Use of imidazolidinyl vanillic acid ether derivatives in the treatment of diseases associated with fibrosis

Abstract

The present invention discloses the use of a compound of formula (I) or an ester or pharmaceutically acceptable salt thereof in the treatment of diseases associated with fibrosis. [Formula 1] TIFF2025533120000015.tif22170

Description

[Technical Field]

[0001] The present invention relates to the field of medicinal chemistry, and in particular to the use of imidazolidinyl vanillic acid ether derivatives in the treatment of diseases associated with fibrosis. [Background technology]

[0002] Systemic fibrotic diseases, particularly systemic sclerosis (SSc), also known as scleroderma, are rare autoimmune diseases affecting multiple organs. The pathological characteristics are fibrosis of the skin and internal organs, accompanied by vascular lesions. Clinical symptoms include Raynaud's phenomenon, digital ulcers, pulmonary arterial hypertension, and scleroderma nephropathy. The global incidence of systemic sclerosis ranges from 8 to 56 per million, with a higher female incidence rate than males (4:1). There is a lack of clear epidemiological data in China.

[0003] Although the pathogenesis of systemic sclerosis is not entirely clear, significant progress has been made in recent years in research into the abnormal immune activation, signaling pathway conduction, and cytokine release involved in the fibrosis of systemic sclerosis. Chronic vascular injury, endothelial activation, and immune activation are all believed to be key factors in the development of secondary fibroblast activation and associated fibrosis (Nat Rev Rheumatol. 2019;15(4):208-24). Vascular injury is an important component of the pathogenesis of systemic sclerosis, activating the blood coagulation cascade, activating platelets, and increasing the production of factors such as thrombin, thromboxane, and platelet-derived growth factor, which have a strong profibrotic effect.

[0004] Due to the rarity and clinical heterogeneity of systemic sclerosis, there are currently no effective therapeutic drugs. Clinical treatments often focus on improving organ function, primarily through anti-inflammatory and immune regulation, improving circulation, and inhibiting fibrosis. Therapeutic drugs are classified as immunomodulators and anti-fibrotic drugs, which can slow the progression of the disease but cannot improve the symptoms (Expert Opin Investig Drugs. 2021; 30(6): 635-652). Therefore, there is a need to develop new therapeutic drugs for systemic sclerosis in clinical practice. Summary of the Invention

[0005] The present invention aims to provide use of a compound represented by formula (I) or an ester or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating a disease associated with fibrosis.

[0006] [ka] wherein R is selected from C1-C6 alkyl and n is selected from the integers 0, 1, 2, 3, 4, 5, and 6.

[0007] In some embodiments of the invention, the disease associated with fibrosis comprises systemic sclerosis.

[0008] In some embodiments of the present invention, systemic sclerosis includes, but is not limited to, diffuse systemic sclerosis (e.g., diffuse cutaneous systemic sclerosis), localized systemic sclerosis (e.g., localized cutaneous systemic sclerosis), overlapping systemic sclerosis, and various lesions associated with systemic sclerosis.

[0009] In some embodiments of the present invention, various lesions associated with systemic sclerosis include skin fibrosis, fibrosis of internal organs (e.g., kidney, intestine, lung, blood vessels, etc.), fibrotic skin lesions of renal origin, systemic fibrosis of renal origin, keloid formation and systemic sclerosis-associated CRET syndrome (calcinosis, esophageal dysfunction, sclerosis, and telangiectasia), vascular lesions, etc.

[0010] In some embodiments of the present invention, vascular lesions associated with systemic sclerosis include, but are not limited to, vascular occlusive disease vasculitis, microvascular and macrovascular lesions, Raynaud's phenomenon, digital ischemic lesions, digital ulcers, digital necrotic lesions, gangrene, and digital loss.

[0011] In some embodiments of the present invention, diseases associated with fibrosis also include fibrosis of the skin, intestine, liver, lung, heart, bladder, prostate, blood vessels, or other localized or systemic fibrosis in tissues due to collagen excess (regardless of etiology, for example, autoimmune disease, chronic graft-versus-host disease, diabetes, poisoning, surgery, radiation therapy, etc.). Specifically, other fibrotic diseases include collagen excess liver fibrosis, liver cirrhosis, pulmonary fibrosis, endomyocardial fibrosis, glomerulonephritis, renal interstitial fibrosis, diabetic fibrotic lesions, bone marrow fibrosis and similar fibrotic diseases, hypertrophic scarring (including after surgery), etc. [Effects of the Invention]

[0012] The compounds of the present invention have a good effect in treating diseases accompanied by fibrosis. The compounds of the present invention can achieve the desired therapeutic effect even when administered at a low dose, and the administration safety of the compounds of the present invention is good, and when used as a drug, they have obvious clinical advantages. [Brief explanation of the drawings]

