Furan compounds with anti-inflammatory and hepatoprotective activity, their preparation and use

Abstract

The application belongs to the technical field of medicine, and discloses preparation of furan new compounds 1 and 2 and pharmaceutically acceptable salts thereof, and application of a pharmaceutical composition thereof in preparation of anti-inflammatory liver-protecting drugs. Cell level experiments prove that the compounds 1 and 2 have good anti-inflammatory liver-protecting effects, and are expected to become therapeutic drugs for diseases related to liver injury.

Description

Technical Field

[0001] The present invention relates to a class of furan compounds and pharmaceutically acceptable salts thereof, their preparation methods, pharmaceutical compositions containing the compounds, and applications of the compounds in anti-inflammatory and liver protection, belonging to the field of medical technology. Background Art

[0002] The liver is one of the most important organs in the human body and is also the largest substantial organ in the human body. It has important functions such as biological transformation, detoxification, protein secretion, blood sugar regulation, and participation in blood coagulation. At the same time, the liver is also an organ that is often invaded by various pathogenic factors or diseases. Abnormal metabolism, drugs, microorganisms, etc. can all cause liver damage. In recent decades, liver disease has become one of the diseases with the highest mortality rate in the world, including acute hepatitis, liver cancer, etc., accounting for 4% of all deaths worldwide. [1] my country is a country with a high incidence of liver disease, with more than one-fifth of the population affected by some form of liver disease, particularly hepatitis B virus (HBV) and hepatitis C virus (HCV) infection, cirrhosis, liver cancer, non-alcoholic fatty liver disease (NAFLD), alcohol-related liver disease (ALD) and drug-induced liver injury (DILI). [2] According to statistics, direct economic losses from chronic hepatitis (including cirrhosis and liver cancer in later stages) in my country reach 900 billion RMB annually. In recent years, the incidence of liver-related diseases has also been increasing year by year. Therefore, the development of new anti-inflammatory and liver-protecting drugs is urgent and has become one of the main research areas in the current drug research and development field.

[0003] As we all know, liver disease is a progressive disease. There is a variety of evidence suggesting that liver inflammation is seen in almost all causes of liver disease and often runs throughout the course of liver disease. In 2014, the Expert Consensus Committee on Liver Inflammation and Its Prevention and Treatment of the Infectious Diseases Branch of the Chinese Medical Association published the Expert Consensus on Liver Inflammation and Its Prevention and Treatment. [3] The "Consensus" points out: "Liver inflammation and the resulting liver fibrosis, cirrhosis and liver failure are the main pathophysiological and pathological histological basis of liver disease progression; anti-inflammatory and liver-protecting treatment is an important part of the comprehensive treatment of liver inflammation. For liver inflammation, regardless of whether there is an effective etiological treatment, anti-inflammatory and liver-protecting treatment should be considered; for some patients who lack effective etiological treatment or are temporarily unable to undergo etiological treatment, anti-inflammatory and liver-protecting treatment should be considered." Anti-inflammatory and liver-protecting drugs are a class of drugs that have the effects of improving liver function, promoting liver cell regeneration and / or enhancing liver detoxification function. They can reduce liver damage caused by pathogenic factors such as hepatitis viruses, drugs, toxins and alcohol, and are an important part of the comprehensive treatment of liver disease. However, for a long time, the progress in anti-inflammatory and liver-protecting treatment has not been ideal. The clinically available anti-inflammatory and liver-protecting drugs and means are limited, and the main varieties are limited to licorice products. [4] , silymarin[5] , glutathione [6] and bicyclic alcohol [7] There are even fewer varieties that are original to China or have independent intellectual property rights in China. Therefore, the development of new anti-inflammatory and liver-protecting drugs with independent intellectual property rights in my country has important strategic significance and medical value.

[0004] References

[0005] [1] P. Byass, The global burden of liver disease: a challenge for methods and for public health. BMC Med, 2014, 12, 159.

[0006] [2]F.-S.Wang,J.-G.Fan,Z.Zhang,B.Gao,H.-Y.Wang,Hepatology.2014,60,2099-2108.

[0007] [3] Li Lanjuan, Wang Yuming. Expert consensus on liver inflammation and its prevention and treatment. Chinese Journal of Hepatology, 2014, 22, 94-103.

[0008] [4]

[0009] [5] S. Pradhan, C. Girish, Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine. Indian J Med Res, 2006, 124, 491-504.

