Composition for improving muscle function, muscle strength or muscle loss containing β-conglycinin, a soybean-derived 7S globulin, as an active ingredient

Abstract

The present invention relates to a composition for improving muscle function, muscle strength, or muscle loss containing β-conglycinin, a soybean-derived 7S globulin, as an active ingredient, and a method for manufacturing the same. The composition containing β-conglycinin as an active ingredient according to the present invention and the method for preparing the same can effectively extract β-conglycinin from soybeans and, by having effects such as promoting muscle cell differentiation, protecting muscle cells, regenerating muscles, and increasing muscle mass, can be usefully used in the manufacture of foods, pharmaceuticals, etc. for improving muscle function, improving muscle strength, improving muscle loss, and preventing, improving, or treating muscle diseases.

Description

Technology Field

[0001] The present invention relates to a composition for improving muscle function, muscle strength, or muscle loss containing β-conglycinin, a soybean-derived 7S globulin, as an active ingredient, and a method for manufacturing the same. Background Technology

[0002] Muscle is an important component of the human body and is a tissue derived from mesenchymal stem cells. Muscles account for approximately 40% of our body weight; they are supported by bones and tendons and consist of bundles of muscle fibers that move together and change cell size to induce contraction. Muscles are classified into skeletal, cardiac, and visceral muscles; each generates force and induces movement at its respective location, while also playing a role in protecting bodily organs such as bones, joints, and internal organs. Furthermore, muscles possess regenerative capabilities; if damaged, they can be regenerated into muscles with their original contractile and relaxation abilities after undergoing degeneration through satellite cells and the surrounding environment.

[0003] Muscle diseases are caused by congenital genetic or environmental factors, and diseases related to muscle loss are on the rise in recent years due to the trend of an aging society and increased life expectancy. Human muscle mass decreases by more than 1% annually starting from age 40, and by age 80, it has declined to 50% of its peak mass; consequently, muscle loss in old age is recognized as the most significant cause of declining overall physical function. These muscle diseases are on the rise globally compared to the past.

[0004] Sarcopenia occurs when the breakdown of muscle fiber proteins exceeds protein synthesis and is characterized by a decrease in muscle fiber diameter or density. In particular, age-related sarcopenia affects muscle strength by selectively reducing Type II fast-twitch muscles, which contract quickly, over Type I slow-twitch muscles, which contract slowly.

[0005] Korea is already an aging society, and as aging accelerates and the elderly population continues to increase, aging will be regarded as a serious problem for our society, and muscular diseases will also be a challenge that needs to be addressed.

[0006] Consistent exercise is necessary to prevent sarcopenia, a representative disease associated with aging. However, according to the 2019-2021 National Health Statistics, the volume of aerobic and strength training among the Korean population tends to decrease starting in the late 30s, and declines sharply, particularly after the age of 65. This decrease in exercise volume with increasing age exacerbates muscle loss in the elderly, having a significant negative impact on their quality of life.

[0007] In addition, according to the National Health and Nutrition Survey, as age increases, the proportion of plant-based food intake rises from 73.46% to 80.74%, while the proportion of animal-based food intake decreases from 26.54% to 19.26%. The intake of meat, a major source of protein, decreases from 198.54g to 60.28g, which can lead to muscle loss due to insufficient protein intake in the elderly. Although the intake of legumes also increases from 28.08g to 53.88g with age, protein intake among the elderly remains insufficient.

[0008] Meanwhile, soybeans, one of the representative legume foods, contain approximately 40% protein, of which β-conglycinin, a 7S globulin, and glycinin, an 11S globulin, account for more than 80%. β-conglycinin has good processability due to its high surface hydrophilicity and emulsion stability, and is known to have effects on improving hypercholesterolemia and hyperlipidemia, as well as effects on the absorption and metabolism of low-density lipoprotein (LDL). Prior art literature

[0009] Republic of Korea Registered Patent No. 10-0492142 The problem to be solved

[0010] The present invention aims to solve the aforementioned problem and other related problems.

[0011] One exemplary objective of the present invention is to provide a food composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient.

[0012] Another exemplary objective of the present invention is to provide a health functional food containing β-conglycinin as an active ingredient.

[0013] Another exemplary objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of muscle diseases containing β-conglycinin as an active ingredient.

[0014] Another exemplary objective of the present invention is to provide a quasi-drug composition for preventing or improving muscle diseases containing β-conglycinin as an active ingredient.

[0015] Another exemplary objective of the present invention is to provide a feed composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient.

[0016] Another exemplary objective of the present invention is to provide a method for preparing a composition for improving muscle function, improving muscle strength, improving muscle loss, or preventing or treating muscle diseases, comprising β-conglycinin as an active ingredient. means of solving the problem

[0017] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in this application may 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. Furthermore, the scope of this application should not be considered limited by the specific descriptions provided below.

