KR20140141871A - Composition comprising germinated soybean peptides for anti-obesity - Google Patents

Abstract

The present invention relates to a composition for preventing and alleviating or treating obesity comprising germinated soybean peptides as active ingredients. According to the present invention, the germinated soybean peptides show excellent effects of inhibiting cell differentiation of 3T3-L1 preadipocyte cells and accumulation of fat in mature fat cells, thereby being effective in preventing, alleviating, or treating obesity. Also, the germinated soybean peptides have very high health safety as natural substances, thereby being highly useful.

Description

The present invention relates to an anti-obesity composition comprising germinated soybean peptides as active ingredients.

The present invention relates to a composition for preventing or ameliorating or treating obesity comprising germinated soybean peptides as an active ingredient.

Obesity is a chronic metabolic disease caused by excessive accumulation of fat in the body due to the accumulation and consumption imbalance of energy, and it is known that it is highly likely to cause various complications such as diabetes, hypertension, myocardial infarction, cancer, arthritis and stroke. According to the National Health and Nutrition Survey, the obesity rate was 32.7% in 2001, 34.9% in 2005, and 19.5% in 2001 and 20.3% in 2005, As the intake of high-calorie foods increases, the obesity population is increasing.

Various methods such as diet, exercise, behavior modification therapy, and drug and surgery have been developed and tried to overcome obesity. Among them, Xenical (Roche Korea) and Reductil (Eixin New Drug), which are approved by the US FDA and marketed domestically and overseas, are the representative drug therapies. In addition, non-prescription medicines developed in France include green tea catechol (Exxon Pharmaceuticals Co., Ltd.). However, Xenical causes gastrointestinal side effects such as lowering of fat, gas production, and lowering of fat-soluble vitamin. In the case of reductil, it causes side effects such as headache, anorexia, insomnia or constipation by increasing the concentration of serotonin and noradrenaline in the sympathetic nervous system There is a limit to the treatment of obesity without side effects. In addition, aminophylline, known to have a lipid-lowering effect, is known to exhibit a wide range of side effects from the nervous system, circulatory system, and digestive system. Penfluramine, dexfenfluramine, topiramate, ephedrine and isoproterenol, And it is currently prohibited from being sold by the Korea Food and Drug Administration.

Due to the above-mentioned side effects due to the pharmacotherapy, there is a growing interest in anti-obesity natural substances using food materials. For example, mulberry leaves that lower the absorption rate of fats and sugars, fermented red peppers that inhibit pancreatic lipase activity, and foods that enhance fat burning are being marketed as dietary health foods. In addition, foods such as brown rice, cereals, herbs and dairy products have been reported to have obesity inhibitory activity (Jin-Nyoung Ho, Mi-Eun Son, Won-chul Lim, Seung Taik Lim, Antiobesity effects of germinated brown rice extracts on down-regulation of lipogenic genes in high fat diet-induced obese mice, Biosci. Biotechnol. Biochem., 76 (6), 1068-1074, 2012; Kristel Diepvens, Klaas R. Westerterp, and Margriet S. Westerterp-Plantenga, Obesity and thermogenesis related to caffeine, ephedrine, capsaicin, and green tea, Am. J. Physiol. Regul. Integr. Comp. Physiol. 292: 77-85, 2007).

Accordingly, it is an object of the present invention to provide a composition for the prevention, improvement and treatment of obesity, which includes natural plant material having pharmacological and physiological activity on obesity and no side effects on the human body.

As a means for solving the above problems, the present invention provides a composition for preventing, improving and treating obesity comprising germinated soybean peptide as an active ingredient.

The soybean consists of 35-40% of protein, 15-20% of fat, 35% of carbohydrate and 5% of other fat, and isoflavone, oligosaccharide, saponin, lecithin, , Lutein, and unsaturated fatty acids, fibrin, vitamins and minerals. In particular, unsaturated fatty acids contain linoleic acid (linoleic acid) 52% and linolenic acid (11%) in large amounts and are known as excellent sources of essential fatty acids. In the germination process, nutrient changes such as qualitative changes of storage proteins, increase of micronutrients, increase of digestibility of constitutional carbohydrates in the body and decrease of the reflecting quantitative factors are carried out in the germination process. In the germination process, It is known that the water absorption is improved and the permeability is improved. Thus, researches on the application to various functional food materials have been actively conducted. However, germinated soy peptides have never been used as active ingredients until now.

