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

Abstract

The present invention relates to the use of a compound represented by formula (I) or an ester or a pharmaceutically acceptable salt thereof in the treatment of diseases accompanied by fibrosis. # imgabs0 #

Description

Use of imidazolidinyl vanillic acid ether derivatives in treating diseases associated with fibrosis Technical Field

[0001] The present invention relates to the field of medicinal chemistry, and in particular to use of an imidazolidinyl vanillic acid ether derivative in treating diseases accompanied by fibrosis. Background Art

[0002] Systemic fibrosis, particularly systemic sclerosis (SSc), also known as "scleroderma," is a rare, multi-organ autoimmune disease characterized by fibrosis of the skin and internal organs accompanied by vascular lesions. Clinical manifestations include Raynaud's phenomenon, digital ulcers, pulmonary hypertension, or scleroderma renal crisis. The worldwide prevalence of systemic sclerosis ranges from 8 to 56 per 1 million people, with a higher incidence in women than in men (4:1). Clear epidemiological data are lacking in my country.

[0003] Although the etiology of systemic sclerosis is still not fully understood, in recent years, research on abnormal immune activation, signaling pathway conduction, and cytokine release involved in systemic sclerosis fibrosis has made great progress. Chronic vascular injury, endothelial activation, and immune activation are all considered to be key to secondary fibroblast activation and related fibrosis (Nat Rev Rheumatol. 2019; 15(4): 208-24). Among them, vascular injury is an important component of the pathogenesis of systemic sclerosis. It can activate the coagulation cascade, activate platelets, and increase the production of factors such as thrombin, thromboxane, and platelet-derived growth factor, which has a strong pro-fibrotic effect.

[0004] Due to the rarity and clinical heterogeneity of systemic sclerosis, there is currently no specific treatment. Clinical treatment mainly focuses on improving organ function, with main measures including anti-inflammatory and immune regulation, improving circulation, and inhibiting fibrosis. Therapeutic drugs are divided into immunomodulators and anti-fibrotic drugs. These drugs can alleviate disease progression but cannot improve the condition (Expert Opin Investig Drugs. 2021; 30(6): 635-652). New drugs for the treatment of systemic sclerosis are still needed in clinical practice.

[0005] Summary of the Invention

[0006] The present invention aims to provide the use of a compound represented by formula (I) or an ester or pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating diseases associated with fibrosis.

[0007] Wherein, R is selected from C1-C6 alkyl; n is selected from integers of 0, 1, 2, 3, 4, 5, and 6.

[0008] In certain embodiments of the present invention, the disease associated with fibrosis comprises systemic sclerosis.

[0009] In certain 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.

[0010] In certain embodiments of the present invention, various lesions associated with systemic sclerosis include skin fibrosis, fibrosis of internal organs (such as kidneys, intestines, lungs, blood vessels, etc.), nephrogenic fibrosing skin lesions, nephrogenic systemic fibrosis, keloid formation, and systemic sclerosis-related CRET syndrome (calcification, esophageal dysfunction, sclerosis and capillary dilation), vascular lesions, etc.

[0011] In certain embodiments of the invention, vascular disorders associated with systemic sclerosis include, but are not limited to, vaso-occlusive disease vasculitis, micro- and macrovascular disease, Raynaud's phenomenon, digital ischemic lesions, digital ulcers, digital necrotic lesions, gangrene, and digital loss.