[0018] As one embodiment for achieving the above objective, the present invention provides a food composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient.

[0019] In the present invention, 'β-conglycinin' is also called 7S globulin and corresponds to a lysine-rich protein isolated from soybeans.

[0020] In the present invention, the 'β-conglycinin' may be isolated from soybeans, but if the material of β-conglycinin according to the present invention is the same, it is included within the scope of the present invention without limitation of chemical synthesis known in the art, or obtaining from other sources or manufacturing processes.

[0021] In the present invention, the 'β-conglycinin' can be isolated from defatted soy flour. Specifically, the method comprises: (a) mixing defatted soy flour and a protein extraction buffer in a ratio of 1:10 to 1:20, stirring the mixture for extraction, and then centrifuging to recover the supernatant; (b) mixing an acidity regulator with the supernatant recovered in step (a) to adjust the pH to 6.0 to 6.4; (c) letting the mixture stand for at least 1 hour and then centrifuging to recover the supernatant; and (d) mixing an acidity regulator with the supernatant recovered in step (c) to adjust the pH to 4.8 to 5.2 and then centrifuging to recover the precipitate. and (e) a step of drying the precipitate obtained in step (d) to obtain β-conglycinin; thereby β-conglycinin can be isolated from soybeans, and a composition for improving muscle function, improving muscle loss, improving muscle strength, preventing or treating muscle diseases containing β-conglycinin as an active ingredient may be prepared.

[0022] In the present invention, the stirring extraction of step (a) may be performed at 20 to 30°C for 1 to 3 hours, and the centrifugation of step (a) may be performed at 20 to 30°C at 7000 to 9000 rpm for 20 to 40 minutes. The acidity regulator of step (b) or (d) may be hydrochloric acid (HCl), specifically hydrochloric acid with a normal concentration (N) of 1 to 3. Additionally, the settling of step (c) may be performed at 2 to 10°C for 1 to 3 hours, and the centrifugation of step (c) may be performed at 20 to 30°C at 7000 to 9000 rpm for 20 to 40 minutes. The centrifugation of step (d) above may include (i) a step of washing the precipitated β-conglycinin with distilled water after centrifuging for 10 to 30 minutes; and (ii) a step of centrifuging for 3 to 7 minutes, and the centrifugation of (i) and (ii) may be performed at 7000 to 9000 rpm.

[0023] In the present invention, the term "active ingredient" refers to a component that exhibits the intended activity alone, or can exhibit activity together with a carrier that is inactive itself.

[0024] In the present invention, 'muscle' comprehensively refers to tendons, muscles, and tendons, and 'muscle function' is a function that a muscle can perform, which may include all functions such as joint fixation, posture maintenance, exercise, and heat generation.

[0025] In the present invention, 'muscle strength' refers to the ability to exert force through muscle contraction, and said muscle strength is proportional to muscle mass. 'Muscle weakness' refers to a state in which the strength of one or more muscles is reduced, and said muscle weakness may be limited to a single muscle, one side of the body, the upper or lower limbs, etc., or may occur throughout the entire body.

[0026] In the present invention, "improvement of muscle strength" refers to differentiating root cells into muscle cells to increase muscle mass, and improving physical performance, maximal endurance, enhancing muscle recovery, reducing muscle fatigue, and improving energy balance. Furthermore, such muscle strength is related to the body's "mobility" or "exercise performance." "Exercise performance" refers to the ability to perform physical movements in daily life or sports quickly, strongly, for a long time, and skillfully, and "improvement of exercise performance" refers to the effect of improving muscle functions such as muscle strength and muscle endurance.

[0027] In the present invention, 'muscle loss' may collectively refer to a continuous decrease in skeletal muscle mass, loss of muscle mass, functional impairment, etc.

[0028] The composition of the present invention can improve muscle function or muscle strength and improve muscle loss through, for example, activities that promote muscle cell differentiation, protect muscle cells, regenerate muscle cells, and increase muscle mass, and has efficacy in preventing, improving, and treating diseases or conditions related to muscle function or muscle loss, such as sarcopenia, muscle loss, muscle atrophy, and muscle degeneration.

[0029] The food composition of the present invention may contain the β-conglycinin in various amounts as long as it has the effect of improving muscle function, improving muscle strength, or improving muscle loss.

[0030] In addition to β-conglycinin, the food composition of the present invention may further include any compound or natural extract known in the art to have its safety verified and to possess the corresponding activity in order to enhance convenience of taking or consuming.