Meanwhile, the inventors of the present invention confirmed that the germinated soybean peptides exhibit the proliferation inhibitory activity of lipid precursor cells and the inhibitory activity of lipid accumulation in mature adipocytes in the process of extracting substances having pharmacological and physiological activities of obesity from natural products, . As can be seen in the following examples, the germinated group has superior inhibitory effect on lipid progenitor cell proliferation and lipid accumulation in matured adipocytes, and is superior to general germinated soybean extract (germinated soybean peptide) Suggesting that the proliferation inhibitory effect of lipid precursor cells and the fat accumulation inhibitory effect in mature adipocytes are superior.

In the present invention, the germinating soybean peptide comprises (a) germinating soybeans; (b) obtaining a germinated soybean extract from the germinated soybean; And (c) separating the germinated soybean peptides from the germinated soybean extract (see FIG. 1).

The method of germinating the soybeans in step (a) may be according to a conventional method. However, the present invention is not limited to this, but the soybeans are immersed in water in a state where the temperature and humidity are maintained. Preferably, the immersion is carried out by dipping in water at a temperature of 18 to 35 캜 for 24 to 72 hours, sprinkling gently in a dark state in which the temperature is maintained at 20 to 35 캜 and the relative humidity is maintained at 50 to 100%. The intermittent spraying may be done by spraying at intervals of 4 to 6 hours for 1 to 10 minutes, and cultivation is convenient when a device equipped with a timer is used. The germination is preferably germinated until the bud of the bean grows to 1 to 3 cm. The soybean is applicable to any seed of a plant belonging to the soybean family.

The method of obtaining the germinated soybean extract of step (b) may be according to a conventional method. But it is not limited thereto, and can be obtained by pulverizing the germinated soybean after drying and heat treatment, and immersing it in water or an organic solvent. Preferably, the germinated soybean extract may be a hot-water extract.

The method of obtaining the germinated soybean peptide of step (c) may be according to a conventional method. But is not limited to, can be obtained by treating with ordinary proteolytic enzymes. As can be seen from the examples, the germinated soybean peptides obtained by treating the supernatant obtained by centrifuging the germinated soybean extract of step (b) above with proteolytic enzymes were obtained from germinated soybean peptides obtained by centrifuging after proteolytic enzyme treatment The proliferation inhibitory effect of lipid precursor cells and the fat accumulation inhibitory effect in mature adipocytes were better. Therefore, it is preferable to prepare by treating the proteolytic enzyme after centrifugation, drying with hot air and pulverizing.

The term 'obesity' in the present invention refers to a state in which fat is excessively accumulated, and in particular, a state in which a white fat tissue, which plays a role of storing in the form of triglyceride as a result of excessive energy absorption My fat accumulation type refers to obesity.

The type of obesity can be classified according to the cause, the number and size of fat cells, and the area. Especially, according to the number and size of adipocytes, it can be divided into fat-growing obesity and fat-accumulation-type obesity. Fat-proliferative obesity is an obesity that increases the number of fat cells by congenital factors. Is a form in which adipose cells are filled with fat due to acquired factors and the size of adipocytes is increased.

The adipocytes play a central role in the synthesis, storage and degradation of energy in the form of triglycerides (TG). That is, when the amount of calories stored in the form of triglyceride in the case of excessive calories is large, it is decomposed and consumed as energy. Thus, many different enzymes and regulatory factors involved in lipogenesis and lipolysis are required for fat cells to maintain energy balance. That is, obesity is caused by a failure to maintain balance in the body by the above-mentioned enzymes, which causes excessive accumulation of adipose tissue.

The adipose tissue can be classified into white adipose tissue (WAT) and brown adipose tissue (BAT) according to anatomical location, morphological structure, function and regulation. These two adipose tissues have a common tendency to store energy in the form of triglycerides, but in the case of white fats, this energy is released as needed, and in the case of brown fats, this energy source is converted into heat. Brown fat is distributed in many blood vessels compared to white fat, and it is brown due to cytochrome pigment, and it plays an important role especially in hibernating hibernating animals and controlling the temperature of newly born mammals. In contrast, white fat plays a pivotal role in maintaining energy homeostasis in the body. Specifically, it stores energy in the form of triglycerides when the amount of energy absorption is greater than consumption, and controls the use of energy sources in case of lack of energy I am responsible. Thus, the distribution and function of the two adipocytes are different, and white adipocytes, commonly referred to as adipocytes, are distributed predominantly in the subcutaneous tissues and organs and are more closely related to obesity as they function as reservoirs of fuel.