[0012] In certain embodiments of the present invention, diseases associated with fibrosis also include excess collagen (independent of etiology, such as autoimmune diseases, chronic graft-versus-host disease, diabetes, poisoning, surgery, radiotherapy, etc.) leading to fibrosis of the skin, intestines, liver, lungs, heart, bladder, prostate, blood vessels, or other local or systemic fibrosis in tissues. Specifically, other fibrotic diseases include liver fibrosis, cirrhosis, pulmonary fibrosis, endomyocardial fibrosis, glomerulonephritis, interstitial renal fibrosis, fibrotic lesions caused by diabetes, myelofibrosis and similar fibrotic diseases, hypertrophic scars (including after surgery), etc. Beneficial effects

[0013] The compounds of the present invention have good therapeutic effects on diseases associated with fibrosis. Even when administered at low doses, the compounds of the present invention can achieve the desired therapeutic effects, indicating that the compounds of the present invention have good drug safety and have significant clinical benefits when used as drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figures 1A and 1B show the effects of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate (HY-2) of the present invention on skin thickening in model mice (mean ± SD, n = 8); ## P < 0.01, vs blank group; * P<0.05, **P<0.01 vs model group (bleomycin (BLM)).

[0015] FIG2 shows the effect of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on lung tissue pathology (HE staining) in model mice (mean ± SD, n = 8).

[0016] 3A and 3B show the effects of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition (Masson staining) in skin tissue of model mice (mean ± SD, n = 8); ## P < 0.01, vs blank group; ** P<0.01vs BLM.

[0017] 4A and 4B show the effects of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate according to the present invention on collagen deposition (Masson staining) in lung tissue of model mice (mean ± SD, n = 8); ## P < 0.01, vs blank group; ** P<0.01vs BLM. DETAILED DESCRIPTION

[0018] definition

[0019] As used in this specification, the following words and phrases are generally intended to have the meanings set forth below, unless the context in which they are used indicates otherwise.

[0020] As used herein, the term "alkyl" refers to a monovalent group having a straight or branched saturated hydrocarbon chain of 1 to 6 carbon atoms, more typically 1 to 5 carbon atoms, 1 to 4 carbon atoms, or 1 to 3 carbon atoms. This term is exemplified by 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.

[0021] As used herein, the term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness and properties of a given compound and are not biologically or otherwise undesirable. Pharmaceutically acceptable salts can be acid addition salts and / or base addition salts. Acid addition salts can be prepared from inorganic acids and organic acids. Salts derived from inorganic acids include hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, bisulfate, hydrogenphosphate, dihydrogenphosphate, bicarbonate, etc.; 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, pamoate, trifluoroacetate, etc. Base 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, etc.), piperazine, piperidine, morpholine, tromethamine, choline and the like.

[0022] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0023] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to effect treatment, as defined below, when administered to a mammal in need of such treatment. The therapeutically effective amount will vary depending on the subject and disease condition being treated, the weight and age of the subject, the severity of the disease condition, the mode of administration, etc., and can be readily determined by one of ordinary skill in the art.

[0024] As used herein, the term "subject" is typically a human subject, but 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" can be applied to animals including, but not limited to, mice, rats, monkeys, dogs, pigs, and rabbits; as well as domestic swine (pigs and hogs), ruminants, horses, poultry, felines, bovines, rodents, canines, and the like.

[0025] As used herein, the term "treating" includes preventing the disease from occurring in an animal that may be susceptible to the disease but does not yet experience or show symptoms of the disease (prophylactic treatment), inhibiting the disease (slowing or arresting its development), alleviating the symptoms or side effects of the disease (including palliative treatment), and remediating the disease (causing regression of the disease).

[0026] The term "parenteral" as used herein includes subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques.

[0027] Active compound

[0028] The compound of the present invention is a compound represented by formula (I)

[0029] Wherein, R is selected from C1-C6 alkyl; n is selected from integers of 0, 1, 2, 3, 4, 5, and 6.

[0030] In certain embodiments of the present invention, the esters of the compound represented by formula (I) include C1-C6 alkyl esters, and the pharmaceutically acceptable salts of the compound represented by formula (I) include acid addition salts and base addition salts.

[0031] In certain embodiments of the present invention, the acid addition salts of the compound shown in formula (I) include inorganic acid salts and organic acid salts. In a preferred embodiment of the present invention, inorganic acid salts include but are not limited to hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, bisulfate, hydrogen phosphate, dihydrogen phosphate, bicarbonate; 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, pamoate, trifluoroacetate.

