Application of TRPC4 inhibitor in preparation of medicine for preventing or treating inflammatory bowel disease

Abstract

The invention discloses a novel application of a transient receptor potential cation channel subgroup C member 4 (TRPC4) inhibitor in preparation of a medicine for preventing or treating inflammatory bowel disease (IBD). The research finds that in a colitis mouse model, the enteritis symptom of the colitis mouse can be obviously improved by systemic gene knockout of TRPC4 or application of a TRPC4 inhibitor. Therefore, the TRPC4 is related to the disease progress of the colitis, and the TRPC4 can be used as a biomarker for screening patients and predicting the curative effect.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the use of TRPC4 (transient receptor potential cation channel subfamily C member 4) inhibitors in the preparation of drugs for preventing or treating inflammatory bowel disease (IBD). Background Art

[0002] Inflammatory bowel disease (IBD), which includes ulcerative colitis and Crohn's disease, is characterized by chronic disorders of the mucosal layer of the gastrointestinal system. Ulcerative colitis is a recurrent, chronic, nonspecific immune disease involving poorly characterized genetic factors, immune disorders, and environmental factors. The colon of patients with ulcerative colitis typically shows increased inflammatory cell infiltration, edema, crypt structural damage, and loss of the epithelial barrier, with colonic ulcers developing later in the disease. Not only is the etiology of ulcerative colitis complex, its pathological development is also driven by the interaction and mutual response of multiple cells. The integrity of the mucus layer and the integrity of the epithelial cells in the mucosal layer are important foundations for the normal physiological function of the intestine. Loss of the mucus layer and dysfunction of colonic epithelial cells can cause damage to the epithelial barrier, leading to increased permeability of the colonic mucosal layer, allowing pathogens in the intestinal lumen to enter the intestinal wall. Due to the invasion of these exogenous antigens, the intestinal immune response is activated.

[0003] The main medications for the treatment of inflammatory bowel disease include 5-aminosalicylic acid (5-ASA), prednisolone, and thiopurines. 5-ASA is commonly used for patients with mild to moderate ulcerative colitis, but this medication may cause adverse reactions such as leukopenia, hepatitis, and nausea, vomiting, abdominal pain, and bloating. Prednisolone is limited to patients with moderate to severe ulcerative colitis or those who do not respond to conventional treatments, but it can be addictive and should not be taken long-term. Thiopurines, such as azathioprine and mercaptopurine, have been used to alleviate ulcerative colitis without steroids. However, thiopurines can cause certain adverse reactions in people with thiopurine S-methyltransferase (TPMT) deficiency. Currently, many therapeutic targets are being explored, including sphingosine-1-phosphate receptor modulators (such as ozanimod and etrasimod), JAK inhibitors (such as upadacitinib), leukocyte inhibitors (such as ustekinumab), monoclonal antibodies (such as mrikizumab), and fecal microbiota transplantation. These novel targets provide new development directions for the drug treatment of ulcerative colitis, but their clinical efficacy and safety remain to be tested. Therefore, there is still an urgent clinical need to develop more stable, safe, and effective drugs for the treatment of inflammatory bowel disease.

[0004] Transient receptor potential cation channels (TRPs) are an important family of non-selective cation channels located on cell membranes and expressed in nearly all cells across all organs of the human body. TRP channels are divided into six subfamilies: TRPC (cannonical, TRPC1-7), TRPV (vanilloid, TRPV1-6), TRPM (melastatin, TRPM1-8), TRPP (polycystin, TRPP2, TRPP3, TRPP5), TRPML (mucolipin, TRPML1-3), and TRPA (ankyrin, TRPA1). These channels are involved in functions including vision, hearing, smell, taste, and somatic sensations (such as pain, mechanical stimulation, and temperature).

[0005] TRPC has the highest protein sequence similarity to Drosophila TRP channels. The TRPC subfamily contains 7 members, TRPC1-7. Based on sequence homology, TRPC channels are further divided into four subfamilies: TRPC1, TRPC2, TRPC4 / 5 and TRPC3 / 6 / 7 subfamilies. TRPC2 channels are not expressed in humans. Similar to Drosophila TRPs, TRPCs are localized to the plasma membrane and activated in a GPCR-PLC-dependent manner. TRPCs can also be activated by receptor tyrosine kinases, lysophospholipids, hypotonic solutions and mechanical stimulation. Activated TRPCs allow Ca 2+ and monovalent cations flow into the cell cytoplasm, leading to cell depolarization and intracellular Ca 2+ TRPCs are expressed in a variety of mammalian cell types and participate in numerous physiological and pathological processes. The development of gene knockout / knockin animal models and the use of drug tools are the primary strategies for studying the physiological and pathophysiological functions of TRPCs. Using these biological and pharmacological approaches, studies have shown that TRPC channels play an important role in the mammalian cardiovascular system, skeletal muscle, pancreatic β cells, neurons, bone, salivary gland cells, immune system, and many other organ systems.

[0006] However, there are currently no reports on the correlation between TPRC4 and IBD, no studies have proposed listing TRPC4 as a therapeutic target for IBD, and there are no drugs under development that target and regulate TRPC4 to treat IBD. Summary of the Invention

[0007] Purpose of the invention: The purpose of the present invention is to address the deficiencies of the prior art and provide a TRPC4 (transient receptor potential cation channel subfamily C member 4) inhibitor for use in the preparation of a drug for preventing or treating inflammatory bowel disease (IBD).

[0008] The present invention has found through systematic and in-depth research that inhibiting the function and / or expression of TRPC4 may be an effective strategy for preventing or treating IBD, especially ulcerative colitis (UC).

[0009] Technical solution: The purpose of the present invention is achieved through the following technical solution:

[0010] The present invention provides use of a TRPC4 inhibitor in preparing a medicament for preventing or treating inflammatory bowel disease (IBD).

[0011] Through data analysis of gene expression datasets, the present invention found that TRPC4 mRNA expression levels in the colon tissue of UC patients were significantly higher than in healthy controls. Furthermore, the present invention also found that in a colitis mouse model, systemic knockout of TRPC4 or administration of a TRPC4 inhibitor to colitis mice downregulated TRPC4 mRNA and protein expression in colon tissue, reduced the disease activity index, improved intestinal pathology, and suppressed the expression of inflammatory factors in colon tissue. Therefore, TRPC4 inhibitors have potential application in the preparation of drugs for the prevention or treatment of IBD.

