KR20190130241A - Pharmaceutical composition for treating or preventing fatty liver containing cromolyn or pharmaceutically acceptable salts thereof as an active ingredient - Google Patents

Abstract

The present invention is a pharmaceutical composition for preventing or treating fatty liver containing cromolyn or a pharmaceutically acceptable salt thereof as an active component. More specifically, the present invention relates to uses for preventing and treating nonalcoholic fatty liver of cromolyn or the pharmaceutically acceptable salt thereof, wherein cromolyn or the pharmaceutically acceptable salt thereof has an effect of inhibiting the accumulation of fatty fat in the liver and has an effect of inhibiting liver fibrosis by inhibiting the expression of α-SMA, TNF-a, TGF-β, and IL-1β in the liver, thereby being able to be usefully used for preventing and treating fatty liver, in particular, non-alcoholic fatty liver.

Description

PHARMACEUTICAL COMPOSITION FOR TREATING OR PREVENTING FATTY LIVER CONTAINING CROMOLYN OR PHARMACEUTICALLY ACCEPTABLE SALTS THEREOF AS AN ACTIVE INGREDIENT}

The present invention is a pharmaceutical composition for preventing or treating fatty liver containing chromoline or a pharmaceutically acceptable salt thereof as an active ingredient, and for preventing and treating nonalcoholic fatty liver of chromoline or a pharmaceutically acceptable salt thereof. It is about.

The liver is a large organ located in the right and upper abdominal cavity of the body, weighing about 1.5 kg in adults. The normal liver is visually reddish and the surface is smooth. The shape and size of the liver will change as the liver develops. As a representative example, if you drink a lot of alcohol to accumulate oil in the liver, the size of the liver is increased, and the oily color is yellowish. Conversely, as cirrhosis develops, the size of the liver decreases and the surface becomes rugged. The liver is a very important organ that has several functions. First, the liver functions to properly process the metabolism of various nutrients in our body. Foods absorbed from the intestine are properly changed in the liver for use in various tissues of our bodies, and nutrients are used in the various tissues, and the remaining wastes are transported back to the liver for disposal. Second, the liver functions to store some of the nutrients our bodies need. Third, bile acids, which are essential for the absorption of nutrients from the intestine, are made and released into the intestine through the biliary tract. If these bile acids are not produced, they will not be absorbed by various nutrients, resulting in malnutrition. Fourth, our body functions to make substances that are absolutely necessary for proper function (albummin, coagulation protein, cholesterol, etc.). Finally, alcohol (alcohol), drugs, and detoxification of various toxins in our body (Eugene Brainward, et al. Harrison's principles of internal medicine. 15th edition. 2001).

Fat liver refers to a condition in which abnormal fat is accumulated in liver cells, and a medical condition refers to a pathological condition in which the neutral lipid content exceeds 5% of the total liver weight. In general, fatty liver can be divided into alcoholic fatty liver disease (ALD) caused by persistent and excessive drinking and nonalcoholic fatty liver with little alcohol intake but showing liver tissue similar to alcoholic fatty liver. have. Fatty liver can be divided into alcoholic fatty liver disease (alcoholic fatty liver disease) due to excessive drinking and nonalcoholic fatty liver disease (NAFLD) due to obesity, diabetes, hyperlipidemia, drugs and the like. Alcoholic fatty liver is caused by the ingestion of alcohol in the liver to promote fat synthesis and lack of normal energy metabolism, and may develop to hepatitis or cirrhosis depending on the degree of alcohol intake. In addition, non-alcoholic fatty liver can be caused by the causes of obesity, diabetes, hyperlipidemia, drugs, etc., regardless of drinking alcohol, hepatic inflammatory reaction (steatosis) and hepatocytes (hepatocellular) that do not have an inflammatory response depending on progress It refers to a wide range of diseases including non-alcoholic steatohepatitis (NASH), advanced fibrosis and cirrhosis.

Nonalcoholic fatty liver refers to a simple fatty liver that does not accompany the inflammatory response seen in patients who have not consumed excessive alcohol, and thereby develops a broad spectrum of diseases, including inflammatory responses, hepatic fibrosis and cirrhosis of the hepatocytes. Nonalcoholic fatty liver usually occurs when the amount of triglyceride synthesis in the liver exceeds the amount of triglyceride catabolic, such as increased free fatty acid absorption in the liver or increased biosynthesis of triglycerides from carbohydrate metabolism (Birkenfeld AL et al., 2014). Free fatty acids, which are the source of triglyceride synthesis in the liver, are freed from diet, biosynthesis in the body and adipose tissue and introduced into the liver (Musso G et al., 2009). Chronic inflammation in adipocytes due to prolonged malnutrition results in increased lipolysis resulting in increased fatty acid and triglycerides in the blood, while macrophages infiltrate adipocytes and secrete a representative inflammatory cytokine, TNF-α. This results in increased fatty acid inflow into skeletal muscle, resulting in increased fat mass in skeletal muscle and insulin resistance in muscle (Guilherme A et al., 2008).

Nonalcoholic fatty liver is divided into primary and secondary depending on the cause. Primary is caused by hyperlipidemia, diabetes, or obesity, which is characteristic of metabolic syndrome (Szczepaniak LS et al., 2005). , Intestinal bypass surgery), various drugs, toxic substances (poison mushrooms, bacterial toxins), metabolic causes and other factors. The incidence of nonalcoholic fatty liver associated with diabetes and obesity, an important feature of the primary metabolic syndrome, is known to occur in about 50% of diabetic patients, about 76% of obese patients, and almost all of obese diabetic patients (Gupte P). et al., 2004). In addition, biopsies in diabetic and obese patients with elevated levels of alanine aminotransferase (ALT) showed 18-36% incidence of fatty hepatitis (Braillon A et al., 1985).

