KR20120134939A - A composition for treating or preventing liver failure comprising 4-hydroxy tamoxifen analog or pharmaceutically acceptable salts thereof as an effective ingredient - Google Patents

Abstract

PURPOSE: A composition containing a 4-hydroxy tamoxifen analog for preventing or treating liver diseases is provided to reduce GOT(glutamyl oxaloacetic transaminase) and GPT(glutamyl pyruvate transaminase) level and to prevent and treat liver diseases. CONSTITUTION: A pharmaceutical composition for preventing or treating liver diseases contains a 4-hydroxy tamoxifen analog of chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition additionally contains a pharmaceutically acceptable carrier. The liver diseases are non-alcoholic or alcoholic fatty liver. The pharmaceutical composition reduces fat accumulation in the liver. The non-alcoholic fatty liver is non-alcoholic steatohepatitis or late phase fibrosis liver diseases. A food composition for preventing or treating liver diseases contains 4-hydroxy tamoxifen analog of chemical formula 1 or sitologically acceptable salt thereof as an active ingredient.

Description

A composition for treating or preventing liver failure comprising 4-hydroxy tamoxifen analog or pharmaceutically acceptable salts according as an active ingredient. effective ingredient}

The present invention is a composition for the prevention or treatment of liver disease comprising a 4-hydroxy tamoxifen analog or a pharmaceutically acceptable salt thereof as an active ingredient, a method for treating or preventing liver disease using the composition and a liver comprising the composition It relates to a food composition for the treatment and prevention or improvement of diseases.

Liver is the organ that plays a central role in nutrient metabolism. Normal human liver weighs about 1,500 g and abnormal liver function causes problems in nutrient metabolism in the body, making glucose into glycogen or protein in albumin. Abnormalities in liver function, such as switching to or dissolving unnecessary ones and delivering them to the bile. In case of mental rest, hepatic cells can be restored in the case of minor liver damage, but liver disease is aggravated because busy modern society cannot afford to rest. In general, hepatitis, which causes inflammation of the liver, accounts for most of liver diseases, and according to aspects, it can be divided into acute hepatitis, chronic hepatitis, and viral hepatitis, alcoholic hepatitis, and drug hepatitis depending on the cause. Liver diseases caused by these abnormalities include fatty liver, hepatitis, cirrhosis, liver cancer. Healing liver disease in combination with exercise, alcoholism, diet, and drug treatment, but fundamentally difficult to completely heal the development of effective and improved liver disease treatment or protection is needed.

In addition, due to the recent improvement in the standard of living due to economic development, the hygienic environment is improved and the calorie intake is rapidly increased due to the improvement of diet, while the lack of exercise and the amount of calories consumed are low, resulting in excessive accumulation of nutrients in the body. The incidence of various adult diseases is increasing. Among these diseases, especially fatty liver is caused by the accumulation of fat in the liver due to excessive fat or alcohol intake, increased liver fat synthesis, reduced triglycerides and burning, etc. In general, fatty liver has a fat percentage of 5% or more. Is defined.

Most of the fat accumulated in fatty liver is triglyceride, and fatty liver can be divided into alcoholic fatty liver due to excessive drinking and non-alcoholic fatty liver due to obesity, diabetes, hyperlipidemia or drugs. Alcoholic fatty liver is caused by excessive intake of alcohol, which promotes fat synthesis in the liver and does not undergo normal energy metabolism. Some people think that fatty liver is simply the accumulation of fat in the liver, but considering that 50% of patients who have been diagnosed as alcoholic fatty liver and 30% of patients who are identified as nonalcoholic fatty liver develop liver cirrhosis, fatty liver is very serious. It should be considered one of the liver diseases.

In particular, alcoholic fatty liver may progress to chronic liver disease. Among patients with alcoholic fatty liver, 10-35% are reported to progress to hepatitis, and 8-20% to cirrhosis, and alcoholic fatty liver patients are compared to nonalcoholic fatty liver patients. Alcoholic fatty liver is a more serious problem because it has more cirrhosis and is more likely to develop hepatitis.