[0012] In the present invention, the inflammatory bowel disease (IBD) refers to a non-specific chronic inflammatory disease of the intestine, mainly including ulcerative colitis (UC) and Crohn's disease (CD). The former mainly damages the colon and rectum, and the latter can damage any part of the gastrointestinal tract from the mouth to the anus, with the terminal small intestine and colon being more common. The cause of IBD is still unclear and is related to abnormal intestinal immune response. Common risk factors include environmental factors, dietary structure, genetic factors, intestinal flora imbalance, etc. In addition, it may also include intestinal inflammatory diseases caused by infection, ischemia, bleeding, diverticulitis, radiation, drug damage, etc.

[0013] In a preferred embodiment, the inflammatory bowel disease (IBD) is ulcerative colitis (UC).

[0014] In the present invention, the "inhibitor" can also be expressed as "antagonist" or "blocker", which have equivalent meanings in the present invention and refer to a compound or mixture that can be used to reduce or inhibit biological activity, or reduce gene or / and protein expression. Therefore, the TRPC4 inhibitor of the present invention refers to a compound or mixture that can inhibit or block the activity of the TRPC4 ion channel, or reduce the TRPC4 gene or / and protein expression. Such substances include but are not limited to small molecules, polypeptides, antibodies, small interfering RNA, anti-oligonucleotides, etc.

[0015] In a preferred embodiment, the TRPC4 inhibitor is a small molecule.

[0016] In a preferred embodiment, the TRPC4 inhibitor inhibits or blocks the activity of TRPC4, or / and downregulates the expression level of the TRPC4 gene or / and protein.

[0017] In a preferred embodiment, the TRPC4 inhibitor is a compound having a structure represented by the following general formula (I), a stereoisomer thereof, a deuterated product thereof, or a pharmaceutically acceptable salt thereof:

[0018]

[0019] in,

[0020] R 1 、R 2 Each is independently H, C1-C6 alkyl, C1-C6 alkoxy, azido, cyano, hydroxyl, amino, C1-C6 alkylamino, or R 1 、R 2 Together with the carbon atoms to which they are attached, The C1-C6 alkyl group or C1-C6 alkoxy group is unsubstituted or optionally substituted with one or more substituents selected from C3-C8 cycloalkyl group, C6-C10 aryl group, halogenated C6-C10 aryl group, cyano group, deuterium atom or halogen;

[0021] R a 、R b Each is independently H, C1-C6 alkyl or cyano;

[0022] R 1 、R 2 Not at the same time H;

[0023] Each R 3 are independently H, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, cyano or nitro;

[0024] R 4 is H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or cyano;

[0025] m=1, 2, 3 or 4.

[0026] In another preferred embodiment, the TRPC4 inhibitor is a compound having a structure as shown in general formula (Ia), a stereoisomer thereof, a deuterated product thereof, or a pharmaceutically acceptable salt thereof:

[0027]

[0028] in,

[0029] Each R 3 are independently H, halogen, C1-C3 alkyl, C1-C3 haloalkyl or nitro;

[0030] R 5 is a C1-C3 alkyl group or a C1-C3 deuterated alkyl group;

[0031] m=1, 2 or 3.

[0032] Further preferably, the TRPC4 inhibitor is compound B1T1, its stereoisomer, its deuterated product, or a pharmaceutically acceptable salt thereof as shown in the following formula:

[0033]

[0034] In the present invention, the compound B1T1 is equivalent to the compound E21 disclosed in patent application CN202310462302X. The patent application discloses that compound E21 inhibits the activity of TRPC4 and TRPC5 channels and the medical use of E21 in treating kidney disease and liver disease. The present invention found that in the dextran sodium sulfate (DSS)-induced colitis mouse model, compound B1T1 has a significant effect on improving the disease condition of colitis, which is specifically manifested in the ability to inhibit weight loss in mice, reduce the soft layer of stool and occult blood symptoms, increase colon length, reduce intestinal tissue inflammation levels and pathological scores, etc. At the same time, B1T1 can also downregulate the expression level of the TRPC4 gene.

[0035] In the present invention, the TRPC4 inhibitors also include other compounds that inhibit or block TRPC4 activity that have been disclosed in the literature, such as, but not limited to, CN2023100836239, WO2014143799, WO2020191056, WO2022166817, etc. It should be understood that based on the application of inhibiting TRPC4 in the treatment of colitis disclosed in the present invention, the TRPC4 inhibitors disclosed in the literature are also expected to have the effect of improving IBD disease conditions. Therefore, they also have potential application value in the preparation of drugs for preventing or treating IBD.

[0036] In a preferred embodiment, the TRPC4 inhibitor is a compound having a structure represented by general formula (Ib), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0037]

[0038] in,

[0039] R 1 is a C1-C6 alkyl group;

[0040] Each R2 are independently H or a halogen atom;

[0041] Each R 3 independently selected from H, deuterium, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 5-8 membered heteroaryl, 4-8 membered heterocyclic group, halogen atom, cyano, hydroxyl, -NR 4 R 5 ; or two R 3 Together with the carbon atoms connected to the cyclohexyl group, it forms a 4-8 membered heterocyclic ring;

[0042] R 4 、R 5 Each independently represents H, C1-C6 alkyl;

[0043] n=1, 2, 3 or 4.

[0044] The compound with the structure represented by the general formula (Ib) is selected from CN2023100836239.

[0045] Further preferably, the TRPC4 inhibitor is selected from all compounds shown in claim 7 of CN2023100836239.

[0046] Furthermore, the TRPC4 inhibitor is selected from compounds E21 and E45 shown in claim 7 of CN2023100836239:

[0047]

[0048] In a preferred embodiment, the TRPC4 inhibitor is selected from the compound represented by formula (III) in WO2014143799 or a pharmaceutically acceptable salt thereof:

[0049]

[0050] in,

[0051] R 2 For 1-3 R 6 Substituted C1-C6 alkoxy or C6-C10 aryloxy;

[0052] R 3 is C1-C6 heteroalkyl or C2-C6 hydroxyalkyl;

[0053] R 4 is a C1-C6 alkyl group;

[0054] Each R 6are independently C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 haloalkoxy or C1-C6 alkoxy;

[0055] Each R a are independently C1-C6 alkyl, C1-C6 haloalkyl or halogen;

[0056] n = 1 or 2;

[0057] m=1, 2 or 3.