Nonalcoholic fatty liver has been considered a benign disease that has not progressed, but it has been shown to exhibit a histological pattern of inflammation and liver fibrosis similar to alcoholic hepatitis, despite being a non-drinker, and is known to be a disease of poor prognosis. In particular, recently, the metabolic syndrome (Metabolic Syndrome) with the background of obesity, diabetes, etc. has attracted attention, the spread of the idea that nonalcoholic steatohepatitis (NASH) is one of his syndrome. However, the mechanism is unclear, and no effective treatment or therapeutic drug has been established. To date, there are no established therapies for nonalcoholic fatty liver, as nonalcoholic fatty liver appears to be associated with a variety of factors including diabetes, obesity, coronary artery disease, lifestyle and the like. Several reports have been reported on the effects of fatty liver disease on the treatment of diabetes or obesity. Orlistat, which is used as an oral obesity drug, has been shown to induce hepatic histological improvement in patients with fatty liver (Hussein et al., 2007) reported that metformin reduced blood liver enzyme levels, hepatic necrotic inflammation and fibrosis in nonalcoholic fatty liver patients without diabetes (Bugianesi et al., 2005). In addition, thiazolidinedione (TZD) family of agonists of peroxisome proliferatoractivated receptors (PPARs) inhibits fat accumulation in liver and muscle, and directly antifibrotic action against liver in nonalcoholic fatty liver animal models. Has been reported (Galli A et al., 2002). However, since there are few drugs available to treat fatty liver pharmacologically and only exercise and diet are recommended, development of new fatty liver drugs is required.

Therefore, until now, there is almost no pharmacological agent that can treat fatty liver, and therefore, there is an urgent need to develop an appropriate therapeutic agent.

An object of the present invention is to provide chromoline or a pharmaceutically acceptable salt thereof as a prophylactic and therapeutic substance for fatty liver.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of fatty liver containing chromoline or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a dietary supplement for preventing and improving fatty liver including chromoline or a pharmaceutically acceptable salt thereof.

The chromoline or a pharmaceutically acceptable salt thereof of the present invention has an effect of inhibiting fatty fat accumulation in the liver, inhibiting inflammation by inhibiting expression of α-SMA, TNF-a, TGF-β and IL-1β in the liver and Since there is an inhibitory effect of liver fibrosis, it can be usefully used for the prevention and treatment of fatty liver, in particular, non-alcoholic fatty liver.

1 is a diagram showing the weight change by chromoline administration.
2 is a diagram showing the change in diet by chromoline administration.
Figure 3 is a diagram showing a photograph of the liver tissue extracted from a non-alcoholic fatty liver animal model administered chromoline.
Figure 4 is a graph showing the histopathological findings of liver tissue fragment staining and non-alcoholic fatty liver animal model administered chromoline.
FIG. 5 is a diagram illustrating blood biochemistry up to 8 weeks of diet in a nonalcoholic fatty liver animal model administered chromoline.
Figure 6 is a diagram showing the blood biochemistry after the diet and chromoline administration of non-alcoholic fatty liver animal model administered chromoline.
Figure 7 is a diagram showing the fat accumulation color area in the liver tissue in a non-alcoholic fatty liver animal model by the administration of chromoline.
Figure 8 is a diagram showing the relative weight of liver in a non-alcoholic fatty liver animal model by administration of chromoline.
9 is a diagram confirming the change of triglyceride accumulation in liver tissue in the non-alcoholic fatty liver animal model by the administration of chromoline as a triglyceride and total cholesterol in the tissue.
10 is a diagram confirming the immunohistochemical changes of liver tissue in a non-alcoholic fatty liver animal model by the administration of chromoline.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the following embodiments are presented by way of illustration of the present invention, and if it is determined that the detailed description of the well-known technology or construction well known to those skilled in the art may unnecessarily obscure the subject matter of the present invention, the detailed description thereof may be omitted. However, the present invention is not limited thereto. The invention is susceptible to various modifications and applications within the scope of the following claims and the equivalent scope thereof.

Also, the terminology used herein is a term used to properly express a preferred embodiment of the present invention, which may vary depending on a user, an operator's intention, or customs in the field to which the present invention belongs. Therefore, the definitions of the terms should be made based on the contents throughout the specification. Throughout the specification, when a part is said to "include" a certain component, it means that it can further include other components, without excluding other components unless specifically stated otherwise.

All technical terms used in the present invention, unless defined otherwise, are used in the meaning as commonly understood by those skilled in the art in the related field of the present invention. Also described herein are preferred methods or samples, but similar or equivalent ones are within the scope of the present invention. The contents of all publications described herein by reference are incorporated into the present invention.

In one aspect, the present invention relates to a pharmaceutical composition for preventing and treating fatty liver containing chromoline or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment, the chromoline may be a compound represented by Formula 1:

In one embodiment, the fatty liver can be nonalcoholic fatty liver (NALCOD), any one of simple steatosis, non-alcoholic steatohepatitis (NAS) and cirrhosis It may contain the above.

In one embodiment, preventing or treating fatty liver may consist of inhibiting fatty fat accumulation in the liver, inhibiting inflammation and inhibiting liver fibrosis.

In one embodiment, chromoline or a pharmaceutically acceptable salt thereof may inhibit the expression of α-SMA, TNF-a, TGF-β and IL-1β in the liver.

In one embodiment, the pharmaceutically acceptable salt of chromoline may be selected from the group consisting of alkali metal salts, alkaline earth metal salts, salts with inorganic acids, salts with organic acids and salts with acidic amino acids, Most preferred.

In one embodiment, chromoline or a pharmaceutically acceptable salt thereof of the present invention may be administered at 50 mg / kg to 1 g / kg.

The present invention includes not only the chromoline represented by the formula (1), but also pharmaceutically acceptable salts thereof, and possible solvates, hydrates, racemates or stereoisomers which can be prepared therefrom.

The chromoline represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and acid salts formed by the pharmaceutically acceptable free acid are useful as salts. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, diaidogen phosphate, metaphosphate, pyrophosphate chloride, bromide and iodide. Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suverate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Oxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesul Nate, Xylene Sulfonate, Phenyl Acetate, Phenylpropionate, Phenyl Butyrate, Citrate, Lactate, Hydroxybutyrate, Glycolate, Maleate, Tartrate, Methanesulfonate, Propanesulfonate, Naphthalene-1-sulfonate , Naphthalene-2-sulfonate or mandelate.

