KR20240105014A - Composition for preventing, improving or treating fatty liver disease comprising Rhynchosia nulubilis - Google Patents

Abstract

The present invention relates to a composition for preventing, improving, or treating fatty liver disease, which contains an extract of the black bean as an active ingredient.
The composition containing the black bean extract according to the present invention inhibits the accumulation of fat in the blood and liver due to a high-fat diet and exhibits a liver-protecting effect, making it a food and food product for the prevention, improvement or treatment of fatty liver disease. It can be useful in pharmaceutical manufacturing.

Description

Composition for preventing, improving or treating fatty liver disease comprising Rhynchosia nulubilis}

The present invention relates to a composition for preventing, improving, or treating fatty liver disease, comprising an extract of the black bean.

The liver is an important organ in our body that is responsible for various metabolic processes, detoxification, decomposition, synthesis and secretion. (i) energy metabolism management function, (ii) synthesis, storage and distribution function of enzymes, albumin, phospholipids, etc., (iii) ) It has the function of excreting various metabolites into the duodenum through the bile duct and (iv) detoxification and decomposition functions. If there is an abnormality in the function of the liver, which plays this role, it causes problems in the body's nutrient metabolism. Liver diseases that cause abnormal liver function include fatty liver, hepatitis, cirrhosis, and liver cancer.

Among them, fatty liver refers to a symptom in which a greater amount of fat is accumulated in the liver than the proportion of fat in the normal liver (5%). Fatty liver can be broadly divided into alcoholic fatty liver caused by excessive drinking and non-alcoholic fatty liver caused by obesity, diabetes, hyperlipidemia, drugs, etc. In alcoholic fatty liver, when alcohol is consumed in large quantities, fat synthesis is promoted in the liver and normal energy metabolism is disrupted. Non-alcoholic fatty liver disease occurs due to causes other than alcohol. It is known that non-alcoholic fatty liver disease frequently accompanies adult diseases that cause abnormalities in fat metabolism, and in particular, non-alcoholic fatty liver disease (NAFLD) is one of the most common liver diseases, and is characterized by fatty liver disease. This accumulation occurs, and this disease makes the liver more sensitive to inflammation and reduces liver function.

Specifically, non-alcoholic fatty liver disease is non-alcoholic simple fatty liver disease without inflammatory response in patients who do not consume excessive amounts of alcohol, non-alcoholic steatohepatitis (NASH), and the progression of this disease to liver cell damage. It refers to a wide range of diseases including inflammatory response, liver fibrosis, and cirrhosis. Non-alcoholic fatty liver disease is divided into primary and secondary depending on the cause. Primary is known to be caused by hyperlipidemia, diabetes, or obesity, which are characteristics of metabolic syndrome, and secondary is known to be caused by nutritional causes such as rapid weight loss, starvation, various drugs, metabolic causes, and other factors.

In the case of alcoholic fatty liver disease, symptoms can be expected to be alleviated or improved by suppressing alcohol intake, whereas in the case of non-alcoholic fatty liver disease, the clear cause of the disease is not known, making treatment relatively difficult. Regarding the treatment of non-alcoholic fatty liver disease, the effect of administering several diabetes or obesity treatment drugs on fatty liver disease has been reported. In the case of Orlistat, which is used as an oral obesity treatment, there is a report that it led to histological improvement of the liver in patients with steatohepatitis (Hussein et al. , 2007), and it is an agonist of PPAR (peroxisome proliferator-activated receptor). ), the thiazolidinedione (TZD) class of drugs has been reported to inhibit fat accumulation in the liver and muscles and to have a direct anti-fibrotic effect on the liver in an animal model of non-alcoholic fatty liver disease (Galli A et al. , 2002). Currently, liver disease is being treated through exercise, abstinence from drinking, diet, etc. in combination with drug treatment, but since it is difficult to completely cure the disease, there is a continued need for the development of effective and improved liver disease treatments or liver protectors.

Meanwhile, studies examining the effect of improving liver function using natural products include fermented milk useful for maintaining and improving liver function and its manufacturing method (Korean Patent Registration No. 10-0461580), and liver containing extract of Hovenia tree fruit as an active ingredient. Functional food composition with functional improvement effect and hangover improvement effect (Korean Patent Publication No. 10-2003-0027615), Hepatitis B treatment containing extract of Jinju plant, and method for manufacturing the same (Korean Patent Publication No. 10-2003- 0063308) etc. have been developed. However, there is still a need for a technology derived from natural products that is commercially effective in preventing or treating fatty liver disease and has minimal side effects.

