 min, ATP, GTP, pyrophosphatase and HT-DNA (control group without HT-DNA) were added to the transparent 384 plate, and the total reaction system was 40 μL (10 mM HEPES, 140 mM NaCl, 0.01% Tween-20, 5 mM MgCl2, pH 7.5), and co-incubation was performed for 60-90 min. 40 μL of EDTA and 20 μL of phosphoric acid developing solution were added to the above reaction system, mixed, and incubated for 20 min, and then the OD 620 absorbance value was read by an enzyme marker.

[0459] 2. Experimental results

[0460] Inhibition activity of the compound on h-cGAS FL enzyme level (IC 50 ) value is shown in Table 1. The IC 50 value reported ranges: A indicates <10 μM, B indicates 10-50 μM, and C indicates >50 μM. The experimental results show that the IC 50 values of most of the compounds for human cGAS inhibition are less than 50 μM, and the IC 50 values of some compounds are less than 10 μM, indicating that the compounds provided by the present application generally have good inhibition activity on h-cGAS FL .

[0461] Table 1

[0462] Example 2: Inhibition activity of the compound of the present application on mouse cGAS enzyme level

[0463] 1. Experimental method

[0464] (1) Protein expression

[0465] The nucleic acid sequence (synthesized by GenScript) of the mouse catalytic domain protein (m-cGAS CD ) (amino acids 147-507, Uniprot ID: Q8C6L5) was cloned into the pET-15b vector with N-terminal 6xHis-SUMO2, wherein a serine was introduced between the SUMO2 and the N-terminal of m-cGAS CD . Then the constructed plasmid was transformed into E. coli strain BL21 (DE3) pLysS (Bi Yun Tian), and cultured in 2x YT medium at 37°C, and when the OD 600When the expression level reached 0.8-1.0, the culture temperature was lowered to 17°C, and protein expression was induced by adding 0.4 mM IPTG. After induction for 8-10 hours, cells were collected by centrifugation at 5000 rpm for 30 min. Cells were resuspended in lysis buffer (20 mM HEPES, 400 mM NaCl, pH 7.5) and lysed using an autoclave. After centrifugation at 22000 rpm for 30 min, the supernatant was collected and loaded onto a manually packed Ni-NTA affinity column (GE Healthcare). The column was washed with wash buffer (20 mM HEPES, 1 M NaCl, 50 mM imidazole, pH 7.5), and the recombinant protein was eluted with elution buffer (20 mM HEPES, 400 mM NaCl, 300 mM imidazole, pH 7.5). To remove the 6×His-SUMO2 tag, SENP2 protease was added to the eluted protein, and the cells were incubated at 4°C for 4 hours. Unlabeled cGAS was further fractionated using a Heparin column (GE Healthcare) followed by gel filtration using size exclusion chromatography (HiLoad 16 / 600 Superdex 200pg, GE Healthcare). This was used for m-cGAS. CD The buffer solution is 20 mM Tris, 300 mM NaCl, 1 mM DTT, pH 7.5.

[0466] (2) Enzyme activity assay by pyrophosphate coupling method

[0467] The compound (DMSO in the positive group) was mixed with m-cGAS CD After co-incubating for 20 min, ATP, GTP, pyrophosphatase, and HT-DNA (no HT-DNA added to the control group) were added to a clear 384 plate. The total reaction volume was 40 μL (10 mM HEPES, 140 mM NaCl, 0.01% Tween-20, 5 mM MgCl2, pH 7.5), and co-incubated for 60–90 min. Then, 40 μL of EDTA and 20 μL of phosphate chromogenic solution were added to the above reaction volume, mixed well, and incubated for 20 min. The OD values ​​were then read using a microplate reader. 620 Absorbance value.

[0468] 2. Experimental Results

[0469] Preferred compounds for m-cGAS CD Inhibitory activity at enzyme level (IC50) 50 The values ​​are shown in Table 2. IC 50 The reported value ranges are: A represents <10 μM, B represents 10-50 μM, and C represents >50 μM. Experimental results show that the compound provided by this invention is effective against m-cGAS. CDAlso very good inhibitory activity.

[0470] Table 2

[0471] Example 83: Evaluation of the cellular activity of the compounds of the application on THP1-dual cells

[0472] 1. Experimental method

[0473] THP1-dual cells (InvivoGen) were seeded at a density of 5 x 10 5 cells per well in 24-well plates, incubated for 1 h with 20 μΜ of compound, then transfected with HT-DNA using Lipofectamine 2000 (Invitrogen) at the indicated titration and incubated for 24 h. Supernatants were collected and 20 μΐ, of supernatant per well was mixed with 50 μΐ, of QUANTI-Luc luciferase reagent (InvivoGen). Luciferase luminescence was immediately measured using a microplate reader (BioTek Synergy H1) with 1 s integration time per well.