The term "mature adipocyte" as used herein means a form of complete adipocyte produced in the differentiation process of adipocyte. Specifically, the term " mature adipocyte " means that the stem cell is preadipocytes, It is produced through a process in which precursor cells mature into complete adipocytes.

The term "lipid precursor cell" in the present invention means a cell having the ability to differentiate into adipocytes, which is first produced from stem cells during the differentiation stage of adipocytes. When lipid precursors grow to 100% confluency Growth is stopped and differentiation into adipocytes progresses slowly. The process by which lipid precursor cells are differentiated into adipocytes is a phenomenon caused by differentiation inducers in the serum added to the culture medium. Therefore, the addition of differentiation-promoting factors (insulin, dexamethasone, 3-isobutyl-1-methylxanthine) to the culture medium can further promote differentiation of cells. On the other hand, differentiation inhibitors (actinomycin D, tumor necrosis factor α, bromo deoxyuridine, etc.) may be added to inhibit differentiation. Finally, the differentiated lipid precursor cells become mature adipocytes such as accumulation of intracellular fat, and the activation of enzymes inducing adipocyte-specific gene expression and fat accumulation occurs. As a representative example of promoting adipocyte differentiation, insulin-like growth factor-I (IGF-I) and the like have been known so far (Hwang, Hyejung, et al., Effect of alginate on differentiation of 3T3- , Journal of the Korean Fisheries Society 33 (6), 541-545, 2000).

As one embodiment of the present invention, the present invention provides a food composition for improving obesity comprising germinated soybean peptides as an active ingredient.

The term " food composition " in the present invention refers to a food prepared by adding the germinated soybean peptide to food materials such as beverage, tea, spices, gum and confectionery, or encapsulated, powdered or suspended, Unlike general medicines, it has the advantage that it does not have side effects that can occur when a drug is taken for a long time by using the food as a raw material.

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

Since the food composition according to the present invention thus obtained can be routinely ingested, a high anti-obesity effect can be expected, which is very useful.

When the germinated soybean peptide is used as a food additive, the germinated soybean peptide may be directly added or used together with other food or other food ingredients, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed may be within a range that does not impair the original taste of the food, and is usually in the range of 0.01 to 50 parts by weight, preferably 0.1 to 20 parts by weight, based on 100 parts by weight of the whole composition. In the case of foods in the form of granules, tablets or capsules, they may be added in an amount of usually 0.1 to 100 parts by weight, preferably 0.5 to 80 parts by weight. However, in the case of long-term intake intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

In addition, the food composition according to the present invention may further contain various flavors or natural carbohydrates. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau Martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. In addition to the above, the food composition according to the present invention may further contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the food composition of the present invention.

According to another embodiment of the present invention, there is provided a pharmaceutical composition for improving obesity comprising germinated soybean peptides as an active ingredient.

The sprout peptides according to the present invention may be in the form of pharmaceutically acceptable salts.

The salt may be an acid addition salt formed using an organic acid or an inorganic acid. Examples of the organic acid include formic acid, acetic acid, propionic acid, lactic acid, butyric acid, isobutyric acid, trifluoroacetic acid, malic acid, maleic acid, malonic acid, But are not limited to, lactic acid, succinic acid, succinic monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, salicylic acid, anthranilic acid, dichloroacetic acid, And inorganic acids include, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid-based salts. Preferably in the hydrochloride or acetate form, more preferably in the hydrochloride form.

Separately, additionally saltable forms include, but are not limited to, the salts of gabapentin, gabapentin, pregabalin, nicotinate, adipate, hemimarate, cysteine, acetylcysteine, methionine, arginine, lysine, Aspartate and the like.

The pharmaceutical composition according to the present invention may further comprise a pharmaceutically acceptable carrier. The carrier may contain various components such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salt and polysorbate.

The content of the germinating soybean peptides in the composition may be appropriately adjusted according to the symptoms of the disease, the progress of the symptoms, the condition of the patient, the administration method, the form of the preparation, etc. For example, Preferably 0.001 to 1 part by weight, based on the weight of the composition. The content ratio is a value based on the dried amount from which the solvent is removed.

The pharmaceutical composition according to the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, , Extenders, binders, wetting agents, disintegrating agents, and surfactants.

Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like. Such solid preparations can be prepared by adding to the pharmaceutical composition of the present invention at least one or more excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like can be prepared.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of liquid formulations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are commonly used simple diluents.

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

The germinated soybean peptides according to the present invention show excellent effects on inhibition of cell differentiation and inhibition of fat accumulation in mature adipocytes of 3T3-L1 adipocyte precursor cell line, thus being effective for prevention, improvement or treatment of obesity, As the safety of human body is very high, the utilization value is very high.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing a process for producing an unrooted soybean peptide. FIG.
FIG. 2 is a graph showing the inhibitory activity of germinating soybean peptides on lipid progenitor cell proliferation.
3 is a chart showing the fat accumulation inhibitory activity of germinated soybean peptides in mature adipocytes.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

[Production Example 1]

Process for the production of germinated peptides or germinated peptides

The process of manufacturing germinated peptides adds to the germination process of the beans. The above germination was carried out in a germ capable of intermittently spraying at an interval of about 1 hour after immersing in water at 25 캜 for 4 hours at a temperature of 25 캜 and a light-shielded environment at 25 캜. The germination was germinated until the bud of the bean grew to 1 cm or more, and it took about 2-3 days.

The germinated soybean peptides and germinated soybean peptides are prepared as follows. The germinated soybeans and germinated soybeans were immersed in water for 4 hours, respectively, and then they were removed and melted. Thereafter, about 10 times as much water as the weight of the soybeans removed and melted was added, and the mixture was heated to 100 캜 for 15 minutes. Thereafter, the mixture was pulverized at 7,000 rpm for 3 minutes using Turrex, and further heated to 100 ° C for 15 minutes to obtain undiluted soybean extract (SM) and germinated soybean extract (GSM).

In the step of obtaining peptides from the germinated soybean extract, the germinated soybean extracts were treated with proteolytic enzymes (BSM and BGSM, respectively) after centrifugation, and the germinated (germinated) soybean extract (BWSM, BWGSM, respectively) were separated by proteolytic enzymes. The proteolytic enzyme is hydrolyzed by treatment with 0.2% (w / v) of proteolytic enzyme for 2, 4 and 6 hours. The centrifugation is carried out by centrifuging at 10,000 rpm for 15 minutes and then taking the supernatant. The supernatant was obtained by centrifuging the above germinated soybean extracts (SM, GSM, respectively) after proteolytic enzyme treatment, and the germinated soybean extract (SM, GSM, respectively) The enzyme was treated for 10 minutes at 100 ° C to inactivate the proteolytic enzyme. After centrifugation at 10,000 rpm for 15 minutes, the supernatant was taken out and dried in hot air at 70 ° C. and pulverized to prepare germinated soybean peptides (BSM and BGSM, respectively) and germinated soybean peptides (BWSM and BWGSM, respectively).

[Example 1]

Inhibition of lipoprotein proliferation of germinated soybean peptides

3T3-L1 (mouse embryo fibroblast cell line) was cultured in a 96 well plate at a concentration of 1 × 10 ⁴ cells / well for 24 hours. Then, the original medium was removed and 10% calf serum (CS) The germinated soybean peptides and germinated soybean peptides were added at 1 mg / ml and 100 μl respectively to the DMEM medium. 100 μl of distilled water was added to the control, 1 μg / ml of caffeine was added to the positive control, 100 μl of the caffeine was added, and the cells were incubated at 37 ° C. for 48 hours in a CO ₂ incubator. After 48 hours, the original medium was removed and 100 μl of 10% MTT solution (5 mg / ml) was added to the DMEM medium. The reaction was carried out at 37 ° C. for 4 hours in a CO ₂ incubator. Progenitor cell proliferation was measured. The inhibitory effect of lipid proliferation was determined by the following equation.

Cell viability (%) = {(Sample OD - Blank OD) / (Control OD - Blank OD)} 100

As shown in FIG. 2, when the proliferative potential of the lipoprotein of the control was 100%, the positive control caffeine was 77%, the germinated soybean extract (SM) was 105% The percentage of germinated soybean peptides (BGSM) was 102%, 98%, and 79%, respectively, for the extracts (GSM) and the germinated soybean peptides (BSM) treated with protease after centrifugation were 96%, 91% (BWSM) were 96%, 91% and 86%, respectively. The percentage of germinated soybean peptides (BWSM) was 108%, 105% and 94%, respectively.

