KR20200082262A - Composition for prevention, improvement or treatment of obesity comprising extract of germinated soy germ - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating, or treating obesity containing a germinated soybean germ extract. More specifically, the present invention relates to a pharmaceutical composition for preventing or treating obesity, a food composition for preventing or alleviating obesity, a livestock feed composition for preventing or alleviating obesity. The germinated soybean germ extract according to the present invention exhibits an effect of inhibiting differentiation into adipocytes, reduces the expression of genes involved in adipogenesis, increases the expression of genes related to energy metabolism, and exhibits an effect of inhibiting weight gain when administered to an actual animal model, thereby being usefully used for preventing, alleviating or treating obesity.

Description

Composition for prevention, improvement or treatment of obesity comprising extract of germinated soy germ

The present invention relates to a composition for preventing, improving or treating obesity comprising germinated soybean germ extract, and more specifically, a pharmaceutical composition for preventing or treating obesity, a food composition for preventing or improving, a composition for preventing or improving livestock feed will be.

Obesity refers to an excess of body fat and can be defined as a health risk condition that causes a number of diseases, including heart disease caused by excess body fat. Obese people have a greater risk of developing various diseases, including type 2 diabetes, hyperlipidemia, arthritis and apnea, compared to people with normal weight, and are known to cause various types of cancer and heart disease.

It is difficult to define that overweight and obesity originate from one cause, but basically it is caused by eating more calories than the body consumes.

There are three main approaches to treatment with obesity drugs (Cooke and Bloom, 2006; Shi and Burn, 2004). The first approach is to suppress appetite. Diethylproprion, phendimetrazine, and phentermine are used to treat obesity by stimulating the brain's nerves to make them feel full. The second approach is to suppress the absorption of fat intake. Orlistat is typical, and this drug acts on the digestive system and prevents absorption of fat, so it can be used for a long time. The third approach is to induce weight loss through energy consumption in vivo. A typical drug is sibutramine, which increases energy consumption through nerve control. However, these drugs need to be used with caution as side effects such as side effects of nerve drugs or interference with the absorption of fat-soluble vitamins. In particular, it is not recommended to use obesity treatments for children, pregnant women, or those with high blood pressure. However, most obesity requires a long-term treatment because weight increases again after a short-term weight loss, so it is necessary to find new alternatives.

Currently, attempts are being made to find a composition for preventing or treating obesity and overweight worldwide, and research into separating natural compositions from plants is currently underway.

However, prior to the present invention, there has been no report on the effect of preventing or treating obesity of germinated soybean germ extract.

Accordingly, the inventors of the present invention have excellent body fat reduction activity and have been exploring natural substances that can be safely applied. As a result, germinated soybean germ extract shows the effect of inhibiting differentiation into fat cells, reduces the expression of genes involved in fat production, and metabolizes energy. The present invention was completed by confirming that it increases the expression of related genes and inhibits weight gain when administered to an actual animal model.

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity, including germinated soybean germ extract as an active ingredient.

In addition, another object of the present invention is to provide a food composition for preventing or improving obesity comprising germinated soybean germ extract as an active ingredient.

Another object of the present invention is to provide a composition for preventing or improving obesity, including germinated soybean germ extract as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating obesity comprising germinated soybean germ extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a food composition for preventing or improving obesity comprising germinated soybean germ extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a composition for preventing or improving obesity, including germinated soybean germ extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating obesity comprising germinated soybean germ extract as an active ingredient.

The pharmaceutical composition of the present invention may be a composition comprising germinated soybean germ extract as an active ingredient, or may be a composition consisting of germinated soybean germ extract as an active ingredient, or germinated soybean germ extract as an active ingredient essentially consisting of It may be a composition.

In the present specification, the term'comprising' is used in the same sense as'including' or'characterized by', and in the composition or method according to the present invention, specifically It does not exclude additional component or method steps not mentioned. In addition, the term'consisting of' means excluding additional elements, steps, or ingredients not separately described. The term'essentially consisting of' means that, in the scope of a composition or method, it may include substances or steps that do not materially affect the basic properties thereof in addition to the substances or steps described. .