[0058] Further preferably, the TRPC4 inhibitor is selected from Compound 1 to Compound 648 shown in Table A of WO2014143799.

[0059] Furthermore, the TRPC4 inhibitor is compound 31 or compound 260 shown in Table A of WO2014143799:

[0060]

[0061] In a preferred embodiment, the TRPC4 inhibitor is selected from the compound represented by the general formula (II) in WO2020191056 or a pharmaceutically acceptable salt thereof:

[0062]

[0063] in,

[0064] R 11 is chlorine, -CF3, -CHF2 or -CH3;

[0065] R 12 is hydrogen or fluorine; and

[0066] R 13 It is hydrogen, -NH2, -CH2OH or CH(OH)-CH2OH.

[0067] Further preferably, the TRPC4 inhibitor is selected from all compounds shown in claim 6 of WO2020191056;

[0068] Furthermore, the TRPC4 inhibitor is compound 100 shown in claim 6 of WO2020191056:

[0069]

[0070] In a preferred embodiment, the TRPC4 inhibitor is selected from compound L015 shown in WO2022166817, its prodrug compounds 1 to 6, or pharmaceutically acceptable salts thereof;

[0071] Further preferably, the TRPC4 inhibitor is compound L015 or prodrug compound 1 shown in WO2022166817:

[0072]

[0073] The TRPC4 inhibitor has a strong inhibitory effect on the activity of TRPC4 channels, and therefore can be used to prepare drugs for preventing or treating IBD, in particular for preparing drugs for preventing or treating UC.

[0074] TRPC family proteins exist not only in the form of homotetramers, but also often in the form of heterotetramers. Taking TRPC4 as an example, it not only constitutes TRPC4:C4 homotetramers, but also constitutes TRPC4:C5, TRPC1:C4, TRPC1:C4:C5 and other heterotetramers. Therefore, the TRPC4 inhibitors described in the present invention not only inhibit TRPC4 homotetramer channels, but also inhibit TRPC4 heterotetramer channels.

[0075] By analyzing data from a gene expression collection compiled by the National Center for Biotechnology Information (NCBI), the present invention found that TRPC4 mRNA expression levels in the colon tissue of UC patients were significantly higher than in healthy controls. Studies in disease models also revealed that TRPC4 mRNA expression in the colon tissue of mice with DSS-induced colitis was much higher than in normal mice. Furthermore, administration of a TRPC4 inhibitor significantly reduced TRPC4 mRNA levels while improving colitis symptoms. These findings suggest that TRPC4 is associated with the progression of colitis and may serve as a biomarker for patient screening and therapeutic efficacy prediction.

[0076] Therefore, the drug prepared by the TRPC4 inhibitor of the present invention can be used in the following scheme to obtain a better therapeutic response when used for treatment: (1) measuring the expression level of TRPC4 mRNA or protein in the intestinal tissue of IBD patients; (2) selecting individuals with TRPC4 mRNA or protein expression levels higher than normal levels and administering the drug prepared by the TRPC4 inhibitor of the present invention.

[0077] In the present invention, the method for determining the mRNA or protein expression level can be achieved by techniques and devices known in the art, such as but not limited to quantitative PCR, microarray, Northern blotting, next generation sequencing, protein blotting, ELISA, immunofluorescence or mass spectrometry.

[0078] The drug for preventing or treating inflammatory bowel disease of the present invention comprises the compound of the present invention, its stereoisomers, its deuterated products, or its prodrugs and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients.

[0079] The excipients include pharmaceutically acceptable carriers, excipients, sustained-release agents, odorants, flavoring agents, etc.

[0080] In the medicine, the compound of the present invention serves as the active ingredient, and its weight accounts for 0.1 to 99.9% of the total weight of the medicine, and the rest are pharmaceutically acceptable excipients; the preferred ratio of the compound of the present invention to the excipients is: the compound of the present invention serves as the active ingredient and accounts for more than 60% of the total weight, and the rest accounts for 0-40% of the total weight, and the weight of the rest is preferably 1-20%, and most preferably 1-10%.

[0081] The drug of the present invention can be prepared into various dosage forms, such as tablets, capsules, powders, syrups, solutions, suspensions, sprays, creams, ointments, gels, transdermal patches, etc., based on conventional processes in the field of pharmaceutical preparations, and can be present in suitable solid or liquid carriers or diluents. The drug of the present invention can also be stored in suitable sterile equipment for injection or infusion.

[0082] The drug of the present invention can be used in mammals, including humans and animals. The routes of administration include oral administration, nasal inhalation, topical administration to the skin, intravenous injection, intramuscular injection, subcutaneous injection, etc. The preferred route of administration is oral administration.

[0083] The solid dosage forms of the medicament of the present invention for oral administration include capsules, tablets, pills, powders, and granules. Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, and kaolin, while liquid carriers include sterile water, polyethylene glycol, nonionic surfactants, and edible oils (such as corn oil, peanut oil, and sesame oil), as long as they are suitable for the characteristics of the active ingredient and the specific administration method required. Adjuvants commonly used in the preparation of medicaments may also be advantageously included, such as flavorings, pigments, preservatives, and antioxidants such as vitamin E, vitamin C, BHT, and BHA.

[0084] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain inert diluents conventionally employed in the art, such as water or other solvents, solubilizing agents and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc. In addition to these inert diluents, the medicine may also contain adjuvants, such as wetting agents, emulsifying agents and suspending agents, sweeteners, flavoring agents and spices.

[0085] In addition to the active ingredient, the suspension may contain various solubilizers, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0086] When used as a pharmaceutical formulation, the compounds of the present invention are preferably administered in a unit dose, each dose containing 0.1 mg to 2000 mg of the active ingredient, preferably 20 mg to 500 mg, for single or divided administration. Regardless of the method of administration, the optimal individual dosage will be determined based on the specific treatment. Typically, a low dose is started and gradually increased until the optimal dosage is found.