[0032] In certain embodiments of the present invention, the base addition salts of the compound shown in formula (I) include salts formed with inorganic bases and salts formed with organic bases. In a preferred embodiment of the present invention, the 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; the salts formed with organic bases include but are not limited to salts formed with various primary amines, secondary amines, and tertiary amines, preferably but not limited to salts formed with ethylamine, diethylamine, n-propylamine, isopropylamine, diethanolamine, meglumine, amino acids (such as lysine, arginine, glycine, etc.), piperazine, piperidine, morpholine, tromethamine, and choline.

[0033] In certain embodiments of the present invention, R is selected from C1-C3 alkyl; preferably, R is selected from methyl and ethyl.

[0034] In certain embodiments of the present invention, n is selected from 0, 1 or 2; preferably, n is selected from 1.

[0035] In certain embodiments of the present invention, the compound represented by formula (I) is selected from 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, meglumine salt of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid, arginine salt of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid, hydrochloride of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid, fumarate of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid.

[0036] Pharmaceutical composition

[0037] The imidazolidinyl vanillic acid ether derivatives of the present invention can be administered by any pharmaceutically effective route, for example, they can be formulated into corresponding preparations for oral administration, intranasal administration, rectal administration, vaginal administration, sublingual administration, buccal administration, parenteral administration, or transdermal administration.

[0038] In certain embodiments, the imidazolidinyl vanillic acid ether derivatives of the present invention can be formulated into pharmaceutical oral dosage forms, including but not limited to oral solid dosage forms and oral liquid dosage forms. Oral solid dosage forms can include but are not limited to tablets, capsule bases, caplets, powders, powders, pills, granules, and any combination thereof. As needed, these oral solid dosage forms can be formulated as immediate release formulations, controlled release formulations, sustained (extended) release formulations, or modified release formulations. The oral solid dosage forms of the present invention can also contain pharmaceutical excipients, such as fillers, diluents, lubricants, surfactants, glidants, adhesives, dispersants, suspending agents, disintegrants, tackifiers, film-forming agents, granulation aids, flavorings, sweeteners, coatings, solubilizers, and combinations thereof. Depending on the desired release characteristics, the oral solid dosage forms of the present invention can contain an appropriate amount of controlled release agents, extended release agents, modified release agents, etc. 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 formulated with any pharmaceutically acceptable excipient known to those skilled in the art for preparing liquid dosage forms, such as water, glycerol, syrup, alcohol, and any combination thereof.

[0039] In certain embodiments of the present invention, the imidazolidinyl vanillic acid ether derivatives of the present invention can be formulated into dosage forms suitable for parenteral use, including but not limited to lyophilized powders, solutions, suspensions (e.g., reservoir suspensions). In other embodiments, the compounds of the present invention can be formulated into dosage forms for topical use, including but not limited to patches, gels, pastes, creams, emulsions, liniments, balms, lotions, ointments, etc.

[0040] Drug administration

[0041] In certain embodiments of the present invention, there is provided a method for preventing and / or treating a disease accompanied by fibrosis by administering a therapeutically effective amount of an imidazolidinylvanillic acid ether derivative of the present invention to a subject in need thereof.

[0042] The administration of the imidazolidinyl vanillic acid ether derivatives of the present invention can be achieved by any therapeutically useful route of administration, including but not limited to oral administration, intranasal administration, rectal administration, vaginal administration, sublingual administration, buccal administration, parenteral administration or transdermal administration. The dosage administered should be adjusted according to the age, weight and condition of the subject, as well as the route of administration, dosage form and regimen and the desired result. In order to obtain the desired plasma concentration of the compounds of the present invention for the treatment or prevention of diseases with fibrosis, the daily dose of the compounds of the present invention is preferably in the range of about 0.1 mg / day to about 5000 mg / day. In certain embodiments, the dosage of the compounds of the present invention is preferably in 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.

