Triazole fused ring compound as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention discloses a triazole fused ring compound as well as a preparation method, a pharmaceutical composition and application thereof. The structure of the compound is shown as a formula I, and the compound also comprises pharmaceutically acceptable salts thereof. The compound disclosed by the invention has the characteristics of high efficiency and low toxicity, the inhibition rate on cell fibrosis at a 10 micromole concentration level is even higher than 80%, cells even can completely survive, and the compound has a good application prospect in preparation of medicines for treating heart, lung, liver and kidney fibrosis. The preparation method is simple and convenient, high in universality and beneficial to expansion of various structures. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a triazole ring compound and a preparation method, a pharmaceutical composition and application thereof, and in particular to a triazole ring compound which can be prepared as an anti-fibrosis drug and a preparation method, a pharmaceutical composition and application thereof. Background Art

[0002] Excessive accumulation of extracellular matrix (ECM) components such as collagen and fibronectin is a prominent feature of fibrosis. During the development of fibrosis, ECM components continue to accumulate, leading to tissue structural destruction, organ dysfunction, and ultimately organ failure. Therefore, fibrosis is a pathological process in many chronic inflammatory diseases or injuries.

[0003] Fibrosis affects almost all tissues in the body and leads to high morbidity and mortality in many diseases, such as idiopathic pulmonary fibrosis (IPF). Despite a deep understanding of the pathobiology of fibrosis, there are few effective treatment strategies or therapeutic agents. Currently available drugs, pirfenidone and nintedanib, can slow the progression of pulmonary fibrosis, but they have serious side effects in clinical applications, such as nausea, diarrhea, and in a few cases, they can cause more serious allergic reactions. Therefore, there is an urgent need to develop new safe and effective anti-fibrotic drugs.

[0004] Summary of the invention

[0005] Purpose of the invention: Purpose of the invention: The first purpose of the present invention is to provide a triazole ring compound, the second purpose is to provide a method for preparing the compound, the third purpose is to provide a pharmaceutical composition containing the compound, and the fourth purpose is to provide an application of the compound and its pharmaceutical composition in the preparation of anti-fibrosis drugs.

[0006] Technical solution: The triazole ring compound of the present invention has a structure of Formula I, and also includes a pharmaceutically acceptable salt thereof:

[0007]

[0008] in:

[0009] R is selected from C1-C4 alkyl, 3-6 membered cycloalkyl, -M-(6-12 membered aryl), -M-(6-12 membered heteroaryl containing 1-2 nitrogen atoms), M is connected to the 6-12 membered aryl, the 6-12 membered heteroaryl containing 1-2 nitrogen atoms by chemical bonds or cyclization;

[0010] M is selected from -(CH 2 ) n-, 4-7 membered cycloalkyl;

[0011] n is an integer selected from 0 to 4;

[0012] The 6-12 membered aryl group and the 6-12 membered heteroaryl group containing 1-2 nitrogen atoms are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, hydroxyl, amino, nitro, and cyano.

[0013] Preferably, in the structure:

[0014] R is selected from C1-C4 alkyl, 3-5 membered cycloalkyl, -M-phenyl, -M-pyridyl;

[0015] M is selected from -(CH 2 ) n -, M is connected to the phenyl group or pyridyl group by a chemical bond; when M is selected from a 4-6 membered cycloalkyl group, M is connected to the phenyl group or pyridyl group by a ring;

[0016] n is an integer selected from 1 to 3;

[0017] The phenyl group and the pyridyl group are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

[0018] Further preferably, in the structure:

[0019] R is selected from methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyclopropyl, cyclopentyl, -(CH 2 ) n -phenyl, -(CH 2 ) n -pyridyl, cyclopentylphenyl;

[0020] n is an integer selected from 1 to 3;

[0021] The phenyl group and the pyridyl group are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

[0022] More preferably, in the structure:

[0023] The phenyl group is substituted with at least one substituent selected from any of the following: hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy; the pyridyl group is substituted with hydrogen; wherein when the phenyl group is monosubstituted, the substituent is located at the ortho position, meta position or para position, and when the phenyl group is disubstituted, the substituent is located at the meta position.

[0024] Most preferably, in the structure, R is selected from any of the following structures:

[0025]

[0026] Preferably, the pharmaceutically acceptable salt is a salt formed by the compound and an acid selected from any one of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, and ferulic acid.

