KR20150130128A - Composition for anti-obesity comprising extract from young barley leaves - Google Patents

Abstract

The present invention relates to a composition for inhibiting adipocyte differentiation, containing lutonarin or saponarin; a method for inhibiting adipocyte differentiation using the composition; a pharmaceutical composition for preventing or treating obesity, comprising an extract of barley sprouts containing lutonarin or saponarin or an fraction of the extract; a food composition for preventing or alleviating obesity; and a pharmaceutical composition and a food composition for preventing or treating obesity, containing lutonarin, saponarin, or a pharmaceutically acceptable salt thereof. By using the composition of the present invention, adipocyte differentiation, which is the cause of obesity, is inhibited so the level of cholesterol contained in the generated adipocytes as well as the number of adipocytes can be reduced, thereby being able to be widely used in effective treatment for obesity.

Description

TECHNICAL FIELD The present invention relates to a composition for inhibiting obesity,

More particularly, the present invention relates to a composition for inhibiting the differentiation of adipocyte comprising rutonin or saponin, a composition for suppressing differentiation into adipocytes using the composition, , A pharmaceutical composition for the prevention or treatment of obesity comprising rosemaryin or a saponin-containing shoot barley extract or a fraction thereof, a food composition for the prevention or amelioration of obesity, rutonin, saponin, or a pharmaceutically acceptable salt thereof The present invention relates to a pharmaceutical composition and a food composition for preventing or treating obesity.

Generally, obesity generally means that the fat tissue is excessively present in the body, and the energy consumed by the food is not balanced with the energy consumed by the physical activity, so that the surplus energy accumulates in the body fat. If the body fat is abnormally increased due to energy imbalance over a long period of time, diabetic, hyperlipidemia, heart disease, stroke, atherosclerosis, fatty liver and various metabolic diseases such as fatty liver and adult diseases are induced. This is a serious social problem in Korea as well as in the West .

There is a multidisciplinary study on the development of obesity drugs worldwide. Drugs for the treatment of obesity can be broadly divided into the inhibition of fat absorption, the promotion of fat decomposition and heat generation, the regulation of appetite and satiety, the inhibition of protein metabolism, and the emotional regulation associated with the ingestion of food. Representative anti-obesity drugs include Xenical ™, which inhibits fat absorption by using orlistat, and Reductil ™, which stimulates the sympathetic nervous system by using sibutramine as the main ingredient . However, Xenical ™ has been reported to have adverse effects such as nausea, abdominal pain, vomiting, itching, and liver damage. Reductin ™ has serious side effects such as headache, anorexia, insomnia and constipation as well as serious cardiovascular side effects There has been controversy such as the recent use standard being strengthened.

In addition to pharmacotherapy through these obesity drugs, there are also dietary therapies to limit the intake of foods and exercises to increase energy consumption, psychotherapy, behavioral therapy, and surgical therapy.

The preferred method of treating obesity is to promote energy consumption through exercise and to treat obesity drugs with few side effects as the most safe and effective method. However, in the case of drugs for the treatment of obesity, serious side effects such as the above-mentioned examples of Xenical ™ and Reductil ™ have been reported and no clear trust in safety has been supported. Therefore, there is a demand for the development of a substance which shows excellent efficacy of anti-obesity against the human body while ensuring safety.

On the other hand, barley leaves have long been used as medicines and have been known to be rich in nutrients such as vitamins, enzymes and chlorophyll. In recent years, interest in health functional raw material crops has been increasing, and it has been appreciated for livestock feeds and the like, as the effect of excellent nutritional ingredients contained in barley, a balanced diet of health functional substances, and improvement of constitution are known.

A barley is a young plant that grows about 15cm in length and has been sprinkled on a barley for about a day. The barley is usually called water for a day, then drained and laid on a suitable size plastic baskets or styrofoam newspapers, It is covered with paper until it is blossomed, and if it shoots, it is removed after 7-10 days when it is removed and the moisture is retained. It is known that the germinated barley contains vegetable fiber, carotene, vitamin C and the like in a much higher content than other vegetables and fruits. In addition, from the viewpoint of pharmacological activity, it is known that the above-mentioned shoot barley has effects of cholesterol lowering, prevention of constipation, prevention of arteriosclerosis, antioxidant activity, prevention of aging, prevention of osteoporosis, improvement of insomnia and prevention of anemia Antioxidant, anticancer, anti-diabetic and whitening effect, Global Food and Food Culture, 2010; Korean Patent Registration No. 1174497; Korean Patent Publication No. 2013-0051181). In particular, it is known that rutheorin contained in the bud of barley has remarkably superior antioxidant activity than vitamin C and genistein, which are known as antioxidants, and rhetherin is known to exhibit superior antidiabetic activity than acabos, which is a treatment for diabetes, In particular, since it is known that barley barley has a cholesterol-lowering effect, it is expected to be used to treat obesity, but it is not yet known whether barley barley is effective for obesity treatment.