[0013] [Figure 1A] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate (HY-2) according to the present invention on skin thickening in model mice (mean ± SD, n = 8) is shown, where ##P < 0.01, vs. blank group; *P < 0.05, **P < 0.01, vs. model group (bleomycin (BLM)). [Figure 1B] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate (HY-2) according to the present invention on skin thickening in model mice (mean ± SD, n = 8) is shown, where ##P < 0.01, vs. blank group; *P < 0.05, **P < 0.01, vs. model group (bleomycin (BLM)). [Figure 2] 1 shows the pathological (HE staining) effects of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on lung tissue in model mice (mean value ±SD, n=8). [Figure 3A] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition in skin tissue (Masson staining) in model mice (mean ± SD, n = 8), where ##P < 0.01 vs. blank group; **P < 0.01 vs. BLM. [Figure 3B] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition in skin tissue (Masson staining) in model mice (mean ± SD, n = 8), where ##P < 0.01 vs. blank group; **P < 0.01 vs. BLM. [Figure 4A] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition in lung tissue (Masson staining) in model mice (mean ± SD, n = 8), where ##P < 0.01 vs. blank group; **P < 0.01 vs. BLM. [Figure 4B] The effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition in lung tissue (Masson staining) in model mice (mean ± SD, n = 8), where ##P < 0.01 vs. blank group; **P < 0.01 vs. BLM. DETAILED DESCRIPTION OF THE INVENTION

[0014] definition As used herein, the following words and phrases are generally intended to have the meanings set forth below, unless their context indicates otherwise.

[0015] As used herein, the term "alkyl" refers to a monovalent group having a straight or branched saturated hydrocarbon chain of 1 to 6 (more typically 1 to 5, 1 to 4, or 1 to 3) carbon atoms. This term is illustrative of groups such as methyl, ethyl, 1-propyl (n-propyl), 2-propyl (isopropyl), 1-butyl (n-butyl), 2-methyl-1-propyl (isobutyl), 2-butyl (sec-butyl), 2-methyl-2-propyl (tert-butyl), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, and the like.

[0016] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness and properties of a given compound, and is not a salt that is biologically or otherwise undesirable. Pharmaceutically acceptable salts may be acid addition salts and / or base addition salts. Acid addition salts can be prepared from inorganic and organic acids. Salts derived from inorganic acids include hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, hydrogensulfate, hydrogenphosphate, dihydrogenphosphate, bicarbonate, etc., and salts derived from organic acids include formate, acetate, propionate, glycolate, pyruvate, oxalate, malate, malonate, succinate, maleate, fumarate, tartrate, citrate, benzoate, cinnamate, mandelate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate, lactate, nicotinate, lauryl sulfate, naphthalenesulfonate, camphorsulfonate, gluconate, glucuronate, oleate, palmitate, stearate, bishydroxynaphthoate, trifluoroacetate, etc. Alkali addition salts can be formed with inorganic or organic bases. Salts derived from inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, lithium, barium, aluminum salts, and the like; salts derived from organic bases include salts formed with various primary, secondary, and tertiary amines, such as ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, amino acids (e.g., lysine, arginine, glycine, and the like), piperazine, piperidine, morpholine, tromethamine, choline, and the like.

[0017] As used herein, the term "pharmaceutically acceptable" indicates that the substance or composition must be chemically and / or toxicologically compatible with other components of the formulation and / or mammal being treated therewith.

[0018] As used herein, the term "therapeutically effective amount" means an amount sufficient to affect treatment as defined below when administered to a mammal in need of such treatment. The therapeutically effective amount varies depending on the subject and disease state to be treated, the subject's weight and age, the severity of the disease state, the mode of administration, etc., and can be easily determined by one skilled in the art.

[0019] As used herein, the term "subject" typically refers to a human subject, although it should be understood that the methods and / or uses described herein are also effective for other animals, such as mammals and vertebrate species. More specifically, the term "subject" refers to animals including, but not limited to, mice, rats, monkeys, dogs, pigs, and rabbits; as well as swine (pigs and pork), ruminants, horses, poultry, felines, bovines, murines, canines, etc.

[0020] As used herein, the term "treatment" includes preventing a disease, preventing a disease from occurring in an animal that may be affected by the disease but has not yet experienced or shown symptoms of the disease (prophylactic treatment), suppressing a disease (alleviating or preventing its progression), alleviating symptoms or side effects of a disease (including palliative treatment), and relieving a disease (attenuation of the disease).

[0021] The term "parenteral" as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection, or infusion techniques.

[0022] active compound The compound of the present invention is a compound represented by formula (I).

[0023] [ka] wherein R is selected from C1-C6 alkyl and n is selected from the integers 0, 1, 2, 3, 4, 5, and 6.

[0024] In some embodiments of the present invention, esters of compounds of Formula (I) include C1-C6 alkyl esters, and pharmaceutically acceptable salts of compounds of Formula (I) include acid addition salts and alkali addition salts.

