JP2013523810A - 用途 New use of yellow or yellow extract - Google Patents

Abstract

The present invention relates to a novel use of yellow or yellow extract for the prevention and treatment of obesity or metabolic syndrome, and more particularly, prevention of obesity, obesity complications or metabolic syndrome containing yellow or yellow extract. And a novel pharmaceutical or food composition for treatment, obesity, obesity complications or a novel use of yellow extract for producing a preparation for the prevention and treatment of obesity complications or metabolic syndrome, and obesity, obesity combination using the extract The present invention relates to a method for treating symptom or metabolic syndrome. Scarlet yellow or yellow extract extracts adipocyte differentiation and maintains SIRT1 at a high level to reduce body weight, abdominal fat and glucose tolerance, and is useful for the prevention and treatment of obesity, obesity complications or metabolic syndrome can do.

Description

  This application claims Korean patent application No. 10-2010-0032921 filed on April 09, 2010 and Korean patent application No. 10-2010-0095987 filed on October 01, 2010 as priority. The entire specification is a reference for the present invention.

  The present invention relates to a novel use of an extract of Polygonoatum falcatum or Polygonoatum sibiricum, and more particularly, maintains intracellular SIRT1 protein at a high level, and increases body weight, abdominal fat and sugar tolerance. Pharmaceutical or food composition for the prevention and treatment of obesity, obesity complications or metabolic syndrome comprising reduced yellow jade or yellow spirit as an active ingredient, use and treatment of obesity, obesity complications or metabolic syndrome of said extract Regarding the method.

  Obesity is a typical adult-type metabolic disease, and the ratio is greatly increased due to the extension of life expectancy due to medical development and improvement of living standards and the reduction of exercise amount due to the increase in the population living in the city center. Patients with fatal diseases such as hypertension, abnormal lipid metabolism and hyperglycemia due to increased insulin resistance, resulting in type 2 diabetes and its complications, myocardial infarction due to arteriosclerosis, stroke, etc. The number tends to increase.

  Metabolic syndrome is a conceptualization of a phenomenon in which various cardiovascular diseases and risk factors for type 2 diabetes, etc., gather together in one disease group. It is a useful concept that can comprehensively explain insulin resistance and many complicated and diverse metabolic abnormalities and clinical aspects associated with it. Having a metabolic syndrome increases the risk of developing cardiovascular disease or type 2 diabetes. This symptom has been known for a long time, but in 1988 Reaven claimed that a common cause of these symptoms was insulin resistance in which the body's insulin action was not smooth, and X (X) syndrome, insulin Named resistance syndrome. In 1998, the World Health Organization decided to call it “metabolic syndrome” instead of the term “insulin resistance syndrome” because there is no confirmation that insulin resistance can explain all these factors.

  The cause of metabolic syndrome is that even if there is insulin in the body, the high blood sugar level is not improved by resistance, but only the insulin concentration is increased. Insulin over-secreted to lower blood sugar cannot regulate blood glucose normally, and instead induces vascular cell proliferation to thicken the blood vessel wall, or promotes sodium reabsorption from the kidney to increase blood pressure, Or to increase the neutral fat in the blood by accelerating lipolysis and lowering high-density cholesterol (HDL-cholesterol), and at the same time, causing the disease by general action such as promoting the storage of fat components in the viscera This includes diabetes due to abnormal blood glucose metabolism, increased neutral fat due to abnormal lipid metabolism, high density cholesterol, hypertension due to increased sodium component, ventilation due to increased uric acid, and the like. The reason for this insulin resistance has not yet been clarified, but the biggest factor affecting it is known to be abdominal obesity, and excessive stress and aging are also known to be the cause. ing. In addition, in the past, insulin resistance has been considered as a single etiology of metabolic syndrome, but recently, in addition to insulin resistance, obesity and fat cell disorders, as well as multiple effects such as metabolic syndrome factors have been added. It is estimated as three major etiologies.

  The determination of metabolic syndrome is based on the following criteria. According to the NCEP (NationalCholesterol Education Program) standards published in 2001, waist circumference is 40 inches (102 cm) for men, abdominal obesity for women 35 inches (88 cm) or more, triglycerides 150 mg / dL or more HDL cholesterol is 40 mg / dL for men, 50 mg / dL or less for women, blood pressure is 130/85 mmHg or more, fasting glucose is 110 mg / dL or more, and one patient has 3 or more risk factors. When shown, it comes to be determined as metabolic syndrome. In the case of Orientals, if the waistline is 90 cm for men and 80 cm for women, it has been adjusted slightly to abdominal obesity, and applying this rule, about 25% of the total population in Korea has metabolic syndrome symptoms. There are also research reports.