Acid addition salts according to the invention are dissolved in conventional methods, for example, chromoline represented by the formula (1) in an excess of aqueous acid solution, and the salts are water miscible organic solvents such as methanol, ethanol, acetone or aceto. It can be prepared by precipitation using nitrile. In this mixture, the solvent or excess acid may be evaporated to dryness, or the precipitated salt may be prepared by suction filtration. Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. In this case, as the metal salt, it is pharmaceutically suitable to prepare sodium, carpal or calcium salts. Corresponding silver salts are also obtained by reacting alkali or alkaline earth metal salts with a suitable negative salt (eg, silver nitrate).

In the present invention, the terms "non-alcoholic fatty liver" and "non-alcoholic fatty liver disease (NAFLD)" and "non-alcoholic steatohepatitis (NASH)" can be used in combination. have.

The pharmaceutical composition of the present invention may further include a known fatty liver therapeutic agent in addition to chromoline or a salt thereof as an active ingredient, and may be combined with other known treatments for the treatment of these diseases.

In the present invention, the term "prevention" means any action that inhibits or delays the development, spread and recurrence of the interhepatic by administration of the pharmaceutical composition according to the present invention, and "treatment" refers to the chromoline or pharmaceutical of the present invention. By the administration of a composition containing the same, or a salt thereof, which is acceptable, it means any action that improves or beneficially changes the symptoms of fatty liver. Those skilled in the art to which the present invention pertains can refer to the data presented by the Korean Medical Association, etc., to know the exact criteria of the disease in which the composition of the present invention is effective, and to determine the extent of improvement, improvement and treatment. will be.

The term "therapeutically effective amount" as used in combination with an active ingredient in the present invention means an amount effective for preventing or treating a target disease, and the therapeutically effective amount of the composition of the present invention may be various factors, for example, a method of administration. It may vary depending on the purpose, location of the patient and the condition of the patient. Therefore, when used in humans, the dosage should be determined in an appropriate amount in consideration of both safety and efficiency. It is also possible to estimate the amount used in humans from an effective amount determined through animal testing. Such considerations when determining the effective amount include, for example, Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment and not to cause side effects, and the effective dose level refers to Factors including health status, type of fatty liver, cause of fatty liver, severity, drug activity, drug sensitivity, method of administration, time of administration, route of administration and rate of release, duration of treatment, combination or simultaneous use of drugs and other medicines It may be determined according to factors well known in the art. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered in single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one skilled in the art.

The pharmaceutical compositions of the present invention may include carriers, diluents, excipients, or combinations of two or more thereof conventionally used in biological preparations. As used herein, the term "pharmaceutically acceptable" means to exhibit a characteristic that is not toxic to cells or humans exposed to the composition. The carrier is not particularly limited so long as it is suitable for in vivo delivery of the composition, for example, Merck Index, 13th ed., Merck & Co. Inc. Compounds, saline solution, sterile water, Ringer's solution, buffered saline solution, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more of these components can be mixed and used as needed. Conventional additives can be added. In addition, diluents, dispersants, surfactants, binders and lubricants may be additionally added to formulate into main dosage forms, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, it may be preferably formulated according to each disease or component by a suitable method in the art or using a method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990).

In one embodiment, the pharmaceutical composition may be one or more formulations selected from the group consisting of oral formulations, external preparations, suppositories, sterile injectable solutions and sprays, with oral or injectable formulations being more preferred.

As used herein, the term "administration" means providing a given substance to an individual or patient in any suitable manner, and according to the method desired, non-oral administration (eg, intravenous, subcutaneous, intraperitoneal). Or topically injectable formulations) or orally, and the dosage varies depending on the weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and severity of the patient. Liquid preparations for oral administration of the composition of the present invention include suspensions, solvents, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are commonly used simple diluents. And the like may be included together. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories, and the like. The pharmaceutical composition of the present invention may be administered by any device in which the active substance may migrate to the target cell. Preferred modes of administration and preparations are intravenous, subcutaneous, intradermal, intramuscular, drip and the like. Injections include non-aqueous solvents such as aqueous solvents such as physiological saline solution and ring gel solution, vegetable oils, higher fatty acid esters (e.g., oleic acid), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.). Stabilizers (e.g. ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH control, to prevent microbial growth Preservatives (eg, mercury nitrate, chimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.) may be included.

As used herein, the term "individual" means monkeys, cows, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, mice, rabbits, or humans, including humans who may or may not develop the fatty liver. All animals, including guinea pigs, can be effectively prevented or treated by administering the pharmaceutical composition of the present invention to an individual. The pharmaceutical composition of the present invention can be administered in parallel with existing therapeutic agents.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, Lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, oppadry, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, stearic acid Calcium, sucrose, dextrose, sorbitol, talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included 0.1 parts by weight to 90 parts by weight based on the composition, but is not limited thereto.

In one aspect, the present invention relates to a dietary supplement for preventing and improving fatty liver comprising chromoline or a pharmaceutically acceptable salt thereof.

In one embodiment, the chromoline may be a compound represented by Formula 1:

[Formula 1]

.

.

In one embodiment, the fatty liver can be nonalcoholic fatty liver (NALCOD), any one of simple steatosis, non-alcoholic steatohepatitis (NAS) and cirrhosis It may contain the above.

In one embodiment, preventing or ameliorating fatty liver may consist of inhibiting fatty fat accumulation in the liver, inhibiting inflammation and inhibiting liver fibrosis.

In one embodiment, chromoline or a pharmaceutically acceptable salt thereof may inhibit the expression of α-SMA, TNF-a, TGF-β and IL-1β in the liver.

In one embodiment, the pharmaceutically acceptable salt of chromoline may be selected from the group consisting of alkali metal salts, alkaline earth metal salts, salts with inorganic acids, salts with organic acids and salts with acidic amino acids, Most preferred.