[0031] These compounds or extracts may include compounds or extracts listed in pharmacopoeias of each country (the 'Korean Pharmacopoeia' in Korea), health functional food codes of each country (the 'Standards and Specifications for Health Functional Foods' notified by the Ministry of Food and Drug Safety in Korea), compounds or extracts that have received product approval in accordance with the laws of each country regulating the manufacture and sale of pharmaceuticals (the 'Pharmaceutical Affairs Act' in Korea), and compounds or extracts whose functionality has been recognized in accordance with the laws of each country regulating the manufacture and sale of health functional foods (the 'Act on Health Functional Foods' in Korea).

[0032] The term 'food' in the present invention includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, nutritional supplements, functional foods, food additives, health functional foods, and health foods, and includes all foods in the conventional sense.

[0033] The aforementioned "health food" refers to a food that has active effects of maintaining or promoting health compared to general food, and "health supplement food" refers to a food intended for the purpose of health support. Depending on the context, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0034] The term "health functional food" in this invention is synonymous with "food for special health use (FosHU)," and refers to a food with high medical or therapeutic effects that is processed to efficiently exhibit bio-regulatory functions in addition to nutritional supply. Here, "functionality" refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological actions regarding the structure and function of the human body. The food of this invention can be manufactured by methods commonly used in the industry, and during such manufacturing, raw materials and ingredients commonly added in the industry may be added. Furthermore, the formulation of the food can be manufactured without restriction as long as it is a formulation recognized as a food. The food composition of this invention can be manufactured in various forms of formulations. Unlike general pharmaceuticals, it has the advantage of being made from food ingredients, thus avoiding side effects that may occur with long-term use of pharmaceuticals. Additionally, due to its excellent portability, the food of this invention can be consumed as an adjuvant for improving muscle function or preventing or improving muscle diseases.

[0035] Specifically, the above-mentioned health functional food is a food made by adding β-conglycinin to food materials such as beverages, teas, spices, gum, and confectionery, or by manufacturing it into pills, tablets, capsules, powders, or suspensions, and means that consuming it brings about specific health effects. However, unlike general medicines, it has the advantage of not having side effects that may occur when taking medicine for a long period of time because it is made from food.

[0036] The food composition of the present invention is very useful because it can be consumed on a daily basis, allowing for the expectation of effects such as improved muscle function, improved muscle strength, or improved muscle loss.

[0037] The food composition of the present invention may be manufactured in any form, for example, as beverages such as tea, juice, carbonated beverages, and isotonic drinks; processed dairy products such as milk and yogurt; food products such as chewing gum, rice cakes, Korean confectionery, bread, confectionery, and noodles; and formulations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and bars. Furthermore, the food composition of the present invention may take on any product classification in terms of legal and functional classification, as long as it complies with the regulations in effect at the time of manufacture and distribution. For example, it may be a health functional food under the Korean 'Act on Health Functional Foods', or a confectionery, tea, beverage, or special dietary food according to each food type in the Food Code of the Korean 'Food Sanitation Act' (Notification of the Ministry of Food and Drug Safety 'Standards and Specifications for Food').

[0038] In addition, the food composition of the present invention may include food additives in addition to its active ingredients. Food additives can generally be understood as substances added to, mixed with, or permeated into food during its manufacture, processing, or preservation; since they are consumed daily and over a long period along with food, their safety must be guaranteed. In terms of use, such food additives are classified into sweeteners, flavor enhancers, preservatives, emulsifiers, flavorings, etc.

[0039] The above sweeteners are used to impart a suitable sweetness to food, and may be natural or synthetic. For example, sweeteners such as neotame, lactitol, D-ribose, mannitol, D-maltitol, and sodium saccharin may be used.

[0040] The above flavor enhancers are food additives that enhance the taste or aroma of food, and both natural and synthetic ones may be used. For example, flavor enhancers may include disodium 5'-guanylate, L-glutamic acid, glycine, betaine, etc.

[0041] Sodium metabisulfite, sulfur dioxide, sorbic acid, benzoic acid, grapefruit seed extract, etc. may be used as the above preservatives.

[0042] The above 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.

[0043] The above flavorings are food additives that impart a unique aroma to food or reinforce the original aroma of food lost during the manufacturing process, and may include ethyl vanillin, ethyl octanoate, linalyl acetate, allyl caproate, paramethylacetophenone, etc.

[0044] In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and guaranteed to be safe as food additives for the purpose of supplementing and reinforcing functionality and nutritional value. Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, etc. Examples of such minerals include calcium preparations such as calcium citrate, magnesium preparations such as magnesium stearate, iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc.

[0045] The food composition of the present invention may include the aforementioned food additives in an appropriate amount to achieve the purpose depending on the product type, and regarding other food additives that may be included in the food composition of the present invention, reference may be made to the food codes or food additive codes of each country.