Under this background, the present inventors have made extensive research efforts to develop a composition that can prevent or treat fatty liver disease, and as a result, it has been found that the extract of the black bean can be used as a material for food compositions and pharmaceutical compositions for preventing, improving, or treating fatty liver disease. was confirmed and the present invention was completed.

Korean Society for Biotechnology and Bioengineering Journal 30(6): 339-344 (2015)

The present invention aims to solve the above-described problems and other problems associated therewith.

The purpose of the present invention is to provide a food composition for preventing or improving fatty liver disease, which contains a soybean extract as an active ingredient.

Another object of the present invention is to provide a health functional food containing the above food composition.

Another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient.

Another object of the present invention is to provide a quasi-drug composition for the prevention or treatment of fatty liver disease, containing an extract of the rat's eye bean as an active ingredient.

Another object of the present invention is to provide a feed composition for preventing or ameliorating fatty liver disease, which contains a soybean extract as an active ingredient.

This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Additionally, the scope of the present application cannot be considered limited by the specific description described below.

In one aspect for achieving the above object, the present invention provides a food composition for preventing or improving fatty liver disease, which contains an extract of black soybean as an active ingredient.

The term ' Rhynchosia nulubilis ' in the present invention is a type of black bean, so named because it looks like a rat's eyeball, and is also called Seomoktae (鼠目太). They are characterized by having much smaller kernels than black beans called seoritae. Although they are very small, they are rich in nutrients and are known to help strengthen bones and maintain youth when consumed regularly.

The term 'extract' in the present invention refers to an extract, such as an extract obtained by extraction treatment of the above-described black bean, a diluted or concentrated liquid of the extract, a dried product obtained by drying the extract, a crude product or purified product of the extract, or a mixture thereof. It includes extracts of all formulations that can be formed on their own and using extracts.

The method for extracting the extract of the present invention is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, cold immersion extraction, solvent extraction, steam distillation, elution, compression, ultrasonic extraction, filtration, and reflux extraction, which are performed alone or using a combination of two or more methods. It can be performed by doing this.

The extract of the present invention is extracted from soybean using an appropriate solvent, and may include, for example, a crude extract, a polar solvent-soluble extract, or a non-polar solvent-soluble extract. The type of extraction solvent used to prepare the extract is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the extraction solvent include water; lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, propanol, isopropanol, butanol, propyl alcohol, and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; Hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane;　Or a mixture thereof can be used. Specifically, the extraction solvent may be ethanol. Additionally, a solvent extract can be prepared by extracting the extract one or more times using the solvent.

Specifically, in the present invention, the soybean extract may be obtained by adding 5 to 15 times the weight of the soybean distilled water to the soybean and extracting it under reflux cooling at a temperature of 20 to 80°C several times for 1 to 5 hours. More specifically, distilled water may be added to the soybeans in an amount of 8 to 12 times the weight of the soybeans, and the extract may be refluxed and cooled several times for 2 to 4 hours at a temperature of 40 to 60°C.

The extract of the present invention is filtered to remove floating solid particles, for example, by filtering the particles using nylon, etc., or filtered using filter paper, cryofiltration, etc., and then used as is, or concentrated or dried (e.g., freeze-dried, It can be used by hot air drying, spray drying, etc.). Additionally, the extract may further undergo a conventional fractionation process or may be purified using a conventional purification method.

Specifically, in the present invention, the soybean can be extracted by adding distilled water, then concentrated in a vacuum concentrator and freeze-dried for use, but is not limited to this.

The term 'active ingredient' in the present invention refers to an ingredient that exhibits the desired activity alone or can exhibit activity in combination with a carrier that is not active on its own.

In the composition of the present invention, the active ingredient may be included in any amount (effective amount) depending on the specific use, formulation, purpose of formulation, etc., as long as it can exhibit fatty liver disease prevention or treatment activity, liver protection activity, or liver function improvement activity. A typical effective amount can be determined within the range of 0.01% by weight to 99.9% by weight based on the total weight of the composition. The 'effective amount' refers to the intended functional and pharmacological effects, such as prevention or treatment of fatty liver disease, when the composition of the present invention is administered to an individual to whom the composition of the present invention is applied during the administration period recommended by a person skilled in the art. It refers to the amount of active ingredient contained in the composition of the present invention that can be expressed. This effective amount can be determined experimentally by a person skilled in the art within the scope of his or her ordinary ability.