Under these circumstances, the inventors of the present invention have conducted extensive research to confirm whether or not obesity can be prevented or treated by using the bud barley. As a result, it has been confirmed that rutonaline and saponin derived from the bud barley inhibit the differentiation into adipocytes, It has been confirmed that the germanium barley extract or its fraction containing rutonaline or saponin can be used as an active ingredient of a composition for the prevention or treatment of obesity and the present invention has been completed.

One object of the present invention is to provide a composition for inhibiting the differentiation into adipocytes comprising lutonin or saponin.

Another object of the present invention is to provide a method for inhibiting differentiation into adipocytes using the composition for inhibiting differentiation into adipocytes.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating obesity, which comprises a budurox extract of Bud or a fraction thereof containing losartan or saponin.

It is another object of the present invention to provide a food composition for preventing or ameliorating obesity, which comprises a budorberry extract or a fraction thereof containing lutonin or saponin.

It is another object of the present invention to provide a pharmaceutical composition for preventing or treating obesity comprising lutonin or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a food composition for preventing or ameliorating obesity comprising lutonin or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating obesity comprising saponin or a pharmaceutically acceptable salt thereof.

It is still another object of the present invention to provide a food composition for preventing or ameliorating obesity comprising saponin or a pharmaceutically acceptable salt thereof.

The inventors of the present invention found that the effect of shoot barley extract on the differentiation into adipocytes was inhibited by inhibiting the differentiation of adipocytes into the adipocyte precursor cells, Respectively. In order to identify the components exhibiting the effect of inhibiting lipid differentiation contained in the above-mentioned germanium barley extract, 10 compounds were isolated from the above-mentioned germanium barley extract, and among them, rutonaline and saponin On the differentiation of adipocytes. First, it was confirmed that rutonaline and saponin were treated to lipid precursors and induced differentiation into adipocytes. As a result, it was confirmed that adipocyte differentiation was effected in a concentration-dependent manner, And the cytotoxicity of saponin. As a result of evaluating the effect of various kinds of factors on the expression level of various factors using the saponin, the expression of the adipocyte differentiation marker was suppressed, the expression of the enzyme involved in lipogenesis was inhibited, It was confirmed that the expression of hydrolyzing enzyme can be increased and the content of cholesterol contained in adipocytes can be reduced.

As described above, the saponin and the lutonin contained in the barley extract of the buds showed the effect of inhibiting the differentiation into the adipocyte, so that the saponin or the lutonaline as well as the saponin or the lutonaline, It was found that the fraction could be used as an effective ingredient of a composition showing the effect of inhibiting or treating obesity by inhibiting differentiation into adipocytes.

As one embodiment for achieving the above object, the present invention provides a composition for inhibiting the differentiation into adipocytes comprising rutonin or saponin.

The term " lutonarin " of the present invention means a compound derived from bud of barley and having a molecular weight of 610.52 and represented by the following formula (1). The rutonaline may be isolated from a natural source such as barley barley, but is not limited thereto, and may be chemically synthesized by a method known in the art, or a commercially available material may be used.

The term "saponarin " of the present invention means a compound of flavone glucoside, which has a molecular weight of 594.52 derived from bud of barley and is represented by the following formula (2). The saponin can be separated from a natural source such as barley barley, but is not limited thereto, and may be chemically synthesized by a method known in the art, or a commercially available material may be used.

Said rutonarin or saponin can be isolated and purified from the extract of rosemary barley or its fraction, so that the composition can be a rosemary extract or its fraction containing roto-marin or saponin.

The term "sprout barley" of the present invention means a young plant that has grown on barley to about 15 cm in length. Normally, the bud barley can be obtained by dipping the barley in water for one day, then removing water and germinating under the condition of preventing loss of water, and then growing for 7 to 10 days.

In the present invention, the above-mentioned germanium barley contains the aforementioned lutonin or saponin which has an effect of inhibiting differentiation into adipocytes, and the extract of the germanium barley or a fraction thereof can be used to inhibit the differentiation into adipocytes.

The term "extract " of the present invention means a liquid substance obtained by immersing a desired substance in various solvents and then extracting the substance at room temperature or a constant temperature for a certain period of time, a solid matter obtained by removing the solvent from the liquid substance, it means. In addition, in addition to the above-mentioned results, the present invention can be broadly interpreted as including the diluted solution, the concentrate thereof, the controlled preparation thereof, and the purified product thereof.

In the present invention, the above-mentioned extract means a barley extract. The above-mentioned germanium barley extract can be obtained by extracting the entire plant of the above-mentioned germanium bar, the dried product thereof, the processed product thereof, etc., alone or in combination thereof, with water or various organic solvents. Here, the organic solvent to be used is not particularly limited as long as it can obtain an extract containing lutonin or saponin which exhibits an effect of inhibiting differentiation into adipocytes, but preferably water, a polar solvent or a nonpolar solvent And more preferably water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol, etc.), a mixed solvent thereof, etc. Most preferably, ethanol or a mixed solvent thereof can be used have. Also, the method for obtaining the above extract is not particularly limited as long as it is possible to obtain an extract containing rutonaline or saponin which has an effect of inhibiting differentiation into adipocytes, but preferably, A freeze extraction method in which the whole, a dried product thereof, a workpiece and the like are immersed in the solvent and extracted at a room temperature of 10 to 25 DEG C, a heat extraction method in which heating is performed at 40 to 100 DEG C, an ultrasonic extraction method in which ultrasonic waves are applied, And a reflux extraction method.