[0046] When the above-mentioned β-conglycinin is used as a food additive, it may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the mixture of active ingredients may be appropriately determined according to the purpose of use.

[0047] The term 'improvement' in the present invention refers to any act of using the composition in the present invention to improve or benefit the condition of symptoms caused by muscle function, muscle strength, or muscle loss.

[0048] In another embodiment for achieving the above objective, the present invention provides a pharmaceutical composition for the prevention or treatment of muscle diseases containing β-conglycinin as an active ingredient.

[0049] The terms 'β-conglycinin', 'active ingredient', and 'composition' of the present invention are as described above.

[0050] In the present invention, "muscle disease" refers to a disease or condition reported in the art as a muscle disease resulting from decreased muscle function, muscle atrophy, muscle wasting, or muscle degeneration. The said muscle wasting or degeneration occurs due to genetic factors, acquired factors, aging, etc., and muscle wasting is characterized by the gradual loss of muscle mass and the weakening and degeneration of muscles, particularly skeletal or voluntary muscles and cardiac muscles. Examples of related muscle diseases include muscle loss, sarcopenia, muscle degeneration, and sarcopenia. The composition of the present invention has the effect of increasing muscle mass or improving muscle strength, and the type of muscle is not limited.

[0051] In the present invention, "prevention" refers to any act of administering the composition of the present invention to an individual to suppress or delay muscle diseases and related symptoms.

[0052] In the present invention, "treatment" refers to any act of administering the pharmaceutical composition of the present invention to an individual to improve, alleviate, or benefit the degree of muscle disease and related symptoms.

[0053] The pharmaceutical composition of the present invention may further comprise pharmaceutically acceptable carriers, excipients, or diluents according to conventional methods. Pharmaceutically acceptable carriers are known in the art according to the route of administration or formulation, and specifically, reference may be made to the pharmacopoeias of each country, including the Korean Pharmacopoeia. Carriers, excipients, and diluents that may be included in the composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition, carriers, excipients, and diluents that may be included in the composition of the present invention may be non-natural carriers, but are not limited thereto.

[0054] The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as external preparations, suppositories, or sterile injectable solutions, according to conventional methods. Specifically, when formulating, it may be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, and surfactants that are commonly used. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules, and such solid formulations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc., with the above compounds. In addition, lubricants such as magnesium stearate and talc may also be used in addition to simple excipients. Liquid formulations for oral administration include suspensions, oral liquids, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as humectants, sweeteners, flavorings, and preservatives, may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. As non-aqueous solvents and suspensions, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As bases for suppositories, Witepsol, Macrogol, Tween 61, cocoa dough, laurin dough, glycerogelatin, etc., may be used. Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to literature such as [Remington's Pharmaceutical Sciences (19th ed., 1995)]. The said literature is considered to be part of this specification.

[0055] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment and that does not cause adverse effects. The effective dose level may be determined based on factors including the patient's health status, type and severity of the disease, drug activity, sensitivity to the drug, method of administration, time of administration, route of administration and elimination rate, duration of treatment, drugs used in combination or concurrently, 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.

[0056] The pharmaceutical composition of the present invention may be administered to mammals such as rats, dogs, cats, cattle, horses, pigs, and humans via various routes, and may be preferred in humans. Any mode of administration may be anticipated and may be administered, for example, orally, intravenously, intramuscularly, or by subcutaneous injection, but is not limited thereto.

[0057] As another embodiment for achieving the above objective, the present invention provides a quasi-drug composition for preventing or improving muscle diseases containing β-conglycinin as an active ingredient.

[0058] The terms 'β-conglycinin', 'active ingredient', 'muscle disease', 'prevention', 'improvement', and 'composition' of the present invention are as described above.

[0059] In the present invention, "quasi-drug" refers to an article used for the purpose of diagnosing, treating, alleviating, managing, or preventing diseases in humans or animals, excluding instruments, machines, or devices, and an article used for the purpose of having a pharmacological effect on the structure and function of humans or animals, excluding instruments, machines, or devices. One embodiment may include, but is not limited to, an oral formulation. The formulation method, dosage, method of use, and components of the quasi-drug may be appropriately selected from the ordinary technology known in the technical field.

[0060] The quasi-drug composition of the present invention may further include, in addition to the above components, a pharmaceutically acceptable carrier, excipient, or diluent as needed. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effects of the present invention, and may include, for example, fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, sweeteners, fragrances, preservatives, etc.

[0061] As one embodiment for achieving the above objective, the present invention provides a feed composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient.

[0062] The terms 'β-conglycinin', 'active ingredient', 'muscle', 'muscle function', 'muscle strength', 'muscle reduction', 'improvement', and 'composition' of the present invention are as described above.