[0043] Drug combination

[0044] The imidazolidinyl vanillic acid ether derivatives of the present invention can be administered in combination with one or more additional therapeutic agents for the prevention and / or treatment of diseases associated with fibrosis. In certain embodiments of the present invention, the additional therapeutic agent can be a corticosteroid, an antifibrotic drug, an immunosuppressant, a proton pump inhibitor, an angiotensin-converting enzyme inhibitor, an endothelin receptor antagonist, a prostacyclin derivative, a type 2 cannabinoid receptor antagonist, or an IL-17 pathway inhibitor.

[0045] In certain embodiments of the invention, the adrenocortical steroid includes, for example, prednisolone.

[0046] In certain embodiments of the invention, anti-fibrotic agents include, for example, pirfenidone and nintedanib.

[0047] In certain embodiments of the invention, immunosuppressive agents include, for example, cyclophosphamide, mycophenolate mofetil, cyclosporine, tacrolimus, azathioprine, mizoribine, and methotrexate.

[0048] In certain embodiments of the invention, proton pump inhibitors include, for example, omeprazole, lansoprazole, rabeprazole, and esomeprazole.

[0049] In certain embodiments of the invention, angiotensin converting enzyme inhibitors include, for example, captopril, enalapril, alacepril, imidapril, and temocapril.

[0050] In certain embodiments of the invention, endothelin receptor antagonists include, for example, bosentan, ambrisentan, and macitentan.

[0051] In certain embodiments of the present invention, the prostacyclin derivative includes, for example, iloprost, beraprost, treprostinil, epoprostenol, or clinprost.

[0052] In certain embodiments of the invention, the cannabinoid receptor type 2 antagonist includes, for example, Lenabasum.

[0053] In certain 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 berdalumab, secukinumab, ixekizumab, nitakinumab, bimegilide, ustekinumab, tirazumab, risankizumab, migilide, brevitzumab, or guselkumab.

[0054] The compound of the present invention and one or more additional therapeutic agents may be administered simultaneously or at different times; they may be administered together in the same formulation or separately in different formulations.

[0055] The present invention is further illustrated below by way of specific examples, which are not intended to be limiting thereof. The following specific examples utilize sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate as a representative imidazolidinylvanillic acid ether derivative of the present invention to evaluate the biological activities of the compounds of the present invention.

[0056] Example

[0057] Example 1 Synthesis of Compound 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate (HY-2)

[0058] Compound 1 (200 mg, 0.76 mmol) and water (2 mL) were added to a 25 mL single-necked flask. NaOH (31 mg, 0.76 mmol) was added under an ice-water bath and allowed to react at room temperature for 2 h. The reaction solution was slowly added dropwise to acetone (20 mL), resulting in the precipitation of a white solid. After the addition, the mixture was stirred at room temperature for 1 h. Filtered, washed with acetone (2 mL x 3), and dried to afford 190 mg of the compound as a white solid, with a yield of 87.7%.

[0059] 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).

[0060] Example 2 Synthesis of the compound 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate lithium

[0061] Compound 1 (200 mg, 0.76 mmol) and water (2 mL) were added to a 25 mL single-necked flask. LiOH·H₂O (36 mg, 0.77 mmol) was added under an ice-water bath and allowed to react at room temperature for 2 h. The reaction solution was slowly added dropwise to acetone (20 mL), resulting in the precipitation of a white solid. After the addition, the mixture was stirred at room temperature for 1 h. The mixture was filtered, washed with acetone (2 mL x 3), and dried to afford 140 mg of the compound as a white solid, with a yield of 68.5%.

[0062] 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).