The term 'fatty liver disease' in the present invention refers to a symptom in which a greater amount of fat is accumulated in the liver than the proportion of fat in the normal liver (5%). The fatty liver includes alcoholic fatty liver caused by excessive drinking; Alternatively, it may include non-alcoholic fatty liver disease caused by obesity, diabetes, hyperlipidemia, or drugs. Alcoholic fatty liver disease occurs when excessive alcohol intake promotes fat synthesis in the liver and normal energy metabolism does not occur, while non-alcoholic fatty liver disease occurs due to causes other than alcohol. It is known that it frequently accompanies adult diseases that cause abnormalities.

The fatty liver disease of the present invention may be non-alcoholic fatty liver disease, but is not limited thereto.

The term 'Non-alcoholic fatty liver disease (NAFLD)' of the present invention refers to a disease in which fat accumulates in the liver due to causes other than alcohol. Non-alcoholic fatty liver disease includes both primary and secondary non-alcoholic fatty liver disease, but preferably refers to non-alcoholic fatty liver disease resulting from primary hyperlipidemia, diabetes, or obesity. For example, non-alcoholic fatty liver disease includes simple steatosis disease, nutritional fatty liver disease, starvation fatty liver disease, obese fatty liver disease, diabetic fatty liver disease, steatohepatitis, and liver fibrosis caused by the advancement of these diseases. It may include liver fibrosis and liver cirrhosis.

The composition of the present invention may reduce the level of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) in the blood.

The term 'alanine aminotransferase (ALT)' of the present invention is mainly found in tissues such as the liver and kidneys, and is involved in cellular nitrogen metabolism, amino acid metabolism, and gluconeogenesis in the liver within the body. As a related enzyme, it is transported from skeletal muscle in precursor form and performs its function depending on physiological and pathological conditions. Alanine aminotransferase converts the amino-group of alanine into L- during the reaction that changes alanine and α-ketoglutarate into L-glutamate and pyruvate. It has aminotransferase activity that transfers it to glutamate, and it sequentially carries out the process of converting pyruvate into L-lactic acid. In addition to the above reversible reaction, alanine aminotransferase also converts amino groups in the reverse reaction. It acts as a moving medium. In addition, the process of gluconeogenesis that occurs in the liver involves the transamination reaction of alanine aminotransferase, so a very large amount of ALT enzyme is present in the liver. In fact, ALT enzyme activity in the liver is known to be approximately 3000 times higher than that in the blood (Lott and Wolf, 1986). Therefore, alanine aminotransferase, which exists in particularly high concentrations in liver tissue, is released into the blood and circulates when the liver tissue is damaged by various factors such as toxic substances, viral infection, alcohol, obesity, etc. The alanine aminotransferase activity increases. Therefore, the diagnosis of liver disease can be inferred by measuring alanine aminotransferase activity in the blood.

The term 'Aspartate aminotransferase (AST)' of the present invention is similar to ALT in that it is another enzyme associated with cells of liver tissue, and AST increases when there is acute liver damage.

Additionally, the composition of the present invention may reduce the levels of lactate dehydorgenase (LDH), triglyceride (TG), total cholesterol, or low-density lipoprotein (LDL) in the blood. there is.

The composition of the present invention may prevent or improve the fatty liver disease by inhibiting fat production in liver tissue or inhibiting fat accumulation in the liver.

The food composition of the present invention may contain the above-mentioned soybean extract in various amounts as long as it has the effect of preventing or improving fatty liver disease.

In addition to the black bean extract, the food composition of the present invention may further include any compounds or natural extracts whose safety has already been proven in the art and known to have the corresponding activity in order to improve the convenience of taking or ingesting. .

These compounds or extracts include compounds, extracts, and medicinal products listed in compendiums such as the Pharmacopoeia of each country ('Korean Pharmacopoeia' in Korea) and the Code of Health Functional Foods of each country ('Standards and Specifications for Health Functional Foods' notified by the Ministry of Food and Drug Safety in Korea). The laws of each country governing the manufacture and sale of compounds, extracts, and health functional foods that have received product approval in accordance with the laws of each country that govern the manufacture and sale ('Pharmaceutical Affairs Act' in Korea) (the 'Act on Health Functional Foods' in Korea) Accordingly, compounds or extracts with recognized functionality may be included.