At present, there are few drugs that are useful for pharmacological treatment of fatty liver, and only exercise and diet are recommended, but in reality, the treatment efficiency of fatty liver by such a method is very low, and an effective therapeutic agent is required. It has been reported that some hypoglycemic agents, such as metformin, are effective in treating fatty liver, although fatty liver has been found to be associated with insulin resistance in cells observed in diabetes and obesity, but the drug has side effects such as liver toxicity or lactic acidosis. There is a problem that causes. In addition, alternative drugs, such as betaine, glucuronate, methionine, choline and lipotrophic agents, may be used as supplements, It is not fully proven. Therefore, the development of a safe fatty liver treatment that does not cause side effects with excellent effects is urgently needed.

In view of the above, the present inventors have no side effects, and while searching for an effective substance for liver disease, the GSK5182, which is a 4-hydroxy tamoxifen analog, has GOT (glutamyl oxaloacetic transaminase) and GPT (glutamyl pyruvate transaminase), which are liver damage markers. The present invention has been completed by confirming that the level can be reduced and effective in the prevention and treatment of liver disease.

It is an object of the present invention to provide a pharmaceutical composition for the prevention or treatment of liver disease, comprising a 4-hydroxy tamoxifen analog or a pharmaceutically acceptable salt thereof as an active ingredient.

It is another object of the present invention to provide a food composition for preventing or ameliorating liver disease, including a 4-hydroxy tamoxifen analog or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a method for preventing or treating liver disease, comprising administering the pharmaceutical composition for treating or preventing liver disease to an individual having or possibly developing liver disease.

As one embodiment for achieving the above object, the present invention provides a composition for the prevention or treatment of liver disease comprising a 4-hydroxy tamoxifen analog of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. .

In the present invention, the 4-hydroxy tamoxifen analog is (Z) -4- (1- {4- [2- (dimethylamino) ethoxy] phenyl} -5-hydroxy-2-phenylpent-1-enyl ) Phenol [(Z) -4- (1- {4- [2- (dimethylamino) ethoxy] phenyl} -5-hydroxy-2-phenylpent-1-enyl) phenol]. The compound of Formula 1 is also called GSK5182. Although GSK5182 is known as an inhibitor of ERRγ (estrogen-related receptor gamma), it has not been disclosed for the treatment of diseases of liver disease, and the inventors have found the first use of the treatment of liver disease.

In the present invention, GSK5182 may be synthesized by a known synthesis method, or may be commercially available.

In the present invention, GSK5182 may be prepared with pharmaceutically or food acceptable salts and solvates according to methods conventional in the art.

As pharmaceutically or food acceptable salts, acid addition salts formed by free acid are useful. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, and hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, and the like may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, and trifluoro may be used as the organic acid. Acetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, Galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid, hydroiodic acid and the like can be used.

Bases may also be used to prepare pharmaceutically or food acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

The composition GSK5182 of the present invention may inhibit the expression of sterol regulatory element binding protein 1c (SREBP1c; sterol regulatory element binding protein 1c) as an ERRγ-specific inverse agonist to inhibit fat synthesis in hepatocytes.

The composition of the present invention can reduce fat accumulation in the liver, and preferably can suppress fat accumulation in alcoholic fatty liver or non-alcoholic fatty liver.

The composition GSK5182 of the present invention can reduce the levels of GOT (glutamyl oxaloacetic transaminase) and GPT (glutamyl pyruvate transaminase).

The composition comprising GSK5182 of the present invention is characterized by inhibiting sterol regulatory element binding protein 1c (SREBP1c) in the liver to treat or prevent liver disease.

As used herein, the term “liver disease” includes without limitation any liver disease that can be treated by the composition of the present invention, for example, non-alcoholic fatty liver, alcoholic fatty liver, non-viral chronic hepatitis, cirrhosis, chronic liver disease, Liver cancer and the like. Preferably non-alcoholic fatty liver, or alcoholic fatty liver.