When the composition of the present invention is used as a food composition, the chromoline or a pharmaceutically acceptable salt thereof may be added as it is, or used together with other food or food ingredients, and may be appropriately used according to a conventional method. The composition may include a food acceptable additive in addition to the active ingredient, the amount of the active ingredient may be appropriately determined according to the purpose of use (prevention, health or therapeutic treatment).

The term "food supplement" used in the present invention means a component that can be added to food supplements, and can be appropriately selected and used by those skilled in the art as being added to prepare a health functional food of each formulation. Examples of food additives include flavors such as various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH regulators, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like, but is not limited to the kind of food additives of the present invention by the above examples.

The food composition of the present invention may include a health functional food. As used herein, the term "health functional food" refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, 'functional' means to obtain a useful effect for health purposes such as nutrient control or physiological action on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food can also be prepared without limitation as long as the formulation is recognized as a health functional food. Food composition of the present invention can be prepared in various forms of formulation, unlike the general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug as a raw material, and excellent portability, the present invention Dietary supplements are available as supplements to enhance the effectiveness of anticancer drugs.

In addition, there is no restriction on the kind of health foods in which the composition of the present invention can be used. In addition, a composition comprising the chromoline of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient may be prepared by mixing a known additive with an appropriate other auxiliary ingredient that may be contained in a dietary supplement according to the choice of a person skilled in the art. . Examples of foods that can be added include meat, sausages, breads, chocolates, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products, including ice cream, various soups, beverages, teas, drinks, alcoholic beverages and Vitamin complexes, and the like, can be prepared by adding the extract according to the present invention as a main ingredient juice, tea, jelly and juice.

The present invention will be described in more detail with reference to the following examples. However, the following examples are only intended to embody the contents of the present invention, and the present invention is not limited thereto.

Example 1 Establishment of a Non-Alcoholic Fatty Liver Disease (NAFLD) Model and Chromoline Administration

Based on the body weight of 58 male SD (Sprague-Dawley) rats (Oriental Bio Co., Ltd.) about 7 weeks old, group separation was performed to minimize the weight variation of each group so as to distribute the weight evenly among the groups. All experimental animals except the control diet group (G1) were allowed to freely consume a high fat diet (G2-G4). Five weeks after the diet, the G2-G4 group was reconstituted with ALT values obtained from blood collection (Table 1). The test substance (D.W or chromoline sodium) was orally administered once a day at a dose of 10 ml / kg body weight for 5 weeks (35 days) with the diet. Blood was collected from the jugular vein before and after the high fat diet and 5 and 8 weeks after the diet. Blood was collected from the abdominal vein on the day of necropsy. In addition, general symptoms and mortality / mortality of animals were observed once a day for 5 days a week during the test period, and records were maintained only for individuals showing unusual symptoms (not recorded if normal).

As a result, animals with specific symptoms due to chromoline administration were not observed during the test period.

Example 2 Measurement of Changes in Weight and Diet

The non-alcoholic fatty liver animal model established in Example 1 was measured by body weight once a week from the time of acquisition to necropsy. Body weight measurements during the test period were measured using the same scale on the same day of the week and at the same time to minimize errors in each measurement. Dietary measurements were determined by dividing the daily diet per cage by the number of digits from the beginning of the diet to the week before necropsy.

As a result, no significant weight difference was observed between the groups at the time of acquisition, separation of the group, and 2 weeks before the administration of the test substance, but after 3 weeks, the average weight of G2 (high fat diet) was G1 (control diet). It was significantly increased (p <0.05, p <0.01), and after 5 weeks of diet, 9.3% increased significantly compared to G1 (control group) (p <0.01). In addition, after 5 weeks of diet, the mean body weight of the test animals when regrouped with serum ALT values was G1 (control diet, D.W-administered group): 508.5 ± 20.2 g; G2 (high fat diet, D.W administration group): 556.7 ± 53.3 g; G3 (high fat diet, 100 mg / kg dose group): 558.9 ± 35.7 g; And G4 (high fat diet, 500 mg / kg dose group): 551.7 ± 39.2 g. G2 (high fat diet, DW-administered group) was significantly higher than G1 (control diet, DW-administered group) (p <0.05), and no significant weight difference was observed between groups in G2-G4 (high fat diet group). . In addition, the mean body weight of G2 (high fat diet, D.W-administered group) increased significantly compared to G1 (control diet, D.W-administered group) during the test substance administration period (p <0.05). At necropsy, the average body weight of the test animals was G1 (control diet, DW group): 611.0 ± 34.9 g, G2 (high fat diet, DW group): 691.8 ± 86.4 g: G3 (high fat diet, 100 mg / kg group): 696.7 ± 66.5 g; And G4 (high fat diet, 500 mg / kg dose group): 688.7 ± 52.9 g. That is, G2 (high fat diet, DW-administered group) was significantly increased by 13.2% compared to G1 (control diet, DW-administered group) (p <0.05), G2 (high fat diet, No statistically significant body weight difference was observed as compared to the DW group) (Table 2 and FIG. 1).

In addition, as a result of measuring the amount of diet during the test period, G2 (high fat diet group) was 14.4%, 23.6%, 21.6%, 21.3%, 17.8 compared to G1 (control group) at 1-5 weeks before the administration of the test substance. % Was statistically significantly lower (p <0.01). In addition, the average dietary amount of G2 (high fat diet, DW group) was 12.6%, 11.8%, 28.2%, 14.3 compared to G1 (control group, DW group). % And 10.1% were significantly lower (p <0.05, p <0.01). That is, no significant dietary difference was observed in G3-G4 (chromoline sodium administration group) compared to G2 (high fat diet, D.W administration group) (Table 3 and Fig. 2).

In other words, there was no statistically significant weight gain in the high-fat diet group compared to the control diet group, and no weight change was observed by the administration of chromoline sodium, and in the high-fat diet group compared to the control diet group. The significant decrease in the amount of dietary intake was due to the high content of fat in the diet.