The term 'food' in the present invention refers to dairy products including meat, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gum, and ice cream, various soups, beverages, tea, drinks, alcoholic beverages, There are vitamin complexes, nutritional supplements, functional foods, food additives, health functional foods, and health foods, and include all foods in the conventional sense.

The above-mentioned health food refers to food that has a more active health maintenance or promotion effect compared to general food, and health supplement food refers to food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement are used interchangeably.

The term 'health functional food' in the present invention is the same term as food for special health use (FoSHU), and refers to food with high medical and medical effects that has been processed to efficiently exhibit bioregulatory functions in addition to supplying nutrients. it means. Here, ‘function’ means controlling nutrients for the structure and function of the human body or obtaining useful effects for health purposes, such as physiological effects. The food of the present invention can be manufactured by a method commonly used in the industry, and can be manufactured by adding raw materials and ingredients commonly added in the industry. Additionally, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various dosage forms and, unlike general drugs, has the advantage of not having side effects that may occur when taking the drug for a long period of time as it is made from food as a raw material, and is excellent in portability, making the composition of the present invention Food can be consumed as a supplement to improve fatty liver disease.

Specifically, the above-mentioned health functional food is a food made by adding the extract of black bean to food ingredients such as beverages, teas, spices, gum, confectionery, etc., or manufactured into pills, tablets, capsules, powders, suspensions, etc., and consuming it may cause health problems. It means that it brings about a specific effect, but unlike general drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time because it is made from food.

The food composition of the present invention is very useful because it can be consumed on a daily basis and can be expected to improve fatty liver disease.

The food composition of the present invention can be manufactured in any form, for example, beverages such as tea, juice, carbonated beverages, and electrolyte drinks, processed oils such as milk and yogurt, gums, rice cakes, Korean snacks, bread, It can be manufactured into foods such as snacks and noodles, and preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, etc. In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it complies with the enforcement laws and regulations at the time of manufacture and distribution. For example, it is a health functional food according to Korea's 'Act on Health Functional Foods', or confectionery, tea, beverages, and special foods according to each food type according to the food code of Korea's 'Food Sanitation Act' (Ministry of Food and Drug Safety Notification 'Food Standards and Specifications'). It may be food for use, etc.

Additionally, the food composition of the present invention may contain food additives in addition to the active ingredients. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated into food when manufacturing, processing, or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be guaranteed. In terms of use, these food additives are divided into sweeteners, flavor enhancers, preservatives, emulsifiers, and flavorings.

The sweetener is used to impart an appropriate sweetness to food, and can be either natural or synthetic. For example, sweeteners such as neotame, lactitol, D-ribose, mannitol, D-maltitol, and sodium saccharin can be used.

The flavor enhancer is a food additive that enhances the taste or aroma of food, and both natural and synthetic agents can be used. For example, flavor enhancers may include disodium 5'-guanylate, L-glutamic acid, glycine, betaine, etc.

As the preservative, sodium metabisulfite, sulfurous anhydride, sorbic acid, benzoic acid, grapefruit seed extract, etc. may be used.

The emulsifier is a food additive that homogeneously mixes or maintains two or more immiscible phases such as water and oil, and may include sodium gluconate, glycerin fatty acid ester, lecithin, magnesium stearate, alginic acid, etc.

The flavoring agent is a food additive that gives a unique flavor to food or reinforces the original flavor of food lost during the manufacturing process. Ethyl vanillin, ethyl octanoate, linalyl acetate, allyl caproate, paramethylacetophenone, etc. may be used. .

In addition to the food additives described above, the food composition of the present invention may contain bioactive substances or minerals known in the art and whose safety is guaranteed as food additives for the purpose of supplementing and reinforcing functionality and nutrition. The bioactive substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, and dibenzoylthiamine. Minerals include calcium preparations such as calcium citrate and stearic acid. Magnesium preparations such as magnesium, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are included.

The food composition of the present invention may contain the above-described food additives in an appropriate amount to achieve the purpose depending on the product type, and with respect to other food additives that may be included in the food composition of the present invention, each country's food code or You can refer to the Food Additives Code.

When using the black bean extract as a food additive, the black bean extract can be added as is or used together with other foods or food ingredients, and can be used appropriately according to conventional methods. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use.

The term 'prevention' of the present invention refers to any act of suppressing or delaying the development of fatty liver disease through the composition of the present invention, and the term 'improvement' of the present invention refers to improving or beneficially improving fatty liver disease through the composition of the present invention. It means all actions that make it possible.