  Metabolic syndrome is rapidly increasing in morbidity and morbidity due to the aging of the population and changes in the environment. Not only is the mortality rate associated with it very high, but the incidence of complications is also extremely high, resulting in enormous economic and social losses. However, most of the currently used drugs have low efficacy, have a high incidence of side effects when taken for a long time, and the development of therapeutic drugs is urgently necessary to improve the quality of the raw food itself. .

  Currently, the obese population in the West, including the United States, Europe and Japan, is estimated to be over 300 million, and in developing countries it is increasing by more than 1% annually, and the proportion of obese population is increasing worldwide. It is. Obesity, even defined by the World Health Organization (WHO) as a “disease that needs treatment,” is the second most common cause of death after smoking in the United States, resulting in more than $ 110 billion annually. Costs are high, the economic burden is high, and the social seriousness is becoming extremely serious.

  Worldwide patent applications related to anti-obesity agents have been increasing from 1999 to 2004, especially in the United States, where there was a sharp increase at this time, but gradually decreased from 2005 This is related to the fact that the development of substantial therapeutic agents for obesity has not been visualized. About 90% of obesity-related patents are related to appetite suppression, diet control and fat metabolism, but recently there has been a gradual increase in the share of patents related to fat growth regulation and energy metabolism. This is because the limitations of existing anti-obesity agents such as xenical and lidactyl have begun to be recognized and the need to develop anti-obesity agents as a new mechanism to improve this.

  SIRT1 is a mammalian homologue of Sirtuins (Sir2-related enzymes) protein (~ 110kD) known to induce life-span-extending effect from yeast, and class histone deacetylase that requires NAD + in cofactor Belonging to a class. There are seven types of Sirtuins that have been discovered in animals from SIRT1 to SIRT7, and they differ from each other in terms of their location and function. Among them, SIRT1 is activated in the nucleus and is metabolized, inflammatory response and degenerative brain. Modulates the activity of various substrates associated with disease.

  When dietary restriction increases SIRT1 expression and activity from various tissues such as liver, fat, and pancreas in the body, transcription factors such as PPARγ and FOXO1, and energy metabolism of mitochondria such as UCP1, UCP2, and UCP3 Inhibition of the activity in the liver by direct deacetylation of proteins related to glycine or indirect activity regulation by deacetylation of core regulators such as p300, NcoR, PGC1α, etc. Not only increases insulin sensitivity in the pancreas, but also causes weight loss by increasing energy metabolism from muscle tissue.

  Considering the fact that the demand for therapeutic agents is decreasing in the current situation where the obese population is increasing rapidly, this is a new mechanism that can overcome this, because it is a dissatisfaction with the side effects and efficacy of existing drugs. The development of the drug is in earnest.

  Therefore, if a drug that can replace the increased expression and activity of SIRT1 due to dietary restrictions such as small meals in vivo is developed, it can be used as a novel therapeutic agent for obesity and obesity complications.

  While the inventors have continued research on natural substances that increase the activity of SIRT1, the jaundice or yellow extract of the present invention increases the activity of SIRT1, resulting in obesity and obesity complications or metabolic syndrome. As a result, the present invention was completed.

  Accordingly, an object of the present invention is to provide a novel use of an extract of the genus Amadocoro (Ganono) that is yellow or yellow.

  In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome, which contains a yellow or yellow extract as an active ingredient. A functional composition is provided.

  In order to achieve another object of the present invention, the present invention relates to the prevention and / or prevention of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome, which contains blue japonica or yellow extract as an active ingredient. A mitigating food composition is provided.

  In order to achieve still another object of the present invention, the present invention relates to a preparation for preparing a preparation for the prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome. It provides the use of Amadokoroko plant extract which is yellow spirit.

  In order to achieve still another object of the present invention, the present invention is characterized by administering an extract of Amadocorus plant, which is yellow or yellow, to an individual in need thereof in an effective amount. Provided is a method for preventing and treating one or more diseases selected from the group consisting of complications and metabolic syndrome.

  Hereinafter, the present invention will be described in detail.

  The composition of the present invention contains yellow japonica or yellow extract as an active ingredient, and can be used for the purpose of prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome.