[0027] "Pharmaceutically acceptable salts" refer to salts of compounds prepared from compounds having specified substituents with relatively nontoxic acids or bases. When the compound contains relatively acidic functional groups, base addition salts can be obtained by contacting the free form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compound contains relatively basic functional groups, acid addition salts can be obtained by contacting the free form of such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming carbonate or bicarbonate), phosphoric acid (forming phosphate, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid (forming sulfate or bisulfate), hydroiodic acid, phosphorous acid, and the like; and organic acid salts such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid. Acids such as citric acid, tartaric acid and methanesulfonic acid; organic acid salts also include salts of organic acids such as amino acids (such as arginine, etc.) and glucuronic acid. When certain specific compounds contain basic and acidic functional groups, they can be converted into any base or acid addition salt. Preferably, the salt is contacted with a base or an acid in a conventional manner, and the parent compound is separated to regenerate the free form of the compound. The free form of the compound differs from its various salt forms in certain physical properties, such as different solubility in polar solvents.

[0028] "Pharmaceutically acceptable salts" can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. In general, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid in water or an organic solvent or a mixture of the two. In general, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred.

[0029] The preparation method of the triazole ring compound of the present invention comprises the following steps:

[0030]

[0031] Compound 1 is subjected to reduction, condensation and aminolysis reactions to obtain compound I; that is, compound 1 is dissolved in an ethanol / ethyl acetate (1:1) solution and reduced under 10% Pd / C catalytic hydrogenation to obtain intermediate 2; then 2 is dissolved in dichloromethane and subjected to a condensation reaction in the presence of HATU and DIEA to obtain intermediate 3; finally, 3 is reacted with a substituted amine compound in a tert-butanol solution to obtain the target product 4-31 (specific compound of formula I).

[0032] Wherein, R is as defined above; specifically, the structures of compounds 4 to 31 prepared by the present invention are as follows:

[0033]

[0034]

[0035] The corresponding acid is reacted with the compound I prepared by the above method to form a salt, thereby obtaining a pharmaceutically acceptable salt of the compound.

[0036] The pharmaceutical composition of the present invention comprises the triazole ring compound of the present invention and a pharmaceutically acceptable carrier.

[0037] The pharmaceutical composition also contains a pharmaceutically acceptable carrier.

[0038] "Pharmaceutically acceptable carrier" can be an excipient widely used in the field of drug production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method to dissolve the active ingredient at a desired rate after the subject receives the administration, or promote the effective absorption of the active ingredient after the subject receives the composition. The pharmaceutical excipient can be an inert filler, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipient can include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0039] The pharmaceutical composition of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0040] The pharmaceutical composition of the present invention can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical composition of the present invention can also be a controlled release or sustained release dosage form (e.g., liposomes or microspheres). Examples of solid oral preparations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs, and solutions. Examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of preparations for parenteral administration include, but are not limited to, solutions for injection, dry powder preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalers; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

[0041] The triazole ring compound or the pharmaceutical composition thereof of the present invention is used in preparing anti-fibrosis drugs.

[0042] Preferably, the drug is a drug for preventing cardiac, pulmonary, liver or renal fibrosis.

[0043] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages:

[0044] This type of compound has the characteristics of high efficiency and low toxicity. The inhibition rate of cell fibrosis at a concentration level of ten micromoles is even higher than 80%, and the cells can even survive completely. It has good application prospects in the preparation of drugs for heart, lung, liver, and kidney fibrosis. The preparation method of the compound is simple and versatile, which is conducive to the expansion of various structures. DETAILED DESCRIPTION

[0045] The technical solution of the present invention is further described below in conjunction with embodiments.

[0046] Example 1: Preparation of compounds

[0047] 1. Synthesis of intermediate 2

[0048] 10% Pd / C (17 mg, 0.158 mmol) and compound 1 (100 mg, 0.395 mmol) were added to a three-necked flask, and ethanol / ethyl acetate (1:1) solution was slowly added. Hydrogen was introduced at room temperature for 4 h. After the reaction was completed, the mixture was filtered. The filtrate was concentrated and purified by column chromatography to obtain compound 2 (white solid, 72.1 mg, yield: 71.0%).

[0049] 2. Synthesis of Intermediate 3

[0050] Compound 2 (300 mg, 1.53 mmol) was dissolved in dichloromethane, and ursodeoxycholic acid (900 mg, 2.3 mmol), HATU (874 mg, 2.3 mmol) and DIEA (380 μL, 2.3 mmol) were added in sequence, and the mixture was reacted at room temperature for 1 h. After the reaction was completed, water was added, and the mixture was extracted with dichloromethane for 3 times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to obtain compound 3 (white solid, 516.6 mg, yield: 59.2%).

[0051] 3. Synthesis of Compound 4-31

[0052] Compound 3 (100 mg, 0.175 mmol) and substituted amine (0.875 mmol) were placed in a tert-butyl alcohol solution, and the reaction mixture was heated to 90°C for 2 h. After the reaction was completed, water was added, and the mixture was extracted with dichloromethane three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to obtain the target compound 4-31.