[0087] When the TRPC4 inhibitor of the present invention is used for treatment, it can be used alone or in combination with one or more other drugs known to treat or improve similar symptoms. Compared with the use of any one drug alone, the combination of these drugs may be safer or more effective. The other drugs can be administered simultaneously with the compound of the present invention or before or after the administration through commonly used routes of administration and dosages. When the TRPC4 inhibitor of the present invention is used simultaneously with one or more other drugs, a pharmaceutical composition comprising the other drugs and the compound of the present invention in a unit dosage form is preferred. However, the drug combination may also include a therapy in which the compound of the present invention and one or more other drugs are administered in different overlapping schemes. When used in combination with one or more other active ingredients, the compound of the present invention and the other drugs can be used in a lower dose than when used alone.

[0088] Drugs or active ingredients that can be used in combination with the compounds of the present invention include, but are not limited to, the following drugs for treating IBD: aminosalicylic acids, such as sulfasalazine, mesalazine, olsalazine, etc.; glucocorticoids, such as prednisolone, beclomethasone, budesonide, hydrocortisone, etc.; immunosuppressants, such as methotrexate, azathioprine, mercaptopurine, tacrolimus; anti-TNF-α drugs, such as infliximab, adalimumab, golimumab , certolizumab pegol, etc.; cell adhesion molecule inhibitors, such as natalizumab, vedolizumab, AJM300, etc.; interleukin-12 or 23 antagonists, such as ustekinumab, risankizumab, migizone, etc.; JAK inhibitors, such as tofacitinib, upadacitinib, etc.; S1P receptor modulators, such as ozanimod, etc.; cell therapy, such as darvadstrocel, etc.; broad-spectrum antibiotics, such as metronidazole, ciprofloxacin, etc.; and probiotic therapy.

[0089] In the present invention, unless otherwise specified, the terms used have the general meanings commonly known to those skilled in the art.

[0090] In the present invention, the term "C1-C6" refers to having 1, 2, 3, 4, 5 or 6 carbon atoms, and so on.

[0091] In the present invention, the "alkyl" is a branched or straight-chain hydrocarbon group having a specific number of carbon atoms, and representative examples include but are not limited to methyl, ethyl, n-propyl, and isopropyl.

[0092] In the present invention, the term "alkoxy" refers to an -O-alkyl group. For example, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms, representative examples of which include but are not limited to methoxy, ethoxy, n-propoxy, isopropoxy, and butoxy.

[0093] Similarly, the "alkylamino" refers to -NH-alkyl or -N(alkyl)2. Representative examples include but are not limited to methylamino, dimethylamino, ethylamino, diethylamino, and the like.

[0094] In the present invention, the "haloalkyl" and "haloalkoxy" refer to groups in which the hydrogen atoms in an "alkyl" or "alkoxy" group having a specific number of carbon atoms are partially or completely replaced by "halogen atoms".

[0095] In the present invention, the "hydroxyalkyl" refers to a group formed by replacing one or more hydrogen atoms on an "alkyl" with a specific number of carbon atoms by a hydroxyl group. The substitution can occur at the terminal carbon atom of the alkyl group or at the middle carbon atom of the alkyl group.

[0096] In the present invention, the "heteroalkyl" refers to a group formed by an "alkyl" group with a specific number of carbon atoms being interrupted by one or more heteroatoms selected from N, O, and S, and the interruption position occurs between two carbon atoms, or between a carbon atom and a connection site of the general structure.

[0097] In the present invention, the "cycloalkyl" refers to a non-aromatic cyclic aliphatic hydrocarbon group having a specific number of ring carbon atoms, and a "C3-C8 cycloalkyl" refers to a cyclic aliphatic hydrocarbon group consisting of 3 to 8 ring carbon atoms. The "cycloalkyl" described in the present invention includes not only aliphatic hydrocarbon groups with fully saturated carbon atoms, but also aliphatic hydrocarbon groups with unsaturated bonds in some of the carbon atoms. Examples of the "cycloalkyl" described in the present invention include, but are not limited to:

[0098] In the present invention, "aryl" is defined as a monocyclic or bicyclic ring system consisting of a specific number of carbon atoms and obeying Hückel's rule, including phenyl and naphthyl. The term "aryloxy" includes an aryl group covalently linked to -O-, such as phenoxy.

[0099] In the present invention, "heteroaryl" is defined as a monocyclic or bicyclic ring system having a specific number of ring atoms and containing 1, 2, 3 or 4 heteroatoms (selected from N, O, S) and obeying Hückel's rule; examples of "heteroaryl" include but are not limited to pyridine, pyrrole, imidazole, thiophene, benzimidazole, benzothiophene, benzofuran, etc.

[0100] In the present invention, "heterocyclyl" is defined as a saturated or partially unsaturated non-aromatic monocyclic and bicyclic ring system having a specific number of ring atoms and containing 1, 2, 3 or 4 heteroatoms (selected from N, O, S); examples of "heterocyclyl" include but are not limited to oxetane, dioxolanyl, morpholinyl, piperazinyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, benzotetrahydrofuranyl, etc.

[0101] In the present invention, the "halogen" includes fluorine, chlorine, bromine and iodine.

[0102] As used herein, "substituted" means replacement by one or more groups. Unless a specific atom is specified, the term "substituted" refers to any atom where the number of substituents is not yet saturated. When multiple substituents are selected from the same series, they may be the same or different.

[0103] The term "independently" used in the present invention means that two or more substituents defined herein do not interfere with each other when selected from a series of candidate groups and may be the same or different.

[0104] The "pharmaceutically acceptable salt" of the present invention can be a salt formed by an anion and a positively charged group on the compound. Suitable anions are chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronate, lactate, glutarate, or maleate. Similarly, salts can be formed by cations and negatively charged groups on the compound. Suitable cations include sodium, potassium, magnesium, calcium, and ammonium ions.

[0105] In certain preferred embodiments, "pharmaceutically acceptable salts" refer to salts formed by compounds carrying basic atoms such as nitrogen atoms with inorganic or organic acids, such as but not limited to hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid and isethionic acid. "Pharmaceutically acceptable salts" also refer to salts formed with inorganic bases of compounds carrying acidic groups such as carboxyl groups, such as but not limited to sodium salts, potassium salts, calcium salts, aluminum salts, ammonium salts, etc., or salts formed with organic bases, such as but not limited to methylamine salts, ethylamine salts, ethanolamine salts, etc.