The term “germinated soybean germ” of the present invention means a state in which germ (embryo) germinates among the soybean seeds. More preferably, it means obtained by germination of embryos separated from soybeans. The germination refers to a phenomenon in which seeds, spores, flower pots of plants and buds formed on branches, roots, etc. are generated or start to grow. The soybean is a plant having excellent nutritional value because the essential amino acids are balanced in such a way that it is said to be beef from the field. Soybean seeds are morphologically composed of seed coat, cotyledon, and germule. The proportion of each part of soybean seed is 90-92% of cotyledon, 6-8% of seed, and 2% of embryo. To a degree. Soybeans are high in protein and fat, and contain various functional ingredients, including isoflavone and soyasaponin.

The content of isoflavones and saponins in soybeans varies depending on the part of the seed chamber, and the content of these components in embryos is significantly higher than that of seed and cotyledon. In particular, it has been reported that germination of embryos separated from soybeans increases the content of the physiologically active substances isoflavones and saponins, and changes in components in which sugar content rapidly decreases.

Therefore, it can be expected that the extract of germinated soybean germ increased the content of useful components more than the extract of simply soybean germ.

The pharmaceutical composition for preventing or treating obesity of the present invention may contain one or more active ingredients exhibiting similar functions to the germinated soybean germ extract according to the present invention. The inclusion of additional ingredients may further enhance the effectiveness of treating obesity in the compositions according to the invention. When adding the above components, skin safety, ease of formulation, and stability of active ingredients according to the combined use may be considered.

Germinated soybean germ extract of the present invention may include a pre-treatment process such as a drying process for the raw material in order to increase extraction efficiency in the manufacture of the extract.

Germinated soybean germ extract of the present invention can be extracted by a known natural product extraction method. Preferably, it may be extracted with one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, and a subcritical or supercritical fluid. The organic solvent having 1 to 6 carbon atoms is alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride (methylene) chloride), hexane (hexane), cyclohexane (cyclohexane) and petroleum ether (petroleum ether) may be selected from the group consisting of, but is not limited thereto.

In addition, the germinated soybean germ extract of the present invention can be obtained by extracting and purifying germinated soybean germ using purified water, ethanol and subcritical water or supercritical carbon dioxide suitable for food processing, or from oil obtained by directly compressing germinated soybean germ. It can be obtained by separating and purifying. For example, germinated soybean embryos may be extracted under ultra-high pressure conditions of 100 Mpa or more to obtain an extract. Preferably, it may be an ultra high pressure condition of 100 MPa to 1000 MPa, but is not limited thereto.

The extract of the present invention can be used as a liquid by filtration and/or concentration, and can be solidified through a common drying process such as spray drying or freeze drying. In the drying process, dextrin may be mixed and dried before spray drying or freeze drying.

More preferably, the germinated soybean germ extract of the present invention comprises refluxing germinated soybean germ; Concentrating the extracted extract under reflux under reduced pressure; And spray-driing the concentrated extract.

The reflux extraction is preferably performed for 3 hours or more.

The reduced pressure concentration is preferably performed at a temperature of 10°C to 70°C, more preferably 25°C to 50°C. In one embodiment of the present invention, the extract extracted under reflux was filtered and then concentrated under reduced pressure using a rotary vacuum evaporator.

The pharmaceutical composition for preventing or treating obesity of the present invention may further include a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like. In addition, stabilizers and preservatives may be further included. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-parabens and chlorobutanol. As other pharmaceutically acceptable carriers, reference may be made to those described in the following documents (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition of the present invention can be administered by any method to mammals, including humans. For example, it can be administered orally or parenterally, and the parenteral administration method is not limited thereto, but intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal , Intranasal, intestinal, topical, sublingual or rectal administration.

The pharmaceutical composition of the present invention may be formulated as a formulation for oral administration or parenteral administration according to the administration route as described above. When formulated, one or more buffers (e.g. saline or PBS), antioxidants, bacteriostats, chelating agents (e.g. EDTA or glutathione), fillers, extenders, binders, adjuvants (e.g. aluminum hydroxide) Side), a suspending agent, a thickening agent wetting agent, a disintegrating agent or a surfactant, a diluent or an excipient.