[0053] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthren-17-yl)pentanoyl)-N-ethyl-5,6,7,8-tetrahydro-[1,2,4-triazolo[4,3-a]pyrazine-3-carboxamide (4). Yield: 71.0%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ*5.01(s,0.8H),4.94(d,J=2.3Hz,1.2H),4.49(t,J=5.3Hz,1.2H),*4.38(t,J=5.3H z,0.8H),4.05-3.97(m,2H),3.53-3.43(m,2H),3.40(dd,J=14.4Hz,7.2Hz,2H),2.64-2.53(m ,1H),2.49-2.39(m,1H),2.08-2.00(m,1H),1.94-1.76(m,5H),1.65-1.53(m,4H),1.51-1.41 (m,6H),1.40-1.08(m,11H),1.02-0.99(m,3H),0.97(s,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0054] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-isopropyl-5,6,7,8-tetrahydro-[1,2,4-triazolo[4,3-a]pyrazine-3-carboxamide (5). Yield: 95.5%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ*5.01(s,0.8H),4.94(d,J=2.6Hz,1.2H),4.50(t,J=5.2Hz,1.2H),*4.39(t,J=5 .2Hz,0.8H),4.22-4.11(m,1H),4.04-3.96(m,2H),3.53-3.43(m,2H),2.63-2.52(m,1H), 2.49-2.40(m,1H),2.08-2.00(m,1H),1.94-1.76(m,5H),1.65-1.53(m,4H),1.52-1.42(m ,6H),1.40-1.05(m,14H),1.02-0.99(m,3H),0.97(s,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0055] N-cyclopropyl-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4-triazolo[4,3-a]pyrazine-3-carboxamide (6). Yield: 76.6%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ*5.00(s,0.8H),4.94(d,J=2.5Hz,1.2H),4.50(t,J=5.4Hz,1.2H),*4.39(t,J=5. 4Hz,0.8H),4.05-3.96(m,2H),3.54-3.43(m,2H),2.87-2.81(m,1H),2.63-2.52(m,1H),2. 48-2.38(m,1H),2.10-1.99(m,1H),1.93-1.76(m,5H),1.63-1.53(m,4H),1.51-1.41(m,6H ),1.38-1.05(m,8H),1.03-0.99(m,3H),0.97(s,3H),0.85-0.79(m,2H),0.74-0.66(m,5H).

[0056] N-cyclopentyl-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (7). Yield: 93.5%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ*5.01(s,0.8H),4.94(d,J=3.2Hz,1.2H),4.49(t,J=5.4Hz,1.2H),*4.38(t,J=5 .4Hz,0.8H),4.33-4.24(m,1H),4.04-3.95(m,2H),3.54-3.42(m,2H),2.64-2.52(m,1H), 2.51-2.39(m,1H),2.08-1.97(m,3H),1.93-1.74(m,7H),1.68-1.52(m,8H),1.51-1.41(m ,6H),1.40-1.03(m,8H),1.02-0.98(m,3H),0.97(s,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0057] N-(2,3-dihydro-1H-inden-2-yl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (8). Yield: 88.0%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.24-7.19(m,2H),7.17-7.12(m,2H),*5.00(s,0.8H),4.93(d,J=2.8Hz,1.2H),4.81-4.75(m,1H),4.50(t ,J=5.2Hz,1.2H),*4.39(t,J=5.2Hz,0.8H),4.04-3.96(m,2H),3.53-3.43(m,2H),3.34-3.27(m,2H),3.03(d,J=6.4H z,1.2H),3.00(d,J=6.4Hz,0.8H),2.63-2.51(m,1H),2.50-2.39(m,1H),2.08-2.00(m,1H),1.93-1.77(m,5H),1.64 -1.53(m,4H),1.52-1.42(m,6H),1.37-1.05(m,8H),1.03-0.98(m,3H),0.97(s,3H),0.72(s,1.8H),*0.68(s,1.2H).