[0063] Example 3 Synthesis of calcium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate

[0064] Compound 1 (100 mg, 0.38 mmol), water (2 mL), and calcium hydroxide (14 mg, 0.19 mmol) were added sequentially to a 10 mL single-necked flask under nitrogen atmosphere. The mixture was allowed to react overnight at room temperature. The reaction solution was slowly added dropwise to acetone (10 mL) at room temperature and stirred for 1 h. Filtration and drying afforded 80 mg of the compound as a white solid, a 37.3% yield.

[0065] 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).

[0066] Example 4 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid meglumine salt

[0067] Compound 1 (100 mg, 0.38 mmol), meglumine (74 mg, 0.38 mmol) and methanol (2 mL) were added sequentially into a 25 mL single-necked bottle, reacted at 60° C. for 1 h, and concentrated to obtain 160 mg of a white solid compound with a yield of 92.0%.

[0068] 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).

[0069] Example 5 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid arginine salt

[0070] Compound 1 (100 mg, 0.38 mmol) and methanol (2 mL) were added sequentially to a 25 mL single-necked flask. After complete addition and stirring, a solution of arginine (66 mg, 0.38 mmol) in water (2 mL) was added dropwise to the reaction mixture. After addition, the mixture was stirred at 60°C for 1 h. The solvent was removed by vacuum distillation to obtain a colorless oil. Isopropyl ether (10 mL × 3) was added and evaporated to obtain a white powder. The product was dried under vacuum at 45°C for 5 h to obtain 150 mg of the compound as a white solid, with a yield of 90.4%.

[0071] 1 H 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).

[0072] Example 6 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid hydrochloride

[0073] Compound 1 (200 mg, 0.76 mmol) and ethyl acetate (2 mL) were added to a 25 mL single-necked flask. A 4 N HCl solution in ethyl acetate (1 mL) was added dropwise under an ice-water bath and stirred at room temperature for 1 h. The reaction mixture was filtered, washed with ethyl acetate (2 mL x 3), and dried to afford 220 mg of the compound as a white solid (96.7% yield).

[0074] 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).

[0075] HRMS(ESI):m / z[MH] - C 13 H 15Theoretical value of ClN2O4: 297.0642; Measured value: 297.0632.

[0076] Example 7 Synthesis of 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoic acid fumarate

[0077] To a 25 mL single-necked flask, compound 1 (100 mg, 0.38 mmol), fumaric acid (44 mg, 0.38 mmol), methanol (2 mL), and water (2 mL) were added sequentially. After addition, the mixture was stirred at 60°C for 1 h. The solvent was removed by distillation under reduced pressure to obtain a white powder, which was dried in vacuo at 45°C for 5 h to afford 132 mg of the compound as a white solid, a yield of 92.1%.

[0078] 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).

[0079] Example 8: Effects of Imidazolidinyl Vanillyl Ether Derivatives on Skin Thickening in a BLM-Induced Systemic Sclerosis Mouse Model

[0080] 1.1 Materials and Methods

[0081] Female BALB / c mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. 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 sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate (HY-2). A 1 cm x 1 cm area on the back of each mouse was shaved. Except for the blank group, which was subcutaneously injected with normal saline, all other groups were injected with 100 μL (0.5 mg / mL) of BLM for 28 consecutive days (the injection area was the four corners of the shaved area for the first four injections and the center of the shaved area for the fifth injection, and this cycle was repeated. The skin changes at the subcutaneous injection site and the weight changes of the mice were observed every week). After the injection of BLM, the treatment group was orally (gavage) administered with the corresponding dose of sodium 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate once a day for 28 consecutive days.

[0082] 1.2 Experimental Results

[0083] After the final dose, the animals were sacrificed, and appropriate skin tissue was collected for HE staining and measurement of skin thickening. The results showed that the skin of the model group mice was significantly thickened and subcutaneous fat was lost. Compared with the model group, skin thickening was significantly inhibited in the HY-2 30 and 60 mg / kg groups (P < 0.05, P < 0.01) (see Figures 1A and 1B).