As used herein, the term "non-viral" refers to an illness caused by alcoholic, non-alcoholic (obesity, diabetes, hyperlipidemia) or other (toxic, pollution, drug, neurological, overworked) causes.

In the present invention, the term "fatty liver" refers to a phenomenon in which triglycerides appear abnormally deposited in liver cells unlike normal cases. About 5% of normal liver is composed of fatty tissue, and triglycerides, fatty acids, phospholipids, cholesterol, and cholesterol esters are the main components of fat, but once fatty liver occurs, most of the components are replaced by triglycerides. If more than 5% of the liver weight is diagnosed as fatty liver.

Fatty liver is caused by a fat metabolism disorder in liver cells or a defect in the process of carrying excess fat, and is mainly caused by a fat metabolism disorder in the liver. Most of the fat accumulated in the fatty liver is triglyceride, and fatty liver can be divided into non-alcoholic fatty liver due to obesity, diabetes, hyperlipidemia, drugs, and alcoholic fatty liver due to excessive drinking.

The "non-alcoholic fatty liver" refers to the case of accompanied by fatty liver without a history of alcohol intake, and is known to be associated with metabolic diseases such as obesity, diabetes, hyperlipidemia. These nonalcoholic fatty livers include not only fat accumulation in the liver, but also non-alcoholic steatohepatitis or terminal fibrotic liver disease.

"Alcoholic fatty liver" means that alcohol is consumed to promote fat synthesis, which is caused by lack of normal energy metabolism. Alcohol is not stored in the body and is completely eliminated by oxidation in the liver. Specifically, alcohol in the liver is largely alcohol dehydrogenase (ADH) pathway, microsomal ethanol oxidizing system (MEOS). It is metabolized and converted into acetaldehyde by three pathways, a) and a catalase pathway, which are in turn metabolized to acetitis by dehydrogenase (ALLDH). At this time, acetaldehyde is toxic and can damage liver cells, and also, as a result of the metabolism of alcohol, fatty acids are made up to accumulate fat in the liver. Fatty liver accumulated in the liver due to the above causes is called alcoholic fatty liver. In a preferred embodiment of the present invention, as a result of administering GSK5182, a 4-hydroxy tamoxifen analog, to the alcoholic fatty liver induced mice using a liquid alcohol diet in addition to non-alcoholic fatty liver, it was confirmed that the effect of inhibiting fatty liver (Fig. 2 and Fig. 4).

Fatty liver is mostly associated with obesity, but it can also occur in people who are dry or normal. This may be supported by reports that 40% (32/81) of patients with fatty liver had a normal body mass index (Nucl. Med. Mol. Imaging., 40, 243-248 (2006)). In particular, triglycerides were reported to be a major factor in non-obese groups (J. Clin. Gastroenterol., 40, 745 ~ 752 (2006)). Is statistically significant, indicating that there is a high association between fatty liver and triglyceride in non-obesity cases.

In the present invention, the term "chronic liver disease" includes "chronic hepatitis", "liver cirrhosis" and "liver cancer" and "cirrhosis", also called cirrhosis, is characterized by the loss of a significant amount of hepatocytes, the proliferation of fibrous tissue, and regenerative nodules. All forms of liver disease. The main causes of cirrhosis are hepatitis virus and alcohol, respectively, treated with antiviral therapy or alcohol / nutrition therapy. Complications can lead to death, including ascites (bleeding in the belly), varicose veins bleeding, and hepatic coma.

In the present invention, the term "liver cancer" refers to a disease resulting from patients with chronic liver disease caused by cirrhosis transducer, alcohol and the like. If you have a typical liver mass or if your blood test shows an increase in alpha-fetoprotein (AFP), you can diagnose it as a liver cancer.

The causes of all these liver diseases may vary from patient to patient, but as the disease progresses to hepatitis, liver cirrhosis, and liver cancer, the composition containing GSK5182 is characterized by GOT (glutamyl oxaloacetic transaminase) and GPT ( Reducing the levels of glutamyl pyruvate transaminase may suggest treatment or prevention of liver disease.