[0063] In the present invention, the term "feed" refers to any natural or artificial prescribed food, single meal, etc., for animals to eat, consume, and digest, or the ingredients of said single meal; it may be manufactured in various forms of feed known in the industry, and specifically may include, but is not limited to, concentrate feed, roughage, feed additives, feed aids, pet nutritional supplements, or special feeds.

[0064] Concentrated feeds include, but are not limited to, seed grains such as wheat, oats, and corn; bran, which is a byproduct obtained by refining grains and includes rice bran, wheat bran, and barley bran; oilseed meal, which is a byproduct obtained by extracting oil from soybeans, rapeseed, sesame, flaxseed, and coconut; residues such as residual starch, which is the main component of starch residue remaining after removing starch from sweet potatoes and potatoes; fish meal, fish residue; fish soluble, which is a concentrated fresh liquid obtained from fish; meat meal; blood meal; feather meal; skim milk powder; dried whey, which is the residue obtained when making cheese from milk or casein from skim milk; yeast, Chlorella, and seaweed. Roughage includes raw grass feeds such as wild grass, pasture grass, and green cuts; root vegetables such as feed turnips, feed beets, and a type of turnip called lutea bearger; silage, which is a stored feed made by filling a silo with raw grass, green cut crops, and grains and fermenting them with lactic acid; hay made by cutting and drying wild grass and pasture grass; straw of livestock breeding crops; and leaves of legumes, but is not limited thereto. Special feeds include mineral feeds such as oyster shells and rock salt; urea feeds such as urea or its derivatives such as diuretic isobutane; feed additives, which are substances added in trace amounts to compound feed to supplement components that are prone to being deficient when only natural feed ingredients are mixed, or to improve the shelf life of the feed; and dietary supplements, but are not limited thereto.

[0065] The feed composition of the present invention may further include ingredients added to conventional feeds. Examples of ingredients added to such feeds may include cereal powder, meat powder, and legumes. In the above, the cereal powder may be one or more selected from rice flour, wheat flour, barley flour, and corn flour. In the above, the meat powder may be powdered meat powder obtained by selecting one or more selected from chicken, beef, pork, and ostrich meat. In the above, the legumes may be one or more selected from soybeans, kidney beans, peas, and black beans.

[0066] The feed composition of the present invention may include one or more selected from nutrients and minerals in addition to the cereal powder, meat powder, and legumes, which are components added to conventional feeds mentioned above, in order to increase the nutritional value of the feed, and may include one or more selected from antifungal agents, antioxidants, anticoagulants, emulsifiers, and binders to prevent deterioration of feed quality.

[0067] The feed composition according to the present invention can be prepared by adding β-conglycinin in an appropriate effective concentration range according to various feed manufacturing methods known in the art.

[0068] The feed composition according to the present invention is not particularly limited and can be applied to any individual intended for the purpose of improving muscle function, improving muscle strength, or improving muscle loss. For example, it can be applied to any individual, such as non-human animals like monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cattle, sheep, pigs, goats, birds, and fish.

[0069] As one embodiment for achieving the above objective, the present invention provides a method for preparing a composition for improving muscle function, improving muscle strength, improving muscle loss, or preventing, improving, or treating muscle diseases, comprising β-conglycinin as an active ingredient.

[0070] The terms 'β-conglycinin', 'active ingredient', 'muscle', 'muscle function', 'muscle strength', 'muscle loss', 'improvement', 'muscle disease', 'prevention', 'improvement', 'treatment', and 'composition' of the present invention are as described above.

[0071] In the present invention, the manufacturing method may comprise: (a) a step of mixing defatted soybean flour and a protein extraction buffer in a ratio of 1:10 to 1:20, stirring and extracting, and then centrifuging to recover the supernatant; (b) a step of mixing an acidity regulator with the supernatant recovered in step (a) to adjust the pH to 6.2 to 6.4; (c) a step of settling for at least 1 hour and then centrifuging to recover the supernatant; (d) a step of mixing an acidity regulator with the supernatant recovered in step (c) to adjust the pH to 5.0 to 5.2 and then centrifuging to recover the precipitate; and (e) a step of drying the precipitate obtained in step (d) to obtain β-conglycinin.