The term "fraction " of the present invention means a product obtained by a fractionation method for separating a specific component or a specific group from a mixture containing various constituents.

In the present invention, the fraction may be interpreted as a fraction containing lutonin or saponin, obtained by applying the above-mentioned germanium barley extract to various fractionation methods. The fraction of the extract can be obtained by applying the extract to various fractionation methods. The fractionation method is not particularly limited, but may be a solvent fractionation method which is carried out by treating various solvents, an ultrafiltration membrane having a constant molecular weight cut- , A chromatographic fractionation method in which various chromatographies (which are prepared for separation according to size, charge, hydrophobicity or affinity) are performed, and the like. In particular, the solvent used in the solvent fractionation method is not particularly limited, but a polar solvent or a nonpolar solvent can be used, and a nonpolar solvent can be preferably used. The solvent fractionation method may be carried out by sequentially fractionating the extract from a solvent having a high non-polarity level by using a low solvent. For example, a method of sequentially fractionating the extract using hexane or ethyl acetate may be used .

The extract or fraction thereof of the barley barley contains various compounds other than rutonin or saponin. Examples of the compound contained in the extract of the bud or the fraction thereof include 3-O-Feruloylquinic acid shown below; 3-FQA (Formula 3), Isoorientin (Formula 4), Luteolin-8-glucoside; 7-O- [6-feruloyl] -glucoside (8), Isovitexin-7-O- [6-sinapoyl] -glucoside (7), Isoscoparin- 7-O- [6-cinapoyl] -glucoside (9), and Isovitexin-7-O- [6-feryloyl] -glucoside (10).

According to one embodiment of the present invention, ten compounds were isolated and identified from the 80% ethanol extract of the buds of barley (Fig. 1 and Table 1), and the two compounds, lutonin and saponin, The effects on the differentiation of prisoners were analyzed. As a result, lutonin or saponin may have the effect of reducing the fat content in adipocytes differentiated in a concentration-dependent manner (Figs. 2 (a) to 2 (c)), (Figs. 3A to 3C). In addition, saponin or lutonaline decreased the expression levels of genes encoding aP2 and PPARgamma, which are adipocyte differentiation markers in a concentration-dependent manner (Figs. 4A and 4B), and proteins GLUT4, SREBP-1c, FAS And ACC (Fig. 5A to 5H), the levels of expression of genes encoding ATGL and HSL among the proteins involved in the hydrolysis of fat are decreased, and the expression level of the gene encoding adiponectin is decreased And the expression level of the gene coding for MGL was not greatly changed, but it was confirmed that it showed a slightly increased pattern (FIGS. 6A to 6H), and it was confirmed that the content of cholesterol contained in adipocytes was decreased 7).

Therefore, it was found that saponin and rutonarin derived from bud from barley extract inhibited differentiation into adipocytes.

As another embodiment for achieving the above object, the present invention provides a method for inhibiting differentiation into adipocytes, comprising the step of treating the composition for inhibiting differentiation into adipocytes into stem cell capable of differentiating into adipocyte precursor cells or fat .

As described above, a budorubic extract or a fraction thereof containing rutonaline or saponin can inhibit differentiation into adipocytes. Therefore, a composition comprising the roots or roots of the budonel extract or the fraction thereof containing saponin, When treated with stem cells capable of differentiating into fat precursor cells or fat, the differentiation into adipocytes can be inhibited.

As another embodiment for achieving the above object, the present invention provides a pharmaceutical composition for preventing or treating obesity, comprising a budorberry extract or its fraction containing roto-marin or saponin.

The term "obesity" of the present invention is generally referred to as " obesity " when an excess amount of adipose tissue is present in the body or when the body mass index (body mass index, body weight index divided by the square of height . In individuals with obesity symptoms, the fatty acids and glucose that enter the adipocytes from plasma are usually esterified and accumulate in the form of triglycerides. For the purpose of the present invention, the obesity symptom is used as a target disease which can be prevented, alleviated, ameliorated or treated by administration of the composition of the present invention, but is not particularly limited thereto.

As described above, the extract or fraction thereof of rosemary or saponin-containing rosemary, which is provided in the present invention, does not exhibit toxicity to adipocytes, but inhibits differentiation into adipocytes and is involved in the synthesis of fat Inhibits the expression of the enzyme, improves the expression of the enzyme involved in the hydrolysis of the fat, and decreases the content of the cholesterol contained in the adipocyte, thereby suppressing or treating the obesity by inhibiting the differentiation into the adipocyte May be used as an active ingredient of the composition.