[0010] [6] G. Li, Z. Zhang, C. Yang, The Classification and function of common anti-inflammatory drugs. Chin Prac Med, 2012, 7, 236-238.

[0011] [7]W.Xie,G.Shi,H.Zhang,G.Zhao,Z.Yu,Z.Lang,H.Zhao,J.Yan,J.Cheng,Arandomized,multi-central,controlled study of patients with hepatitis Beantigen-positive chronic hepatitis B treated by adefovir dipivoxil oradefovir dipivoxil plus bicyclol.Hepatol Int,2012,6,441-448. Summary of the Invention

[0012] The technical problem solved by the present invention is to provide a class of polysubstituted furan compounds and pharmaceutically acceptable salts thereof, as well as application of the pharmaceutical composition thereof in the preparation of anti-inflammatory and liver-protecting drugs.

[0013] In order to solve the technical problems of the present invention, the present invention provides the following technical solutions:

[0014] The first aspect of the technical solution of the present invention is to provide compounds shown in the following group:

[0015]

[0016] The second aspect of the technical solution of the present invention is to provide a method for preparing the compound described in the first aspect.

[0017]

[0018] 4-Phenyl-1-toluenesulfonyl-1H-1,2,3-triazole 3 and benzoenaminone 4 under Rh catalysis underwent cyclization reaction to give 7-methyl-2-phenyl-4,5-dihydronaphtho[1,2-b]furan compound 1. 4-(m-tolyl)-1-tolyl-1H-1,2,3-triazole 5 and (E)-6-((dimethylamino)methylene)-6,7,8,9-tetrahydro-5H-benzo[7]enone 6 under Rh catalysis underwent cyclization reaction to give 2-(m-tolyl)-5,6-dihydro-4H-benzo[6,7]cyclohepta[1,2-b]furan compound 2.

[0019] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition containing compounds as shown in structures 1 and 2 and pharmaceutically acceptable salts thereof, wherein the pharmaceutical composition contains compounds as shown in compounds 1 and 2 included in the group and pharmaceutically acceptable salts thereof as active ingredients, and optionally contains a pharmaceutical carrier.

[0020] The pharmaceutical composition of the present application usually contains 0.1-95% by weight of the compound of the present application.

[0021] The pharmaceutical composition of the compound of the present application can be prepared according to the methods well known in the art. For this purpose, the compound of the present application, if desired, can be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to form a suitable administration form or dosage form which can be used as a human or veterinary medicine.

[0022] The compound of the present application or the pharmaceutical composition containing it can be administered in unit dosage form, and the administration route can be enteral or parenteral, such as oral, intramuscular, subcutaneous, nasal, oral mucosal, dermal, peritoneal or rectal, etc., preferably oral.

[0023] The administration route of the compound of the present application or the pharmaceutical composition containing it can be injection administration. Injection includes intravenous injection, intramuscular injection, subcutaneous injection and intradermal injection, etc.

[0024] The administration dosage form can be a liquid dosage form or a solid dosage form. For example, the liquid dosage form can be a true solution, a colloid, a microparticle dosage form, an emulsion dosage form, a suspension dosage form. Other dosage forms such as tablets, capsules, dripping pills, aerosol, pills, powders, solutions, suspensions, emulsions, granules, suppositories, lyophilized powder injections, etc.

[0025] The extract or compound of the present application can be prepared into a general preparation, or a sustained-release preparation, a controlled-release preparation, a targeted preparation and various microparticle administration systems.

[0026] In order to make tablets into unit administration dosage forms, various carriers well known in the art can be widely used. Examples of the carriers are, for example, diluents and absorbents such as starch, dextrin, calcium sulfate, lactose, mannitol, sucrose, sodium chloride, glucose, urea, calcium carbonate, white clay, microcrystalline cellulose, aluminum silicate, etc.; humectants and binders such as water, glycerol, polyethylene glycol, ethanol, propyl alcohol, starch paste, dextrin, sugar syrup, honey, glucose solution, acacia paste, gelatin paste, sodium carboxymethyl cellulose, shellac, methyl cellulose, potassium phosphate, polyvinylpyrrolidone, etc.; disintegrants such as dry starch, alginate, agar powder, brown algae starch, sodium bicarbonate and citric acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, methyl cellulose, ethyl cellulose, etc.; disintegration inhibitors such as sucrose, glycerol triestearate, cocoa butter, hydrogenated oil, etc.; absorption promoters such as quaternary ammonium salt, sodium dodecyl sulfate, etc.; lubricants such as talc, silicon dioxide, corn starch, stearate, boric acid, liquid paraffin, polyethylene glycol, etc. The tablets can be further prepared into coated tablets such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.