[0106] The compounds of the present invention, or pharmaceutically acceptable salts thereof, are obtained by distillation, crystallization, or recrystallization from water or an organic solvent, and may contain molecules of the solvent used. Furthermore, different crystallization conditions may result in different crystalline forms of the compounds. Therefore, compounds of the formula, or pharmaceutically acceptable salts thereof, containing different stoichiometric amounts of the crystallization solvent and all crystalline forms are within the scope of the present invention.

[0107] In the present invention, the "effective therapeutic dose" means that the subject receiving the treatment with the dose is cured, improved, effectively prevented, or the incidence of the disease or side effect is significantly reduced compared with the subject not receiving the treatment with the dose; in addition, it also includes an effective dose for enhancing normal physiological function.

[0108] In the present invention, the term "small molecule" refers to an organic compound with a low molecular weight of ≤1000 Daltons.

[0109] For the purposes of this invention, the term "prodrug" refers to an inactive or partially inactive, chemically modified drug molecule that requires chemical and / or enzymatic biotransformation in vivo to release the active parent drug at or near the site of action, thereby producing a therapeutic effect. Prodrugs are a proven, effective drug development strategy that can improve the physicochemical, biopharmacological, or pharmacokinetic properties of a compound, overcoming various obstacles in drug formulation and delivery, such as poor aqueous solubility, chemical instability, insufficient oral absorption, first-pass metabolism, insufficient brain penetration, toxic reactions, and local irritation.

[0110] If the compounds of the present invention have a chiral center, a potential chiral center, or an unsaturated bond, they may form various stereoisomers, such as racemates, enantiomers, diastereomers, E / Z isomers, cis-trans isomers, tautomers, and the like. Unless otherwise indicated, in the specification and the appended claims, a given chemical formula or name is intended to encompass all forms of stereoisomers, as well as mixtures composed of individual isomers in varying proportions, and pharmaceutically acceptable salts thereof. A person of ordinary skill in the art can use commonly used laboratory separation methods to separate the compounds of the present invention containing asymmetric centers to obtain single isomers.

[0111] Replacing hydrogen atoms with deuterium atoms to change the physical and chemical properties of compounds has become a well-known structural modification method for those skilled in the art. Unless otherwise specified, the present invention intends that the deuterated forms of the compounds are also included in the invention content.

[0112] Beneficial effects:

[0113] The present invention provides a use of a TRPC4 inhibitor in the preparation of a drug for preventing or treating IBD. By performing data analysis on a gene expression collection constructed by the National Center for Biotechnology Information (NCBI), the present invention found that the expression level of TRPC4 mRNA in the colon tissue of UC patients was significantly higher than that in healthy people. The present invention's research on disease models also found that the expression level of TRPC4 mRNA in the colon tissue of DSS-induced colitis mice was much higher than that of normal mice; and taking a TRPC4 inhibitor can significantly reduce the level of TRPC4 mRNA while improving the disease condition of colitis. These evidences suggest that TRPC4 is correlated with the disease progression of colitis, and TRPC4 may be used as a biomarker for screening patients and predicting therapeutic effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0114] Figure 1-4 This is the effect of knocking out the TRPC4 gene on DSS-induced UC model mice in a preferred embodiment of the present invention.

[0115] Figure 1 This is a curve of the Disease Activity Index (DAI) score, which evaluates the severity of the disease based on comprehensive body weight and stool conditions during the 7-day experiment.

[0116] Figure 2 A is the wild-type mouse (WT) and the TRPC4 knockout mouse (Trpc4 - / - ) Colon length image after 7 days of experiment; Figure 2 B is the statistical result of colon length.

[0117] Figure 3A is the wild-type mouse (WT) and the TRPC4 knockout mouse (Trpc4 - / - ) HE staining results of colon tissue 7 days after the experiment; Figure 3 B is the pathological score of HE staining results.

[0118] Figure 4 A is the wild-type mouse (WT) and the TRPC4 knockout mouse (Trpc4 - / - ) Myeloperoxidase (MPO) levels in serum and colon tissue were measured 7 days after the experiment; Figure 4 BD is wild-type mice (WT) and TRPC4 knockout mice (Trpc4 - / - ) The expression levels of inflammation-related factors (TNF-α, IL-1β and IL-6) mRNA in colon tissue.

[0119] Figure 5-8 The present invention provides the effects of administering B1T1 on UC model mice induced by dextran sulfate sodium salt (DSS) in a preferred embodiment.

[0120] Figure 5 Effect of B1T1 on the disease activity index of DSS-induced ulcerative colitis model in mice.

[0121] Figure 6 A is a picture showing the effect of B1T1 on colon length in a DSS-induced mouse ulcerative colitis model; Figure 6 B is the statistical result of colon length.

[0122] Figure 7 A is the HE staining result of colon tissue of DSS-induced ulcerative colitis model in mice by B1T1; Figure 7 B is the pathological score of HE staining results.

[0123] Figure 8 A shows the effect of B1T1 on the expression of MPO in the DSS-induced mouse ulcerative colitis model; Figure 8 BD is the effect of B1T1 on the changes in the mRNA levels of inflammatory factors (TNF-α, IL-1β, IL-6) in the colon tissue of the DSS-induced mouse ulcerative colitis model in the preferred embodiment provided by the present invention. DETAILED DESCRIPTION

[0124] The technical solution of the present invention is described in detail below through specific embodiments, but the protection scope of the present invention is not limited to the embodiments.

[0125] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0126] In the following examples, the effects of drugs on ulcerative colitis were evaluated using a mouse ulcerative colitis model induced by seven consecutive days of free administration of a 3% dextran sulfate sodium salt (DSS) solution (MP biomedicals, USA, Catalog No. 160110). The 3% DSS-induced ulcerative colitis model is currently the most widely used model for evaluating the effects of drugs on the progression of ulcerative colitis.

[0127] In the following examples, the transcription levels of specific genes in tissues were detected by real-time quantitative PCR experiments.

[0128] Example 1

[0129] This example is used to illustrate the application of the present invention in developing a drug for treating UC using TRPC4 as a target.