[0072] Specifically, the stirring extraction of step (a) may be performed at 20 to 30°C for 1 to 3 hours, and the centrifugation of step (a) may be performed at 20 to 30°C at 7000 to 9000 rpm for 20 to 40 minutes. The acidity regulator of step (b) or (d) may be hydrochloric acid (HCl), specifically hydrochloric acid with a normal concentration (N) of 1 to 3. Additionally, the settling of step (c) may be performed at 2 to 10°C for 1 to 3 hours, and the centrifugation of step (c) may be performed at 20 to 30°C at 7000 to 9000 rpm for 20 to 40 minutes. The centrifugation of step (d) above may include (i) a step of washing the precipitated β-conglycinin with distilled water after centrifuging for 10 to 30 minutes; and (ii) a step of centrifuging for 3 to 7 minutes, and the centrifugation of (i) and (ii) may be performed at 7000 to 9000 rpm.

[0073] The method for preparing a composition containing β-conglycinin as an active ingredient according to the present invention can effectively extract β-conglycinin from soybeans, and the composition containing such β-conglycinin can provide excellent effects such as improving muscle function, improving muscle strength, improving muscle loss, preventing, improving, or treating muscle diseases. Effects of the invention

[0074] The composition containing β-conglycinin as an active ingredient according to the present invention and the method for preparing the same can effectively extract β-conglycinin from soybeans and, by having effects such as promoting muscle cell differentiation, protecting muscle cells, regenerating muscles, and increasing muscle mass, can be usefully used in the manufacture of foods, pharmaceuticals, etc. for improving muscle function, improving muscle strength, improving muscle loss, and preventing, improving, or treating muscle diseases. Brief explanation of the drawing

[0075] Figure 1 shows the manufacturing process of soybean-derived 7S globulin (β-conglycinin). Figure 2 shows the electrophoresis results of soybean-derived 7S globulin prepared in the present invention. Figure 3 shows the results of measuring muscle cell viability and creatine kinase activity following treatment with soybean-derived 7S globulin in C2C12 cells. Figure 4 shows the results of comparing muscle cell viability in C2C12 cells after treatment with dexamethasone and soybean-derived 7S globulin. Specific details for implementing the invention

[0076] The present invention will be explained in more detail below through the following examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0078] Example 1: Preparation of Soybean-Derived 7S Globulin (β-conglycinin)

[0079] The overall process for manufacturing the soybean-derived 7S globulin (β-conglycinin) of the present invention is the same as the process diagram shown in Fig. 1.

[0080] Specifically, defatted soybean flour was mixed with a protein extraction buffer (extraction solution) at a ratio of 1:20 (w / v) and stirred for 2 hours at 20°C. The mixture was then centrifuged at 8000 rpm for 30 minutes at 20°C, and the supernatant was collected using a gauze (extraction cloth). The pH of the obtained extract was adjusted to 6.2–6.4 with 2N HCl and left to stand at 4°C for 2 hours. Afterward, the mixture was centrifuged at 8000 rpm for 30 minutes at 20°C, and the supernatant was collected. The pH of the obtained extract was adjusted to 5.0, and the recovered β-conglycinin was centrifuged at 8000 rpm for 20 minutes. The recovered β-conglycinin was washed with distilled water, centrifuged again to be recovered, and dried to obtain 7S globulin. The recovered 7S globulin was identified by electrophoresis, and the electrophoresis results are shown in Figure 2.

[0082] In subsequent experiments, soy-derived 7S globulin was dissolved in phosphate-buffered saline (PBS) and treated to C2C12 cells, while it was orally administered to C57BL / 6J mice as a suspension dissolved in saline. Additionally, statistical analysis of all data from subsequent experiments was performed using SPSS 25.0, and the results were presented as the mean and standard error per experimental group. Significance between treatment groups was tested using one-way analysis variance (ANOVA) at the p<0.05 level.

[0084] Example 2. Confirmation of the effects of soybean-derived 7S globulin on muscle cells

[0085] 2.1. Effects on muscle cell differentiation

[0086] Mouse myoblast cell line (C2C12 cells) were purchased from ATCC (Manassas, VA, USA), and when the density of the C2C12 myoblast cells reached approximately 70% in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% Penicillin / Streptomycin (P / S), 2.5 × 10⁶ cells were placed in a 24-well plate. 4 The cells were seeded at a density of cell / well and cultured at 37°C under 5% CO2 conditions. Subsequently, when the cell density reached 95% or higher, the medium was changed to DMEM supplemented with 2% horse serum (HS) and 1% P / S, and differentiation induction was carried out for 4 to 6 days.

[0087] To induce differentiation of C2C12 cells, soybean-derived 7S globulin prepared in Example 1 (hereinafter referred to as '7S globulin') was treated at concentrations of 25, 50, 100, and 200 μg / mL and cultured for 6 days. Then, to measure cell viability, a mixture of 20 μL of water-soluble tetrazolium 1 (WST-1) reagent and 500 μL of serum-free, phenol red-free DMEM was applied, and absorbance was measured at 450 nm using a UV-visible spectrometer. In addition, the effect of 7S globulin on C2C12 cell differentiation at each concentration was determined by measuring creatine kinase (CK) activity using a creatine kinase activity assay kit (Abcam).