[0084] Example 9: Effects of Imidazolidinyl Vanillyl Ether Derivatives on Lung Tissue Pathology in a BLM-Induced Systemic Sclerosis Mouse Model

[0085] 2.1 Materials and Methods

[0086] Female BALB / c mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. 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 receiving HY-2 at doses of 15 mg / kg, 30 mg / kg, and 60 mg / kg. A 1 cm x 1 cm area on the back of each mouse was shaved. Except for the blank group, which received a subcutaneous injection of normal saline, all other groups received a 100 μL (0.5 mg / mL) injection of BLM for 28 consecutive days (the first four injections were given at the four corners of the shaved area, followed by the fifth injection at the center of the shaved area, and this cycle continued). Skin changes at the subcutaneous injection site and weight changes of the mice were observed weekly. Following BLM injection, each treatment group received the corresponding dose of HY-2 orally (gavage) once daily for 28 consecutive days.

[0087] 2.2 Experimental Results

[0088] After the final dose, the animals were sacrificed, and appropriate lung tissue was obtained for HE staining and preparation to examine lung tissue structure and inflammatory cell infiltration. The results showed that the lung tissue structure of the model group mice was disordered and infiltrated with inflammatory cells. Compared with the model group, the lung tissue structure of mice in all HY-2 dose groups was regular, with less inflammatory cell infiltration (see Figure 2).

[0089] Example 10: Effects of Imidazolidinyl Vanillyl Ether Derivatives on Collagen Deposition in Skin and Lung Tissues of BLM-Induced Systemic Sclerosis Mouse Model

[0090] 3.1 Materials and Methods

[0091] Female BALB / c mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. 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 receiving HY-2 at doses of 15 mg / kg, 30 mg / kg, and 60 mg / kg. A 1 cm x 1 cm area on the back of each mouse was shaved. Except for the blank group, which received a subcutaneous injection of normal saline, all other groups received a 100 μL (0.5 mg / mL) injection of BLM for 28 consecutive days (the first four injections were given at the four corners of the shaved area, followed by the fifth injection at the center of the shaved area, and this cycle continued). Skin changes at the subcutaneous injection site and weight changes of the mice were observed weekly. Following BLM injection, each treatment group received the corresponding dose of HY-2 orally (gavage) once daily for 28 consecutive days.

[0092] 3.2 Experimental Results

[0093] After the last administration, the animals were sacrificed and appropriate skin and lung tissues were taken for Masson staining and preparation to examine the collagen deposition in the skin and lung tissues. The percentage of collagen deposition was quantified using HY-2 Plus software. The results showed that significant collagen deposition was observed in the dermis of the skin and in the alveoli or blood vessels of the lung tissue of the model group mice (P < 0.01). Compared with the model group, collagen deposition in the skin and lung tissue of the mice in each HY-2 dose group was significantly reduced (P < 0.01) (see Figures 3A, 3B, 4A, and 4B).

[0094] 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

[0095] 1.1 Materials and Methods

[0096] Female BALB / c mice, weighing 18-22 g, were purchased from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. and housed in an SPF environment for 1 week with free access to water. They were then randomly divided into nine groups, each consisting of 10 mice: blank, model, ozagrel, dipyridamole, clopidogrel, cilostazol, ticagrelor, aspirin, and HY-2. A 1 cm x 1 cm area on the back of each mouse was shaved. Except for the blank group, which was subcutaneously injected with normal saline, all other groups were subcutaneously injected with BLM 100 μL (0.5 mg / mL) for 28 consecutive days (the injection area was the four corners of the shaved area for the first four injections and the center of the shaved area for the fifth injection, and this cycle was repeated. The skin changes at the subcutaneous injection site and the weight changes of the mice were observed every week). Each treatment group was given sodium ozagrel, dipyridamole, clopidogrel, cilostazol, ticagrelor, aspirin, and HY-2 30 minutes after the injection of BLM, once a day for 28 consecutive days (the dosage and administration method are shown in Table 1).