[0058] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-phenylethyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (9). Yield: 77.4%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureofrotamers*indicates minor amide rotamer δ7.30-7.23(m,4H),7.21-7.16(m,1H),*5.00(s,0.8H),4.93(d,J=2.4Hz,1.2H),4.46(t,J=5.2Hz ,1.2H),*4.36(t,J=5.2Hz,0.8H),4.03-3.95(m,2H),3.59(t,J=7.5Hz,2H),3.53-3.43(m,2H),2.90(t,J=7 .5Hz,2H),2.61-2.52(m,1H),2.48-2.39(m,1H),2.08-2.00(m,1H),1.94-1.76(m,5H),1.65-1.53(m,4H),1 .51-1.41(m,6H),1.38-1.04(m,8H),1.02-0.98(m,3H),0.97-0.94(m,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0059] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-(4-fluorophenylethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (10). Yield: 84.1%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.30-7.24(m,2H),7.03-6.97(m,2H),*5.00(s,0.8H),4.94(d,J=2.5Hz,1.2H),4.47(t,J=5.4H z,1.2H),*4.37(t,J=5.4Hz,0.8H),4.03-3.96(m,2H),3.58(t,J=7.4Hz,2H),3.53-3.43(m,2H),2.89(t,J =7.4Hz,2H),2.61-2.53(m,1H),2.48-2.40(m,1H),2.08-2.01(m,1H),1.94-1.77(m,5H),1.65-1.53(m,4 H),1.51-1.41(m,6H),1.38-1.09(m,8H),1.02-0.99(m,3H),0.96(m,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0060] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-(3-fluorophenylethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (11). Yield: 80.6%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.30-7.25(m,1H),7.09-7.05(m,1H),7.04-6.98(m,1H),6.95-6.87(m,1H),*5.00(s,0.8H),4.93(d,J=1 .9Hz,1.2H),4.47(t,J=5.2Hz,1.2H),*4.36(t,J=5.2Hz,0.8H),4.04-3.95(m,2H),3.60(t,J=7.4Hz,2H),3.53-3.4 3(m,2H),2.92(t,J=7.4Hz,2H),2.61-2.52(m,1H),2.48-2.39(m,1H),2.07-1.99(m,1H),1.93-1.76(m,5H),1.64- 1.53(m,4H),1.51-1.41(m,6H),1.38-1.04(m,8H),1.02-0.98(m,3H),0.96(m,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0061] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-N-(2-fluorophenylethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (12). Yield: 84.3%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamerδ7.29(td,J=7.6Hz,1.4Hz,1H),7.25-7.19(m,1H),7.08(td,J=7.6Hz,0.8Hz,1H),7.06-7.00(m,1H),*5.00(s,0 .8H),4.93(d,J=1.9Hz,1.2H),4.46(t,J=5.3Hz,0.8H),*4.35(t,J=5.3Hz,0.8H),4.04-3.95(m,2H),3.61(t,J=7.5Hz,2H ),3.53-3.43(m,2H),2.96(t,J=7.5Hz,2H),2.63-2.51(m,1H),2.49-2.38(m,1H),2.09-1.99(m,1H),1.92-1.76(m,5H), 1.64-1.53(m,4H),1.50-1.41(m,6H),1.39-1.04(m,8H),1.02-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0062] N-(4-chlorophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (13). Yield: 80.6%. White solid. 1 HNMR (600MHz, CD 3OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ7.29-7.22(m,4H),*5.00(s,0.8H),4.94(d,J=1.7Hz,1.2H),4.46(t,J=5.1Hz,1.2H),*4.3 5(t,J=5.1Hz,0.8H),4.04-3.95(m,2H),3.59(t,J=7.5Hz,2H),3.53-3.43(m,2H),2.89(t,J=7.5Hz,2 H),2.63-2.52(m,1H),2.48-2.38(m,1H),2.07-1.99(m,1H),1.93-1.77(m,5H),1.64-1.53(m,4H),1 .51-1.42(m,6H),1.40-1.04(m,8H),1.02-0.98(m,3H),0.96(s,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0063] N-(3-chlorophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (14). Yield: 82.3%. White solid. 1 HNMR (600MHz, CD 3OD):~60:40mixture ofrotamers*indicates minor amide rotamerδ7.31-7.28(m,1H),7.27-7.23(m,1H),7.22-7.17(m,2H),*5.00(s,0.8H),4.93(d,J=1.5Hz,1.2H),4 .46(t,J=5.3Hz,1.2H),*4.35(t,J=5.3Hz,0.8H),4.03-3.95(m,2H),3.60(t,J=7.2Hz,2H),3.53-3.44(m,2H), 2.90(t,J=7.2Hz,2H),2.62-2.53(m,1H),2.48-2.40(m,1H),2.07-2.00(m,1H),1.93-1.78(m,5H),1.64-1.53 (m,4H),1.51-1.42(m,6H),1.38-1.05(m,8H),1.02-0.98(m,3H),0.97(s,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0064] N-(2-chlorophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (15). Yield: 84.0%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamerδ7.37-7.34(m,1H),7.33-7.31(m,1H),7.23-7.17(m,2H),*5.00(s,0.8H),4.93(d,J=2.0Hz,1.2H),4 .46(t,J=5.1Hz,1.2H),*4.35(t,J=5.1Hz,0.8H),4.02-3.96(m,2H),3.63(t,J=7.5Hz,2H),3.53-3.43(m,2H), 3.06(t,J=7.5Hz,2H),2.61-2.53(m,1H),2.48-2.40(m,1H),2.06-2.00(m,1H),1.92-1.77(m,5H),1.65-1.53 (m,4H),1.51-1.41(m,6H),1.36-1.10(m,8H),1.02-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0065] N-(4-bromophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (16). Yield: 77.3%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureofrotamers*indicates minor amide rotamer δ7.41(d,J=8.2Hz,2H),7.18(d,J=8.2Hz,2H),*5.00(s,0.8H),4.93(d,J=1.4Hz,1.2H),4.45(t,J=5 .4Hz,1.2H),*4.35(t,J=5.4Hz,0.8H),4.03-3.95(m,2H),3.59(t,J=7.2Hz,2H),3.53-3.44(m,2H),2.88(t, J=7.2Hz,2H),2.61-2.52(m,1H),2.48-2.39(m,1H),2.07-2.00(m,1H),1.93-1.77(m,5H),1.64-1.53(m,4H) ,1.51-1.41(m,6H),1.37-1.05(m,8H),1.02-0.98(m,3H),0.97-0.94(m,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0066] N-(3-bromophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (17). Yield: 78.9%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.46-7.43(m,1H),7.37-7.34(m,1H),7.26-7.22(m,1H),7.20(t,J=7.8Hz,1H),*5.00(s,0.8H),4.94(d,J= 1.8Hz,1.2H),4.46(t,J=5.4Hz,1.2H),*4.36(t,J=5.4Hz,0.8H),4.03-3.96(m,2H),3.59(t,J=7.5Hz,2H),3.52-3.44 (m,2H),2.90(t,J=7.5Hz,2H),2.61-2.53(m,1H),2.48-2.39(m,1H),2.07-2.00(m,1H),1.93-1.78(m,5H),1.64-1.5 3(m,4H),1.51-1.42(m,6H),1.37-1.05(m,8H),1.02-0.98(m,3H),0.97-0.95(m,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0067] N-(2-bromophenethyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (18). Yield: 79.7%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.54(dd,J=8.0Hz,1.1Hz,1H),7.33(dd,J=7.6Hz,1.6Hz,1H),7.25(td,J=7.6Hz,1.1Hz,1H),7.20(td,J=7.8Hz,1.6H z,1H),*5.00(s,0.8H),4.93(d,J=2.1Hz,1.2H),4.56(t,J=5.1Hz,1.2H),*4.35(t,J=5.1Hz,0.8H),4.03-3.96(m,2H),3.63(t, J=7.2Hz,2H),3.53-3.43(m,2H),3.06(t,J=7.2Hz,2H),2.62-2.52(m,1H),2.48-2.39(m,1H),2.06-1.99(m,1H),1.93-1.77(m ,5H),1.65-1.53(m,4H),1.50-1.41(m,6H),1.37-1.08(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0068] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-N-(4-methoxyphenethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (19). Yield: 82.8%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture ofrotamers*indicates minor amide rotamer δ7.15(d,J=8.7Hz,2H),6.81(d,J=8.7Hz,2H),*4.99(s,0.8H),4.93(d,J=1.9Hz,1.2H),4.45(t,J=4.9H z,1.2H),*4.34(t,J=4.9Hz,0.8H),4.02-3.94(m,2H),3.74(s,3H),3.55(t,J=7.6Hz,2H),3.52-3.43(m,2H),2. 82(t,J=7.6Hz,2H),2.61-2.52(m,1H),2.47-2.39(m,1H),2.06-1.99(m,1H),1.93-1.76(m,5H),1.64-1.52(m,4 H),1.51-1.41(m,6H),1.38-1.04(m,8H),1.01-0.98(m,3H),0.97-0.94(m,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0069] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-(4-methylphenethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (20). Yield: 87.1%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.13(d,J=8.2Hz,2H),7.08(d,J=8.2Hz,2H),*4.99(s,0.8H),4.93(d,J=2.1Hz,1.2H),4.46(t,J=5.3H z,1.2H),*4.35(t,J=5.3Hz,0.8H),4.03-3.94(m,2H),3.57(t,J=7.5Hz,2H),3.52-3.43(m,2H),2.85(t,J=7.5H z,2H),2.62-2.52(m,1H),2.48-2.38(m,1H),2.28(s,3H),2.08-1.99(m,1H),1.93-1.76(m,5H),1.65-1.53(m,4 H),1.51-1.41(m,6H),1.40-1.03(m,8H),1.02-0.98(m,3H),0.97-0.94(m,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0070] 7-(((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthren-17-yl)pentanoyl)-N-(4-(trifluoromethoxy)phenethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (21). Yield: 87.0%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.36(d,J=8.0Hz,2H),7.19(d,J=8.0Hz,2H),*5.00(s,0.8H),4.94(d,J=2.3Hz,1.2H),4.47(t,J=5 .0Hz,1.2H),*4.36(t,J=5.0Hz,0.8H),4.04-3.93(m,2H),3.60(t,J=7.5Hz,2H),3.53-3.43(m,2H),2.94(t, J=7.5Hz,2H),2.62-2.52(m,1H),2.49-2.38(m,1H),2.08-2.00(m,1H),1.92-1.76(m,5H),1.64-1.53(m,4H) ,1.51-1.41(m,6H),1.40-1.03(m,8H),1.02-0.98(m,3H),0.97-0.93(m,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0071] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-N-(2-(pyridin-4-yl)ethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (22). Yield: 65.5%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ8.45-8.40(m,2H),7.42-7.33(m,2H),*5.00(s,0.8H),4.94(d,J=1.5Hz,1.2H),4.46(t,J=5.3Hz ,1.2H),*4.35(t,J=5.3Hz,0.8H),4.05-3.93(m,2H),3.67(t,J=7.2Hz,2H),3.53-3.43(m,2H),2.98(t,J=7 .2Hz,2H),2.62-2.52(m,1H),2.49-2.39(m,1H),2.07-1.99(m,1H),1.94-1.76(m,5H),1.65-1.52(m,4H),1 .50-1.40(m,6H),1.39-1.04(m,8H),1.01-0.98(m,3H),0.97-0.94(m,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0072] N-Benzyl-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadienyl[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4-triazolo[4,3-a]pyrazine-3-carboxamide (23). Yield: 62.1%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture ofrotamers*indicates minor amide rotamer δ7.38-7.33(m,2H),7.33-7.29(m,2H),7.27-7.22(m,1H),*5.00(s,0.8H),4.94(d,J=2.6Hz, 1.2H),4.55(s,2H),4.49(t,J=5.3Hz,1.2H),*4.38(t,J=5.3Hz,0.8H),4.02-3.95(m,2H),3.52-3.44( m,2H),2.62-2.52(m,1H),2.48-2.38(m,1H),2.07-2.01(m,1H),1.92-1.77(m,5H),1.65-1.54(m,4H), 1.51-1.42(m,6H),1.39-1.08(m,8H),1.02-0.98(m,3H),0.96(s,3H),0.72(s,1.8H),*0.68(s,1.2H).