[0089] As a result, as shown in Figure 3, when 7S globulin was applied, there was no effect on cell viability compared to the control group (CON) that was not treated with the substance, and by confirming that creatine kinase activity increased compared to the control group, it was confirmed that 7S globulin does not exhibit cytotoxicity and has activity that promotes the differentiation of muscle cells.

[0091] 2.2. Inhibitory effect on the decrease in muscle cell viability

[0092] On the 4th day of C2C12 cell differentiation induction, cells were treated with 7S globulin at concentrations of 25, 50, 100, and 200 μg / ml and cultured for 2 days. Subsequently, the medium was changed to serum-free DMEM, and 100 μM of dexamethasone (DEX) was added for 24 hours. Afterward, a mixture of 20 μL of water-soluble tetrazolium 1 (WST-1) reagent and 500 μL of serum-free, phenol red-free DMEM was added. After 2 hours, absorbance was measured at 450 nm using a UV-visible spectrometer to assess cell viability.

[0094] As a result, as shown in Figure 4, cell viability decreased by about 20% compared to the control group upon dexamethasone treatment, and it was confirmed that treatment with 7S globulin inhibited the decrease in muscle cell viability in a concentration-dependent manner.

[0096] Example 3. Confirmation of the effects of soy-derived 7S globulin on improving muscle loss and muscle strength in a sarcopenia-induced animal model

[0097] 3.1. Experimental Design

[0098] 5-week-old male C57BL / 6J mice free of specific pathogens were obtained from Jung-Ang Lab. Animal, Inc. (Seoul, Korea). After an acclimatization period, the animals were classified into 6 experimental groups using a randomized block design (n = 8 / group).

[0099] Specifically, the groups were divided into a normal control group ('CON'), a dexamethasone-induced sarcopenia group ('DEX'), a sarcopenia + 25 mg / kg 7S globulin administration group ('25'), a sarcopenia + 50 mg / kg 7S globulin administration group (7SG50), a sarcopenia + 100 mg / kg 7S globulin administration group ('100'), and a sarcopenia + 200 mg / kg 7S globulin administration group ('200').

[0100] To induce sarcopenia (muscle atrophy), dexamethasone 1 mg / kg was administered via intraperitoneal injection for 20 days, and the test substance (7S globulin) was orally administered for 30 days starting 10 days after the intraperitoneal injection of dexamethasone. Both the test substance and dexamethasone were dissolved in physiological saline and administered once daily. During the study period, the body weight of the experimental animals was measured once a week at a fixed time, and food and water were allowed to be consumed freely.

[0102] Experimental animals were housed in polycarbonate rearing boxes (278×420×200 mm, Three Shine Co., Ltd., Geumsan) at a rate of 4 to 4 animals per box in a rearing environment set with a temperature of 20.9 to 22.6℃, relative humidity of 45 to 55%, ventilation rate and method of 10 to 15 times / hour, total exhaust method, lighting time of 12 hours (08:00~20:00), and illuminance of 150 to 300 Lux.

[0104] 3.2. Body Weight Change and Muscle Weight Measurement

[0105] The effects of 7S globulin on body weight gain and muscle regeneration were confirmed using dexamethasone-induced sarcopenic mice. Body weight was measured once a week, and muscle weight was measured by dissection after the end of the study period.

[0106] The results of the weight change based on the above measurements are shown in Table 1 below (values ​​with different superscripts in Table 1 indicate significant differences at p<0.05).

[0107] Specifically, at the start of the experiment, there was no difference in body weight among all test groups, but the body weight of the sarcopenic group ('DEX') decreased due to the induction of dexamethasone. This is believed to be a phenomenon caused by the induction of sarcopenia after the injection of dexamethasone. Meanwhile, the body weight showed a trend of increasing again with the administration of 7S globulin, and it was confirmed that the body weights of the '100' and '200' groups increased significantly compared to the 'DEX' group.

[0108] [Table 1]

[0109]

[0111] In addition, the results of muscle weight measurements of sarcopenic mice after the end of the experiment period are shown in Table 2 below (values ​​with different superscripts in Table 2 indicate significant differences at p<0.05).

[0112] Specifically, compared to the control group ('CON'), the weights of the quadriceps femoris, gastrocnemius, soleus, extensor digitorum longus, and tibialis anterior decreased in the 'DEX' group; however, compared to the 'DEX' group, the weights of the gastrocnemius and tibialis anterior significantly increased in the '100' group, and the weights of the quadriceps femoris, gastrocnemius, extensor digitorum longus, and tibialis anterior significantly increased in the '200' group.