Example 3. Histopathological Confirmation

After the test, animal models were anesthetized (Ketamine / xylazine) to take blood, and then bleed, and liver tissue was extracted for each individual. The weight of the extracted liver tissue was measured and photographed (FIG. 3). After the liver tissue was fixed in 10% buffered neutral formalin, the fixed tissue was cut to a predetermined thickness, and paraffin was embedded through general tissue treatment to prepare 4-5 μm tissue sections. . Thereafter, hematoxylin & Eosin staining (H & E stain) was performed, and histopathological findings were observed.

Observed findings in liver tissue were described in Liang W et al., PLoS One. Scored for Microvesicular steatosis, Macrovesicular steatosis, Hypertrophy and Inflammation according to 2014 Dec 23; 9 (12): e115922. In sipodiosis, G1 (control diet, DW group): 0.0 ± 0.0, G2 (high fat diet + DW group): 2.1 ± 0.8, G3 (high fat diet, 100 mg / kg group): 2.0 ± 0.5 and G4 (high fat diet, 500 mg / kg dose group): 2.0 ± 0.6. There was a statistically significant increase in G2 (high fat diet, DW group) than G1 (control diet, DW group) (p <0.01), and all chromoline sodium groups were significantly higher than G2 (high fat diet, DW group). No difference was observed. In megaphobic steatosis, G1 (control diet, DW group): 0.1 ± 0.3, G2 (high fat diet, DW group): 0.3 ± 0.6, G3 (high fat diet, 100 mg / kg group): 0.1 ± 0.3 and G4 (high fat) Diet, 500 mg / kg administration group): 0.3 ± 0.5, there was no significant difference in all the test groups. In hypertrophy, G1 (control diet, DW dose group): 0.0 ± 0.0, G2 (high fat diet, DW dose group): 0.5 ± 0.5, G3 (high fat diet, 100 mg / kg dose group): 0.4 ± 0.5 and G4 (high fat diet, 500 mg / kg administration group): 0.5 ± 0.5. G2 (high fat diet, DW group) was significantly higher than G1 (control diet, DW group) (p <0.01), G3-G4 (chromoline sodium group) was G2 (high fat diet, DW group) No significant difference was observed. In inflammation, G1 (control diet, DW group): 0.0 ± 0.0, G2 (high fat diet + DW group): 0.2 ± 0.4, G3 (high fat diet, 100 mg / kg group): 0.1 ± 0.2 and G4 (high fat diet, 500 mg / kg administration group): 0.2 ± 0.4, there was no significant difference in all the test groups. The total score was calculated by adding the above-mentioned microvesicular steatosis, megaphobic steatosis, hypertrophy and inflammation scores. G1 (control diet, DW administration group): 0.1 ± 0.3, G2 (high fat diet, DW administration group): 3.0 ± 1.2, G3 (high fat diet, 100 mg / kg dose group): 2.6 ± 0.9 and G4 (high fat diet, 500 mg / kg dose group); It was measured as 3.0 ± 1.2. In other words, G2 (high fat diet, DW group) was significantly higher than G1 (control diet, DW group) (p <0.01), G3-G4 (chromoline sodium group) was G2 (high fat diet, DW) No significant difference was observed as compared to the administration group) (Table 4 and FIG. 4).

That is, in the high-fat diet group, symptoms of hepatic microvesicle steatosis and hypertrophy were observed.

Example 4 Biochemical Measures

Blood was collected from test animals and centrifuged at 3000 rpm, 4 ° C for 10 minutes to obtain serum from the upper layer. Aspartate transaminase (AST), Alanine transaminase (ALT), Alkaline phosphate (ALP), Total bilirubin (T -Bil), Cholesterol (Chol), Triglyceride (TG), HDL-Cholesterol (HDL-C), LDL-Cholesterol (LDL-C), Total protein (TP) and Albumin (ALB) were measured. .

As a result, as shown in Table 5 and Figure 5, the blood biochemical test measured before the high fat diet showed no significant difference in all the test groups.

In addition, as shown in Table 5 and Figure 5, the results of blood biochemistry when reconstituting the group with the ALT value after a five-week diet, significant differences in AST, Chol, TG, HDL-C, LDL-C, TP and ALB Although not observed, ALT (U / L) and ALP (U / L) of G2 (high fat diet, DW group) were 1.5 and 2.1 times higher than G1 (control diet, DW group). <0.01), TBil (mg / dL) was observed to be significantly reduced by 71.4% (p <0.01).

In addition, as shown in Table 5 and Figure 5, after 8 weeks diet blood biochemical test results ALT (U / L) was G1 (control diet, DW administration group): 35.8 ± 4.1, G2 (high fat diet, DW administration group): 50.9 ± 8.4, G3 (high fat diet, 100 mg / kg dose group): 53.1 ± 6.4 and G4 (high fat diet, 500 mg / kg dose group): measured at 51.7 ± 4.8, G2 (high fat diet, DW dose group) was G1 (control) There was a statistically significant increase of 42.2% (p <0.01) compared to diet, DW group) (p <0.01), and G3-G4 (chromoline sodium group) showed a statistically significant difference compared to G2 (high fat diet, DW group). It wasn't. In addition, ALP (U / L) is G1 (control diet, DW administration group): 202.9 ± 34.9, G2 (high fat diet, DW administration group): 418.6 ± 112.1, G3 (high fat diet, 100 mg / kg administration group): 386.4 ± 71.8 And G4 (high fat diet, 500 mg / kg dose group): measured at 379.3 ± 46.3, G2 (high fat diet, DW dose group) was observed to be statistically significantly increased, 2.1 times that of G1 (control diet, DW dose group). (p <0.01), the chromoline sodium group decreased 7.7% in G3 (high fat diet, 100 mg / kg group) and 9.4% in G4 (high fat diet, 500 mg / kg group) than G2 (high fat diet, DW group). Was observed. In addition, TG (mg / dL) is G1 (control diet, DW administration group): 322.1 ± 109.8, G2 (high fat diet, DW administration group): 176.0 ± 54.4, G3 (high fat diet, 100 mg / kg administration group): 128.7 ± 44.8 And G4 (high fat diet, 500 mg / kg dose group): 139.2 ± 28.3, indicating that G2 (high fat diet, DW dose group) was significantly decreased by 45.4% compared to G1 (control diet, DW dose group) (p < 0.01), the chromoline sodium group decreased 26.9% in G3 (high fat diet, 100mg / kg test group) and 20.9% in G4 (high fat diet, 500mg / kg group) than G2 (high fat diet, DW group). It became. In addition, AST, T-Bil, Chol, HDL-C, LDL-C, T.P and ALB were not significantly different in all test groups.