[0474] 2. The experimental results are shown in Table 3.

[0475] Table 3

[0476] Example 84: Evaluation of the cellular activity of the preferred compounds on RAW-ISG cells

[0477] 1. Experimental method

[0478] RAW-ISG cells (InvivoGen) were seeded at a density of 5 x 10 5 cells per well in 24-well plates, incubated for 1 h with 20 μΜ of compound, then transfected with HT-DNA using Lipofectamine 2000 (Invitrogen) at the indicated titration and incubated for 24 h. Supernatants were collected and 20 μΐ, of supernatant per well was mixed with 50 μΐ, of QUANTI-Luc luciferase reagent (InvivoGen). Luciferase luminescence was immediately measured using a microplate reader (BioTek Synergy H1) with 1 s integration time per well.

[0479] 2. The experimental results are shown in Table 4.

[0480] Table 4

[0481] Example 85

[0482] The compound A46 of the present application has a unique mechanism of action, can produce a synergistic effect with other cGAS inhibitors, enhance the cell inhibition activity of other cGAS inhibitors, and thus achieve the effect of reducing the dosage. Therefore, the compound of the present application can be used in combination with other cGAS inhibitors to enhance the efficacy.

[0483] G150 is a reported cGAS inhibitor, and its mechanism of action is mainly to inhibit the activity of cGAS by binding to the active site of the catalytic domain of cGAS. This embodiment aims to illustrate the synergistic effect of the compound A46 of the present application and the known compound G150 in inhibiting cGAS at the cellular level.

[0484] Experimental method:

[0485] The synergistic effect of A46 and G150 was tested using the method of Example 83. A46 and G150 were combined at a molar ratio of 10:1, 5:1, 2.5:1, 1:1, 1:2.5, 1:5 and 1:10, with a final total concentration of 20 μM. The combined compounds were diluted by 2-fold gradient to obtain 5 concentration gradients. At the same time, A46 and G150 of the same concentration were prepared separately as controls. After THP-1 Dual cells were inoculated, the cells were treated with the compounds and transfected with double-stranded DNA. The luciferase activity was measured according to the protocol of Example 83. The combination index (CI) value was calculated using CompuSyn software when the maximum inhibition effect was 90% and 70% (ED90 and ED75). CI <1 indicates synergistic effect, CI =1 indicates additive effect, and CI >1 indicates antagonistic effect. The data in Figure 1 are expressed as the mean ± standard deviation of three independent experiments.

[0486] Experimental results:

[0487] The results are shown in Figure 1. When the proportion of A46 is higher (A46:G150 is 10, 5, 2.5), there is a significant synergistic effect (synergy coefficients are 0.469, 0.625, 0.711, respectively), and the synergistic effect is significantly enhanced as the proportion of A46 increases. On the contrary, when G150 is the main component, the synergistic effect is weakened.

[0488] Experimental conclusion:

[0489] The above results demonstrate that the compound A46 can produce a significant synergistic effect with the known cGAS inhibitor G150. It is shown that the combination of A46 with other cGAS inhibitors has good cell efficacy.

[0490] Example 86

[0491] This embodiment explores the protective effect of the compound A02 of the present application on acute pancreatitis induced by cerulein in mice.

[0492] cGAS knockout can effectively alleviate the condition of acute pancreatitis induced by cerulein in mice, therefore, this embodiment tests the protective effect of A02 on acute pancreatitis induced by cerulein in mice. As shown in Figure 2A, C57BL / 6J mice were randomly divided into three groups, 10 in each group. One group was given normal saline as a control group; two groups were induced by intraperitoneal injection of 50 μg / kg of cerulein every hour for 0-6h, respectively, as a modeling group and a drug administration group, and the drug administration group was intraperitoneally injected with a dose of 5 mg / kg of A02 twice at-1h and 6h, respectively. The mice were euthanized 12h after the last administration, and their body weight and pancreas after stripping were weighed, the same part of the pancreas was taken for formalin for hematoxylin-eosin staining, and the serum was taken for testing the amylase activity in the serum.

[0493] The results are shown in Figure 2, cerulein successfully induced acute pancreatitis in mice, the pancreas weight ratio of the modeling group was significantly higher than that of the control group, and the amylase in the serum was also significantly increased, and HE staining of pancreatic tissue sections showed that the pancreas had significant vacuolization, tissue dispersion, and more inflammatory cell infiltration, and the pathological score was significantly higher than that of the control group. The pancreas weight ratio of the drug administration group was significantly improved compared with the modeling group (p=0.0038), the amylase activity in the serum was reduced, and the tissue sections also showed that the tissue damage and inflammatory cell infiltration were significantly improved compared with the modeling group, and the pathological score was significantly lower than that of the modeling group (p=0.0005).