Solid preparations for oral administration include tablets, pills, powders, granules, liquids, gels, syrups, slurries, suspensions, capsules, etc., and these solid preparations include, for example, at least one excipient in the pharmaceutical composition of the present invention. For example, starch (including corn starch, wheat starch, rice starch, potato starch, etc.), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol (mannitol), xylitol, erythritol maltitol, cellulose, methyl celulose, sodium carboxymethyl cellulose and hydroxypropylmethyl-cellulose or gelatin It can be prepared by mixing the back. For example, tablets or tablets of sugar can be obtained by blending the active ingredient with a solid excipient and then grinding it and adding a suitable adjuvant to the granule mixture.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions or syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances or preservatives, in addition to water or liquid paraffin, a simple diluent commonly used. .

In addition, if necessary, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant, and may further include an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, and a preservative. .

When administered parenterally, the pharmaceutical composition of the present invention may be formulated according to methods known in the art in the form of injections, transdermal administrations, and nasal inhalants together with suitable parenteral carriers. The injections must be sterile and protected from contamination of microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media comprising water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and/or vegetable oils. Can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) with ethanol amine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. Isotonic solutions such as can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid, thimerosal, etc. may be additionally included. . In addition, the injection may further include isotonic agents such as sugars or sodium chloride in most cases.

In the case of transdermal dosage forms, ointments, creams, lotions, gels, external solutions, pasta agents, linen agents, aerosols, and the like are included. In the above,'transdermal administration' means that the pharmaceutical composition is topically applied to the skin to deliver an effective amount of the active ingredient contained in the pharmaceutical composition into the skin.

For inhalation dosages, the germinated soybean germ extract of the present invention may be prepared using a suitable propellant, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas, pressurized pack or It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of pressurized aerosols, the dosage unit can be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges used in inhalers or insufflators can be formulated to contain a powder mixture of a compound and a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975.Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, a prescription generally known to all pharmaceutical chemistries.

The pharmaceutical composition for preventing or treating obesity of the present invention may provide a desirable obesity prevention and treatment effect when germinated soybean germ extract is contained in an effective amount. In the present specification, the term'effective amount' refers to an amount that exhibits a higher response than the negative control group, and preferably refers to an amount sufficient to alleviate muscle atrophy or improve muscle function. Germinated soybean germ extract may be included in the pharmaceutical composition of the present invention in an amount of 0.01 to 99.99%, and the remaining amount may be occupied by a pharmaceutically acceptable carrier. The effective amount of germinated soybean germ extract contained in the pharmaceutical composition of the present invention will vary depending on the form in which the composition is commercialized.

'Treatment' as used herein refers to a clinical procedure to change the natural course of an individual or cell being treated, and may also be performed for the prevention of clinical pathology. Desirable effects of treatment include suppressing the occurrence or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, reducing the rate of disease progression, improving the condition of the disease, improving, alleviating, or improving prognosis. Includes. The term'prevention' also means any action that suppresses the onset of disease or delays its progression.

The total effective amount of the pharmaceutical composition of the present invention can be administered to a patient in a single dose, and can be administered by a fractionated treatment protocol that is administered for a long time in multiple doses. The pharmaceutical composition of the present invention can vary the content of the active ingredient according to the degree of disease. When parenterally administered, it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per kg of body weight per day, based on germinated soybean germ extract, and when administered orally, based on germinated soybean germ extract. It can be divided into 1 to several times to be administered in an amount of preferably 0.01 to 100 mg, more preferably 0.01 to 10 mg per 1 kg of body weight per day. However, the dose of the germinated soybean germ extract is effective dosage for the patient considering various factors such as the patient's age, weight, health status, sex, disease severity, diet and excretion rate, as well as the route and frequency of administration of the pharmaceutical composition. Considering this, considering this point, a person skilled in the art will be able to determine the appropriate effective dosage according to the specific use for preventing and treating the germinated bean germ extract. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, route of administration and method of administration as long as it shows the effects of the present invention.