[0073] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-N-(3-phenylpropyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (24). Yield: 86.7%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.27-7.22(m,2H),7.22-7.18(m,2H),7.17-7.12(m,1H),*5.00(s,0.8H),4.94(d,J=2.8Hz,1.2H),4.48(t ,J=5.3Hz,1.2H),*4.37(t,J=5.3Hz,0.8H),4.03-3.95(m,2H),3.53-3.43(m,2H),3.40(t,J=7.5Hz,2H),2.68(t,J=7 .5Hz,2H),2.61-2.52(m,1H),2.48-2.39(m,1H),2.07-1.99(m,1H),1.96-1.91(m,2H),1.90-1.76(m,5H),1.65-1.52 (m,4H),1.51-1.41(m,6H),1.40-1.04(m,8H),1.02-0.98(m,3H),0.97-0.93(m,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0074] N-(3,5-dichlorophenyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthrene-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (25). Yield: 85.3%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixtureof rotamers*indicates minor amide rotamer δ7.35-7.31(m,3H),*5.01(s,0.8H),4.94(d,J=1.9Hz,1.2H),4.52(s,2H),4.50(t, J=5.3Hz,1.2H),*4.39(t,J=5.3Hz,0.8H),4.03-3.96(m,2H),3.52-3.44(m,2H),2.60-2.53( m,1H),2.47-2.40(m,1H),2.06-2.00(m,1H),1.91-1.77(m,5H),1.63-1.53(m,4H),1.50-1.4 2(m,6H),1.37-1.05(m,8H),1.07-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0075] 7-(((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthren-17-yl)pentanoyl)-N-(4-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (26). Yield: 88.2%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.62(d,J=7.9Hz,2H),7.54(d,J=7.9Hz,2H),*5.01(s,0.8H),4.95(d,J=2.1Hz,1.2H),4. 63(s,2H),4.92(t,J=5.3Hz,1.2H),*4.38(t,J=5.3Hz,0.8H),4.04-3.95(m,2H),3.54-3.43(m,2H) ,2.61-2.52(m,1H),2.48-2.39(m,1H),2.06-1.99(m,1H),1.93-1.76(m,5H),1.64-1.53(m,4H),1. 51-1.41(m,6H),1.38-1.03(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.68(s,1.2H).