In another aspect for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating fatty liver disease, comprising a soybean extract as an active ingredient.

The terms 'eye bean', 'extract', 'active ingredient', 'fatty liver disease', 'prevention', and 'composition' of the present invention are as described above.

The term 'treatment' of the present invention refers to any action in which fatty liver disease is improved, alleviated, or beneficially changed through administration of the pharmaceutical composition of the present invention.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent according to a conventional method. Pharmaceutically acceptable carriers are well known in the art depending on the administration route or dosage form, and for specifics, each country's pharmacopoeia, including the 'Korean Pharmacopoeia', can be referred to. Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, Examples include, but are not limited to, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. Additionally, carriers, excipients, and diluents that may be included in the composition of the present invention may be unnatural carriers, but are not limited thereto.

The pharmaceutical composition of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, or sterile injectable solutions according to conventional methods. . In detail, it can be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations are made by adding at least one excipient to the compound, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. It can be prepared by mixing. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, they contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. You can. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, wethepsol, macrogol, Tween 61, cacao, laurin, glycerogelatin, etc. can be used. Regarding the specific formulation of pharmaceutical compositions, it is known in the art, and references can be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). The above documents are considered part of this specification.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount that is sufficient to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment and does not cause side effects. The effective dose level refers to the patient's health status, type and severity of the disease, It can be determined based on factors including the activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, drugs combined or used simultaneously, and other factors well known in the medical field. The dosage and frequency of administration do not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, dogs, cats, cows, horses, pigs, and humans through various routes, and humans may be preferable. All modes of administration are contemplated and may include, but are not limited to, oral, intravenous, intramuscular or subcutaneous injection.

In another aspect for achieving the above object, the present invention provides a quasi-drug composition for preventing or treating fatty liver disease, comprising an extract of the pea extract as an active ingredient.

The terms 'eye bean', 'extract', 'active ingredient', 'fatty liver disease', 'prevention', 'treatment' and 'composition' of the present invention are as described above.

In the present invention, 'quasi-drugs' are articles used for the purpose of diagnosing, treating, alleviating, treating or preventing diseases in humans or animals, but are not instruments, machines, or devices, and are not intended to have a pharmacological effect on the structure and function of humans or animals. It refers to products used for the purpose, excluding those that are not instruments, machines, or devices. One specific example may include preparations for internal use, but is not limited thereto, and the formulation method, dosage, usage method, and components of quasi-drugs etc. can be appropriately selected from conventional techniques known in the technical field.

In addition to the above ingredients, the quasi-drug composition of the present invention may further include pharmaceutically acceptable carriers, excipients, or diluents as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effect of the present invention, and includes, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc. It can be included.

In another aspect for achieving the above object, the present invention provides a feed composition for preventing or improving fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient.

The terms 'eye bean', 'extract', 'active ingredient', 'fatty liver disease', 'prevention', 'improvement', and 'composition' of the present invention are as described above.

In the present invention, the term 'feed' refers to any natural or artificial diet, meal, etc., or a component of the meal, for animals to eat, consume, and digest, and can be manufactured with various types of feed known in the art. Specifically, it may include enriched feed, roughage, feed additives, or special feed, but is not limited thereto.

Concentrated feed includes seeds and fruits including grains such as wheat, oats, and corn, bran including rice bran, wheat bran, and barley bran as by-products obtained from refining grains, soybeans, soybeans, sesame seeds, linseed, coco palm, etc. It is a concentrate of residues such as residual starch, which is the main component of the starch residue that is left after removing starch from seed cakes, sweet potatoes, and potatoes, which are by-products of oil extraction, fishmeal, fish residue, and fresh liquid obtained from fish. Animal feed such as fish soluble, meat meal, blood meal, feather meal, skim milk powder, dried whey, which is the remaining whey when producing cheese from milk, and casein from skim milk, yeast, etc. , chlorella, and seaweed, but are not limited to these. Forage includes raw grass feed such as wild grass, grass, and green cuttings, turnips for feed, beets for feed, root vegetables such as rutterbearger, a type of turnip, raw grass, green cuttings, and grains, etc., in silos. These include, but are not limited to, silage, which is stored feed that has been filled and fermented with lactic acid, field grass, hay made by cutting and drying grass, straw from crops for breeding stock, and leaves from legumes. Special feeds include mineral feeds such as oyster shells and rock salt, urea feeds such as urea and its derivative diureide isobutane, and mixed feeds to supplement ingredients that are likely to be lacking when mixing only natural feed ingredients, or to increase the storability of the feed. Substances added in trace amounts include feed additives and dietary supplements, but are not limited to these.