In addition, as shown in Table 5 and Figure 6, after the end of diet and chromoline administration blood biochemical test results AST (U / L) is G1 (control diet, DW administration group): 115.3 ± 15.7, G2 (high fat diet, DW administration group ): 133.2 ± 29.2, G3 (high fat diet, 100 mg / kg dose group): 128.1 ± 24.6 and G4 (high fat diet, 500 mg / kg dose group): measured at 140.7 ± 21.6, G2 (high fat diet, DW dose group) A 15.5% increase over G1 (control diet, DW group) was observed, while chromoline sodium group showed a 3.8% decrease in G3 (high fat diet, 100 mg / kg group) compared to G2 (high fat diet, DW group), while G4 (high fat diet) , 500 mg / kg). In addition, ALT (U / L) is G1 (control diet, DW administration group): 32.1 ± 5.6, G2 (high fat diet, DW administration group): 41.9 ± 7.6, G3 (high fat diet, 100 mg / kg administration group): 43.4 ± 6.2 And G4 (high fat diet, 500 mg / kg dose group): measured at 43.9 ± 4.4, G2 (high fat diet, DW dose group) showed a 30.5% statistically significant increase over G1 (control diet, DW dose group) (p <0.01), the chromoline sodium administration group increased 3.6% in G3 (high fat diet, 100 mg / kg administration group) and 4.8% in G4 (high fat diet, 500 mg / kg administration group) than G2 (high fat diet, DW administration group). In addition, ALP (U / L) is G1 (control diet, DW administration group): 148.5 ± 27.7, G2 (high fat diet, DW administration group): 297.8 ± 86.1, G3 (high fat diet, 100 mg / kg administration group): 283.1 ± 65.1 And G4 (high fat diet, 500 mg / kg dose group): measured at 294.2 ± 55.9, G2 (high fat diet, DW dose group) was 2.0 times higher than G1 (control diet, DW dose group). (p <0.01), the chromoline sodium group decreased 4.9% in G3 (high fat diet, 100mg / kg group) and 1.2% in G4 (high fat diet, 500 mg / kg group) than G2 (high fat diet, DW group). appear. In addition, T-Bil (mg / dL) G1 (control diet, DW administration group): 0.14 ± 0.03, G2 (high fat diet, DW administration group): 0.14 ± 0.02, G3 (high fat diet, 100 mg / kg administration group): 0.14 ± 0.03 and G4 (high fat diet, 500 mg / kg dose group): measured at 0.13 ± 0.02, no significant differences between groups were observed in all test groups. In addition, Chol (mg / dL) G1 (control diet, DW administration group): 93.9 ± 21.9, G2 (high fat diet, DW administration group): 91.4 ± 20.7, G3 (high fat diet, 100 mg / kg administration group): 85.7 ± 25.0 , G4 (high fat diet, 500 mg / kg dose group): measured at 85.6 ± 15.9, no significant differences between groups were observed in all test groups. In addition, TG (mg / dL) is G1 (control diet, DW administration group): 125.0 ± 29.6, G2 (high fat diet, DW administration group): 49.9 ± 14.8, G3 (high fat diet, 100 mg / kg administration group): 44.0 ± 14.9 And G4 (high fat diet, 500 mg / kg dose group): measured as 39.2 ± 12.6, G2 (high fat diet, DW dose group) was found to be 60.1% significantly lower than G1 (control diet, DW dose group) (p < 0.01), the chromoline sodium group showed a decrease of 11.8% in G3 (high fat diet, 100 mg / kg group) and 21.4% in G4 (high fat diet, 500 mg / kg group) than G2 (high fat diet, DW group). . In addition, HDL-C (mg / dL) is G1 (control diet, DW administration group): 61.1 ± 15.1, G2 (high fat diet, DW administration group): 62.2 ± 16.8, G3 (high fat diet, 100 mg / kg administration group): 58.6 ± 19.2 and G4 (high fat diet, 500 mg / kg dose group): measured at 59.0 ± 10.6, no significant differences between groups were observed in all test groups. In addition, LDL-C (mg / dL) G1 (control diet, DW administration group): 18.3 ± 5.8, G2 (high fat diet, DW administration group): 21.4 ± 4.7, G3 (high fat diet, 100 mg / kg administration group): 19.6 ± 4.5 and G4 (high fat diet, 500 mg / kg dose group): measured at 19.6 ± 5.6, no significant differences between groups were observed in all test groups. In addition, TP (g / dL) is G1 (control diet, DW administration group): 6.39 ± 0.49, G2 (high fat diet, DW administration group): 6.33 ± 0.51, G3 (high fat diet, 100 mg / kg administration group): 6.13 ± 0.44 And G4 (high fat diet, 500 mg / kg dose group): 6.17 ± 0.32, so no significant differences between groups were observed in all test groups. In addition, ALB (g / dL) is G1 (control diet, DW administration group): 3.27 ± 0.25, G2 (high fat diet, DW administration group): 3.27 ± 0.27, G3 (high fat diet, 100 mg / kg administration group): 3.14 ± 0.24 And G4 (high fat diet, 500 mg / kg dose group): 3.16 ± 0.16, so no significant differences between groups were observed in all test groups.

In other words, blood biochemistry measured before high fat diet showed no significant difference in all groups, but blood biochemistry measured 5 weeks after diet showed significant increase in ALT and ALP in high fat diet group compared to control diet group. A significant decrease was observed in T-BIL, and a significant increase in ALT and ALP was observed after 8 weeks of diet. In addition, the blood biochemistry measured at autopsy showed a significant increase in ALT and ALP in the high-fat diet group compared to the control diet group, and a significant decrease in TG.