Meanwhile, the pharmaceutical composition for preventing or treating obesity of the present invention may further comprise a suitable carrier, excipient or diluent conventionally used in the production of a pharmaceutical composition. Specifically, the pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method . In the present invention, the carrier, excipient and diluent which may be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The content of the extract or fraction thereof in the pharmaceutical composition of the present invention is not particularly limited but may be in the range of 0.0001 to 50% by weight, more preferably 0.01 to 20% by weight, based on the total weight of the final composition .

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount " of the present invention means a therapeutic or prophylactic treatment of a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention And the effective dose level refers to the level of the disease to be treated, the severity of the disease, the activity of the drug, the age, body weight, health, sex, sensitivity of the patient to the drug, Duration, duration of administration, factors involved in combination with or contemporaneously with the composition of the present invention, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with another therapeutic agent, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered singly or multiply. It is important to take into account all of the above factors and administer an amount that will achieve the maximum effect in the least amount without side effects.

The dosage of the pharmaceutical composition of the present invention can be administered, for example, to a mammal including a human in an amount of 1 to 20 mg / kg, more preferably 1 to 10 mg / kg, for a day, , The frequency of administration of the composition of the present invention is not particularly limited, but it may be administered once a day or divided into several doses.

In another aspect of the present invention, the present invention provides a method for preventing or treating obesity, which comprises administering the pharmaceutical composition to a subject having or likely to have obesity in a pharmaceutically effective amount do.

As described above, the extract or fraction thereof of the present invention can be used as an active ingredient of a pharmaceutical composition for preventing or treating obesity, so that the composition can be used for preventing or treating obesity.

The term "individual" as used herein includes, without limitation, mammals, including rats, livestock, humans, etc., who are at risk for developing obesity.

In the method of treating obesity of the present invention, the route of administration of the pharmaceutical composition may be administered through any conventional route as long as it can reach the target tissue. The pharmaceutical composition of the present invention may be administered intraperitoneally, intravenously, intramuscularly, subcutaneously, intradermally, orally, intranasally, intracorporally, or rectally, as desired, though not particularly limited thereto. However, the oral composition should be formulated so as to protect the active drug from being coated or decomposed on the top, since the extract or fraction thereof of the germinated barley may be denatured by gastric acid upon oral administration. In addition, the composition may be administered by any device capable of transferring the active agent to the target cell.

As another embodiment for achieving the above object, the present invention provides a food composition for preventing or improving obesity comprising an extract of roots or a fraction thereof containing roponin or saponin.

The sprouts are used as food resources for a long time and their extracts have been used for a long time and their safety has been proved so that they can be prepared and taken in the form of foods which are common but can prevent or ameliorate obesity. At this time, the content of the extract or the fraction thereof in the food is not particularly limited, but may be 0.01 to 100% by weight, more preferably 1 to 80% by weight based on the total weight of the food composition . When the food is a beverage, it may be contained in a proportion of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may contain additional ingredients which are commonly used in food compositions and which can improve odor, taste, vision and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid and the like. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn) and copper (Cu) It may also include amino acids such as lysine, tryptophan, cysteine, valine, and the like. In addition, it is also possible to use antiseptics (such as potassium sorbate, sodium benzoate, salicylic acid, and sodium dehydroacetate), disinfectants (such as bleaching powder and highly bleached white powder, sodium hypochlorite), antioxidants (butylhydroxyanilide (BHA), butylhydroxytoluene BHT), etc.), coloring agents (such as tar pigments), coloring agents (such as sodium nitrite and sodium acetic acid), bleaching agents (sodium sulfite), seasoning (such as MSG sodium glutamate), sweeteners (such as hypoglycemia, , Food additives such as flavorings (vanillin, lactones, etc.), swelling agents (alum, potassium hydrogen D-tartrate), emulsifiers, emulsifiers, thickeners, encapsulating agents, gum bases, foam inhibitors, ) Can be added. The additives are selected according to the type of food and used in an appropriate amount.

On the other hand, a functional food for preventing or ameliorating obesity can be produced by using the food composition containing the extract or fraction thereof of roots or barley containing saponin.

As a specific example, the food composition can be used to produce processed foods that can prevent or improve obesity. Such processed foods include, for example, confectionery, drinks, liquor, fermented foods, canned foods, processed milk products, processed marine foods, noodles and the like. The sweets include biscuits, pies, cakes, breads, candies, jellies, gums, cereals (including dinner utensils such as cereal flakes). Drinks include drinking water, carbonated beverages, functional ionic beverages, juices (such as apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.) and sikhye. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned products include canned products (eg, tuna, mackerel, saury, canned fish, etc.), canned products (canned beef, pork, chicken, turkey canned food) and canned products (corn, peach and canned pineapple). Milk processed foods include cheese, butter, yogurt and the like. Meat processed foods are pork cutlet, beef cutlet, chicken cutlet, sausage. Sweet and sour pork, nuggets, and nubani. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.