As used herein, the term "prevention" refers to any action that inhibits or delays the development of liver disease by administration of a composition according to the invention.

As used herein, the term "treatment" refers to any action by which administration of a composition according to the invention improves or advantageously alters the symptoms of the disease.

In one embodiment of the present invention, GSK5182 is a kind of liver disease by inhibiting the expression of sterol regulatory element binding protein 1c (SREBP1c; sterol regulatory element binding protein1c) as an ERRγ-specific inverse agonist, inhibiting fat synthesis in hepatocytes. It was confirmed that it can be effective in the treatment or prevention of fatty liver (Fig. 1).

In particular, in the embodiment of the present invention experiments using a mouse model to confirm the role of GSK5182 in non-alcoholic fatty liver and alcoholic fatty liver, it was confirmed that significantly inhibit the fatty liver compared to the control (Fig. 2 and 4). In addition, the analysis of GOT and GPT in blood for hepatotoxicity following administration of GSK5182 revealed that GSK5182 was significantly lower than the control group and lowered to almost normal levels in the GSK5182 administration group, resulting in GSK5182 induced by non-alcoholic and alcoholic fatty liver. It was confirmed that there is a marked improvement in the liver toxicity (Fig. 3 and 5).

In a preferred embodiment of the present invention, the composition of the present invention is a pharmaceutical composition.

When the composition of the present invention is a pharmaceutical composition, the composition may include a pharmaceutically acceptable carrier. The composition comprising a pharmaceutically acceptable carrier can be of various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or lactose ( lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, and syrups. In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.

The pharmaceutical composition may be in the form of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations and suppositories It can have one formulation.

The composition of the present invention is administered in a pharmaceutically effective amount.

As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to the type and severity of the subject, the severity, age, sex and activity of the drug. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts. However, for the desired effect, the 4-hydroxy tamoxifen analog of Formula 1 or a salt thereof is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 10 mg / kg.

The liver disease prophylactic or therapeutic agent of the present invention may be administered daily or intermittently, and the number of administrations per day can be administered once or divided into two or three times. The frequency of administration in the case where the two active ingredients are single drugs may be the same or different times. In addition, the composition of the present invention can be used alone or in combination with other drug treatments for the prevention or treatment of liver disease diseases. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art.

In a preferred embodiment of the present invention, the composition of the present invention is a food composition.

That is, GSK5182 or a salt thereof may be added to the food composition for the purpose of preventing or ameliorating liver disease. When GSK5182 or its salt is used as a food additive, GSK5182 or its salt can be added as it is or used with other food or food ingredients, and can be suitably used according to conventional methods. The mixed amount of the active ingredient can be determined suitably according to the purpose of use (prevention, health or therapeutic treatment). In general, GSK5182 or its salt is added in an amount of 0.0001 to 1% by weight, preferably 0.001 to 0.1% by weight in the raw material composition in the manufacture of food or beverage. However, in the case of prolonged ingestion for health and hygiene purposes or for health control purposes, the amount may be used below the above range.

There is no particular limitation on the kind of the food. Examples of the food to which the substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates, and the like as additional components, as in general beverages. The above-mentioned natural carbohydrates are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g, per 100 compositions of the present invention.

In addition to the above, the composition of the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, A carbonating agent used in a carbonated beverage, and the like. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention. In addition, the composition of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. The proportion of such pulp is not critical, but is generally selected in the range of 0.01 to 10 parts by weight per 100 parts by weight of the composition of the present invention. These components can be used independently or in combination.

As another aspect, the present invention is a liver disease comprising administering an effective amount of the 4-hydroxy tamoxifen analogue represented by the formula (1) or a pharmaceutically acceptable salt thereof to a subject having or possibly developing liver disease Provide a method of treatment.

As used herein, the term "individual" means all animals, including humans, who have already developed or may have a liver disease, and can effectively prevent and treat such diseases by administering to a subject a composition comprising GSK5182 or a salt thereof. .