The pharmaceutical composition for preventing or treating obesity of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormonal therapy, chemotherapy, or biological response modifiers.

The pharmaceutical composition for preventing or treating obesity of the present invention may also be provided as a formulation of an external preparation containing germinated soybean germ extract as an active ingredient.

When the pharmaceutical composition for preventing or treating muscle diseases of the present invention is used as an external preparation for skin, fat substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents ), fragrance, surfactant, water, ionic emulsifier, nonionic emulsifier, filler, metal ion blocker, chelating agent, preservative, vitamin, blocker, wetting agent, essential oil, dye, pigment, hydrophilic active agent, lipophilic active agent Or it may contain an adjuvant commonly used in the field of skin science, such as any other ingredients commonly used in external preparations for skin such as lipid vesicles. In addition, the ingredients may be introduced in an amount commonly used in the field of skin science.

When the pharmaceutical composition for preventing or treating obesity of the present invention is provided as an external preparation for skin, the present invention is not limited thereto, and may be a formulation such as ointment, patch, gel, cream or spray.

In addition, the present invention provides a food composition for preventing or improving muscle disease comprising germinated soybean germ extract as an active ingredient.

The food composition according to the present invention can be used to prevent or improve obesity due to increased body fat.

The food composition of the present invention includes all forms of functional foods, nutritional supplements, health foods, food additives, etc., and targets animals including humans or livestock. Is done. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.

Food compositions of this type can be prepared in various forms according to conventional methods known in the art. Beverages (including alcoholic beverages), fruits and processed foods thereof (e.g. canned fruits, canned foods, jams, marmalades, etc.), fish, meat and processed foods (e.g. ham, sausages) Corn beef, etc.), breads and noodles (e.g. udon, buckwheat noodles, ramen, spagate, macaroni, etc.), juice, various drinks, cookies, syrup, dairy products (e.g. butter, cheese, etc.), edible plant maintenance, margarine , Plant protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the germinated soybean germ extract. In addition, it is not limited to the nutritional supplement, but may be prepared by adding germinated soybean germ extract to capsules, tablets, pills, and the like. In addition, as a health functional food, but not limited to, for example, the germinated soybean germ extract itself is prepared in the form of tea, juice, and drink to be liquefied, granulated, encapsulated, and powdered for drinking (healthy beverage). Can be consumed. In addition, in order to use the germinated soybean germ extract in the form of food additives, it can be prepared and used in powder or concentrate form. In addition, it can be prepared in the form of a composition by mixing with germinated bean germ extract and known active ingredients known to have an effect of improving obesity.

When the food composition of the present invention is used as a health drink composition, the health drink composition may contain various flavoring agents or natural carbohydrates, etc., as additional ingredients, such as conventional beverages. The natural carbohydrates described above include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin and cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as thaumatin and stevia extracts; Synthetic sweeteners such as saccharin and aspartame can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g per 100 mL of the composition of the present invention, preferably about 0.02 to 0.03 g.

When the germinated soybean germ extract according to the present invention is contained as an active ingredient in a food composition for preventing or improving obesity, the amount is not particularly limited to an amount effective to achieve the effect of improving obesity, but it is 0.01 to about the total weight of the total composition It is preferably 100% by weight. The food composition of the present invention can be prepared by mixing with germinated soybean germ extract together with other active ingredients known to have an effect of improving obesity.

When the food composition of the present invention is used as a health food in addition to the above, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH It may contain a regulator, stabilizer, preservative, glycerin, alcohol or carbonic acid. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage or vegetable beverage. These ingredients can be used independently or in combination. The proportion of these additives is not critical, but is generally selected from 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the present invention provides a composition for preventing or improving obesity, including germinated soybean germ extract as an active ingredient.

The composition for livestock feed according to the present invention can be used for the prevention or improvement of obesity due to the increase in body fat of livestock. The livestock may be one selected from the group consisting of cow, pig, chicken, duck, goat, sheep and horse.