[0494] In summary, the inhibitor A02 targeting cGAS has obvious protective effect on acute pancreatitis induced by cerulein in mice.

[0495] All the documents mentioned in the present application are cited as references in the present application, just as each document is cited as a reference. In addition, it should be understood that those skilled in the art can make various modifications or modifications to the present application after reading the above teachings of the present application, and these equivalent forms also fall within the scope defined by the claims attached to the present application.

Claims

1. A compound of Formula (I), or a stereoisomer, tautomer, deuterated isotope, solvate, hydrate, prodrug, metabolite, pharmaceutically acceptable salt thereof, or a combination thereof, wherein, R1is selected from the group consisting of H, C1-C 10 alkyl, C3-C 10 cycloalkyl, 5-12 membered heteroaryl, C6-C 14 aryl, 4-12 membered heterocyclyl, C6-C 10 cycloalkenyl; said alkyl, cycloalkyl, aryl, heteroaryl, cycloalkenyl and heterocyclyl are optionally substituted with one or more substituents selected from the group consisting of hydroxy, thiol, nitro, halogen, cyano, amino, oxo (=0), sulfonate (-SO3Na), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester (-COO-C1-C6alkyl), C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl) substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl); wherein, said alkylene, alkoxy and alkyl are substituted by carboxyl, ester (-COO-C1-C6alkyl), oxo; R2 is selected from the following group: H, D, halogens, C1-C 10 Alkyl, C1-C 10 Alkoxy, C3-C8 cycloalkyl, C1-C 10 Haloalkyl, C1-C 10 Halogenated alkoxy groups; R3and R4are each independently selected from the group consisting of compounds of formula II: wherein, X is selected from the group consisting of N, O or S; Y is selected from the group consisting of CH, C or N; ring B is selected from the group consisting of five-membered heteroaryl and five-membered heterocyclyl; R5 is selected from the following group of substituted or unsubstituted groups: H, D, halogen, hydroxyl, amino, amine, mercapto, cyano, nitro, -C(O)R', C1-C 10 Alkyl, C1-C 10 Haloalkyl, C1-C 10 Alkoxy, C1-C 10 Halogenated alkoxy groups, C1-C 10 Alkylthio, C3-C 10 Cycloalkyl, 4-11 membered heterocyclic groups, C3-C8 cycloalkyl-O-, C3-C8 cycloalkyl-S-, C1-C 10 Alkyl-(C=O)-O-, C1-C 10 Alkyl-(C=S)-O-; R' is selected from the group consisting of substituted or unsubstituted H, C1-C 10 alkyl, C1-C 10 haloalkyl, C3-C 10 cycloalkyl; R6 is selected from the following group (substituted or unsubstituted): H, D, halogen, hydroxyl, amino, amine, mercapto, cyano, nitro, acyl, sulfonyl, amide, sulfonamide, C1-C 10 Alkyl, C1-C 10 Alkoxy, C3-C 10 Cycloalkyl, 4-11 membered heterocyclic, C6-C10 aryl, 5-11 membered heteroaryl, C3-C8 cycloalkyl-O-, C3-C8 cycloalkyl-S-; ring A is selected from the group consisting of substituted or unsubstituted 8-14 membered bicyclic or tricyclic heteroaryl, 8-14 membered saturated bicyclic or tricyclic heterocyclyl, 8-14 membered partially unsaturated bicyclic or tricyclic heterocyclyl, wherein the bicyclic or tricyclic rings are connected by spiro, bridging or fusing; said substitution means that one or more hydrogens on the group are replaced by a group selected from the group consisting of hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), sulfonate (-SO3Na), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester (-COO-C1-C6alkyl), urea, sulfone, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl-substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl)-substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl), -(C1-C6alkylene)-5-6 membered heteroaryl, -(C1-C6alkylene)-C3-C6cycloalkyl, -(C1-C6alkylene)-4-7 membered heterocyclyl, -(C1-C6alkylene)-phenyl, wherein the alkyl, alkylene, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, phenyl, heteroaryl are optionally substituted by a group selected from the group consisting of hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=0), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl; said heteroaryl or heterocyclyl contains 1-3 heteroatoms each independently selected from N, O and S.

2. The compound of claim 1, wherein R3and R4are each independently selected from the structure shown in Formula II-A: wherein, ring A, R5 and R6 are as described in claim 1.