Example 5. Confirmation of Lipid Formation

Oil red-O staining was performed to observe the lipids induced during fatty liver formation. Liver tissue was fixed by treatment with 10% formalin and washed with 60% isopropanol. The degree of lipid formation was confirmed by staining with Oil red O working solution. Oil red O staining results were obtained by calculating the color development area by taking a 400x image of a constant cross section (Zen 2.3 blue edition, Carl Zeiss, Germany).

As a result of calculating the mean and standard error of fat accumulation color in liver tissue, G1 (control diet, DW group): 8.36 ± 5.07, G2 (high fat diet, DW group): 32.34 ± 8.18, G3 (high fat) Diet, 100 mg / kg dose group: 37.96 ± 8.32 and G4 (high fat diet, 500 mg / kg dose group): measured at 27.00 ± 4.12 (Table 6 and Figure 7). G2 (high fat diet, DW group) was 3.9 times higher than G1 (control diet, DW group), and statistically significant increase (p <0.01), and chromoline sodium group was higher than G2 (high fat diet, DW group). G3 (high fat diet, 100 mg / kg dose group) increased by 17.4%, but G4 (high fat diet, 500 mg / kg dose group) decreased by 16.5% (p <0.01). The fat accumulation color area measurement result was statistically significant only in the high-fat diet 500 mg / kg group, compared to the high-fat diet alone group, but on the Oil Red O staining slide, Oil Red O stained granules with increasing concentration of chromoline sodium. The size was reduced and the degree of granule staining was reduced.

In other words, the accumulation of fat in liver tissue by high fat diet increased significantly in the high fat diet group compared to the control diet group, and the accumulation of fat in liver tissue was statistically increased by administration of 500 mg / kg chromoline sodium. Significantly reduced, the size and staining of fat granules decreased.

Example 6 Determination of Liver Relative Weight, Triglyceride (TG) and Total Cholesterol (TCHO) in Tissue

By measuring the relative weight of liver to weight and the amount of triglyceride and total cholesterol of liver tissue, the effect of reducing the relative weight of liver and triglyceride by chromoline was confirmed. Specifically, liver tissue was ground with a homogenizer (Homogenizer) and then chloroform: methanol (2: 1) was added and centrifuged for 10 minutes at 4 ° C. at 3000 rpm to separate the organic solvent layer containing lipids. The separated lipids were dried for 12 hours in a 50 ℃ dryer to dry the organic solvent. Sufficiently suspend 100% EtOH in a tube containing only lipids, and then use a kit reagent for measuring triglycerides (AM157S-K, Asan, Korea) and a kit reagent for measuring total cholesterol (AM 202-K, Asan, Korea). Measured.

As a result of calculating the average and standard error by measuring the relative weight of liver, as shown in Table 7 and FIG. 8, G1 (control diet, DW administration group): 3.26 ± 0.22, G2 (high fat diet, DW administration group): 2.84 ± 0.29 , G3 (high fat diet, 100 mg / kg dose group): 2.80 ± 0.22 and G4 (high fat diet, 500 mg / kg dose group): 2.82 ± 0.20, so G2 (high fat diet, DW dose group) was G1 (control diet, 12.9% significantly lower than the DW group) (p <0.01), G3-G4 (chromoline sodium group) G2 (high fat diet, DW group) was not statistically significant.

In addition, TG (mg / g tissue) measurement results to determine the change in triglyceride accumulation in liver tissue G1 (control diet, DW administration group): 11.1 ± 4.8, G2 (high fat diet, DW administration group): 18.7 ± 2.5, G3 ( High fat diet, 100 mg / kg dose group: 18.2 ± 2.9 and G4 (high fat diet, 500 mg / kg dose group): measured as 19.5 ± 2.3 (Table 8 and FIG. 9), G2 (high fat diet, DW dose group) (P <0.01), and chromoline sodium group was 2.7% higher than G2 (high fat diet, DW group) compared with G2 (high fat diet, 100 mg / kg group). It was found to decrease, and increased 4.3% in the G4 (high fat diet, 500 mg / kg administration group). In addition, the result of measuring TCHO (mg / g tissue) content in liver tissue G1 (control diet, DW administration group): 1.8 ± 0.4, G2 (high fat diet, DW administration group): 2.7 ± 0.3, G3 (high fat diet, 100 mg / kg Administration group): 2.5 ± 0.5 and G4 (high fat diet, 500 mg / kg administration group): measured as 2.7 ± 0.3 (Table 8 and Figure 9), G2 (high fat diet, DW administration group) is G1 (control diet, DW administration group) 50.0% increased significantly (p <0.01), and the chromoline sodium group showed a 7.4% decrease in G3 (high fat diet, 100 mg / kg group) than G2 (high fat diet, DW group), and G4 (high fat diet). , 500 mg / kg).

In other words, the results of TG and TCHO measured for changes in triglyceride and total cholesterol in liver tissue were significantly higher in the high-fat diet group than in the control diet group.

Example 7. Confirmation of Immunohistochemical Change

To observe the immunohistochemical changes in liver tissues, liver tissues immobilized in 10% neutral buffered formalin solution were made into thin slices, attached to coating slides, deparaffinized and stained, and then α-SMA (α) on each slide. Immunohistochemical staining (Immunohistochemistry: IHC, n) using smooth muscle actin, TNF-α (Tumor necrosis factor-α), TGF-β (Transforming growth factor-β) and IL-1β (Interleukin-1β) antibodies = 2 / Group). The cross sections of liver tissues were then taken with a 100-fold image to calculate the color development area (Zen 2.3 blue edition, Carl Zeiss, Germany).