[0130] (1) Construction of TRPC4 knockout mice

[0131] TRPC4 KO mice have been established by the Model Animal Center of Nanjing Medical University using the following method:

[0132] TRPC4 KO mice were engineered by targeting exon 3 of the TRPC4 gene. A guide RNA (gRNA) was designed and fused to a transactivating RNA (tracrRNA) to construct a clustered regularly interspaced short palindromic repeat (CRISPR) sequence. This sequence and the Cas9 gene were inserted into an expression vector, transformed into competent cells, amplified, and transfected into embryonic stem cells (ESCs). ESCs expressing the CRISPR / Cas9 system were microinjected into C57BL / 6 mouse blastocysts, which were then transplanted into pseudopregnant mouse uteri to generate F0 offspring. Homozygous offspring were obtained through multiple crosses.

[0133] (2) DSS-induced UC mouse model

[0134] The animal experiment process complies with the animal ethics standards of China Pharmaceutical University. In Example 1 (1), SPF-grade TRPC4·KO mice were bred, 8-10 weeks old, half male and half female, weighing 20-25g. The mice were raised in an independent barrier system with a temperature of 23±2°C, a relative humidity of 40-70%, and a 12 / 12h day and night light cycle, with free access to water and food. After a week of breeding to adapt to the environment, the mice were randomly divided into groups. The mice in the model group were free to drink 3% DSS solution for 7 consecutive days, with DSS changed every 2 days. The mice in the control group were free to drink ultrapure water. The weight of the mice was recorded every day, and feces were collected for fecal occult blood test. The disease severity assessment (DAI) was evaluated based on the comprehensive weight and fecal conditions. The results are as follows Figure 1 As shown, after TRPC4 knockout, the UC symptoms of mice were alleviated to a certain extent. On the 7th day, a sample of 1-2 cm from the end of the colon was taken, fixed with 4% paraformaldehyde as a colon specimen for histopathological examination, and a sample 3-4 cm from the anus was taken for subsequent index detection. Blood was collected from the orbital venous plexus using a capillary tube into a 1.5 ml EP tube, placed at room temperature for 30 minutes, and then centrifuged at 3500r / min to obtain the supernatant to obtain mouse serum. After the experiment, the mice were killed by cervical dislocation and the abdominal dissection was performed on the mice. The colon part and ileocecal valve were separated from the anus to the ileum, and the appearance and length changes of the colon were observed and recorded. The results are shown in Figure 2 A. Figure 2 As shown in Figure B, TRPC4 gene knockout can significantly improve the shortening of colon length caused by DSS. Note: Quantitative data are expressed as mean ± standard deviation (SEM). In the analysis results, ** indicates P < 0.01.

[0135] (3) Evaluation of clinical symptoms in mouse colitis model

[0136] Trpc4 - / - The clinical symptoms of colitis damage in mice and wild-type mice were scored using the DAI scoring system, which is as follows:

[0137] a) Percentage of weight loss: 0 = 0%, 1 = 1%-5%, 2 = 5%-10%, 3 = 10%-15%, 4 = >15%; b) Stool consistency: 0 = normal, 2 = loose stools, 4 = diarrhea; c) Stool bleeding: 0 = normal, 2 = occult blood, 4 = overt bleeding. A comprehensive score is calculated for all three conditions, and the total score is the DAI value.

[0138] The results are as follows Figure 1 Note: The measurement data are expressed as mean ± standard deviation (mean ± SEM) and tested by one-way analysis of variance. In the analysis results, **P < 0.01, relative to the wild-type control group; ## P<0.01, relative to the wild-type model group.

[0139] Giving 3% DSS in drinking water resulted in weight loss, loose stools, and bloody stools in mice, which was significantly different from that in TRPC4 knockout mice ( Figure 1 ).

[0140] (4) Histopathological examination of mouse UC model

[0141] Steps:

[0142] Fixed colon specimens were embedded in paraffin and cut into 5-μm sections. Sections were dewaxed twice in xylene and washed with graded alcohols. After staining with hematoxylin for 5 minutes, the sections were separated with 0.5% hydrochloric acid-ethanol for 30 seconds and then stained with eosin for 3 minutes. After dehydration, transparency, and mounting with neutral resin, the sections were digitally photographed and scored using a NanoZoomer 2.0RS (Hamamatsu Photonics, Hamamatsu, Japan).

[0143] The histological scoring was graded as follows: a) severity of inflammation: 0 = none; 1 = mild; 2 = moderate; 3 = severe; 4 = very severe; b) depth of lesion: 0 = none; 1 = mucosal layer; 2 = submucosa; 3 = muscle layer; 4 = transmural; c) crypt damage: 0 = none; 1 = basal 1 / 3 damaged; 2 = basal 2 / 3 damaged; 3 = only the surface epithelium intact; 4 = loss of the entire crypt and epithelium; d) extent of lesion: 1 = 1-25%; 2 = 26-50%; 3 = 51-75%; 4 = 76-100%.

[0144] Experimental results: From the HE-stained colon tissue sections, we can see that Trpc4 - / - The degree of colon tissue damage in mice was significantly lower than that in wild-type mice (see Figure 3 A, B). Measurement data are expressed as mean ± standard deviation (mean ± SEM) and tested using one-way analysis of variance. ** indicates P < 0.01.

[0145] (5) Detection of MPO content in mice

[0146] The MPO levels in mouse serum and colon tissue were detected according to the instructions of the MPO kit (Nanjing Jiancheng Bioengineering Institute, China). **P<0.01.

[0147] Figure 4 A is the wild-type mouse (WT) and the TRPC4 knockout mouse (Trpc4 - / - ) Results of myeloperoxidase (MPO) levels in serum and colon tissue after 7 days of the experiment. As shown in the figure, this indicates that knocking out the TRPC4 gene can inhibit MPO levels in serum and tissues.