3. The compound of claim 1, wherein Ring A is selected from the following group, either substituted or unsubstituted: C6-C 10 The compounds are aryl, five-membered heteroaromatic rings and six-membered heteroaromatic rings, benzo[5-membered heteroaromatic rings], benzo[6-membered heteroaromatic rings], and six-membered heteroaromatic rings and six-membered heteroaromatic rings, wherein the six-membered heteroaromatic ring is selected from the group consisting of pyridine, pyridazine, pyrimidine, pyrazine, and pyranone; and the five-membered heteroaromatic ring is selected from the group consisting of imidazole, thiazole, oxazole, furan, pyrazole, thiophene, and pyrrole. Preferably, ring A is selected from the group consisting of substituted or unsubstituted benzimidazole, benzothiazole, benzoxazole, benzofuran, benzopyrazole, benzothiophene, benzopyrrole, pyridoimidazole, pyridopyrazole, purine, quinoline, isoquinoline, indole. wherein the substitution means that one or more (e.g., 2, 3, 4, or 5) hydrogens on the group are replaced with a group independently selected from the group consisting of hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=O), sulfonate (-SO3Na), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester (-COO-C1-C6alkyl), urea, sulfone, C1-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl, C1-C6hydroxyalkyl, C1-C6carboxyalkyl, 5-6 membered heteroaryl, C3-C6cycloalkyl, 4-7 membered heterocyclyl, phenyl, -NHCOO-(C1-C6alkyl), carboxyl-substituted C1-C6alkoxy, ester (-COO-C1-C6alkyl)-substituted C1-C6alkoxy, -(C1-C6alkylene)-COO-(C1-C6alkyl), -(C1-C6alkylene)-5-6 membered heteroaryl, -(C1-C6alkylene)-C3-C6cycloalkyl, -(C1-C6alkylene)-4-7 membered heterocyclyl, -(C1-C6alkylene)-phenyl, wherein the alkyl, alkylene, alkenyl, alkynyl, alkoxy, alkylthio, cycloalkyl, heterocyclyl, phenyl, heteroaryl groups are optionally substituted with a group selected from the group consisting of hydroxyl, thiol, nitro, halogen, cyano, amino, oxo (=O), acyl, sulfonyl, sulfonamide, amine, carboxyl, amide (-CONH2), ester, C1-C6alkyl, C1-C6alkoxy, C1-C6alkylthio, C1-C6haloalkyl; More preferably, ring A is selected from the group consisting of:

4. The compound of claim 1, wherein The compound is selected from the group consisting of:

5. A method of making a compound of claim 1, or a stereoisomer, tautomer, deuterated isomer, solvate, hydrate, prodrug, metabolite, pharmaceutically acceptable salt thereof, comprising, comprising the steps of: The compound of formula IV is reacted with a compound of formula V in an inert solvent to obtain a compound of formula I; wherein ring A, ring B, X, Y, R1, R2, R5, and R6 are as described in claim 1.

6. A pharmaceutical composition, characterized by, A pharmaceutical composition comprising, as an active ingredient, a compound of formula I according to claim 1, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof; and a pharmaceutically acceptable carrier.

7. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises: a compound of formula I according to claim 1, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a pro-drug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, as a first active ingredient; a second active ingredient; and optionally a pharmaceutically acceptable carrier.

8. A compound of claim 1 of Formula I, or a stereoisomer, a tautomer, a deuterated isotope, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt thereof, or a combination thereof, or a pharmaceutical composition of claim 6, or a use of a pharmaceutical composition of claim 7, wherein, for the preparation of (a) a cGAS inhibitor; (b) a medicament for preventing, treating and / or adjuvant treatment of a disease associated with abnormal levels of activity and / or expression of cGAS. ​ 9. Use according to claim 8, characterized in that, The disease is selected from the group consisting of infectious diseases, inflammatory diseases, autoimmune diseases, neurodegenerative diseases, metabolic diseases, rare diseases, tumors, radiation damage; The disease is selected from the group consisting of infectious diseases, inflammatory diseases, autoimmune diseases, neurodegenerative diseases, metabolic diseases, rare diseases, tumors, radiation damage; wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus, ankylosing spondylitis, scleroderma, rheumatoid arthritis, multiple sclerosis, or a combination thereof; the neurodegenerative disease is selected from the group consisting of Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, Huntington's disease, or a combination thereof.

10. A method of inhibiting cGAS activity or expression, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-9. Comprising: administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I of claim 1, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt, or a combination thereof, or a pharmaceutical combination of claim 6, or a pharmaceutical composition of claim 7.

11. A method of enhancing the efficiency of DNA delivery into cells or in vivo, characterized by, Comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I of claim 1, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt, or a combination thereof, or a pharmaceutical combination of claim 6, or a pharmaceutical composition of claim 7.

12. A method of improving the manufacturing efficiency of chimeric antigen receptor T cells, characterized in that, Comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I of claim 1, or a stereoisomer, a tautomer, a deuterated analog, a solvate, a hydrate, a prodrug, a metabolite, a pharmaceutically acceptable salt, or a combination thereof, or a pharmaceutical combination of claim 6, or a pharmaceutical composition of claim 7.

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