In order to observe the α-SMA change in liver tissue, the area of α-SMA staining was measured to calculate the mean and standard error. G1 (control diet, DW group): 0.282 ± 0.141, G2 (high fat diet, DW group) ): 0.466 ± 0.239, G3 (high fat diet, 100 mg / kg dose group): 0.370 ± 0.134 and G4 (high fat diet, 500 mg / kg dose group): measured as 0.934 ± 0.033, G2 (high fat diet, DW dose group) A 65.2% increase was observed compared to G1 (control diet, DW group), while G2 (high fat diet, 100 mg / kg group) was 20.6% lower than G2 (high fat diet, DW group) in the chromoline sodium group. Diet, 500 mg / kg administration group) was shown to increase 2.0 times compared to G2 (high fat diet, DW administration group) (Table 9 and Figure 10).

In addition, the average area and the standard error were calculated by measuring the color development area of TNF-α staining. G1 (control diet, DW administration group): 4.711 ± 0.849, G2 (high fat diet, DW administration group): 8.483 ± 0.841, G3 (high fat Diet, 100 mg / kg dose group: 4.430 ± 0.514 and G4 (high fat diet, 500 mg / kg dose group): measured at 5.083 ± 0.680, G2 (high fat diet, DW dose group) was higher than G1 (control diet, DW dose group) An increase of 80.1% was observed in the chromoline sodium-administered group, 47.8% in G3 (high-fat diet, 100 mg / kg) and 40.1% in G4 (high-fat diet, 500 mg / kg) It was found to decrease (Table 9 and FIG. 10).

In addition, as a result of measuring the color development area of TGF-β staining and calculating the average and standard error, G1 (control diet, DW administration group): 21.429 ± 7.443, G2 (high fat diet, DW administration group): 45.683 ± 1.206, G3 (high fat) Diet, 100 mg / kg dose group: 33.540 ± 1.892 and G4 (high fat diet, 500 mg / kg dose group): measured as 40.211 ± 12.289, G2 (high fat diet, DW dose group) was higher than G1 (control diet, DW dose group) A 2.1-fold increase was observed in the chromoline sodium group, 26.6% in G3 (high fat diet, 100 mg / kg) and 22.0% in G4 (high fat diet, 500 mg / kg) It was found to decrease (Table 9 and FIG. 10).

In addition, as a result of measuring the color development area of IL-1β staining, G1 (control diet, DW administration group): 4.010 ± 1.833, G2 (high fat diet, DW administration group): 18.569 ± 2.954, G3 (high fat) Diet, 100 mg / kg dose group: 15.647 ± 4.101 and G4 (high fat diet, 500 mg / kg dose group): measured at 8.514 ± 0.011, G2 (high fat diet, DW dose group) was higher than G1 (control diet, DW dose group) 4.6-fold increase, 15.7% in G3 (high fat diet, 100 mg / kg group) and 54.1% in G4 (high fat diet, 500 mg / kg group) than chromoline sodium group. It was found to decrease (Table 9 and FIG. 10).

That is, the expression of α-SMA, TNF-α, TGF-β, and IL-1β protein was higher in the high fat diet group than the control diet group, and in particular, 4.6-fold increase in IL-1β was observed. In addition, in the 100 mg / kg chromoline sodium-administered group, the expression level of all proteins was observed to be decreased compared to the high-fat diet group, but in the high-dose 500 mg / kg group, TNF-α and TGF-β decreased compared to the high-fat diet group. It was observed that IL-1β was shown to be reduced than the 100 mg / kg administration group. On the other hand, α-SMA was observed to be expressed higher than the high fat diet group.

Example 8 Statistical Processing

Each test result was analyzed by Boxn plot analysis using the commercial statistics program (IBM SPSS statistics version 19.0), excluding abnormal and extreme values for each group and statistically analyzed by Mann-Whitney analysis.

Thus, the expression of TNF-a, TGF-β and IL-1β associated with decreased liver fat accumulation and inflammation in the 500 mg / kg chromoline sodium group was reduced compared to the high fat diet group, especially in the 100 mg / kg group. Decreased α-SMA expression associated with the degree of hepatic fibrosis and inhibition of TNF-a, TGF-β and IL-1β expression associated with inflammation resulted in the administration of chromoline sodium in liver tissues in a high-fat diet-induced fatty liver rat animal model. Since it is believed to have an effect of inhibiting excessive accumulation of fat in the liver and hepatoprotective effect, chromoline may be used for inhibiting fatty liver and thereby protecting liver damage.

Claims (11)

Pharmaceutical composition for the prevention and treatment of fatty liver containing cromolyn or a pharmaceutically acceptable salt thereof as an active ingredient. The pharmaceutical composition for preventing and treating fatty liver of claim 1, wherein the chromoline is a compound represented by Formula 1 below:
[Formula 1]

. The method of claim 1, wherein the fatty liver is nonalcoholic fatty liver, non-alcoholic fatty liver, a pharmaceutical composition for preventing and treating fatty liver. 4. The pharmaceutical composition for preventing and treating fatty liver of claim 3, wherein the non-alcoholic fatty liver is any one or more of simple steatosis, non-alcoholic steatohepatitis (NASS) and cirrhosis. The pharmaceutical composition for preventing and treating fatty liver of claim 1, wherein the prevention or treatment of fatty liver consists of inhibition of fatty fat accumulation in the liver, inhibition of inflammation, and inhibition of liver fibrosis. The pharmaceutical composition for preventing and treating fatty liver of claim 1, which contains chromoline sodium as an active ingredient. The pharmaceutical composition for preventing and treating fatty liver according to claim 1, wherein chromoline or a pharmaceutically acceptable salt thereof inhibits expression of α-SMA, TNF-a, TGF-β and IL-1β in the liver. . A dietary supplement for preventing and improving fatty liver, comprising chromoline or a pharmaceutically acceptable salt thereof. The health functional food of claim 8, wherein the chromoline is a compound represented by Formula 1 below:
[Formula 1]

. The dietary supplement for preventing and improving fatty liver according to claim 8, comprising chromoline sodium. The dietary supplement for preventing and improving fatty liver according to claim 8, wherein the fatty liver is non-alcoholic fatty liver.

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