  In particular, the composition of the present invention can contain, as an active ingredient, an extract of Polygonatum falcatum or Polygonatum sibiricum, more preferably an extract of the rhizome site of Yellow Yellow or Yellow. Polygonatum sibiricum (Polygonatum sibiricum) described in the present invention is also called yellow (Polygonatum sibiricum Delar), naruko lily (Polygonatum sibiricum Redout), or yellow (Polygonatum sibiricum Redt). Including. On the other hand, the extract of the present invention is also a yellow spirit extract. The yellow spirit according to the present invention corresponds to the yellow rhizome or the rhizome part of yellow spirit, and more preferably, the yellow spirit according to the present invention corresponds to the rhizome of Polygonatum falcatum, which is referred to as “yellow coral”.

  The yellow and yellow extract of the present invention can be extracted by a known solvent extraction method. For example, blue to yellow or yellow to water, ethanol, alcohol, methanol, propanol, isopropanol, but 1-6 alcohols such as butanol, acetone, ether, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether Extraction can be performed using any one or a mixed solvent selected from organic solvents such as diethyl ether and benzene. Preferably, one or more solvents selected from the group consisting of ethanol, alcohol, acetone, ethyl acetate or methylene chloride are used, or these solvents are extracted by mixing with other known solvents.

  The yellow extract or yellow extract of the present invention can be used as it is by the solvent extraction method, or fractionated by a known method in order to remove impurities and increase the concentration of the active ingredient be able to. For the efficiency of fractionation, the extract can be concentrated under reduced pressure. Solvents for fractionation include water, alcohols having 1 to 6 carbon atoms such as ethanol, alcohol and methanol, ethyl acetate, methylene chloride, Any one or a mixed solvent selected from organic solvents such as dichloromethane, chloroform, n-hexane, diethyl ether, acetone and benzene can be used, preferably water. , N-butanol, ethanol and ethyl acetate, or a mixture thereof. The fraction can be used as is or can be re-fractionated, and in the case of re-fractionation, preferably ethyl acetate can be used. In particular, in order to perform the extraction process more economically, it is preferable to perform extraction once with a single or mixed solvent. However, water, alcohols having 1 to 6 carbon atoms such as ethanol, alcohol and methanol, ethyl acetate It is preferable to use any one selected from organic solvents such as methylene chloride, dichloromethane, chloroform, n-hexane, diethyl ether, acetone and benzene, or a mixed solvent thereof. Furthermore, ethanol, alcohol, acetone, ethyl acetate and methylene chloride can be preferably used. In particular, when alcohol is used as an extraction solvent, it is most preferable because the separation process of the extract can be shortened.

  In one embodiment of the present invention, 10 g of each sample is cut into small pieces, and then the extraction solvent, which is soaked in 30 ml of 100% methanol and extracted repeatedly three times at room temperature for 24 hours, is filtered by a filtration device and then reduced by a decompression device. Methanol was removed to obtain a methanol extract. After adding 20 ml each of ethyl acetate and water to the methanol extract obtained by the above process and mixing, the upper layer was concentrated to obtain the ethyl acetate extract. As a result of measuring the SIRT1 expression increasing efficacy of the produced fraction, it was confirmed that the expression of SIRT1 was high from the ethyl acetate fraction of the present invention (see Example 1). In yet another example of the present invention, high expression of SIRT1 was also confirmed from an extracted solution obtained by directly immersing each sample in 80% ethanol (see Example 5).

  The composition of the present invention has excellent efficacy for preventing and treating obesity, obesity complications or metabolic syndrome.

  Obesity means a condition with excessive body fat. Obesity complications indicate metabolic syndrome that results from long-term obesity status, which is associated with risk factors such as hypertriglyceridemia, hypercholesterolemia, hypertension, glucose metabolism disorder, blood coagulation disorder and obesity It means a syndrome that appears together.

  Such efficacy of the present invention has been demonstrated through in vivo and in vitro experiments.

  On the other hand, the composition of the present invention has the effect of increasing the level of in vivo SIRT1. SIRT1 is a mammalian homologue (~ 110kD) of Sirtuins (Sir2-related enzymes) protein known to induce a life extension effect from yeast, and is a class of histone deacetylases that require NAD + as a cofactor. Belongs.

  In one example of the present invention, the level-increasing efficacy of SIRT1 by the composition of the present invention was confirmed through in vivo experiments. As a result of measuring the amount of SIRT1 protein from muscle, liver and adipocytes from mice administered the composition of the present invention and control group mice, it was confirmed that the amount of SIRT1 was increased in fat and muscle (see Example 2). ).