The feed composition may include feed additives. The feed additive of the present invention corresponds to the supplementary feed under the feed management law.

In the present invention, the term, "feed" can mean any natural or artificial diet, one meal or the like, or a component of the one meal, for the animal to eat, ingest, and digest.

The type of the feed is not particularly limited, and a feed commonly used in the art may be used. Non-limiting examples of the feed, vegetable feed, such as grains, muscles, food processing by-products, algae, fiber, pharmaceutical by-products, fats and oils, starches, peels or grain by-products; And animal feeds such as proteins, inorganics, oils and fats, oils and fats, single cell proteins, animal planktons and food. These may be used alone or in combination of two or more.

In addition, the feed additive may additionally contain a carrier acceptable to the animal unit. In the present invention, the feed additive may be added as it is or a known carrier, stabilizer, etc., and various nutrients such as vitamins, amino acids, minerals, antioxidants, and other additives may be added as necessary, and the shape thereof may be added. Powder, granule, pellet, suspension, and the like. When the feed additive of the present invention is supplied, it can be supplied to the unit animal alone or by mixing it in the feed.

Therefore, the present invention shows the effect of inhibiting germination bean germ extracts into adipocyte differentiation, reducing the expression of genes involved in fat production, increasing the expression of energy metabolism related genes, and suppressing weight gain when administered to an actual animal model. Having confirmed that, the germinated soybean germ extract may be useful for preventing, improving or treating obesity.

1 is a result of confirming a change in fat production by treating germinated soybean embryo extract (described as GSE) by concentration when differentiating adipocytes into adipocytes by concentration.
Figure 2 is a graph showing the results of treating the germinated soybean embryo extract by concentration and staining them when the adipocytes differentiate into adipocytes.
3 is a result of confirming the expression of fat-forming or energy metabolism-related genes after treating germinated soybean embryo extract on fat cells.
4 is a result of confirming the change in weight after administration of germinated soybean embryo extract by concentration to an animal model ingesting a high-fat diet.
5 is a result of confirming the weight of the internal organs after administration of germinated soybean embryo extract by concentration to the animal model ingesting the high-fat diet.
6 is a result of confirming the distribution of white fat by dyeing its adipose tissue after administration of germinated soybean embryo extract by concentration to an animal model ingesting a high-fat diet.
7 is a result of measuring the level of triglycerides in the blood after administration of germinated soybean embryo extract by concentration to an animal model ingesting a high-fat diet.
8 is a result of confirming the expression of genes related to fat production-related genes or energy metabolism by separating mRNA from fat after administering germinated soybean embryo extract by concentration to an animal model ingesting a high-fat diet.

Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

Example 2: Component analysis of germinated soybean germ extract

The content of Soyasaponin, an indicator of germinated soybean germ extract, in the extract obtained by the above method was measured using Agilent Technologies 1200 Series high-speed liquid chromatography. The indicator substance soya saponin was purchased from chengduo and used. For the preparation of the standard solution, the content of the index component was calculated as in Equation 1 below.

First, about 5 to 10 mg, which is the content calculated according to Equation 1 above, is added to the indicator material standard material precisely in 20 mL of water, and then 80 mL of methanol (or 10 mL of water and then 40 mL of methanol) is added. Exactly 100 mL (or 50 mL) was used as the standard solution. The solution was shaken to dissolve and diluted with 80% methanol (or 0.5% HCl in 100% methanol) to make a standard solution. (Example: 10, 50, 100 µg/mL) For the preparation of the test solution, a specific amount of the germinated soybean germ extract is precisely weighed and 80% methanol (or 0.5% HCl in 100% methanol) is added and extracted (if necessary) Ultrasonic extraction), accurately set to 100 mL, and filtered with a 0.45 μm PTFE membrane filter to prepare a test solution. High-speed liquid chromatography analysis was performed under the conditions shown in Table 1 below using the standard solution and the test solution prepared as described above.