[0027] For example, to prepare the dosing unit into a pill, various carriers known in the art can be widely used. Examples of carriers include diluents and absorbents, such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, polyvinyl pyrrolidone, gelucire, kaolin, talc, etc.; binders, such as gum arabic, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or flour paste, etc.; disintegrants, such as agar powder, dry starch, alginate, sodium lauryl sulfate, methylcellulose, ethylcellulose, etc.

[0028] For example, to prepare a dosing unit in the form of a capsule, the active ingredient extract or compound of the present invention is mixed with the various carriers described above, and the resulting mixture is placed in a hard gelatin capsule or soft capsule. Alternatively, the active ingredient compound of the present invention may be formulated into microcapsules and suspended in an aqueous medium to form a suspension, which may be encapsulated in a hard capsule or formulated as an injection.

[0029] For example, the extracts or compounds of the present invention can be formulated into injectable formulations, such as solutions, suspension solutions, emulsions, or lyophilized powder injections. These formulations can be aqueous or non-aqueous and may contain one or more pharmacologically acceptable carriers, diluents, binders, lubricants, preservatives, surfactants, or dispersants. For example, diluents can be selected from water, ethanol, polyethylene glycol, 1,3-propylene glycol, ethoxylated isostearyl alcohol, polyoxygenated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and the like. Furthermore, to prepare isotonic injections, appropriate amounts of sodium chloride, glucose, or glycerol can be added to the injectable formulation. Conventional cosolvents, buffers, and pH adjusters can also be added. Cosolvents can include poloxamer, lecithin, and hydroxypropyl-β-cyclodextrin; pH adjusters can include phosphates, acetates, hydrochloric acid, and sodium hydroxide; and osmotic pressure regulators can include sodium chloride, mannitol, glucose, phosphates, and acetates. For lyophilized powder injections, mannitol or glucose can also be added as support agents.

[0030] In addition, if necessary, colorants, preservatives, perfumes, flavorings, sweeteners or other materials may be added to the pharmaceutical preparations.

[0031] To achieve the purpose of medication and enhance the therapeutic effect, the drug or pharmaceutical composition of the present invention can be administered by any known method of administration.

[0032] The fourth aspect of the technical solution of the present invention provides the use of the furan compounds 1 and 2 of the present invention and pharmaceutically acceptable salts thereof in the preparation of anti-liver injury, anti-inflammatory and liver-protecting drugs.

[0033] When the furan derivatives and pharmaceutically acceptable salts thereof or the composition of the present invention are used to treat the above-mentioned diseases, the dosage thereof can refer to the dosage used when furan derivatives are used for treatment.

[0034] The dosage of the compounds and pharmaceutical compositions of the present invention depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, personality and individual response of the patient or animal, the route of administration, the number of administrations, and the purpose of treatment. Therefore, the therapeutic dose of the present invention can vary widely. Generally speaking, the dosage of the pharmaceutical ingredients of the present invention is well known to those skilled in the art. It can be appropriately adjusted according to the actual amount of the drug contained in the final preparation of the compound composition of the present invention to achieve the requirements of its therapeutically effective amount and complete the prevention or treatment purpose of the present invention. The daily suitable dosage range of the compounds of the present invention is 0.001-150 mg / kg body weight, preferably 0.01-100 mg / kg body weight, more preferably 0.01-60 mg / kg body weight, and most preferably 0.1-10 mg / kg body weight. The above dosage can be administered in a single dose form or divided into several, such as two, three or four dose forms. This is limited by the clinical experience of the administering physician and the administration regimen including the use of other treatment means.

[0035] The total dose required for each treatment can be divided into multiple doses or administered as a single dose. The compounds and compositions of the present invention can be taken alone or in combination with other therapeutic drugs or symptomatic drugs with adjusted dosages.