From the above results, it can be confirmed that the proteolytic enzyme treatment after centrifugation shows better inhibition of lipid progenitor cell proliferation (BSM, BGSM). In addition, the inhibition of proliferative proliferation of the progenitor cells was improved by increasing the protease digestion time, and proteolytic enzyme treatment for 6 hours showed inhibition of proliferation similar to that of caffeine, which is a positive control.

As the time for treatment with proteolytic enzymes increases, the inhibition of proliferative proliferation of the progenitor cells means that the peptide content is increased. This means that the germination (germination) soybean peptide of the general germination Which means that the cell proliferation inhibiting ability is excellent.

[Example 2]

Inhibition of mature fat cell fat accumulation in germinated soybean peptides

3T3-L1 (mouse embryo fibroblast cell line) was cultured in a 6 well plate at a concentration of 2 × 10 ⁵ cells / well. After reaching 100% confluency, the cells were further cultured for 2 days and then differentiated into mature adipocytes The original medium was removed and incubated with 0.5 mM 3-isobutyl-1-methylxanthine, IBMX, 0.1 uM dexamethasone, DEX, 1 ug / ml insulin and 10% fetal bovine serum 1 mg / ml and 100 μl of germinated soybean peptide and germinated soybean peptide were added to DMEM medium supplemented with fetal bovine serum (FBS), and 100 μl of distilled water was added to the control, 1 mg / ml of caffeine was added to the positive control and the cells were cultured in a CO ₂ incubator at 37 ° C for 48 hours. After 48 hours, the culture medium was removed again, and DMEM medium supplemented with 10% fetal bovine serum (FBS) was incubated at 37 ° C for 48 hours in a CO ₂ incubator. After the culture solution was completely removed, a phosphate buffer (phosphate buffer, PBS), and the cells were fixed by adding 400 μl / well of 10% formalin solution for 1 hour. After that, a phosphate buffer (phosphate buffer, PBS), and oil red O working solution was added per 400 μl / well to stain fat in the adipocyte differentiated for 2 hours. The oil red O working solution was removed, and the oil red O working solution on the wall of the well was completely washed using secondary distilled water, and then dried in a drier for 5 minutes and 500 μl of isopropyl alcohol was added to the well . The lipid-accumulating ability was measured by absorbance at 490 nm using a microplate reader (Model 680 microplate reader, Bio-Rad, USA). Fat accumulation ability was obtained by the following equation.

Lipid accumulation (%) = {(Sample OD - Blank OD) / (Control OD - Blank OD)} 100

As shown in FIG. 3, the fat accumulation capacity of caffeine as a positive control was 68%, that of non-germinated soybean extract (SM) was 98% as compared with that of the control (control) , 92% of germinated soybean extract (GSM), 95%, 92% and 86% of germinated soybean peptides (BSM) treated with proteolytic enzyme after centrifugation, 99% of germinated soybean peptides (BGSM) 98%, 94% and 87% of the germinated soybean peptides (BWSM) and 95%, 89% and 76% of the germinated soybean peptides (BWGSM)

From the above results, it can be confirmed that the treatment with proteolytic enzyme after centrifugation shows the best mature fat cell fat accumulation inhibitory ability. In addition, the inhibition of lipid accumulation of mature adipocytes was increased with increasing protease treatment time, and the treatment with proteolytic enzyme for 6 hours showed that lipid accumulation of mature adipocytes was similar to that of caffeine, which is a positive control group.

Increased inhibition of lipid accumulation in mature adipocytes with increasing proteolytic enzyme treatment time means that the peptide content is increased. This means that the germination (germination) of soybean peptides Which means that the fat cell fat accumulation inhibitory ability is excellent.

Claims (6)

A composition for improving obesity comprising germinated soybean peptides as an active ingredient.
The method according to claim 1,
Wherein the germinated soybean peptides are contained in an amount of 0.01 to 50 parts by weight based on 100 parts by weight of the total food composition.
The method according to claim 1,
Wherein said obesity is fat accumulation type obesity.
A pharmaceutical composition for preventing and treating obesity comprising germinated soybean peptides as an active ingredient.
5. The method of claim 4,
Wherein the germinated soybean peptides are contained in an amount of 0.0001 to 10 parts by weight based on 100 parts by weight of the total pharmaceutical composition.
5. The method of claim 4,
Wherein said obesity is obesity which is fat accumulation type obesity.

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