[0025] In some embodiments of the present invention, the acid addition salts of the compound represented by Formula (I) include inorganic acid salts and organic acid salts. In preferred embodiments of the present invention, inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, hydrogensulfate, hydrogenphosphate, dihydrogenphosphate, and bicarbonate, and organic acid salts include, but are not limited to, formate, acetate, propionate, glycolate, pyruvate, oxalate, malate, malonate, succinate, maleate, fumarate, tartrate, citrate, benzoate, cinnamate, mandelate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate, lactate, nicotinate, lauryl sulfate, naphthalenesulfonate, camphorsulfonate, gluconate, glucuronate, oleate, palmitate, stearate, bishydroxynaphthoate, and trifluoroacetate.

[0026] In some embodiments of the present invention, alkali addition salts of the compound of Formula (I) include salts formed with inorganic bases and salts formed with organic bases. In preferred embodiments of the present invention, salts formed with inorganic bases include, but are not limited to, sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, lithium salts, barium salts, and aluminum salts. Salts formed with organic bases include, but are not limited to, salts of various primary amines, secondary amines, and tertiary amines, preferably, but not limited to, ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, amino acids (e.g., lysine, arginine, glycine, etc.), piperazine, piperidine, morpholine, tromethamine, choline, etc.

[0027] In some embodiments of the present invention, R is selected from C1-C3 alkyl, preferably R is selected from methyl, ethyl.

[0028] In some embodiments of the present invention, n is selected from 0, 1 or 2; preferably, n is selected from 1.

[0029] In some embodiments of the present invention, the compound of formula (I) is selected from the group consisting of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, lithium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, calcium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid meglumine salt, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid arginine salt, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid hydrochloride, and 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid fumarate.

[0030] Pharmaceutical Composition The imidazolidinyl vanillic acid ether derivatives of the present invention can be administered by any pharmaceutically effective route, for example, they can be formulated for oral, intranasal, rectal, vaginal, sublingual, buccal, parenteral, or transdermal administration.

[0031] In some embodiments, the imidazolidinyl vanillic acid ether derivatives of the present invention can be prepared into medicinal oral dosage forms, including, but not limited to, oral solid dosage forms and oral liquid dosage forms. Oral solid dosage forms include, but are not limited to, tablets, capsules, capsules, powders, powders, pills, granules, and any combination thereof. If necessary, these oral solid dosage forms can be prepared as immediate-release, controlled-release, sustained (extended)-release, or modified-release formulations. The oral solid dosage forms of the present invention can further include pharmaceutical excipients such as fillers, diluents, lubricants, surfactants, flow aids, adhesives, dispersants, suspending agents, disintegrants, thickeners, film-forming agents, granulation aids, flavoring agents, sweeteners, coating agents, solubilizers, and combinations thereof. The oral solid dosage forms of the present invention can also include appropriate amounts of controlled-release agents, extended-release agents, modified-release agents, etc., depending on the desired release characteristics. Oral liquid dosage forms include, but are not limited to, solutions, emulsions, suspensions, syrups, and any combination thereof. These oral liquid dosage forms can be prepared using any pharmaceutical excipient known to those skilled in the art for preparing liquid dosage forms, such as water, glycerin, syrup, alcohol, and any combination thereof.

[0032] In some embodiments of the present invention, the imidazolidinyl vanillic acid ether derivatives of the present invention can be prepared in dosage forms for parenteral administration, including, but not limited to, lyophilized powders, solutions, and suspensions (e.g., reservoir suspensions). In other embodiments, the compounds of the present invention can be prepared in dosage forms for topical use, including, but not limited to, patches, gels, pastes, creams, emulsions, liniments, shampoos, lotions, ointments, etc.

[0033] Administration In some embodiments of the present invention, there is provided a method for preventing and / or treating a disease associated with fibrosis by administering to a subject in need thereof a therapeutically effective amount of an imidazolidinyl vanillic acid ether derivative of the present invention.

[0034] The administration of the imidazolidinyl vanillic acid ether derivatives of the present invention can be achieved by any route of administration useful in therapy, including, but not limited to, oral, intranasal, rectal, vaginal, sublingual, buccal, parenteral, or transdermal administration. The dosage should be adjusted according to the age, weight, and condition of the subject, as well as the route, dosage form, and regimen of administration, and the desired results. To achieve the desired plasma concentration of the compound of the present invention, the dosage of the compound of the present invention for treating or preventing fibrotic diseases is preferably within the range of about 0.1 mg / day to about 5000 mg / day. In some embodiments, the dosage of the compound of the present invention is preferably within the range of about 0.5 mg / day to about 2000 mg / day; about 1 mg / day to about 1000 mg / day; about 1 mg / day to about 500 mg / day; about 1 mg / day to about 100 mg / day; about 1 mg / day to about 50 mg / day; or about 1 mg / day to about 20 mg / day.