Injection volume 2 μl Mobile phase A: Water with 0.1% acetic acid added
B: Acetonitrile with 0.1% acetic acid added flux 0.7 mL/min Detector wavelength nm Mobile phase gradient Hour (minute) A (%) B(%) 0 90 10 2 90 10 5 80 20 15 70 30 20 70 30 25 60 40 30 50 50 40 30 70 50 0 100 50.1 90 10 60 90 10

As a result, as shown in Table 2 below, it was confirmed that a lot of soya saponin, which is an indicator material of germinated soybean germ extract under water and alcoholic solvent extraction conditions, was contained. In particular, the content of soya saponin Ab, which is effective in inhibiting fat production, was the highest at 210.9 mg/g, and the soya saponin Ba and Bb were also 116.7 and 199 mg/g, respectively.

(mg/g) Soya saponin Ab Ba Bb Total content 210.9 116.7 199 538.5

Example 3: Evaluation of germination bean germ extract efficacy for reducing body fat

In order to check the mechanism of inhibiting the production of germinated soybean germ extract, the concentration of germinated soybean germ extract (GSE (germinated soy germ extract)) when differentiating adipocytes 3T3-L1 cells into adipocytes was determined by concentration (0 , 25, 50, 100 or 200 μg/ml). At this time, 0.5 mM isobutyl methyl xanthine, 0.25 μM dexamethasone, 1 μg/ml insulin (isobutylmethylxanthine, dexamethasone, insulin, MDI) was treated for 2 days, and then 1 μg/ml insulin was further treated for 2 days to differentiate into adipocytes. Then, the cells of the experimental group were stained using Oil-Red-O staining to confirm the change in fat production.

As a result, as shown in Figures 1 and 2, it could be confirmed that the germinated soybean germ extract reduces fat production in a concentration-dependent manner. In particular, it was confirmed that at 100 or 200 μg/ml, fat production was reduced to less than half.

Example 4: Confirmation of the gene expression change related to fat formation or energy metabolism according to the treatment of germinated soybean embryo extract

Real-time PCR (Real-time PCR) confirmed the expression of genes related to fat formation and energy metabolism in adipocytes. CDNA was generated from RNA using a reverse transcriptase of REVERTRA Ace qPCR RT Master Mix from TOYOBO, Japan, and real-time PCR was performed using REVERTRA Ace qPCR RT Master Mix from TOYOBO, Japan. Primer sequences used in the experiment are shown in Table 3 below.

designation direction order Sequence number FAS Forward 5'-GGAGGTGGTGATAGCCGGTAT-3' One Reverse 5'-TGGGTAATCCATAGAGCCCAG-3' 2 ADIPOQ Forward 5'-TGTTCCTCTTAATCCTGCCCA-3' 3 Reverse 5'-CCAACCTGCACAAGTTCCCTT-3' 4 PRDM16 Forward 5'-TGCTGACGGATACAGAGGTGT-3' 5 Reverse 5'-CCACGCAGAACTTCTCGCTAC-3' 6 PGC-1α Forward 5'-AAGTGGTGTAGCGACCAATCG-3' 7 Reverse 5'-AATGAGGGCAATCCGTCTTCA-3' 8 CIDEA Forward 5'-ATCACAACTGGCCTGGTTACG-3' 9 Reverse 5'-TACTACCCGGTGTCCATTTCT-3' 10 UCP-1 Forward 5'-AGCCATCTGCATGGGATCAAA-3' 11 Reverse 5'-GGGTCGTCCCTTTCCAAAGTG-3' 12 PPARγ Forward 5'-TCGCTGATGCACTGCCTATG-3' 13 Reverse 5'-GAGAGGTCCACAGAGCTGATT-3' 14 C/EBPα Forward 5'-CAAGAACAGCAACGAGTACCG-3' 15 Reverse 5'-GTCACTGGTCAACTCCAGCAC-3' 16 β-actin Forward 5'-ACGTCGACATCCGCAAAGACCTC-3' 17 Reverse 5'-TGATCTCCTTCTGCATCCGGTCA-3' 18

The significance evaluation of each experiment was statistically processed into two groups of T-test equivariance assumptions, P values of P<0.05 as * and P<0.01 as **.