The feed composition for preventing or improving fatty liver disease, protecting the liver, or improving liver function according to the present invention can be prepared by adding the black bean extract in an appropriate effective concentration range according to various feed production methods known in the art.

The feed composition according to the present invention is not particularly limited and can be applied to any object as long as it is aimed at preventing or improving fatty liver disease, protecting the liver, or improving liver function. For example, it can be applied to any entity, including non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, and fish.

The composition containing the black bean extract according to the present invention inhibits the accumulation of fat in the blood and liver due to a high-fat diet and exhibits a liver-protecting effect, making it a food and food product for the prevention, improvement or treatment of fatty liver disease. It can be useful in pharmaceutical manufacturing.

Figure 1 is a graph showing the results of analyzing body weight changes following the administration of rat-eye bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001)
Figure 2 is a graph showing the results of analyzing the change in liver weight according to the administration of the soybean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, ***p < 0.001)
Figure 3 is a graph showing the results of analyzing the changes in serum ALT according to the administration of the black bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, **p < 0.01)
Figure 4 is a graph showing the results of analyzing changes in serum AST according to the administration of the rat's eye bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, *p < 0.05, **p < 0.01)
Figure 5 is a graph showing the results of analyzing the changes in serum LDH according to the administration of the black bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, *p < 0.05, **p < 0.01)
Figure 6 is a graph showing the results of analysis of changes in serum triglycerides (TG) following administration of rat-eye bean extract in mice with fatty liver induction by high-fat diet. (##p < 0.01, **p < 0.01, ***p < 0.001)
Figure 7 is a graph showing the results of analyzing the change in serum total cholesterol (TC) following administration of the rat's eye bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, *p < 0.05)
Figure 8 shows the results of H&E staining confirming the histological changes in the liver due to administration of the rat pea extract in mice with fatty liver induction by a high-fat diet.
Figure 9 is a graph showing the results of analyzing the degree of steatosis in liver tissue according to the administration of the black bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, **p < 0.01)
Figure 10 is a graph showing the results of analyzing the degree of expansion of hepatocytes according to the administration of the black bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, **p < 0.01)
Figure 11 is a graph showing the results of analyzing the degree of inflammation following administration of the rat's eye bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, *p < 0.05, *p < 0.05)
Figure 12 is a graph showing the results of analyzing NAFLD activity according to the administration of the black bean extract in mice with fatty liver induction by a high-fat diet. (###p < 0.001, *p < 0.05, **p < 0.01)
Figure 13 shows the results of confirming the degree of fat distribution in the liver tissue according to administration of the soybean extract in mice induced with fatty liver by a high-fat diet through Oil red O staining.
Figure 14 is a graph showing the results of analyzing the degree of fat distribution in the liver tissue according to the administration of the soybean extract in mice induced with fatty liver by a high-fat diet through Oil red O staining. (###p < 0.001, ***p < 0.001)

Hereinafter, the configuration and effects of the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1: Preparation of black bean extract

To prepare the black bean extract of the present invention, it was purchased from the market, ground to a certain density, added with 10 times the weight of water, and extracted twice at 90°C for 3 hours each. Afterwards, it was concentrated using a vacuum concentrator (R-114 Rotavapor system, Buchi Labortechnik AG, Flawil, Switzerland). The obtained concentrate was dried using a freeze dryer (LP 20, Ilshinbiobase, Yangju, Korea), and the obtained dried product was stored at -20°C and used in the following experiment.

Experimental Example 1: Confirmation of the effect of rat eye bean extract on improving liver function in a mouse model in which non-alcoholic fatty liver disease was induced by a high-fat diet

1.1. Animal preparation and sample preparation

To evaluate the efficacy of the rat's eye bean extract in Example 1 for improving liver function, 5-week-old C57BL/6 male mice (20-22 g) were purchased from Orient Bio Co., Ltd. (Seongnam, Korea) and were subjected to an acclimatization period for one week to adapt to the environment. placed. Afterwards, mice were randomly assigned to groups as shown in Table 1 below, and non-alcoholic fatty liver disease was induced through a high-fat diet (60% kcal fat diet; HFD) to evaluate liver function improvement. It was administered orally at concentrations of 100 mg/kg and 200 mg/kg for 12 weeks. Diet and drinking water were allowed to be consumed freely, and distilled water was used as a vehicle for the test substance. The positive control group used milk thistle extract, which is well known to be effective in improving liver function.