[0035] Drug combinations The imidazolidinyl vanillic acid ether derivatives of the present invention can be used in combination with one or more additional therapeutic agents for the prevention and / or treatment of diseases associated with fibrosis. In some embodiments of the present invention, the additional therapeutic agent may be a corticosteroid, an antifibrotic agent, an immunosuppressant, a proton pump inhibitor, an angiotensin-converting enzyme inhibitor, an endothelin receptor antagonist, a prostaglandin derivative, a type 2 cannabinoid receptor antagonist, or an IL-17 pathway inhibitor.

[0036] In some embodiments of the invention, the corticosteroid comprises, for example, prednisolone.

[0037] In some embodiments of the invention, anti-fibrotic agents include, for example, phenobarbital and nintedanib.

[0038] In some embodiments of the invention, immunosuppressants include, for example, cyclophosphamide, mycophenolate mofetil, cyclosporine, tacrolimus, azathioprine, mizoribine, and methotrexate.

[0039] In some embodiments of the present invention, proton pump inhibitors include, for example, omeprazole, lansoprazole, rabeprazole, and esoprazole.

[0040] In some embodiments of the invention, angiotensin-converting enzyme inhibitors include, for example, captopril, enalapril, alacepril, imidapril, and temocapril.

[0041] In some embodiments of the present invention, endothelin receptor antagonists include, for example, bosentan, ambrisentan, and macitentan.

[0042] In some embodiments of the present invention, prostaglandin derivatives include, for example, iloprost, beraprost, treprostinil, epoprostenol, or clinprost.

[0043] In some embodiments of the present invention, the type 2 cannabinoid receptor antagonist comprises, for example, Lenabasum.

[0044] In some embodiments of the invention, the IL-17 pathway inhibitor is an anti-IL-17RA antibody, an anti-IL-17A antibody, an anti-IL-17A / F antibody, an anti-IL-23p40 subunit antibody and / or an anti-IL-23p19 subunit antibody, such as podalumab, sukinumab, ixekizumab, nitakinumab, bimegizumab, ustekinumab, tirazumab, risakizumab, migizumab, breculumab, or guselkumab.

[0045] The compound of the present invention and one or more other therapeutic agents may be administered simultaneously or non-simultaneously, together in the same formulation, or separately in different formulations.

[0046] The present invention will be further described by way of specific examples below, but is not limited thereto. In the following specific examples, sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate was used as a representative compound of the imidazolidinyl vanillic acid ether derivatives of the present invention to evaluate the biological activity of the compounds of the present invention.

[0047] Example Example 1 Synthesis of Compound 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate sodium salt (HY-2) [ka] Compound 1 (200 mg, 0.76 mmol) and water (2 mL) were added to a 25 mL single-neck flask, and NaOH (31 mg, 0.76 mmol) was added in an ice-water bath. The reaction was allowed to proceed at room temperature for 2 hours. The reaction solution was slowly added dropwise to acetone (20 mL), resulting in the deposition of a white solid. After the addition was complete, the mixture was stirred at room temperature for 1 hour. The mixture was filtered, washed with acetone (2 mL x 3), and dried to obtain 190 mg of a white solid compound. The yield was 87.7%.

[0048] 1 H NMR(500MHz,D2O)δ(ppm):7.70(s,1H,ArH),7.47(s,1H,ArH),7.44(d,J=9.0Hz,1H,ArH),7.19( s,1H,ArH),6.97(s,1H,ArH),6.92(d,J=8.5Hz,1H,ArH),4.39(s,4H,CH2CH2),3.82(s,3H,OCH3)

[0049] Example 2 Synthesis of compound lithium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate [ka] Compound 1 (200 mg, 0.76 mmol) and water (2 mL) were added to a 25 mL single-neck flask, and LiOH·HO (36 mg, 0.77 mmol) was added in an ice-water bath. The reaction was allowed to proceed at room temperature for 2 h. The reaction solution was slowly added dropwise to acetone (20 mL), and a white solid precipitated. After the addition was complete, the mixture was stirred at room temperature for 1 h. The mixture was filtered, washed with acetone (2 mL x 3), and dried to obtain 140 mg of a white solid compound. The yield was 68.5%.

[0050] 1 H NMR(400MHz,D2O)δ(ppm):7.73(t,J=1.2Hz,1H,ArH),7.54-7.45(m,2H,ArH),7.24(d ,J=1.4Hz,1H,ArH),7.04-6.96(m,2H,ArH),4.46(br,4H,CH2CH2),3.87(s,3H,OCH3).