[0036] The inventors have discovered that compounds 1 and 2 and their pharmaceutically acceptable salts have significant anti-inflammatory and hepatoprotective effects. Therefore, compounds 1 and 2 and their pharmaceutically acceptable salts also provide methods for treating and ameliorating diseases associated with liver damage. These methods comprise administering a therapeutically effective amount of compounds 1 and 2 or their pharmaceutically acceptable salts, or a pharmaceutical composition thereof, to a patient in need of treatment.

[0037] The present invention shows that compounds 1 and 2 have significant anti-inflammatory and hepatoprotective effects at the cellular level. There are no public reports on compounds 1 and 2 or their pharmaceutically acceptable salts, and there are no public reports on the anti-inflammatory and hepatoprotective activities of compounds 1 and 2.

[0038] Beneficial technical effects

[0039] During their research on furan compounds, the inventors of this invention synthesized two new compounds, 1 and 2, via chemical methods. Using a classic cell model for evaluating hepatoprotective activity, the compounds were evaluated for their protective effects against paracetamol (APAP)-induced hepatocellular damage. The results demonstrated that compounds 1 and 2 possess significant anti-inflammatory and hepatoprotective activity. These compounds are valuable new lead compounds in the development of anti-inflammatory and hepatoprotective drugs. DETAILED DESCRIPTION

[0040] The following examples further illustrate the present invention but are not intended to limit the present invention in any way.

[0041] Example 1: Preparation of 7-methyl-2-phenyl-4,5-dihydronaphtho[1,2-b]furan (1)

[0042]

[0043] 4-Phenyl-1-toluenesulfonyl-1,2,3-triazole 4 (60 mg) was weighed and added to 1.5 mL of DCE solvent with stirring. Bis[(α,α,α′,α′-tetramethyl-1,3-benzenedipropionic acid)rhodium] catalyst (3.1 mg) and (E)-2-((dimethylamino)methylene)-6-methyl-3,4-dihydronaphthalen-1(2H)-enone 5 (22 mg) were added. The mixture was reacted at 90°C for 20 minutes. The organic solvent was removed under reduced pressure and column chromatography was performed to obtain 7-methyl-2-phenyl-4,5-dihydronaphtho[1,2-b]furan 1 (18 mg, 68%) as a white solid. Mp: 81.7-82.3°C; 1 H NMR (400MHz, CDCl3): δ = 7.75 (d, J = 7.5Hz, 2H), 7.54 (d, J = 7.5Hz, 1H), 7.42 (t, J = 7.5Hz, 2H), 7.28 (t, J = 7 .5Hz,1H),6.84-6.83(m,2H),6.64(s,1H),3.85(s,3H),3.00(t,J=7.5Hz,2H),2.78(t,J=7.5Hz,2H)ppm; 13 C NMR (100MHz, CDCl3): δ=158.6,152.5,149.9,136.8,131.3,128.8,127.0,123.5 ,121.5,120.5,119.4,114.6,111.5,106.7,55.4,29.6,21.1ppm; HRMS(ESI):m / z calcd for C 19 H 16 O[M+H] + 260.1290; found 260.1296.

[0044] Example 2: Preparation of 2-(m-tolyl)-5,6-dihydro-4H-benzo[6,7]cyclohepta[1,2-b]furan (2)

[0045]

[0046] 4-(m-tolyl)-1-tolyl-1H-1,2,3-triazole 6 (62.6 mg) was weighed and added to 1.5 mL of DCE with stirring. Bis[(α,α,α′,α′-tetramethyl-1,3-benzenedipropionic acid)rhodium] catalyst (3.1 mg) and (E)-3-(dimethylaminomethylene)chromogen-4-one 7 (22 mg) were added. The reaction was carried out at 90°C for 30 minutes. The organic solvent was removed under reduced pressure and column chromatography was performed to obtain 2-(m-tolyl)-5,6-dihydro-4H-benzo[6,7]cyclohepta[1,2-b]furan 2 (22 mg, 81%) as a white solid. Mp: 92.2-94.1°C. 1 H NMR (400MHz, CDCl3): δ = 8.01 (d, J = 8.0Hz, 1H), 7.58-7.55 (m, 2H), 7.34-7.29 (m, 2H), 7.19-7.14 (m, 2H), 7.11 ( d,J=8.0Hz,1H),6.61(s,1H),2.94(d,J=4.8Hz,2H),2.86(d,J=6.4Hz,2H),2.43(s,3H),2.06-2.01(m,2H)ppm; 13 C NMR (100MHz, CDCl3): δ=152.5,147.6,139.0,138.4,130.8,130.5,129.6,128.7,128.3,126.6,126.4,1 25.1,124.6,124.6,121.2,109.6,36.2,27.9,24.9,21.7ppm; IR(neat):ν=2928,1605,1497,1249,816cm -1 ; HRMS(ESI):m / z calcd for C 20 H 19 O[M+H] + 275.1423; found 275.1430.