division diet administered substance Administration concentration (mg/kg) N.D. normal diet distilled water - HFD 60% kcal fat diet distilled water - RN100 Snow bean extract 100 RN200 Snow bean extract 200 MT milk thistle 100

1.2. Statistical processing

One-way analysis of variance (ANOVA) was performed for statistical analysis of the experimental results below using GraphPad Prism 7 (GraphPad Software, Inc., SanDiego, CA). To test statistical significance, Dunnet's multiple range test was performed to show statistical significance at the level of p<0.05.

1.3. Weight change and liver weight measurement

In order to analyze the change in body weight according to the administration of the black bean extract in the mice induced with fatty liver by a high-fat diet in this example, body weight was measured once a week for 12 weeks, and after the test was completed, liver tissue was removed from the mouse under inhalation anesthesia and measured for weight. .

As a result, as shown in Figures 1 (body weight, kg) and Figure 2 (liver weight, mg), body weight and liver weight in the HFD group were found to be significantly increased compared to the ND group, respectively, and body weight was significantly increased compared to the HFD group. A decreasing trend was observed in the rat eye soybean (RN) administration group. In the case of liver weight, there was a significant decrease of 22% and 25.5%, respectively, when administered at 100 mg/kg and 200 mg/kg in the rat bean extract group compared to the HFD group (p<0.001).

1.4. Serum biochemical evaluation

In this example, serum biochemical changes were analyzed to evaluate the efficacy of the rat's eye bean extract in mice induced with fatty liver by a high-fat diet. On the last day of the test, the mouse was anesthetized by inhalation, blood was collected, and centrifuged at 3000 rpm for 10 minutes using a centrifuge (Centrifuge 5424 R, Eppendorf, Hamburg, Germany) to obtain serum. From the serum, alanine aminotransferase (ALT) and aspartate aminotransferase were obtained. (AST), lactate dehydrogenase (LDH), triglyceride (TG), and total cholesterol (TC) were measured using an automatic blood biochemical test analyzer (Chemistry analyzer AU680, Beckman Coulter, Tokyo, Japan).

As a result, serum ALT in Figure 3, a representative indicator of liver function, showed a significant increase of 5.6 times in the HFD group compared to the ND group (p<0.001), and the concentration of 100 mg/kg in the rat bean extract administration group compared to the HFD group. The administration group showed a decreasing trend, but did not show a significant difference. However, in the group administered 200 mg/kg of rat bean extract, there was a significant decrease of about 43.4% (p<0.01). The results of serum AST in Figure 4 showed a significant increase of about 2.63 times in the HFD group compared to the ND group (p<0.001), and 22.5% in the 100 mg/kg and 200 mg/kg groups, respectively, in the 100 mg/kg and 200 mg/kg groups, compared to the HFD group. (p<0.05) and 25.2% (p<0.01) showed a significant decrease. In addition, the results of serum LDH in Figure 5, a representative indicator of fat metabolism, showed a significant increase of 1.98 times (p<0.001) in the HFD group compared to the ND group, and a 28.5% increase (p<0.001) by administration concentration in the rat eye bean extract group compared to the HFD group. p<0.05) and a significant decrease of about 35.4% (p<0.01), which is expected to help improve liver function.

In addition, the results of serum TG in Figure 6, a representative indicator of fat metabolism, showed a significant increase of 1.54 times in the HFD group compared to the ND group (p<0.01), and the 100 mg/kg and There was a significant decrease of 37.1% (p<0.01) and 46.6% (p<0.001) in each 200 mg/kg administration group, respectively. The results of serum TC in Figure 7 showed a significant increase of 1.98 times in the HFD group compared to the ND group (p<0.001), and a 7.9% (p<0.05) and 7.9% increase by treatment concentration in the rat bean extract administration group compared to the HFD group, respectively. It showed a significant decrease of 10.8% (p<0.05). Therefore, it is believed that administration of rat eye bean extract will help improve fatty liver disease induced by a high-fat diet.