[0148] (6) Detection of mRNA levels of TRPC4 and other inflammatory factors in mice

[0149] Weigh 50 mg of colon tissue, grind into powder in liquid nitrogen, add 1 mL of Trizol (Vazyme, Cat. No. R401-01), and incubate on ice for 10 minutes. Transfer the lysate to a 1.5 mL enzyme-free (RNase-free) centrifuge tube, add 100 μL of chloroform, shake vigorously for 15 seconds, and let it stand on ice for 2 minutes. Centrifuge at 12,000 rpm at 4°C for 10 minutes. Pipette the upper aqueous phase into a new 1.5 mL RNase-free centrifuge tube and add an equal amount of isopropanol to precipitate RNA. Centrifuge again at 12,000 rpm at 4°C for 10 minutes. Discard the supernatant and wash the RNA pellet with 500 μL of 75% ethanol prepared with DEPC water. Centrifuge again at 12,000 rpm at 4°C for 5 minutes. Discard the supernatant, dry the RNA pellet, and dissolve the RNA in 10 μL of DEPC water. 1 μL was taken for concentration detection using a Nano-100 ultra-micro spectrophotometer (Hangzhou Aosheng Instrument Co., Ltd., China, catalog number: AS-11010). 500 ng of RNA was then taken for reverse transcription reaction. The procedure was performed according to the instructions of the II One Step RT-PCR Kit (Vazyme, Cat. No. P611-01). The reverse transcription product was stored at -20°C until use.

[0150] Real-time quantitative PCR reaction was performed according to AceQ qPCR Green Master Mix (Vazyme, catalog number: Q111-01) was performed in a real-time fluorescence quantitative PCR system (QuantStudio3, Thermo Fisher Scientific, USA) according to the instructions. The reaction system consists of 1 μL reverse transcription product, 0.5 μL 10 μM forward primer, 0.5 μL 10 μM backward primer, 8 μL DEPC water and 10 μL dye premix; the reaction conditions are: 95°C pre-denaturation for 10 minutes, 94°C denaturation for 30 seconds, 60°C annealing and extension for 30 seconds (collecting a signal once), denaturation, annealing and extension repeated 40 times; then enter the melting curve reaction: 95°C for 1 minute, 55°C for 1 minute, and then start increasing 0.5°C for each cycle, maintaining for 13 seconds (collecting a signal once), a total of 81 cycles, and ending at 95°C. The data of real-time quantitative PCR are 2 -ΔΔCt Method for processing.

[0151] The primer sequence information of related inflammatory factors is shown in Table 2:

[0152] Table 2 Sequence list of primers related to inflammatory factors

[0153]

[0154] The above primers were from Shanghai Sangon Biotechnology Co., Ltd.

[0155] The experimental results are as follows Figure 4 As shown in BD, in UC model mice with TRPC4 knockout, the expression of inflammatory factors in the colon tissue of mice was significantly reduced compared with that of wild-type mice, which had a certain alleviating effect on the inflammation of UC mice.

[0156] Note: Measurement data are expressed as mean ± standard deviation (mean ± SEM). ## indicates P<0.01; ** indicates P<0.01.

[0157] Example 2

[0158] This example is used to illustrate the application of B1T1 discovered by the present invention in preventing or treating ulcerative colitis.

[0159] (1) B1T1 improves DSS-induced ulcerative colitis model in mice

[0160] Fifty-two 5- to 6-week-old male C57BL / 6 wild-type mice weighing 20-22 g were purchased from the Experimental Animal Center of Yangzhou University. Mice were housed in a quiet, light-free environment for 3 days prior to the experiment to acclimate to the environment. The light-to-dark ratio was 1:1, the room temperature was maintained at 25°C, and the humidity was 55%. The mice had free access to food and water.

[0161] All drugs were prepared as follows:

[0162] 2% (vol / vol) DMSO was added to the drug, followed by 2% (vol / vol) Tween 80 and 0.5% (wt / vol) CMC-Na, and finally ddH2O was added and ultrasonicated for 1 hour to prepare a drug solution of corresponding dosage.

[0163] C57BL / 6 wild-type mice were randomly divided into 6 groups (6-8 mice in each group), namely: control group, 40 mg / kg B1T1 alone treatment group, DSS modeling group, DSS + 20 mg / kg B1T1 treatment group, 3% DSS + 40 mg / kg B1T1 treatment group, and DSS + 200 mg / kg 5-ASA treatment group.

[0164] All mice, except the control group, were given 3% (wt / vol) DSS in drinking water for 8 days, followed by pure drinking water for 3 days. The control group was fed pure drinking water and free access to food for 10 days. The B1T1-treated group received daily oral administration of B1T1 solution at a volume of 0.1 ml / 10 g based on body weight starting on day 4 of the experiment. The control and model groups received the corresponding volume of 0.5% Tween 80 solution at a volume of 0.1 ml / 10 g.

[0165] During the experiment, the colitis damage score and body weight of mice in all groups were measured every day. The results of the effect of B1T1 on the disease activity index of the DSS-induced ulcerative colitis model in mice can be found in Figure 5 .like Figure 5 As shown, the 40 mg / kg B1T1-administered group could significantly alleviate loose stools, bloody stools, and body weight loss caused by DSS.

[0166] After the experiment, the mice were killed by cervical dislocation and the abdomen was dissected. The colon and ileocecal valve were separated from the anus to the ileum, and the appearance and length of the colon were observed and recorded. Figure 6 A. Figure 6 As shown in B, 40 mg / kg B1T1 can significantly restore the shortening of colon length caused by DSS. Note: The measurement data are expressed as mean ± standard deviation (mean ± SEM). ## Indicates P < 0.01, relative to the model group; ** indicates P < 0.01, relative to the control group.

[0167] (2) Histopathological examination of mouse UC model

[0168] The specific experimental steps refer to Example 1 (4), and the results are as follows Figure 7 A. Figure 7 As shown in B, HE-stained colon tissue sections showed that the 40 mg / kg B1T1 administration group significantly improved the structural damage and inflammatory cell infiltration of the colon tissue in UC model mice, indicating that 40 mg / kg B1T1 administration played a certain protective role against DSS-induced UC in mice. ## Indicates P < 0.01, relative to the model group; ** indicates P < 0.01, relative to the control group.

[0169] (3) Detection of MPO content in mouse colon tissue

[0170] The specific experimental steps refer to Example 1 (5), and the results are as follows Figure 8 As shown in A, administration of 40 mg / kg B1T1 can significantly reduce the MPO content in the colon tissue of UC mice.

[0171] (4) B1T1 inhibits the mRNA levels of inflammatory factors related to ulcerative colitis in mice

[0172] The specific experimental steps refer to Example 1 (6), and the results are as follows Figure 8 As shown in BD, DSS caused a significant increase in the mRNA expression levels of inflammatory factors IL-1β, IL-6, and TNF-α in the colon tissue of the model group mice. After administration of 40mg / kg B1T1, the mRNA levels of the above inflammatory factors in the colon tissue were significantly reduced. Note: Quantitative data are expressed as mean ± standard deviation (mean ± SEM). In the analysis results, ## Indicates P < 0.01, relative to the model group; ** indicates P < 0.01, relative to the control group.