The term "functional food " of the present invention is the same term as " food for special health use (FoSHU) ", and includes medical and medical effects The food may be prepared in various forms such as tablets, capsules, powders, granules, liquids, rings and the like in order to obtain a useful effect for prevention or improvement of obesity.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating obesity comprising rutonaline or a pharmaceutically acceptable salt thereof, a pharmaceutical composition for preventing or treating obesity, a composition for preventing or treating obesity in a pharmaceutically effective amount A method for preventing or treating obesity comprising the step of administering to a subject who is or is likely to have obesity, and a food composition for preventing or improving obesity comprising rutonaline or a pharmaceutically acceptable salt thereof. The details of the pharmaceutical composition, the method for preventing or treating obesity, and the food composition are the same as described above.

The term "pharmaceutically acceptable salt " of the present invention means salts in which the cation and the anion are pharmaceutically usable among the salts in which they are bound by an electrostatic attraction. Usually, the salt is a metal salt, an organic base, Salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. For example, the metal salt may be an alkali metal salt (sodium salt, potassium salt, etc.), an alkaline earth metal salt (calcium salt, magnesium salt, barium salt, etc.), an aluminum salt and the like; Examples of salts with organic bases include salts with organic bases such as triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N, And the like; The salt with inorganic acid may be a salt with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid and the like; Salts with organic acids may be salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid and the like; Salts with basic amino acids may be salts with arginine, lysine, ornithine and the like; The salt with an acidic amino acid may be a salt with aspartic acid, glutamic acid and the like.

In another aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating obesity comprising saponin or a pharmaceutically acceptable salt thereof, a pharmaceutical composition for preventing or treating obesity, a composition for preventing or treating obesity in a pharmaceutically effective amount A method for preventing or treating obesity comprising the step of administering to a subject who is or is likely to have obesity, and a food composition for preventing or improving obesity comprising saponin or a pharmaceutically acceptable salt thereof. The details of the pharmaceutical composition, the method for preventing or treating obesity, and the food composition are the same as described above.

By using the composition of the present invention, not only can the number of adipocytes be reduced by suppressing the differentiation into adipocytes, which is a cause of obesity, but also the content of cholesterol contained in the produced adipocytes can be reduced, It can be widely used for treatment.

FIG. 1 is a graph showing the results of screening 10 compounds by applying the extract of bud of barberry to UPLC (Ultra Performance Liquid Chromatography). FIG.
2a is a micrograph showing the level of fat contained in differentiated adipocytes according to the treatment concentration of rutonaline or saponin.
FIG. 2B is a graph showing the results of measuring the level of fat contained in adipocytes according to the treatment concentration of saponin. FIG.
FIG. 2C is a graph showing the results of measuring the level of fat contained in adipocytes according to the treatment concentration of rutonaline. FIG.
FIG. 3A is a graph showing the change in fat cell survival rate according to the treatment concentration of rutonaline or saponin. FIG.
FIG. 3B is a graph showing the results of comparing the levels of NO produced by the treatment of LPS in adipocytes according to the treatment concentration of rutonaline and the differentiation time.
FIG. 3C is a graph showing the results of comparing the levels of NO produced by the treatment of LPS in adipocytes according to the treatment concentration and time of differentiation of saponin.
4A is a graph showing the change in the expression level of the gene encoding aP2 according to the treatment concentration of saponin in the differentiated adipocytes.
4B is a graph showing the change in the expression level of a gene encoding PPARγ according to the treatment concentration of saponin in differentiated adipocytes.
4C is a graph showing the change in the expression level of a gene encoding aP2 according to the treatment concentration of rutonaline in differentiated adipocytes.
FIG. 4D is a graph showing the change in the expression level of a gene encoding PPARγ according to the treatment concentration of lutonaline in differentiated adipocytes. FIG.
5A is a graph showing the change in the expression level of a gene encoding GLUT4 according to the treatment concentration of saponin in differentiated adipocytes.
5B is a graph showing the change in the expression level of the gene encoding SREBP-1c according to the treatment concentration of saponin in the differentiated adipocytes.
FIG. 5C is a graph showing the change in the expression level of a gene encoding FAS according to the treatment concentration of saponin in differentiated adipocytes. FIG.
5D is a graph showing the change in the expression level of ACC-encoding genes according to the treatment concentration of saponin in the differentiated adipocytes.
5E is a graph showing the change in the expression level of a gene encoding GLUT4 according to the treatment concentration of rutonaline in differentiated adipocytes.
FIG. 5f is a graph showing a change in the expression level of a gene encoding SREBP-1c according to the treatment concentration of rutonaline in differentiated adipocytes. FIG.
FIG. 5G is a graph showing the change in the expression level of the gene encoding FAS according to the treatment concentration of rutonaline in the differentiated adipocytes. FIG.
5H is a graph showing the change in the expression level of a gene encoding ACC according to the treatment concentration of rutonaline in differentiated adipocytes.
6A is a graph showing the change in the expression level of the gene encoding adiponectin according to the treatment concentration of saponin in the differentiated adipocytes.
FIG. 6B is a graph showing the change in the expression level of a gene encoding ATGL according to the treatment concentration of saponin in differentiated adipocytes. FIG.
FIG. 6C is a graph showing the change in the expression level of a gene encoding MGL according to the treatment concentration of saponin in differentiated adipocytes. FIG.
FIG. 6D is a graph showing the change in the expression level of HSL-encoding genes according to the treatment concentration of saponin in differentiated adipocytes. FIG.
Fig. 6E is a graph showing the change in the expression level of the gene encoding adiponectin according to the treatment concentration of rutonaline in differentiated adipocytes. Fig.
FIG. 6f is a graph showing the change in the expression level of a gene encoding ATGL according to the treatment concentration of rutonaline in differentiated adipocytes. FIG.
FIG. 6G is a graph showing the change in the expression level of a gene encoding MGL according to the treatment concentration of rutonaline in differentiated adipocytes. FIG.
6H is a graph showing the change in the expression level of HSL-encoding genes according to the treatment concentration of rutonaline in differentiated adipocytes.
7 is a graph showing changes in the content of cholesterol contained in differentiated adipocytes according to the treatment concentration of saponin.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  1: Isolation and identification of active ingredients derived from barley barley