1.5. Liver histological evaluation

In this example, liver histological changes were analyzed to evaluate the efficacy of the rat's eye bean extract in mice induced with fatty liver by a high-fat diet. Liver tissue extracted during mouse autopsy was fixed with 10% neutral formalin to prepare a paraffin block. Histological changes were analyzed by performing hematoxylin and eosin (H&E) staining using tissue sections cut to a thickness of 4 μm. In addition, to analyze the degree of fat accumulation in liver tissue, frozen sections were obtained using frozen tissue and oil red O (ORO) staining was performed. Images were obtained from each stained tissue slide using a digital slide scanner (Motic Easyscan one, Motic, Hong Kong), and liver histological changes were evaluated using image analysis software (Motic DS Assistant, Hong Kong).

The results shown in Figure 8 confirm the symptoms of non-alcoholic fatty liver disease through histological changes through H&E staining, and each change is shown graphically in Figures 9 to 11. As a result of Figure 9, it can be seen that steatosis of liver tissue clearly increased in the HFD group compared to the ND group (p<0.001), showed a tendency to decrease in the RN100 group compared to the HFD group, and was significantly significantly increased in the RN200 group. showed a decrease (p<0.01). In addition, the results of hepatocyte ballooning in Figure 10 showed a significant increase in the HFD group compared to the ND group (p<0.001), and the swelling tended to decrease in the RN100 group, while the RN200 group showed a significant decrease. shown (p<0.01). The inflammation findings in Figure 11 also showed a significant increase in the HFD group compared to the ND group (p<0.001), and a tendency to decrease in the RN100 and RN200 groups compared to the HFD group, especially in the 200 mg/kg concentration group. A significant decrease was observed (p<0.05). Figure 12 shows a significant increase in NAFLD activity (NAS) in the HFD group compared to the ND group as a result of combining the results of Figures 9 to 11 (p<0.001), and RN100 in the rat bean extract administration group compared to the HFD group. and RN200 groups showed significant decreases of 37.3% (p<0.05) and 45.0% (p<0.001) compared to HFD, respectively. Therefore, it is believed that administration of rat eye bean extract may help alleviate the symptoms of non-alcoholic fatty liver disease induced by a high-fat diet.

In addition, Figure 13 shows the results of Oil red O staining, and the graph in Figure 14 shows significant results regarding changes in fat distribution within liver tissue. As a result, there was a significant increase in the HFD group compared to the ND group (p<0.001), and in the HFD group compared to the HFD group, 43.6% in the 100 mg/kg group and 57.8% in the 200 mg/kg group, respectively. There was a significant decrease of about % (p<0.001). Therefore, it is believed that administration of rat eye bean extract can help alleviate the symptoms of fatty liver induced by a high-fat diet.

Claims (17)

A food composition for preventing or improving fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient. According to paragraph 1,
The food composition wherein the black bean extract is extracted with water, a lower alcohol solvent having 1 to 4 carbon atoms, or a mixed solvent thereof. According to paragraph 1,
The food composition, wherein the fatty liver disease is non-alcoholic fatty liver disease. According to paragraph 3,
The non-alcoholic fatty liver disease is selected from the group consisting of simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, non-obese fatty liver disease, diabetic fatty liver disease, steatohepatitis, liver fibrosis, and cirrhosis. According to paragraph 1,
The composition is a food composition that inhibits fat accumulation in the liver. According to paragraph 1,
The composition is a food composition that reduces the level of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) in the blood. A health functional food comprising the food composition of any one of claims 1 to 6. A pharmaceutical composition for the prevention or treatment of fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient. According to clause 8,
A pharmaceutical composition, wherein the black bean extract is extracted with water, a lower alcohol solvent having 1 to 4 carbon atoms, or a mixed solvent thereof. According to clause 8,
A pharmaceutical composition wherein the fatty liver disease is non-alcoholic fatty liver disease. According to clause 10,
The pharmaceutical composition, wherein the non-alcoholic fatty liver disease is selected from the group consisting of simple fatty liver disease, nutritional fatty liver disease, starvation fatty liver disease, non-obese fatty liver disease, diabetic fatty liver disease, steatohepatitis, liver fibrosis, and cirrhosis. According to clause 8,
The composition is a pharmaceutical composition that inhibits fat accumulation in the liver. According to clause 8,
The composition is a pharmaceutical composition that reduces the level of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) in the blood. A quasi-drug composition for the prevention or treatment of fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient. According to clause 14,
The fatty liver disease is a non-alcoholic fatty liver disease, a quasi-drug composition. A feed composition for preventing or improving fatty liver disease, comprising an extract of the rat's eye bean as an active ingredient. According to clause 16,
The feed composition wherein the fatty liver disease is non-alcoholic fatty liver disease.