[0113] [Table 2]

[0114]

[0116] 3.3. Muscle Strength Measurement

[0117] A grip strength test was conducted two days prior to the end of the experiment. Grip strength was measured in kgf (1 kgf = 1000 g) using a small animal grip strength meter (JD-A-22). The experimental animals were induced to grasp the wire mesh of the grip strength meter with their limbs, and their tails were pulled at a constant speed (2 cm / s). The measurement was recorded when the animals released all their limbs. After obtaining three measurements for each animal, the average value was calculated, and the relative grip strength with respect to weight was calculated using the following formula.

[0118] Relative grip strength (g / g) = Grip strength (kgf) / Body weight (g) × 1000

[0120] The results of mouse grip strength measurements are shown in Table 3 below (values ​​with different superscripts in Table 3 indicate significant differences at p<0.05).

[0121] Specifically, compared to the control group (0.303 ± 0.004 kgf), the grip strength of the 'DEX' group decreased to 0.194 ± 0.003 kgf, and upon administration of 7S globulin, the grip strength of the '50', '100', and '200' groups increased to 0.228 ± 0.002 kgf, 0.250 ± 0.002 kgf, and 0.287 ± 0.003 kgf, respectively.

[0122] In terms of relative grip strength, the control group and the 'DEX' group showed the highest and lowest values, with relative grip strengths of 9.03 ± 0.60 g / g and 7.62 ± 0.69 g / g, respectively, while in the 7S globulin administration group, relative grip strength increased to 8.36 ± 0.31 g / g and 8.57 ± 0.24 g / g in the '100' group and the '200' group, respectively.

[0123] [Table 3]

[0124]

[0126] Synthesizing the above experimental results, it was confirmed that the soybean-derived 7S globulin (β-conglycinin) of the present invention has excellent effects in promoting cell differentiation and inhibiting the decrease in viability in C2C12 cells, and also exhibits excellent effects in muscle regeneration and muscle strength improvement in C57BL / 6J mice, suggesting that soybean-derived 7S globulin can be helpful in inhibiting muscle loss and improving muscle strength.

[0128] From the foregoing description, those skilled in the art to which the present invention pertains will understand that the present invention may be implemented in other specific forms without altering its technical concept or essential features. In this regard, the embodiments described above should be understood as illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as including all modifications or variations derived from the meaning and scope of the claims set forth below and their equivalents, rather than from the detailed description above.

Claims

Claim 1 A food composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient, wherein the composition has activity of promoting muscle cell differentiation, regenerating muscle cells, or increasing muscle mass. Claim 2 A food composition according to claim 1, wherein the β-conglycinin is derived from soybeans. Claim 3 delete Claim 4 A health functional food comprising a food composition according to paragraph 1 or 2. Claim 5 A pharmaceutical composition for the prevention or treatment of muscle diseases containing β-conglycinin as an active ingredient, wherein the composition has activity of promoting muscle cell differentiation, regenerating muscle cells, or increasing muscle mass. Claim 6 A pharmaceutical composition according to claim 5, wherein the muscle disease is a muscle disease caused by decreased muscle function, muscle atrophy, muscle wasting, or muscle degeneration. Claim 7 A pharmaceutical composition according to claim 5, wherein the muscle disease is one or more selected from the group consisting of muscle loss, sarcopenia, muscle degeneration, and sarcopenia. Claim 8 A quasi-drug composition for preventing or improving muscle diseases containing β-conglycinin as an active ingredient, wherein the composition has activity of promoting muscle cell differentiation, regenerating muscle cells, or increasing muscle mass. Claim 9 A feed composition for improving muscle function, improving muscle strength, or improving muscle loss, containing β-conglycinin as an active ingredient, wherein the composition has activity of promoting muscle cell differentiation, regenerating muscle cells, or increasing muscle mass. Claim 10 (a) a step of mixing defatted soybean flour and a protein extraction solution (extraction buffer) in a ratio of 1:10 to 1:20, stirring the mixture for extraction, and then centrifuging to recover the supernatant; (b) a step of mixing an acidity regulator with the supernatant recovered in step (a) to adjust the pH to 6.2 to 6.4; (c) a step of settling for at least 1 hour and then centrifuging to recover the supernatant; (d) a step of mixing an acidity regulator with the supernatant recovered in step (c) to adjust the pH to 5.0 to 5.2 and then centrifuging to recover the precipitate; and (e) a step of drying the precipitate obtained in step (d) to obtain β-conglycinin; comprising a method for preparing a composition for improving muscle function, improving muscle strength, or improving muscle loss containing β-conglycinin as an active ingredient, wherein the composition has activity of promoting muscle cell differentiation, regenerating muscle cells, or increasing muscle mass.

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