[0051] Example 3 Synthesis of calcium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate [ka] Compound 1 (100 mg, 0.38 mmol), water (2 mL), and calcium hydroxide (14 mg, 0.19 mmol) were sequentially added to a 10 mL single-neck flask, protected with nitrogen gas, and reacted overnight at room temperature. The reaction solution was slowly added dropwise to acetone (10 mL) at room temperature and stirred for 1 hour. The mixture was filtered and dried to obtain 80 mg of a white solid compound. The yield was 37.3%.

[0052] 1 H NMR(400MHz,D2O)δ(ppm):7.72(s,2H,ArH),7.49-7.46(m,4H,ArH),7.21(s,2H,ArH), 7.00(s,2H,ArH),6.92(d,J=8.4Hz,2H,ArH),4.41(s,8H,CH2CH2),3.84(s,6H,OCH3).

[0053] Example 4 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid meglumine salt [ka] Compound 1 (100 mg, 0.38 mmol), meglumine (74 mg, 0.38 mmol), and methanol (2 mL) were added to a 25 mL single-neck flask, reacted at 60°C for 1 hour, and concentrated to give 160 mg of a white solid compound in a yield of 92.0%.

[0054] 1 H NMR(400MHz,D2O)δ(ppm):7.66(s,1H,ArH),7.42-7.36(m,2H,ArH),7.14(s,1H,ArH),6.92(s,1H,ArH),6.86(d,J=8.3Hz,1H,ArH),4.33(br, 4H,CH2CH2),4.02-3.98(m,1H,CH),3.75(s,3H,OCH3),3.74-3.72(m,1H,CH),3.72-3.71(m,1H,CH),3.70-3.64(m,1H,CH),3.60-3.56(m,1H,H of CH2),3.56-3.53(m,1H,H of CH2),3.15-3.02(m,2H,CH2),2.66(s,3H,NHCH3).

[0055] Example 5 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid arginine salt [ka] Compound 1 (100 mg, 0.38 mmol) and methanol (2 mL) were added sequentially to a 25 mL single-neck flask. After the addition was complete, the mixture was stirred until completely dissolved. A solution of arginine (66 mg, 0.38 mmol) in water (2 mL) was added dropwise to the reaction mixture. After the addition was complete, the mixture was stirred at 60 °C for 1 h. The solvent was removed by vacuum distillation to give a colorless oil. Isopropyl ether (10 mL x 3) was added and evaporated to give a white powder. This was dried in vacuo at 45 °C for 5 h to give 150 mg of a white solid compound. The yield was 90.4%.

[0056] 1H NMR(400MHz,D2O)δ(ppm):7.79(s,1H,ArH),7.37(d,J=8.2Hz,2H,ArH),7.17(s,1H,ArH),6.96(s,1H,ArH),6.82( d,J=8.2Hz,1H,ArH),4.36-4.32(m,2H,NCH2),4.31-4.27(m,2H,OCH2),3.73(s,3H,OCH3),3.65(t,J=6.1Hz,1H,H of CHNH2),3.10(t,J=6.9Hz,2H,NHCH2),1.85-1.74(m,2H,CH2),1.68-1.47(m,2H,CH2).

[0057] Example 6 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid hydrochloride [ka] Compound 1 (200 mg, 0.76 mmol) and ethyl acetate (2 mL) were added to a 25 mL single-neck flask, and a solution of HCl in ethyl acetate (4N, 1 mL) was added dropwise in an ice-water bath. The mixture was stirred at room temperature for 1 h. The reaction mixture was suction filtered, washed with ethyl acetate (2 mL x 3), and dried to obtain 220 mg of a white solid compound. The yield was 96.7%.

[0058] 1 H NMR(400MHz,D2O)δ(ppm):8.83(t,J=1.4Hz,1H,ArH),7.64-7.46(m,4H,ArH),7.08-7. 00(m,1H,ArH),4.70-4.68(m,2H,NCH2),4.57-4.54(m,2H,OCH2),3.81(br,3H,OCH3). HRMS(ESI):m / z[M−H] - ,C 13 H 15 ClN2O4 theoretical value: 297.0642; measured value: 297.0632.

[0059] Example 7 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid fumarate [ka] Compound 1 (100 mg, 0.38 mmol), fumaric acid (44 mg, 0.38 mmol), methanol (2 mL), and water (2 mL) were added sequentially to a 25 mL single-neck flask, followed by stirring at 60°C for 1 h. The solvent was removed by distillation under reduced pressure to obtain a white powder, which was then vacuum dried at 45°C for 5 h to obtain 132 mg of a white solid compound. The yield was 92.1%.

[0060] 1 H NMR(400MHz,D2O)δ(ppm):8.66(t,J=1.5Hz,1H,ArH),7.45(t,J=1.8Hz,1H,ArH),7.39(dd,J1=8.5Hz,J2=2.0Hz,1H,ArH),7.33-7.2 9(m,2H,ArH),6.82(d,J=8.5Hz,1H,ArH),6.52(s,2H,CH=CH),4.54-4.50(m,2H,NCH2),4.37-4.33(m,2H,OCH2),3.67(s,3H,OCH3).