The optimal amount and dosage interval of the individual doses of the compounds of the present invention will be determined by the nature and extent of the disease being treated, the dosage form, the route and site, and the age and health of the particular patient being treated, which will ultimately be used by the physician. It will be appreciated by those skilled in the art that the appropriate dosage will be determined. Such dosing can be repeated as often as appropriate. If side effects occur, the dosage and frequency can be altered or reduced in accordance with normal clinical practice.

The route of administration of the composition may be administered via any conventional route so long as it can reach the target tissue. The composition of the present invention may be administered as desired, but is not limited to intraperitoneal administration, intravenous administration, subcutaneous administration, intradermal administration, oral administration. The composition may also be administered by any device capable of transferring the active agent to the target cell.

The present invention is a 4-hydroxy tamoxifen analog GSK5182 by reducing the levels of liver damage markers GOT (glutamyl oxaloacetic transaminase) and GPT (glutamyl pyruvate transaminase) to confirm that it is effective in the prevention and treatment of liver disease, 4-hydroxy There is an effect that can provide a composition for the prevention and treatment of liver disease comprising tamoxifen analog GSK5182 as an active ingredient. It may also be applied to food compositions for preventing or ameliorating liver disease.

1 shows inhibition of the promoter activity of SREBP1c induced by ERRγ of GSK5182, a 4-hydroxy tamoxifen analog, in a 293T cell line ( a ) and inhibition of protein expression of ERRγ-induced SREBP1c in rat primary cells. b ) graph.
Figure 2 is a 7-week-old db / db male mouse with a non-alcoholic fatty liver intraperitoneally administered vehicle in the control group (n = 5), GSK5182 administration group intraperitoneally for 30 days at the concentration of GSK5182 40 mg / kg Next, hepatic tissue was cut to prepare frozen tissue sections and subjected to Oil-Red O (ORO) staining, and the ratio of red stained areas in ORO was taken from the whole area observed under a microscope using an image analysis program. As a result of the comparative analysis, it is a graph showing that the non-alcoholic fatty liver of db / db male mice is clearly inhibited by the administration of GSK5182.
FIG. 3 shows GOT (glutamyl oxaloacetic transaminase) in the blood of animals administered intraperitoneally for 30 days at a concentration of 40 mg / kg GSK5182 in the control group (n = 5) using 7 weeks old db / db male mice. Comparison of ( a ) and GPT (glutamyl pyruvic transaminase) ( b ) shows a significant decrease in the GSK5182 group.
Figure 4 was adapted to a liquid alcohol diet by concentration using 8-week-old C57BL / 6 male mice, and then fed a 5.0% alcoholic liquid diet for 4 weeks and the control group (n = 5) for the last 2 weeks. ) And vehicle (n = 5) orally administered GSK5182 at a concentration of 40 mg / kg, and then cut the liver tissue to prepare a frozen tissue section to perform Oil-Red O (ORO) staining, A comparative analysis of the percentage of red stained areas in ORO over the whole area observed under a microscope using an image analysis program showed that the alcoholic fatty liver using C57BL / 6 mice was significantly inhibited by the administration of GSK5182. to be.
Figure 5 was adapted to the liquid alcohol diet by concentration using 8-week-old C57BL / 6 male mice, then fed a 5.0% alcoholic liquid diet for 4 weeks, the control group for the last 2 weeks (n = 5 ) is compared to (b) a vehicle, the experimental group (n = 5), the glutamyl oxaloacetic transaminase (I GOT of the animal orally a GSK5182 at a concentration of 40 mg / kg blood) (a) and GPT (glutamyl pyruvic transaminase) As a result, it is a graph showing a significant decrease in the group administered with GSK5182.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for describing the present invention more specifically, and the scope of the present invention is not limited by these examples.

Materials and methods

Example  1. Preparation of Compound

GSK5182 was used in HCl salt form,in vivo For the experiment It was dissolved in a 30% PEG-400 aqueous solution filtered aseptically prepared and used at a concentration of 40 mg / kg.