  Furthermore, in one example of the present invention, a fat-rich obese diet was given to mice to produce an obesity model and the composition of the present invention was administered, and then changes in body weight and abdominal fat were observed. As a result, the group to which the composition of the present invention was administered lost weight despite the fact that there was no difference in the amount of feed intake compared to the group to which only the obese diet was administered, and the size of the abdominal fat was also reduced. It was confirmed that the amount also decreased. In addition, glucose tolerance due to insulin resistance, blood total cholesterol, neutral fat, free fatty acid values and indicators related to tissue metabolic syndrome were measured. As a result, the composition of the present invention improved the glucose tolerance by up to 60% or more (see Example 6), and cholesterol, neutral fat and free fatty acid decreased by 27.8, 27.0 and 38.2% or more, respectively. Confirmed (see Example 3).

  The composition of the present invention has been confirmed to have excellent efficacy in the prevention and treatment of obesity, obesity complications and metabolic syndrome.

  The composition of the present invention not only exhibits the above-mentioned effects, but also contains a herbal extract as an active ingredient, and therefore has fewer side effects on the human body than chemically synthesized pharmaceuticals.

  Therefore, the jade yellow or yellow extract of the present invention inhibits adipocyte differentiation, maintains SIRT1 at a high level, a high fat diet reduces abdominal fat from obesity-induced mice, significantly reduces body weight, It turns out to be effective in preventing and treating obesity, obesity complications or metabolic syndrome.

  Accordingly, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity, obesity complications or metabolic syndrome, which contains coral japonicum or yellow spirit extract as an active ingredient.

  The pharmaceutical composition according to the present invention may comprise a pharmaceutically effective amount of yellow jade or yellow extract alone or in addition to one or more pharmaceutically acceptable carriers. The pharmaceutical composition of the present invention can be administered to a patient in a single dose, or can be administered by a divided treatment method in which multiple doses are administered over a long period of time. In the above, the pharmaceutically effective amount means an amount showing a reaction higher than that in the negative control group, and preferably means an amount sufficient to treat or prevent obesity. An effective amount of yellow jade or yellow extract according to the present invention is 0.001 to 1000 mg / day / kg body weight, but is not limited thereto. However, the pharmaceutically effective amount can vary appropriately depending on a number of factors such as the disease and its severity, the patient's age, weight, health status, sex, route of administration and duration of treatment.

  The composition of the present invention can be formulated into various dosage forms according to the administration route by a method known in the art together with the pharmaceutically acceptable carrier. The administration route is not limited to this, but can be administered orally or parenterally. Parenteral routes of administration include many routes such as transdermal, nasal, abdominal, muscular, subcutaneous or intravenous.

  When the pharmaceutical composition of the present invention is orally administered, the pharmaceutical composition of the present invention, together with a suitable carrier for oral administration, is powder, granule, tablet, pill, sugar-coated tablet, capsule by a method known in the art. , Liquids, gels, syrups, suspensions, wafers and the like. Examples of suitable carriers include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, and starches including corn starch, wheat starch, rice starch and potato starch, cellulose, methylcellulose And fillers such as celluloses including sodium carboxymethyl cellulose and hydroxypropyl methyl cellulose, gelatin, polyvinylpyrrolidone and the like. Further, in some cases, cross-linked polyvinyl pyrrolidone, agar, alginic acid, sodium alginate, or the like can be added as a disintegrant. The pharmaceutical composition may additionally contain anti-aggregating agents, lubricants, wetting agents, perfumes, emulsifiers and preservatives.

  In addition, when administered parenterally, the pharmaceutical compositions of the present invention can be formulated by methods known in the art in the form of injections, transdermal dosages and nasal inhalants with a suitable parenteral carrier. Can be In the case of the injection, it must be sterilized and protected from contamination by microorganisms such as bacteria and bacilli. In the case of injections, examples of suitable carriers include, but are not limited to, solvents, including water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils or It is also a dispersion medium. More preferably, suitable carriers are Hex's solution, Ringer's solution, PBS (phosphate buffered asline) containing triethanolamine or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose. A solution or the like can be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like may be additionally included. In addition, the injection may in most cases additionally contain isotonic agents such as sugars or sodium chloride. In the case of transdermal administration agents, forms such as ointments, creams, lotions, gels, liquids for external use, pastes, liniments, and aerobes are included. In the above, “transdermal administration” means that a pharmaceutical composition is locally administered to the skin and an effective amount of the active ingredient contained in the pharmaceutical composition is transmitted into the skin.

  These dosage forms are described in prescription literature generally well known in medicinal chemistry (Remington's Pharmaceutical Science, 15th Edition, 1975, Mack Publishing Company, Easton, Pennsylvania).

  In the case of inhalation dosages, the compounds used according to the invention are pressurized using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. Conveniently delivered in the form of an aerosol spray from a pack or smoke device. In the case of a pressurized aerosol, 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 the compound and a suitable powder base such as lactose or starch.