[0061] Example 8: Effects of imidazolidinyl vanillic acid ether derivatives on skin thickening in a BLM-induced systemic sclerosis mouse model 1.1 Materials and Methods Female BALB / c mice weighing 18-22 g were purchased from Beijing Charles River and housed in an SPF environment for one week (with free access to water). They were then randomly divided into five groups of eight mice each: a blank group, a model group, and groups treated with 15 mg / kg, 30 mg / kg, or 60 mg / kg of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate sodium (HY-2). A 1 cm x 1 cm area of ​​hair was shaved on the back of each group. Except for the blank group, which received a subcutaneous injection of saline, the remaining groups received 100 μL of BLM (0.5 mg / mL) for 28 consecutive days. (The first four injections were at the corners of the shaved area, and the fifth injection was at the center of the shaved area. Skin changes at the subcutaneous injection sites and changes in mouse weight were monitored weekly.) The treatment group received the corresponding dose of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate orally (intragastric administration) after BLM injection, once daily for 28 consecutive days.

[0062] 1.2 Experimental results After the final administration, the animals were sacrificed, and appropriate skin tissues were collected and HE-stained sections were prepared to measure skin thickening. The results showed significant skin thickening and loss of subcutaneous fat in the model group. Compared with the model group, skin thickening was significantly suppressed in the 30 mg / kg and 60 mg / kg HY-2 groups (P<0.05, P<0.01) (Figures 1A and 1B).

[0063] Example 9: Pathological effects of imidazolidinyl vanillic acid ether derivatives on lung tissue in a BLM-induced systemic sclerosis mouse model 2.1 Materials and Methods Female BALB / c mice weighing 18-22 g were purchased from Beijing Charles River and housed in an SPF environment for 1 week (with free access to water). Then, eight mice were randomly divided into five groups: a blank group, a model group, and HY-2 treatment groups at 15 mg / kg, 30 mg / kg, and 60 mg / kg. A 1 cm x 1 cm area was shaved on the back of each group. Except for the blank group, which received a subcutaneous injection of saline, the remaining groups received 100 μL of BLM (0.5 mg / mL) for 28 consecutive days. (The first four injections were at the four corners of the shaved area, and the fifth injection was at the center of the shaved area. Skin changes at the subcutaneous injection site and changes in mouse weight were monitored weekly.) After the BLM injection, each treatment group received the corresponding dose of HY-2 orally (intragastric administration) once daily for 28 consecutive days.

[0064] 2.2 Experimental results After the final dose, the animals were sacrificed, and appropriate lung tissue was collected and HE-stained sections were prepared to examine the structure of the lung tissue and the infiltration of inflammatory cells. The results showed that the lung tissue structure of the mice in the model group was disordered and infiltrated with inflammatory cells. Compared to the model group, the lung tissue structure of the mice in each HY-2 dose group was more regular and had less infiltration of inflammatory cells (see Figure 2).

[0065] Example 10: Effects of imidazolidinyl vanillic acid ether derivatives on collagen deposition in skin and lung tissues in a BLM-induced systemic sclerosis mouse model 3.1 Materials and Methods Female BALB / c mice weighing 18-22 g were purchased from Beijing Charles River and housed in an SPF environment for 1 week (with free access to water). Then, eight mice were randomly divided into five groups: a blank group, a model group, and HY-2 treatment groups at 15 mg / kg, 30 mg / kg, and 60 mg / kg. A 1 cm x 1 cm area was shaved on the back of each group. Except for the blank group, which received a subcutaneous injection of saline, the remaining groups received 100 μL of BLM (0.5 mg / mL) for 28 consecutive days. (The first four injections were at the four corners of the shaved area, and the fifth injection was at the center of the shaved area. Skin changes at the subcutaneous injection site and changes in mouse weight were monitored weekly.) After the BLM injection, each treatment group received the corresponding dose of HY-2 orally (intragastric administration) once daily for 28 consecutive days.

[0066] 3.2 Experimental results After the final administration, the animals were sacrificed, and appropriate skin and lung tissues were collected. Masson-stained sections were prepared and collagen deposition in the skin and lung tissues was examined. The percentage of collagen deposition was quantified using Image-Pro® Plus software. The results showed significant collagen deposition in the dermis layer of mouse skin and in the alveoli or blood vessels of lung tissue in the model group (P<0.01). Compared with the model group, collagen deposition in mouse skin and lung tissue in each HY-2 dose group was significantly reduced (P<0.01) (see Figures 3A, 3B, 4A, and 4B).