The ginseng barley was germinated, and after 5 days passed, the germanium barley was harvested and lyophilized, which was pulverized to obtain germinated barley powder.

The obtained germanium barley powder was extracted with 80% ethanol and filtered to obtain a liquid component, and the liquid component was freeze-dried to obtain a germanium barley extract.

The obtained germanium barley extract was applied to UPLC (Ultra Performance Liquid Chromatography) to select ten major compounds (Fig. 1). FIG. 1 is a graph showing the results of screening 10 compounds by applying the extracts of barberry barley to UPLC (Ultra Performance Liquid Chromatography). As shown in FIG. 1, it was confirmed that the third compound was the largest amount (1399.5 mg / 100 g) among the ten compounds and the second compound was the next largest amount (230.4 mg / 100 g).

Each of the above-selected compounds was separated and then applied to an NMR (Nuclear Magnetic Resonance) analyzer to identify each compound (Table 1).

Identification of the compounds contained in the barley extract Compound No. designation One
2
3
4
5
6
7
8
9
10 3-O-Feruloylquinic acid; 3-FQA
Lutonarin
Saponarin
Isoorientin
Luteolin-8-glucoside; Orientin
Isovitexin
Isoscoparin-7-O- [6-cinnamoyl] -glucoside
Isoscoparin-7-O- [6-feruloyl] -glucoside
7-O- [6-cinnamoyl] -glucoside < / RTI >
Isovitexin-7-O- [6-feryloyl] -glucoside

Example  2: Effect on the differentiation of adipocytes Rutonarein Saponin  effect

3T3-L1 cells, which are lipid precursor cells, can be subcultured in a medium containing BCS (Bovine Calf Serum), and can be cultured in a suitable medium such as FBS (feral bovine serum), IBMX (3-isobutyl-methylxanthine), dexanmethasone, Etc.), it is known that it can be differentiated into adipocytes. (C / EBP?, PPAR? And the like) are expressed when fat transcription factors (C / EBP ?, C / EBP ?, PPAR? And adipocyte-specific genes (aP2, Adipsin, etc.) are known to be expressed.

The present inventors sought to evaluate the effect of Lutonarin and Saponarin when the 3T3-L1 cells were differentiated into adipocytes.

Example  2-1: On the differentiation of the local taxa Rutonarin / Saponin  effect

3T3-L1 cells were treated with 0, 20, 40, 60 and 100 μM of rutonaline or saponin, and the differentiated cells were stained with oil-red staining to stain fat in adipocytes , And then the levels of stained fat under a microscope were compared (Figures 2a to 2c). At this time, unfractionated lipid precursor cells were used as a negative control, and adipocyte treated with CLA (Conjugated Linoleic Acid), which is known to have a fat reducing effect, was used as a positive control.

2A is a photomicrograph showing the level of fat contained in adipocytes differentiated according to the treatment concentration of lutonin or saponin, FIG. 2B is a graph showing the level of lipid contained in adipocytes according to the treatment concentration of saponin FIG. 2C is a graph showing the results of measuring the level of fat contained in adipocytes according to the treatment concentration of rutonaline. FIG.

As shown in Figs. 2 (a) to 2 (c), it was confirmed that in the cells treated with rutonaline or saponin, the level of fat was decreased and the level of fat was decreased as the concentration of rutonaline or saponin was increased.

Thus, it was found that rutonaline or saponin can reduce fat in a concentration-dependent manner.

Example  2-2: For fat cells Rutonarin / Saponin  Cytotoxicity Assessment

3T3-L1 cells (Korean Cell Line Bank, Korea), a lipid precursor cell, were purchased and added with 10% FBS (fetal bovine serum) and 1% antibiotics (100 U / ml penicillin and 0.1 mg / ml streptomycin) And cultured in a DMEM medium (Dulbecco's modified eagle medium) at 37 ° C under 5% CO ₂ (95% air).