Example  2. SREBP1c  Promoter Activity and Protein Expression Analysis

To investigate the inhibition of sterol regulatory element binding protein1c (SREBP1c) gene promoter activity and inhibition of protein expression, in which GSK5182 plays a central role in liposynthesis, lipofactin (Invitrogen, Carlsbad, CA, USA) was used to transient transfection of plasmid DNA such as ERRγ and pGL4 / hSREBP-luc, and to compare and analyze luciferase expression following GSK5182 treatment. In addition, experiments on the inhibition of SREBP1c protein expression by GSK5182 was inoculated with Ad-GFP or Ad-ERRg while incubating primary hepatocytes from rat liver and extracting proteins from GSK5182 treated and control samples, followed by Western blot method. We compared and analyzed the expression of SREBP1c protein.

Example  3: animal experiment

To investigate the inhibitory effect of GSK5182 on non-alcoholic fatty liver, 7-week-old db / db male mouse, a non-alcoholic fatty liver model with obesity and diabetes, was used in the control group (n = 5). n = 5) was administered intraperitoneally for 30 days at a concentration of GSK5182 40 mg / kg. In addition, to determine the inhibitory effect of GSK5182 on alcoholic fatty liver, the alcoholic liquid diet (Lieber-DeCarli Regular Ethanol) consisting of 36% ethanol, 35% fat, 18% protein and 11% carbohydrate based on 1 kcal / diet, Dyets Inc) was adapted to 10 male 8-week-old male C57BL / 6 mice with increasing alcohol levels in concentration steps from 0.5% to 5.0% for 2 weeks, followed by a 5.0% alcohol liquid diet for 4 weeks. During the last two weeks of feeding, the control group (n = 5) was orally administered to the control group (n = 5) at a concentration of 40 mg / kg to GSK5182.

Example  4: in the blood GOT  And GPT  Concentration measurement

After administration of GSK5182 in nonalcoholic and alcoholic fatty liver mouse models, GOT (glutamyl oxaloacetic transaminase) and glutamyl pyruvic transaminase GPT (GPT), which are representative indicators of liver damage in the blood, were analyzed. Blood was collected using a heparin-treated capillary tube from an animal's retro-orbital sinus, followed by centrifugation at 3,000 rpm for 10 minutes to obtain plasma from the upper layer. Direct analysis was performed using an automated blood analyzer (Hitachi 7150, Japan).

Example  5: histopathologic analysis of fatty liver

In order to analyze histopathologic findings of non-alcoholic and alcoholic fatty liver following administration of GSK5182, each animal was sacrificed, then liver tissue was excised and frozen and frozen (Tissue-Tek, SAKURA). , Japan) were prepared with 10-thick frozen tissue sections, and were observed with an optical microscope subjected to Oil-Red O (ORO) staining and counterstaining with hematoxylin. Fatty liver findings were analyzed by comparing the proportion of ORO-stained sites in the whole area observed under a microscope using an image analysis program (Image Inside, GS media, Korea).

Example  6: statistical analysis

All values are expressed as mean ± standard error (SEM). The significance between the mean values was analyzed by two-tailed unpaired students t- test or Wilcoxon / Kruskal-Wallis test.

result

Experimental Example  One: GSK5182 of SREBP1c  Promoter Activity and Inhibition of Protein Expression

As a result of analyzing the SREBP1c gene promoter activity using the cell line, the activity of the SREBP1c gene promoter was significantly inhibited by GSK5182 treatment (two-tailed unpaired Students t- test, p <0.01) (FIG. 1A). In addition, as a result of examining the effects on SREBP1c protein expression using rat primary hepatocytes, it was confirmed that the expression of SREBP1c protein was significantly inhibited by GSK5182 treatment (two-tailed unpaired students t- test, p <0.01) (FIG. 1B). . These results indicate that GSK5182 inhibits ERRγ as an inverse agonist of ERRγ and inhibits SREBP1c protein expression by inhibiting the activity of the promoter of the SREBP1c gene induced by ERRγ.