As a result, as shown in Figure 3, it was confirmed that the expression of FAS, PPARg, C/EBPa, and ADIPOQ, genes related to fat production, is reduced in a concentration-dependent manner. On the other hand, it was confirmed that the expression of the energy metabolism related genes PGC1a, PRDM16, CIDEA and UCP1 is increased.

Therefore, it was confirmed that the germinated soybean germ extract of the present invention exhibits fat production inhibition and energy metabolism activity in a concentration-dependent manner in adipocytes.

Example 5: Confirmation of changes in body fat production according to administration of germinated soybean germ extract in animal models

Germinated soybean germ extract was administered orally to animal models fed a high-fat diet at a concentration of 1, 5, 15 or 45 mg/kg for 10 weeks, and changes in body fat production were confirmed. As a result, as shown in Figure 4, it was confirmed that the increase in weight is reduced in the experimental group ingesting germinated soybean germ extract than in the group not ingesting. In particular, in the high concentration 45mg/kg experimental group, statistically significant reduction was observed compared to the obese experimental group (High Fat diet, HFD) that did not take germinated soybean embryos from the 4th week of the animal experiment. There was a greater difference over time, and in the last 10 weeks, a p-value of 0.01 or less showed a statistically significant weight difference.

Example 6: Confirmation of internal organ weight change according to administration of germinated soybean germ extract in animal model

Germinated soybean germ extract is administered orally to animal models fed a high-fat diet at a concentration of 1, 5, 15 or 45 mg/kg for 10 weeks, and dissected to dissolve the liver (Liver), kidney fat (Perirenal Adipo tissue) and epididymal fat. After confirming the weight of (Peri-epididymal Adipo tissue), it is shown in FIG. 5. As a result, it was confirmed that the weight of kidney fat was reduced in all groups in which germinated soybean germ extract was ingested. In addition, in the experimental group administered with the highest concentration of 45 mg/kg, it was confirmed that the weight of liver, kidney fat, and epididymal fat was significantly decreased compared to the obese experimental group not consuming germinated soybean embryos.

Example 7: Confirmation of white fat distribution according to administration of germinated soybean germ extract in animal model

Germinated soybean germ extract was orally administered to animal models fed a high-fat diet at a concentration of 1, 5, 15 or 45 mg/kg for 10 weeks, and their fat tissue was thinly sliced and stained through Oil-Red-O staining. Through this, the distribution of white fat was confirmed and shown in FIG. It was confirmed that white fat increased in the fat of the obese test group that did not take germinated soybean embryos, but it was almost invisible in the fat of the experimental group that consumed 45 mg/kg germinated soybean germ extract.

Example 8: Confirmation of triglyceride level change according to administration of germinated soybean germ extract in animal model

Germinated soybean germ extract was orally administered to an animal model ingesting a high-fat diet at a concentration of 1, 5, 15 or 45 mg/kg for 10 weeks, and the levels of triglycerides in the blood were measured and shown in FIG. 7. As a result, it was confirmed that the triglyceride (TC) level in the experimental group in which the germinated soybean germ extract was consumed at 45 mg/kg was significantly reduced, and the total cholesterol (cholesterol, TG) level was also significantly decreased. In addition, it was confirmed that the level of high density lipoprotein (HDL) for TC was also increased by the administration of germinated soybean embryo extract compared to the obese test group that did not consume germinated soybean embryo.

Example 9: Confirmation of changes in gene expression related to fat formation or energy metabolism according to administration of germinated soybean embryo extract in animal models

MRNA was isolated from fat of an animal model ingesting a high-fat diet, and expression of fat-generation-related genes was confirmed through real-time PCR.

As a result, as shown in FIG. 8, in the obese test group not consuming germinated soybean embryos, expression of fat-generating genes such as FAS, PPARg, C/EBP/a, and ADIPOQ increased, but in the experimental group consuming germinated soybean germ extract It was confirmed that it was reduced as an enemy. Conversely, the expressions of genes related to energy metabolism, PGC-1a, PRDM16, CIDEA, and UCP1, were decreased in the obese test group not ingesting germinated soybean embryos, but significantly increased in the experimental group ingested germinated soybean embryo extract. there was.