The cultured 3T3-L1 cells were treated with 0, 20, 40, 60 and 100 μM of rutonaline or saponin and cultured for 72 hours in a medium containing differentiation inducing substances (FBS, IBMX, dexanmethasone and insulin) And the cells were allowed to react at 37 ° C for 4 hours. Then, DMSO was added to the cells to generate The cell viability was measured by measuring the absorbance at 570 nm (Fig. 3A). At this time, adipocytes not treated with the substance corresponding to rutonaline or saponin were used as a control group, and adipocytes treated with CLA at the same concentration were used as a comparative group.

FIG. 3A is a graph showing the change in fat cell survival rate according to the treatment concentration of rutonaline or saponin. FIG. As shown in FIG. 3A, it was confirmed that lutonin or saponin did not decrease the cell survival rate during the differentiation into adipocytes, but rather saponin increased the cell proliferation level in proportion to the treatment concentration.

On the other hand, the cultured 3T3-L1 cells were treated with 0, 20, 40, 60 and 100 μM of rutonin or saponin, cultured in a medium containing differentiation inducing substances for 72 hours to differentiate into adipocytes, At 48 and 72 hours, intracellular levels of NO (nitric oxide) produced by intracellular lipid peroxidation were measured and compared (Fig. 3b and 3c). At this time, the intracellular level of NO was measured by dividing the differentiated adipocytes into each well of 96-well plate at a concentration of 1 x 10 ⁴ cells, culturing for 24 hours, adding LPS (lipopolysaccharide) The result was applied to the nitrate assay kit and the level of NO produced by LPS was measured. As a control group, CLA - treated adipocytes were used.

FIG. 3B is a graph showing the results of comparing the levels of NO produced by the treatment of LPS in adipocytes according to the treatment concentration of rutonaline and the differentiation time. As shown in FIG. 3B, it was confirmed that even when the concentration of rutonaline was changed, the level of NO produced in adipocytes was not significantly changed.

FIG. 3C is a graph showing the results of comparing the levels of NO produced by the treatment of LPS in adipocytes according to the treatment concentration and time of differentiation of saponin. As shown in FIG. 3C, it was confirmed that even when the concentration of saponin was changed, the level of NO produced in adipocytes was not significantly changed.

From the results of FIGS. 3A to 3C, it was found that lutonaline or saponin did not show cytotoxicity on adipocytes.

Example  2-3: adipocytes Of the differentiation marker  For expression Saponin  effect

The lipid produced by differentiation of 3T3-L1 cells treated with saponin or rutonarin in Example 2-2, aP2 (fatty acid binding protein (aP2), which is known to be overexpressed during fat differentiation, and PPARγ Peroxisome Proliferator-Activated Receptor) was measured by real-time PCR at the mRNA level and compared (Figures 4a-4d).

FIG. 4A is a graph showing the change in the expression level of aP2-encoding gene depending on the treatment concentration of saponin in the differentiated adipocytes. FIG. 4B shows the expression of PPARγ in differentiated adipocytes according to the treatment concentration of saponin FIG. 4C is a graph showing a change in the expression level of a gene encoding aP2 according to the treatment concentration of rutonaline in differentiated adipocytes, and FIG. 4D is a graph showing a change in the expression level of the differentiated adipocytes , Which is a graph showing the change in the expression level of the gene encoding PPARy according to the treatment concentration of lutonin. As shown in FIGS. 4A to 4D, it was confirmed that as the treatment concentration of saponin or rutonarin was increased, the expression level of the gene encoding aP2 and PPARγ, which is an adipocyte differentiation marker, decreased.

As described above, since the expression level of the adipocyte differentiation marker was decreased depending on the treatment concentration of saponin or rutonarin, it was found that saponin or rutonarin showed an effect of inhibiting differentiation into adipocytes.

Example  2-4: On the Expression of Fatty Synthetic Enzymes Saponin  effect

The adipocytes produced by differentiating 3T3-L1 cells treated with saponin or rutonarin in Example 2-2 were tested for GLUT4 (glucose transporter 4), SREBP-1c (sterol regulatory) Changes in expression levels of genes encoding element-binding protein 1c, FAS (Fatty Acid Synthase) and ACC (Acetyl-CoA carboxylase) were measured at the mRNA level by Real-time PCR and compared 5h).