[0097] Table 1 Dosage and administration method

[0098] 1.2 Experimental steps and results

[0099] After the last dose, the animals were sacrificed, and dorsal skin tissue was obtained and fixed with 4% paraformaldehyde, dehydrated, paraffin-embedded, and sectioned. Tissue sections were stained with hematoxylin and eosin. Skin histopathological changes and skin thickening were observed microscopically. The results are shown in Table 2. The results showed that skin thickness in the BLM model group was significantly increased compared with the blank group (P < 0.01), indicating successful model establishment. The skin thickness in the HY-2 group was significantly decreased compared with the model group (P < 0.01). Compared with the ozagrel sodium, dipyridamole, clopidogrel, cilostazol, ticagrelor, and aspirin groups, HY-2 significantly improved BLM-induced skin thickness in SSc mice (P < 0.05, P < 0.01). Data were processed using GraphPad Prism 7, and comparisons between experimental groups were performed using the Student's t-test. P < 0.05 or P < 0.01 indicated statistically significant differences between groups.

[0100] Table 2 Effects of compounds on skin thickening in BLM-induced systemic sclerosis mouse model (mean ± SE, n = 8) # P < 0.01 vs blank group; * P < 0.01 vs model group; a P<0.05, aa P<0.01vs HY-2 group.

[0101] Example 12: Comparison of the effects of imidazolidinyl vanillic acid ether derivatives and other antiplatelet drugs on the levels of thromboxane B2 (TXB2) and transforming growth factor β1 (TGF-β1) in the blood of a BLM-induced systemic sclerosis mouse model

[0102] TGF-β1 is the most potent profibrotic mediator known. Originally discovered in platelets, it is typically upregulated following tissue injury and plays a key role in the production of the extracellular matrix. Platelets are an important source of TGF-β1. Following platelet aggregation and activation, platelets activate TGF-β1 and release it into blood vessels. Activated TGF-β1 also enhances platelet aggregation. Activated TGF-β1 further aggregates near fibroblasts and initiates the fibrogenic process, leading to the ultimate fibrosis in systemic sclerosis.

[0103] 1.1 Materials and Methods

[0104] Similar to Example 11. Mouse TXB2 ELISA kit, purchased from Shanghai Jianglai Biological, batch number: 20220715, antibody type: purified mouse TXB2 capture antibody and horseradish peroxidase (HRP)-labeled detection antibody; mouse TGF-β1 ELISA kit, purchased from Shanghai Jianglai Biological, batch number: 20220806, antibody type: purified mouse TGF-β1 capture antibody and horseradish peroxidase (HRP)-labeled detection antibody.

[0105] 1.2 Experimental Principle

[0106] The kit uses a double-antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA). Serum specimens, standards, and horseradish peroxidase (HRP)-labeled detection antibodies are added sequentially to the coated microwells pre-coated with mouse analyte capture antibodies, followed by incubation and thorough washing. The substrate TMB (3,3',5,5'-tetramethylbenzidine) is used for color development. TMB is converted to blue under the catalysis of peroxidase (HRP) and to the final yellow under the action of acid. The depth of the color is positively correlated with the content of the analyte in the serum sample. The absorbance (OD value) is measured at a wavelength of 450nm using an enzyme reader to calculate the sample concentration.