[0076] 7-(((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentyl[a]phenanthren-17-yl)pentanoyl)-N-(3-(trifluoromethyl)benzyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (27). Yield: 77.6%. White solid. 1 H NMR (600MHz, CD 3 OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.68(s,1H),7.63(d,J=7.7Hz,1H),7.56(d,J=7.7Hz,1H),7.52(t,J=7.7Hz,1H),*5.01(s,0.8H), 4.94(d,J=2.1Hz,1.2H),4.62(s,2H),4.50(t,J=5.2Hz,1.2H),*4.39(t,J=5.2Hz,0.8H),4.03-3.95(m,2H), 3.52-3.44(m,2H),2.61-2.53(m,1H),2.48-2.39(m,1H),2.06-2.01(m,1H),1.92-1.77(m,5H),1.64-1.53(m ,4H),1.50-1.42(m,6H),1.38-1.04(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0077] N-(4-(difluoromethoxy)benzyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (28). Yield: 80.3%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.39(d,J=8.6Hz,2H),7.09(d,J=8.6Hz,2H),6.79(t,J=74.3Hz,1H),*5.00(s,0.8H),4.94(d,J =2.0Hz,1.2H),4.53(s,2H),4.49(t,J=5.3Hz,1.2H),*4.38(t,J=5.3Hz,0.8H),4.03-3.95(m,2H),3.53- 3.44(m,2H),2.61-2.52(m,1H),2.48-2.39(m,1H),2.06-2.00(m,1H),1.92-1.77(m,5H),1.64-1.53(m,4 H),1.50-1.42(m,6H),1.37-1.11(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0078] N-(3-(difluoromethoxy)benzyl)-7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (29). Yield: 82.8%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.35(t,J=8.0Hz,1H),7.22(d,J=8.0Hz,1H),7.15(s,1H),7.03(d,J=8.0Hz,2.3Hz,1H),6.81(t,J=74.1Hz ,1H),*5.01(s,0.8H),4.94(d,J=2.0Hz,1.2H),4.56(s,2H),4.49(t,J=5.3Hz,1.2H),*4.39(t,J=5.3Hz,0.8H),4.0 2-3.94(m,2H),3.52-3.44(m,2H),2.61-2.53(m,1H),2.47-2.40(m,1H),2.06-2.00(m,1H),1.92-1.78(m,5H),1.64 -1.53(m,4H),1.50-1.42(m,6H),1.38-1.05(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.72(s,1.8H),*0.69(s,1.2H).