FIG. 5A is a graph showing the change in the expression level of a gene encoding GLUT4 according to the treatment concentration of saponin in differentiated adipocytes. FIG. 5B is a graph showing changes in SREBP-1c FIG. 5C is a graph showing the change in the expression level of the gene encoding FAS according to the treatment concentration of saponin in the differentiated adipocytes, and FIG. 5D is a graph showing the change in the expression level of the gene encoding FAS FIG. 5E is a graph showing changes in the expression level of a gene encoding ACC according to the treatment concentration of saponin in adipocytes. FIG. 5E shows the expression level of a gene encoding GLUT4 according to the treatment concentration of rutonaline in differentiated adipocytes 5f shows a change in the expression level of a gene encoding SREBP-1c according to the treatment concentration of rutonaline in differentiated adipocytes FIG. 5G is a graph showing a change in the expression level of a gene encoding FAS according to the treatment concentration of rutonaline in differentiated adipocytes. FIG. 5H is a graph showing the change in the concentration of rutonaline Gt; ACC < / RTI >

As shown in FIGS. 5A to 5H, it was confirmed that the expression levels of genes encoding GLUT4, SREBP-1c, FAS and ACC, which are proteins involved in fat synthesis, decrease as the treatment concentration of saponin or rutonaline increases.

As described above, since the expression level of the gene coding for the enzyme involved in liposynthesis was reduced in dependence on the treatment concentration of saponin or rutonaline, it was found that saponin or rutonaline suppressed the differentiation into adipocytes .

Example  2-5: Expression of Enzymes Hydrolyzing Fat Saponin  effect

Adiponectin, ATGL (Adipose Triglyceride Lipase), which is a protein involved in hydrolysis of fat, was prepared from adipocytes produced by differentiating 3T3-L1 cells treated with saponin or rutonarin in Example 2-2 above, , HSL (Hormone-Sensitive Lipase) and MGL (Monoglyceride Lipase) were measured by real-time PCR at the mRNA level and compared with each other (FIGS. 6A to 6H).

FIG. 6A is a graph showing the change in the expression level of adiponectin-encoding gene according to the treatment concentration of saponin in differentiated adipocytes. FIG. 6B is a graph showing the change in ATGL in the differentiated adipocytes according to the treatment concentration of saponin FIG. 6C is a graph showing the change in the expression level of a gene encoding MGL according to the treatment concentration of saponin, in differentiated adipocytes, FIG. 6D is a graph showing a change in the expression level of the differentiated adipocytes FIG. 6E is a graph showing the change in the expression level of the gene encoding HSL according to the treatment concentration of saponin, FIG. 6E shows the change in the expression level of the gene encoding adiponectin according to the treatment concentration of rutonaline in differentiated adipocytes FIG. 6F is a graph showing changes in the expression level of ATGL in the differentiated adipocytes according to the treatment concentration of rutonaline FIG. 6G is a graph showing a change in the expression level of a gene encoding MGL according to the treatment concentration of rutonaline in differentiated adipocytes. FIG. 6H is a graph showing the change in the concentration of rutonaline And the expression level of the gene encoding HSL according to the present invention.

As shown in Figs. 6A to 6H, as the treatment concentration of saponin or rutonarin increases, the expression levels of ATGL and HSL-encoding genes are decreased among the proteins involved in hydrolysis of fat, and expression of the gene encoding adiponectin And the level of expression of the gene coding for MGL was not significantly changed, but it was confirmed that the expression level of MGL was slightly increased.

Example  2-6: For Cholesterol Levels Saponin  effect

In Example 2-2, lipid components were extracted from adipocytes produced by differentiating 3T3-L1 cells treated with saponin, and the extracted lipid components were applied to GC / MS (Gas Chromatograph-Mass Spectrometer) The content of cholesterol was measured and compared (Fig. 7).

7 is a graph showing changes in the content of cholesterol contained in differentiated adipocytes according to the treatment concentration of saponin.

As shown in FIG. 7, the amount of cholesterol contained in the differentiated adipocytes was decreased as the treatment concentration of saponin was increased. In the adipocytes treated with 100 μM saponin, the content of cholesterol .

Claims (10)

A composition for inhibiting differentiation into adipocytes comprising rutonarin or saponin.
The method according to claim 1,
Wherein the composition is a germinated barley extract or fraction thereof containing lutonin or saponin.
3. The method of claim 2,
Wherein the extract is obtained by extracting the buds of barley with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
3. The method of claim 2,
Wherein the fraction is a fraction obtained by applying the germanium barley extract to a method selected from the group consisting of a solvent fractionation method, an ultrafiltration fractionation method, a chromatography fractionation method, and a combination thereof.
A method for inhibiting differentiation into adipocytes, comprising treating the composition of any one of claims 1 to 4 to separate stem cells that can be differentiated into adipose precursor cells or fat.
A pharmaceutical composition for the prevention or treatment of obesity, comprising a bud of barley extract or its fraction containing rutonaline or saponin.
The method according to claim 6,
Wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent.
A food composition for preventing or improving obesity comprising, as an active ingredient, a germanium barley extract or a fraction thereof containing rutonaline or saponin.
A pharmaceutical composition for preventing or treating obesity comprising rutonarin, saponin, or a pharmaceutically acceptable salt thereof.
0.0 > a < / RTI > food composition for prevention or amelioration of obesity comprising rutonarin, saponin, or a pharmaceutically acceptable salt thereof.

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