[0047] Pharmacological experiments

[0048] Example 1. Evaluation of the protective effect of compounds 1 and 2 on APAP-induced hepatocellular damage in experimental cell lines

[0049] Human HepG2 hepatocellular carcinoma cells, which retain the characteristics of normal human hepatocytes, were grown in DMEM (100 U / ml penicillin and 100 μg / ml streptomycin) supplemented with 10% fetal bovine serum at 37°C, 5% CO2, and saturated humidity. They were digested and passaged using a solution containing 0.25% trypsin and 0.02% EDTA.

[0050] Experimental methods

[0051] (1) Effects of compounds on HepG2 cell proliferation

[0052] The MTT method was used. HepG2 cells were seeded in a 96-well cell culture plate and cultured for 24 hours. Different concentrations of the test compound were then added. A solvent control group was also established, with three parallel wells for each drug concentration. After 24 hours of drug exposure, the culture medium was discarded, and 100 μl of MTT (0.5 mg / ml) solution was added to each well. The cells were cultured for another 4 hours. The MTT solution was discarded, and 150 μl of DMSO was added to each well. The cells were mixed and shaken, and the absorbance was measured at 570 nm using a microplate reader. Cell viability (%) = (mean OD value of the treated cells / mean OD value of the solvent control cells) × 100%.

[0053] (2) Protective effect of the compound on paracetamol-induced hepatocyte damage in vitro

[0054] The MTT assay was used. HepG2 cells were seeded in a 96-well cell culture plate and cultured for 24 hours. Non-toxic concentrations of the test compound and paracetamol (APAP, final concentration 8 mM) were then added. A positive drug control group (glutathione GSH), a solvent blank control group, and a model group were also established. The cells were treated for a further 24 hours. The culture medium was discarded, and 100 μl of MTT (0.5 mg / ml) solution was added to each well. The cells were cultured for another 4 hours. The MTT solution was discarded, and 150 μl of DMSO was added to each well. The cells were mixed and shaken, and the absorbance was measured at 570 nm using a microplate reader. Cell viability (%) = (mean OD value of the drug group / mean OD value of the solvent control group) × 100%.

[0055] Experimental results

[0056] (1) Cytotoxicity

[0057] Compounds 1 and 2 had no significant cytotoxicity to HepG2 cells at a concentration of 10 μM for 24 h, and the cell survival rate was greater than 90%. They were used in subsequent experiments at a non-toxic concentration of 10 μM.

[0058] (2) Protective effect on APAP-induced hepatocellular damage

[0059] APAP 8mM treatment of HepG2 cells for 24 hours significantly damaged the cells, with cell survival significantly reduced (42.66%) compared to the blank control group. Under the current experimental protocol, compounds 1 and 2 at a concentration of 10μM showed significant protective effects against APAP-induced HepG2 cell damage. The experimental results are shown in Table 1.

[0060] Table 1. Hepatoprotective effects of compounds 1 and 2

[0061]

[0062] *** P < 0.001 compared with the blank group; # P<0.05, ## P<0.01, compared with the APAP model group.

Claims

1. A compound and a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from the following group:

2. The method for preparing the compound according to claim 1, characterized in that: 4-Phenyl-1-p-toluenesulfonyl-1H-1,2,3-triazole 3 and benzo[7]enone 4 under Rh catalysis undergo cyclization reaction to give trisubstituted furan compound 1; 4-(m-tolyl)-1-p-toluenesulfonyl-1H-1,2,3-triazole 5 and (E)-6-((dimethylamino)methylene)-6,7,8,9-tetrahydro-5H-benzo[7]enone 6 under Rh catalysis undergo cyclization reaction to give trisubstituted furan compound 2.

3. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a therapeutically effective amount of the compound shown in claim 1 and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

4. Use of the compound according to claim 1 and its pharmaceutically acceptable salt or the pharmaceutical composition according to claim 3 in the preparation of anti-liver injury, anti-inflammatory and liver-protecting drugs.

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