Experimental Example  2: GSK5182 Fatty liver inhibitory effect and in blood GOT Of GPT  analysis

First of all, the inventors of the present invention have investigated the inhibitory effect of GSK5182 on nonalcoholic fatty liver. After administration of 40 mg / kg GSK5182 intraperitoneally to db / db mice for 30 days, frozen sections of liver tissue were made, stained with ORO staining fat, and histopathologically observed. to be in the liver of control db / db mice, most of the stem cells, while the observed indicated the red color ORO staining severe fatty liver, liver tissue of the administration of GSK5182 db / db mouse is a site that is stained with ORO reduced Observations were made using the imaging program, and it was found to be markedly reduced in the GSK5182 administration group ( p <0.05, Wilcoxon / Kruskal-Wallis tests) (FIG. 2). In addition, to observe the inhibitory effect of GSK5182 on alcoholic fatty liver, 8-week-old C57BL / 6 male mice were fed a 5.0% liquid alcohol diet for 4 weeks after the alcoholic liquid diet for 2 weeks. During the last 2 weeks, vehicle was administered orally in the control group and GSK5182 in the experimental group at a concentration of 40 mg / kg, and frozen sections of liver tissues were prepared as in the non-alcoholic fatty liver experiment. As a result, the liver tissues of the control group were found to be very severe alcoholic fatty liver, whereas the liver tissues of mice treated with GSK5182 showed a significant decrease in fatty liver. Significant reduction was observed ( p <0.05, Wilcoxon / Kruskal-Wallis tests) (FIG. 4).

These results were judged to clearly indicate that GSK5182 has the effect of inhibiting non-alcoholic and alcoholic fatty liver. On the other hand, the blood GOT and GPT concentrations of the mice used in the non-alcoholic and alcoholic fatty liver experiments showed that the non-alcoholic and alcoholic fatty livers were significantly improved compared to the control group in the experimental group administered with GSK5182 ( 3 and 5)

Overall, these results suggest that GSK5182 inhibits ERRγ in hepatic tissues in nonalcoholic and alcoholic fatty liver mouse models, and thus the activity of the SREBP1c gene promoter, a transcription factor that plays a central role in ERRγ-induced fat synthesis. By inhibiting and inhibiting the expression of SREBP1c protein, it suggests that fatty liver, which is a type of liver disease, can be prevented or treated, and that the liver function can be greatly improved, which can be used for the prevention or treatment of various liver diseases.

Claims (9)

A pharmaceutical composition for preventing or treating liver disease, comprising the 4-hydroxy tamoxifen analog of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient.
&Lt; Formula 1 >

According to claim 1, wherein the pharmaceutical composition is a pharmaceutical composition for the prevention or treatment of liver disease, further comprising a pharmaceutically acceptable carrier.
According to claim 1, wherein the liver disease is non-alcoholic or alcoholic fatty liver, pharmaceutical composition for the prevention or treatment of liver disease.
The pharmaceutical composition of claim 3, wherein the pharmaceutical composition reduces fat accumulation in the liver.
According to claim 3, The non-alcoholic fatty liver is non-alcoholic steatohepatitis or terminal fibrotic liver disease, the pharmaceutical composition for preventing or treating liver disease.
According to claim 1, wherein the liver disease is selected from the group consisting of non-viral chronic hepatitis, cirrhosis and liver cancer, pharmaceutical composition for the prevention or treatment of liver disease.
The composition for preventing or treating liver disease according to claim 1, wherein the pharmaceutical composition has one or more of the following characteristics:
(i) decreased levels of GOT (glutamyl oxaloacetic transaminase), a marker of liver damage in blood; And
(ii) decreased levels of GPT (glutamyl pyruvic transaminase).
Food composition for the prevention or improvement of liver disease comprising a 4-hydroxy tamoxifen analog of Formula 1 or a food acceptable salt thereof as an active ingredient.
&Lt; Formula 1 >

The food composition of claim 7, wherein the food composition further comprises a food acceptable carrier.

Images