[0173] The results of the above examples demonstrate that mice given dextran sulfate sodium freely exhibit symptoms such as shortened colon length, inflammatory cell infiltration in colonic tissue, and increased expression of related inflammatory factors. Administration of B1T1 improves DSS-induced weight loss, stool characteristics, and blood in the stool, restores colonic structure, inhibits DSS-induced colon shortening, and reduces DSS-induced inflammatory factor expression. Therefore, B1T1 is an effective treatment for ulcerative colitis.

[0174] Example 3

[0175] This example is used to illustrate the testing method for the inhibitory activity of compounds on TRPC4 channels.

[0176] HEK-293 cells expressing hTRPC4 (ATCC) were seeded into black-bottomed 96-well plates coated with polylysine (PDL), with 2×10 cells per well. 4 After culturing for 8 hours, the original Dulbecco's Modified Eagle Medium (Gbico, MA, USA) was discarded, and 60 μL of Fluo-4 / AM dye with a final concentration of 4 μM was added. The cells were incubated at 37°C for 60 minutes, and then rinsed with calcium flow detection buffer 5 times. The cell plate was placed in a 30°C preheated (Molecular Devices, Sunnyvale, CA, USA), excited at a wavelength of 488 nm, and continuously recorded fluorescence signals at a sampling frequency of 1 s in the range of 515-535 nm. After recording for 60 seconds, calcium flow detection buffer was added as a solvent control, the test compound and the positive inhibitor (compound 31 described in Table 1) dissolved in calcium flow detection buffer were added, and the signal was collected for 300 s. Then, the agonist Englerin A (EA) (MedChemExpress, Shanghai, China, with a final concentration of 0.3 nM) was added and the fluorescence signal was collected for 600 seconds. The fluorescence signal of the trajectory diagram is expressed as F / F0, where F is the fluorescence signal at different time points and F0 is the basic fluorescence signal, i.e., the average value of the fluorescence signal at the first 10 time points. The dose-effect diagram first uses F / F0=1 as the baseline to calculate the area under the curve of the change in fluorescence intensity after the addition of EA. The IC is calculated by combining the area under the curve and the log value of the compound concentration. 50 value.

[0177] The results of the inhibitory effects of the preferred compounds of the present invention on TRPC4 ion channels are shown in Table 1:

[0178] Table 1 Inhibitory effect of compounds on TRPC4 ion channels

[0179]

[0180] The preferred embodiments of the present invention are described in detail above in conjunction with the accompanying drawings. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0181] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. To avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0182] In addition, the various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. Use of TRPC4 inhibitors in the preparation of drugs for preventing or treating inflammatory bowel disease.

2. The use according to claim 1, characterized in that The inflammatory bowel diseases include ulcerative colitis and Crohn's disease.

3. The use according to claim 1, characterized in that The TRPC4 inhibitor is a small molecule, a polypeptide, an antibody, a small interfering RNA or an anti-oligosense nucleotide.

4. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is a compound having a structure represented by the following general formula (I), a stereoisomer thereof, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof: in, R 1 、R 2 Each is independently H, C1-C6 alkyl, C1-C6 alkoxy, azido, cyano, hydroxyl, amino, C1-C6 alkylamino, or R 1 、R 2 Together with the carbon atoms to which they are attached, The C1-C6 alkyl group or C1-C6 alkoxy group is unsubstituted or optionally substituted with one or more substituents selected from C3-C8 cycloalkyl group, C6-C10 aryl group, halogenated C6-C10 aryl group, cyano group, deuterium atom or halogen; R a 、R b Each is independently H, C1-C6 alkyl or cyano; R 1 、R 2 Not at the same time H; Each R 3 are independently H, halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, cyano or nitro; R 4 is H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or cyano; m=1, 2, 3 or 4.

5. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is a compound having a structure as shown in general formula (Ia), a stereoisomer thereof, a deuterated compound thereof, or a pharmaceutically acceptable salt thereof: in, Each R 3 are independently H, halogen, C1-C3 alkyl, C1-C3 haloalkyl or nitro; R 5 is a C1-C3 alkyl group or a C1-C3 deuterated alkyl group; m=1, 2 or 3.

6. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is a compound having a structure represented by general formula (Ib), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: in, R 1 is a C1-C6 alkyl group; Each R 2 are independently H or a halogen atom; Each R 3 independently selected from H, deuterium, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C6-C10 aryl, C3-C8 cycloalkyl, 5-8 membered heteroaryl, 4-8 membered heterocyclic group, halogen atom, cyano, hydroxyl, -NR 4 R 5 ; or two R 3 Together with the carbon atoms connected to the cyclohexyl group, it forms a 4-8 membered heterocyclic ring; R 4 、R 5 Each independently represents H, C1-C6 alkyl; n=1, 2, 3 or 4.

7. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is a compound having a structure represented by general formula (III) or a pharmaceutically acceptable salt thereof: in, R 2 For 1-3 R 6 Substituted C1-C6 alkoxy or C6-C10 aryloxy; R 3 is C1-C6 heteroalkyl or C2-C6 hydroxyalkyl; R 4 is a C1-C6 alkyl group; Each R 6 are independently C1-C6 alkyl, halogen, C1-C6 haloalkyl, C1-C6 haloalkoxy or C1-C6 alkoxy; Each R a are independently C1-C6 alkyl, C1-C6 haloalkyl or halogen; n = 1 or 2; m=1, 2 or 3.

8. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is a compound having a structure represented by general formula (II) or a pharmaceutically acceptable salt thereof: in, R 11 is chlorine, -CF3, -CHF2 or -CH3; R 12 is hydrogen or fluorine; and R 13 It is hydrogen, -NH2, -CH2OH or CH(OH)-CH2OH.

9. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor is selected from the following compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof:

10. The use according to any one of claims 1 to 3, characterized in that The TRPC4 inhibitor inhibits or blocks the activity of TRPC4, or / and downregulates the expression level of the TRPC4 gene or / and protein.

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