As described above, the germinated soybean germ extract according to the present invention shows the effect of inhibiting differentiation into adipocytes, reduces the expression of genes involved in fat production, increases the expression of energy metabolism related genes, and has been administered to actual animal models. Since it shows the effect of suppressing the weight gain at the time, the germinated soybean germ extract can be useful for preventing, improving or treating obesity.

<110> NOVAREX Co., Ltd. <120> Composition for prevention, improvement or treatment of obesity comprising extract of germinated soy germ <130> NP18-0114 <160> 18 <170> KoPatentIn 3.0 <210> 1 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> FAS forward primer <400> 1 ggaggtggtg atagccggta t 21 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> FAS reverse primer <400> 2 tgggtaatcc atagagccca g 21 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> ADIPOQ forward primer <400> 3 tgttcctctt aatcctgccc a 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> ADIPOQ reverse primer <400> 4 ccaacctgca caagttccct t 21 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PRDM16 forward primer <400> 5 tgctgacgga tacagaggtg t 21 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PRDM16 reverse primer <400> 6 ccacgcagaa cttctcgcta c 21 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PGC-1a forward primer <400> 7 aagtggtgta gcgaccaatc g 21 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PGC-1a reverse primer <400> 8 aatgagggca atccgtcttc a 21 <210> 9 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CIDEA forward primer <400> 9 atcacaactg gcctggttac g 21 <210> 10 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> CIDEA reverse primer <400> 10 tactacccgg tgtccatttc t 21 <210> 11 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> UCP-1 forward primer <400> 11 agccatctgc atgggatcaa a 21 <210> 12 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> UCP-1 reverse primer <400> 12 gggtcgtccc tttccaaagt g 21 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> PPAR gamma forward primer <400> 13 tcgctgatgc actgcctatg 20 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> PPAR gamma revesre primer <400> 14 gagaggtcca cagagctgat t 21 <210> 15 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> C/EBPalpha forward primer <400> 15 caagaacagc aacgagtacc g 21 <210> 16 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> C/EBPalpha reverse primer <400> 16 gtcactggtc aactccagca c 21 <210> 17 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> beta-actin forward primer <400> 17 acgtcgacat ccgcaaagac ctc 23 <210> 18 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> beta-actin reverse primer <400> 18 tgatctcctt ctgcatccgg tca 23

Claims (10)

A pharmaceutical composition for preventing or treating obesity comprising germinated soybean germ extract as an active ingredient.
According to claim 1,
The extract is a pharmaceutical composition, characterized in that the extract by one or more solvents selected from the group consisting of water, an organic solvent having 1 to 6 carbon atoms, subcritical fluid, supercritical fluid and mixtures thereof.
According to claim 2,
The organic solvent having 1 to 6 carbon atoms is alcohol having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride (methylene) chloride), hexane, hexane (cyclohexane) and petroleum ether (petroleum ether) pharmaceutical composition characterized in that it is selected from the group consisting of.
According to claim 1,
Germinated soybean germ is a pharmaceutical composition characterized by obtaining germination separated from soybeans.
According to claim 1,
The germinated soybean germ extract comprises refluxing germinated soybean germ;
Concentrating the extracted extract under reflux under reduced pressure; And
Pharmaceutical composition characterized in that the concentrated extract is obtained through a step of spray drying.
The method of claim 5,
The reflux extraction is a pharmaceutical composition characterized in that carried out for 3 hours or more.
According to claim 1,
The reduced pressure concentration is a pharmaceutical composition, characterized in that carried out at a temperature of 10 ℃ to 70 ℃.
Food composition for preventing or improving obesity comprising germinated soybean germ extract as an active ingredient.
A composition for preventing or improving obesity comprising germinated soybean germ extract as an active ingredient.
The method of claim 9,
The livestock is a composition for livestock feed, characterized in that the cattle, pigs, chickens, ducks, goats, sheep, and horses are one type of livestock selected from the group consisting of.

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