[0067] Example 11: Comparison of the effects of imidazolidinyl vanillic acid ether derivatives and other antiplatelet drugs on skin thickening in a BLM-induced systemic sclerosis mouse model 1.1 Materials and Methods Female BALB / c mice weighing 18-22 g were purchased from Charles River, Beijing, and housed in an SPF environment for one week (with free access to water). Afterward, ten mice were randomly assigned to nine groups: blank, model, ozagrel sodium, dipyridamole, clopidogrel, cilostazol, ticagrelor, aspirin, and HY-2. Each group received a 1 cm x 1 cm shaved area on their backs. Except for the blank group, which received a subcutaneous injection of saline, the remaining groups received 100 μL of BLM (0.5 mg / mL) subcutaneously for 28 consecutive days. (The first four injections were at the corners of the shaved area, and the fifth injection was at the center of the shaved area. Skin changes at the injection sites and changes in mouse weight were monitored weekly.) Each treatment group received ozagrel sodium, dipyridamole, clopidogrel, cilostazol, ticagrelor, aspirin, and HY-2 30 minutes after BLM injection. These were administered once daily for 28 consecutive days (dosage and administration method are shown in Table 1).

[0068] [Table 1]

[0069] 1.2 Experimental procedures and results After the final administration, the animals were sacrificed, and dorsal skin tissue was collected. It was fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin, and sectioned. The tissue sections were stained with HE staining and microscopically observed for pathological changes in the skin tissue to assess skin thickening. The experimental results are shown in Table 2. The results showed that the skin thickness of mice in the BLM model group was significantly increased compared to the blank group (P<0.01), indicating successful modeling of the model group. The skin thickness of mice in the HY-2 group was significantly decreased compared to the model group (P<0.01). Compared with the ozagrel sodium group, dipyridamole group, clopidogrel group, cilostazol group, ticagrelor group, and aspirin group, HY-2 showed significant benefits in improving skin thickening in BLM-induced SSc mice (P<0.05, P<0.01). Experimental data were processed using GraphPad Prism 7, and each experimental group was compared using student t-test. P<0.05 and P<0.01 indicate that the difference between groups is statistically significant.

[0070] [Table 2]

[0071] # P<0.01, vs blank group; * P<0.01, vs. model group; a P<0.05, aa P<0.01, vs. HY-2 group.

[0072] Example 12: Comparison of the effects of imidazolidinyl vanillic acid ether derivatives and other antiplatelet drugs on the blood thromboxane B2 (TXB2) and transforming growth factor β1 (TGF-β1) levels in a BLM-induced systemic sclerosis mouse model TGF-β1 is the most potent pro-fibrotic mediator currently known. It was first found in platelets and is usually upregulated after tissue injury, playing an important role in extracellular matrix production. Platelets are an important source of TGF-β1, and after platelet activation, they activate and release TGF-β1 into the bloodstream. The activated TGF-β1 also enhances platelet aggregation. Activated TGF-β1 then aggregates near fibroblasts, initiating the fibrotic process and ultimately leading to the fibrosis of systemic sclerosis.

[0073] 1.1 Materials and Methods The procedures were the same as in Example 11. The mouse TXB2 ELISA kit was purchased from Shanghai JONLNBIO Co., Ltd., lot number: 20220715, and antibody type: purified mouse TXB2 capture antibody and horseradish peroxidase (HRP)-labeled detection antibody. The mouse TGF-β1 ELISA kit was purchased from Shanghai JONLNBIO Co., Ltd., lot number: 20220806, and antibody type: purified mouse TGF-β1 capture antibody and horseradish peroxidase (HRP)-labeled detection antibody.

[0074] 1.2 (Experimental principle) The kit used a double-antibody, one-step sandwich enzyme-linked immunosorbent assay (ELISA). Serum specimens, standards, and horseradish peroxidase (HRP)-conjugated detection antibodies were sequentially added to microwells pre-coated with mouse analyte capture antibodies, incubated, and thoroughly washed. Color development occurred with the substrate TMB (3,3',5,5'-tetramethylbenzidine), which converted to blue by the catalytic action of peroxidase (HRP) and then to yellow by the action of acid. The color intensity correlated positively with the analyte content in the serum sample. The absorbance (OD value) was measured at 450 nm using a microplate reader, and the sample concentration was calculated.

[0075] 1.3 Experimental procedures and results After the final administration, the animals were sacrificed and serum samples were collected. Standard wells were placed on the enzyme-coated plate, and standard solutions of different concentrations were prepared according to the manufacturer's instructions. 50 μL of standard solutions of different concentrations were added to each standard well. Blank and test sample wells were placed on the enzyme-coated plate. 40 μL of sample diluent was added to the test sample well, followed by 20 μL of test serum sample. 100 μL of enzyme-labeled reagent was added to each well, excluding the blank well. After sealing with sealing film, the wells were incubated at 37°C for 60 minutes. The sealing film was removed, the liquid was discarded, and the wells were dehydrated and dried. Each well was filled with washing solution, left to stand for 30 seconds, then discarded. This was repeated five times and then dried. 50 μL of developer A (containing H2O2) and 50 μL of developer B (containing the substrate TMB) were added to each well. The wells were mixed evenly by gently shaking, and allowed to develop for 15 minutes at 37°C, protected from light. The reaction was stopped by adding 50 μL of the termination solution to each well (the blue color immediately turned yellow). The microplate reader zeroed the blank wells and measured the absorbance OD value of each well at a wavelength of 450 nm. Measurements should be performed within 15 minutes after adding the termination solution. Experimental data were processed using GraphPad Prism 7. Comparisons between experimental groups were performed using a student t-test. P<0.05 and P<0.01 indicated statistical significance.