[0107] 1.3 Experimental steps and results

[0108] After the last dose, the animals were killed and serum samples were collected. Standard wells were set on the enzyme-labeled plate, and standard solutions of different concentrations were prepared according to the instructions. 50uL of standard solutions of different concentrations were added to each standard well. Blank wells and test sample wells were set on the enzyme-labeled plate. 40uL of sample diluent was first added to the test sample well, and then 20uL of the serum sample to be tested was added. 100uL of enzyme-labeled reagent was added to each well, except for the blank well. The plate was sealed with a sealing film and incubated at 37°C for 60 minutes. The sealing film was removed, the liquid was discarded, and the plate was dried. Fill each well with wash buffer, let stand for 30 seconds, then discard. Repeat five times and pat dry. First, add 50 μL of chromogen A (containing H₂O₂) to each well, followed by 50 μL of chromogen B (containing the substrate TMB). Gently shake to mix, and develop at 37°C in the dark for 15 minutes. Add 50 μL of stop solution to each well to terminate the reaction (the blue color immediately turns yellow). Zero the microplate reader with the blank well and measure the absorbance of each well at 450 nm. Measurements should be performed within 15 minutes of adding the stop solution. Experimental data were processed using GraphPad Prism 7. Comparisons between experimental groups were performed using the Student's t-test. P < 0.05; P < 0.01 indicates statistically significant differences between groups.

[0109] Table 3 Effects of compounds on serum TXB2 and TGF-β1 in BLM-induced systemic sclerosis mouse model (mean ± SE, n = 8) # 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;

[0110] The results showed that TXB2 and TGF-β1 levels in the BLM model group were significantly elevated compared with the blank control group (P<0.01), indicating that TXB2 and TGF-β1 play a key 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). Although other antiplatelet drugs had some inhibitory effects on serum TXB2, none significantly inhibited TGF-β1 expression. HY-2 significantly inhibited serum TXB2 levels compared with cilostazol (P<0.05). HY-2 also generally suppressed serum TGF-β1 levels better than other drugs, particularly compared with ozagrel sodium and ticagrelor (P<0.05, P<0.01).

[0111] In summary, the present invention uses a BLM-induced systemic sclerosis disease model to demonstrate that HY-2 can effectively improve the symptoms of systemic sclerosis, 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.

[0112] The references mentioned herein are all incorporated herein by reference. It should be understood that many changes and modifications can be made to the technical solutions of the present invention without departing from the spirit and scope of the present disclosure.

Claims

1. A compound represented by formula (I): or its ester or pharmaceutically acceptable salt in the preparation of a medicament for preventing and / or treating a disease associated with fibrosis, wherein R is selected from C1-C6 alkyl; n is an integer selected from 0, 1, 2, 3, 4, 5, and 6.

2. The use according to claim 1, wherein the disease accompanied by 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 skin fibrosis, fibrotic lesions of internal organs, nephrogenic fibrosing skin lesions, nephrogenic systemic fibrosis, keloid formation, systemic sclerosis-related CRET syndrome, vascular occlusive disease vasculitis, microvascular and macrovascular lesions, Raynaud's phenomenon, digital ischemic lesions, digital ulcers, digital necrotic lesions, gangrene and digital loss.

5. The use according to claim 2, wherein the disease accompanied by fibrosis comprises fibrosis of the skin, intestine, liver, lung, heart, bladder, prostate, blood vessels or other local or systemic fibrosis in tissues caused by excessive collagen.

6. The use according to claim 1, wherein the ester of the compound represented by formula (I) comprises a C1-C6 alkyl ester, and the pharmaceutically acceptable salt of the compound represented by formula (I) comprises an acid addition salt and a base addition salt.

7. The method according to claim 6, wherein the acid addition salt comprises an inorganic acid salt and an organic acid salt, wherein the inorganic acid salt comprises hydrochloride, hydrobromide, sulfate, nitrate, phosphate, carbonate, hydrogen sulfate, hydrogen phosphate, dihydrogen phosphate, and hydrogen carbonate, and the organic acid salt comprises 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, pamoate, trifluoroacetate; the base addition salt includes 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, aluminum salts; 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. The method according to claim 1, wherein R is selected from methyl or ethyl.

9. The method according to claim 1, wherein n is an integer selected from 0, 1 or 2.

10. The method according to claim 1, wherein the compound represented by formula (I) is selected from 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, meglumine 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, arginine 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, hydrochloride 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate, fumarate 4-(2-(1H-imidazol-1-yl)ethoxy)-3-methoxybenzoate.

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