[0079] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-N-(4-(trifluoromethoxy)benzyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (30). Yield: 92.6%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.46(d,J=8.6Hz,2H),7.23(d,J=8.6Hz,2H),*5.01(s,0.8H),4.95(d,J=2.6Hz,1.2H),4. 57(s,2H),4.49(t,J=5.5Hz,1.2H),*4.39(t,J=5.5Hz,0.8H),4.03-3.95(m,2H),3.54-3.43(m,2H) ,2.62-2.53(m,1H),2.49-2.38(m,1H),2.07-2.00(m,1H),1.92-1.76(m,5H),1.64-1.53(m,4H),1. 50-1.40(m,6H),1.39-1.03(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0080] 7-((4R)-4-((3R,5S,7S,8R,9S,10S,13R,14S)-3,7-dihydroxy-10,13-dimethylhexahydro-1H-cyclopentadien[a]phenanthren-17-yl)pentanoyl)-N-(3-(trifluoromethoxy)benzyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine-3-carboxamide (31). Yield: 79.1%. White solid. 1 H NMR (600MHz, CD 3OD):~60:40mixture of rotamers*indicates minor amide rotamer δ7.42(t,J=8.0Hz,1H),7.37(d,J=8.0Hz,1H),7.29(s,1H),7.17(d,J=8.0Hz,1H),*5.01(s,0.8H), 4.95(d,J=2.2Hz,1.2H),4.59(s,2H),4.50(t,J=5.3Hz,1.2H),*4.39(t,J=5.3Hz,0.8H),4.03-3.95(m,2H), 3.53-3.44(m,2H),2.61-2.52(m,1H),2.47-2.39(m,1H),2.07-2.01(m,1H),1.92-1.78(m,5H),1.64-1.53(m ,4H),1.50-1.42(m,6H),1.38-1.04(m,8H),1.01-0.98(m,3H),0.96(s,3H),0.71(s,1.8H),*0.69(s,1.2H).

[0081] Example 2: Evaluation of anti-fibrotic activity

[0082] 1. Experimental methods

[0083] CCC-HPF-1 cells were cultured at 2×10 5 Cells were seeded in 96-well plates at a density of 10 cells / mL and allowed to converge for 48 hours. Cells were then starved with serum-free MEM-NEAA for 24 hours. After starvation, the culture medium was replaced with MEM-NEAA containing or not containing 10ng / mL TGF-β1 (Peprotech) and cultured in the presence of various concentrations of compounds. After incubation for 48 hours, the cells were washed with phosphate-buffered saline (PBS) and fixed in 4% paraformaldehyde for 10 minutes. The cells were stained in 0.1% PSR staining solution for 1 hour. The staining solution was then removed, the cells were washed twice with 0.1% acetic acid and once with pure water. 200μL of 0.1mol / L potassium hydroxide solution was added to each well, shaken for 10 minutes to dissolve the dye, and the absorbance was measured at 540nm using an enzyme reader.