[0076] [Table 3]

[0077] # P<0.01, vs blank group; * P<0.05, ** P<0.01, vs. model group; a P<0.05, aa P<0.01, vs. HY-2 group.

[0078] The experimental results showed that TXB2 and TGF-β1 levels in the BLM model group were significantly higher than those in the blank control group (P<0.01), indicating that TXB2 and TGF-β1 play an important role in the BLM-induced systemic sclerosis model. The HY-2 group significantly reduced serum TXB2 and TGF-β1 levels (P<0.05, P<0.01). Other antiplatelet drugs, while having a certain inhibitory effect on serum TXB2, did not significantly inhibit TGF-β1 expression. Here, HY-2's inhibitory effect on serum TXB2 levels was significantly superior to cilostazol (P<0.05). HY-2's inhibitory effect on serum TGF-β1 levels was significantly superior to other drugs, particularly ozagrel sodium and ticagrelor (P<0.05, P<0.01).

[0079] In conclusion, this invention employs a BLM-induced systemic sclerosis model to demonstrate that HY-2 can effectively ameliorate the symptoms of systemic sclerosis disease, inhibit pathological thickening of the skin, protect the structural integrity of lung tissue, and reduce inflammatory cell infiltration, and has great potential for clinical application.

[0080] All references mentioned herein are incorporated herein by reference. It should be understood that the invention of this disclosure is capable of many variations and modifications without departing from the spirit and scope of the disclosure.

Claims

1. Use of a compound represented by formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating a disease associated with fibrosis. 【Chemical 1】 where: The R is C １ -C ６ selected from alkyl, The n is selected from the integers 0, 1, 2, 3, 4, 5, and 6.

2. The use according to claim 1 , wherein the disease associated with fibrosis comprises systemic sclerosis.

3. The use according to claim 2, wherein the systemic sclerosis is selected from diffuse systemic sclerosis, localized systemic sclerosis, overlapping systemic sclerosis, and various lesions associated with systemic sclerosis.

4. The use according to claim 3, wherein the various lesions associated with systemic sclerosis include fibrotic skin lesions, fibrotic lesions of internal organs, fibrotic skin lesions of renal origin, systemic fibrotic lesions of renal origin, keloid formation, systemic sclerosis-associated CRET syndrome, vascular occlusive disease vasculitis, microvascular and macrovascular lesions, Raynaud's phenomenon, ischemic digital injury, digital ulcer, necrotic digital injury, gangrene, and digital loss.

5. The use according to claim 2, wherein the diseases associated with fibrosis include fibrosis of the skin, intestine, liver, lung, heart, bladder, prostate, blood vessels due to excess collagen or other local or systemic fibrosis in tissues.

6. The use according to claim 1, wherein the pharmaceutically acceptable salts of the compound of formula (I) include acid addition salts and alkali addition salts.

7. The acid addition salts include inorganic acid salts and organic acid salts, the inorganic acid salts include hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, hydrogensulfate, hydrogenphosphate, dihydrogenphosphate, and hydrogencarbonate, and the organic acid salts include formate, acetate, propionate, glycolate, pyruvate, oxalate, malate, malonate, succinate, maleate, fumarate, tartrate, citrate, benzoate, cinnamate, mandelate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate, lactate, nicotinate, lauryl sulfate, naphthalenesulfonate, camphorsulfonate, gluconate, glucuronate, oleate, palmitate, and stearate. , bishydroxynaphthoate, and trifluoroacetate, wherein the alkali addition salts include salts formed with inorganic bases and salts formed with organic bases, wherein the salts formed with inorganic bases include sodium salts, potassium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, lithium salts, barium salts, and aluminum salts, and wherein the salts formed with organic bases include salts formed with various primary amines, secondary amines, and tertiary amines, preferably including salts formed with ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, amino acids, piperazine, piperidine, morpholine, tromethamine, and choline.

8. 2. The use according to claim 1, wherein R is selected from methyl or ethyl.

9. The use according to claim 1, wherein n is selected from the integers 0, 1 and 2.

10. The compound represented by the formula (I) is sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, lithium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, calcium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, 2. The use according to claim 1, wherein the hydroxybenzoic acid is selected from the group consisting of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid meglumine salt, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid arginine salt, 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid hydrochloride, and 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid fumarate.

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