[0084] 2. Experimental results

[0085] The results are shown in Tables 1 and 2.

[0086] Table 1. In vitro antifibrotic activity of compounds 4-31

[0087]

[0088] Table 2. IC values ​​of in vitro antifibrotic activity of compounds 22, 23, 26, 28, 29, and 30 50 value

[0089]

[0090] The compound designed by the present invention has an inhibition rate of more than 40% on cell fibrosis at a concentration of 10 μM, and even more than 80%; the inhibition IC 50 The values ​​were lower than the 10 μM concentration level, and even lower than the 5 μM concentration level.

[0091] Example 3: Toxicity Evaluation

[0092] 1. Experimental methods

[0093] Cells were seeded in 96-well plates and treated with compounds (20 μM, dissolved in DMSO) for 48 hours. Afterwards, 10 μL of MTT (5 mg / mL) was added to each well and incubated for 4 hours. Then, the supernatant was discarded and 150 μL of DMSO was added to dissolve the formazan crystals in each well. The absorbance was measured at 490 nm using a microplate reader.

[0094] 2. Experimental results

[0095] The results are shown in Table 3.

[0096] Table 3. Effects of compounds on the survival rate of CCC-HPF-1 cells

[0097]

[0098] The compound designed by the present invention not only has highly effective activity in inhibiting fibrosis, but also has excellent safety, with a cell survival rate of more than 85%, or even 100%.

Claims

1. A triazole ring compound, It is characterized in that Having the structure of formula I, and also including pharmaceutically acceptable salts thereof: in: R is selected from C1-C4 alkyl, 3-6 membered cycloalkyl, -M-(6-12 membered aryl), -M-(6-12 membered heteroaryl containing 1-2 nitrogen atoms), M is connected to the 6-12 membered aryl, the 6-12 membered heteroaryl containing 1-2 nitrogen atoms by chemical bonds or cyclization; M is selected from -(CH 2 ) n -, 4-7 membered cycloalkyl; n is an integer selected from 0 to 4; The 6-12 membered aryl group and the 6-12 membered heteroaryl group containing 1-2 nitrogen atoms are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, hydroxyl, amino, nitro, and cyano.

2. The triazole ring compound according to claim 1, It is characterized in that In the structure: R is selected from C1-C4 alkyl, 3-5 membered cycloalkyl, -M-phenyl, -M-pyridyl; M is selected from -(CH 2 ) n -, M is connected to the phenyl group or pyridyl group by a chemical bond; when M is selected from a 4-6 membered cycloalkyl group, M is connected to the phenyl group or pyridyl group by a ring; n is an integer selected from 1 to 3; The phenyl group and the pyridyl group are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

3. The triazole ring compound according to claim 2, It is characterized in that In the structure: R is selected from methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyclopropyl, cyclopentyl, -(CH 2 ) n -phenyl, -(CH 2 ) n -pyridyl, cyclopentylphenyl; n is an integer selected from 1 to 3; The phenyl group and the pyridyl group are substituted by at least one substituent selected from any of the following: hydrogen, halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy.

4. The triazole ring compound according to claim 3, It is characterized in that In the structure: The phenyl group is substituted by at least one substituent selected from any of the following: hydrogen, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, tert-butyl, methoxy, ethoxy, difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy; the pyridyl group is substituted by hydrogen.

5. The triazole ring compound according to claim 1, It is characterized in that In the structure, R is selected from any of the following structures:

6. The triazole ring compound according to claim 1, It is characterized in that The pharmaceutically acceptable salt is a salt formed by the compound and an acid selected from any one of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, and ferulic acid.

7. A method for preparing the triazole ring compound according to claim 1, It is characterized in that The following steps are involved: Compound 1 is subjected to reduction, condensation and aminolysis to obtain compound I; Wherein, R is defined as in claim 1; The corresponding acid is reacted with the compound I prepared by the above method to form a salt, thereby obtaining a pharmaceutically acceptable salt of the compound.

8. A pharmaceutical composition, It is characterized in that The pharmaceutical composition comprises the triazole ring compound according to claim 1 and a pharmaceutically acceptable carrier.

9. Use of the triazole ring compound according to claim 1 or the pharmaceutical composition according to claim 8 in the preparation of anti-fibrosis drugs.

10. The use according to claim 9, It is characterized in that The medicine is a medicine for resisting cardiac, pulmonary, liver and kidney fibrosis.

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