Other pharmaceutically acceptable carriers can be referred to those described in the following literature (Remington's Pharmaceutical Science, 19th Edition, 1995, Mack
Publishing Company, Easton, Pennsylvania). Preferably, the pharmaceutical composition according to the present invention may comprise 0.001 to 99.999% by weight and a pharmaceutically acceptable carrier 99.999 to 0.001% by weight.

  The pharmaceutical composition of the present invention can be administered in combination with known compounds having the effect of preventing and treating obesity, obesity complications or metabolic syndrome.

  The yellow jade or yellow extract according to the present invention can be provided in the form of a food composition for the purpose of improving obesity, obesity complications or metabolic syndrome. The food composition of the present invention includes all forms such as functional foods, nutritional supplements, health foods and food additives. The food composition of the above type can be manufactured in various forms by methods known in the art.

  For example, for the health food, the yellow or yellow extract itself of the present invention can be produced in the form of tea, juice and drink and used for drinking, or can be ingested into granules, encapsulated and powdered. Furthermore, it can be produced in the form of a composition by mixing with the known yellow or yellow extract of the present invention and known substances or active ingredients known to have an effect of improving obesity, obesity complications or metabolic syndrome. .

  Furthermore, functional foods include beverages (including alcoholic beverages), fruits and processed foods thereof (eg canned fruits, bottling, jam, marmalade, etc.), fish, meat and processed foods thereof (eg ham, sausage corn beef) Etc.), breads and noodles (eg udon, buckwheat, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, strawberries, dairy products (eg butter, cheese, etc.), edible vegetable oils, margarine, vegetable The protein can be produced by adding the yellow or yellow extract of the present invention to protein, retort food, frozen food, various seasonings (eg, miso, soy sauce, sauce, etc.).

  Although the preferable content of the said yellow coral or yellow extract is not limited to this in the food composition of this invention, it can be 0.001-99.999 weight% in the food composition accompanying this invention.

  Furthermore, in order to use the yellow and yellow extract of this invention in the form of a food additive, it can be manufactured and used in the form of a powder or a concentrate.

  Furthermore, the present invention provides a novel extract of the genus Amadokoro, which is yellow or yellow, for producing a preparation for the prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome. One selected from the group consisting of obesity, obesity complications and metabolic syndrome, characterized in that it is administered in an effective amount to an individual in need thereof, and the extract of the genus Amadocoro, which is a yellow or yellow spirit The present invention relates to a method for preventing and treating the above diseases.

  In the present invention, the “effective amount” means that the composition or preparation of the present invention has an effect of preventing or treating obesity, obesity complications and metabolic syndrome in the individual to which the drug is administered. The term “individual” means an animal, preferably a mammal, particularly an animal including a human, and is also a cell, tissue, organ or the like derived from the animal. The individual is also a patient in need of treatment.

  As discussed above, the present invention relates to a novel use of yellow jade or yellow extract, and yellow jade or yellow extract maintains a high level of SIRT1 to reduce body weight, abdominal fat and sugar tolerance, It can be usefully used for the prevention and treatment of obesity, obesity complications or metabolic syndrome.

  FIG. 1A shows the result of treating the HEK293 cell line with a crude drug ethyl acetate extract belonging to the genus Amadokoro and measuring changes in SIRT1 protein by Western blot. FIG. Results of measurement of SIRT1 protein expression at concentration (C: negative control group; ID1216: yellow ethyl acetate extract; ID1215: jade yellow ethyl acetate extract; ID1214: yellow ethyl acetate extract; ID1213: Naryu lily ethyl acetate extraction object).

  Figure 2 shows the expression of SIRT1 in the order of liver, fat, and muscle tissue from the results of Western blotting, which measured whether or not SIRT1 expression increased in each tissue of mice by administration of japonic ethyl acetate extract and yellow ethyl acetate extract. Measured (Vehicle: negative control group).

  FIG. 3 shows the results of testing the anti-obesity effect of the composition of the present invention from an obesity-inducing animal model, and measuring the body weight of mice according to time. Numerical values are mean standard deviation values of 9 to 10 mice, * means p <0.05, ** means p <0.01, which means that it is significant (□: normal diet; ■: High fat diet; ▲: High fat diet + sibutramine (100 mg / kg feed); ●: High fat diet + ID1216 (5000 mg / kg feed); ◆: High fat diet + ID1215 (5000 mg / kg feed)).

  FIG. 4 is a graph showing the results of testing the anti-obesity effect of the composition of the present invention from an obesity-inducing animal model, and measuring the weight of liver and adipose tissue. The numerical values are the mean standard deviation values of 9-10 mice, * means p <0.05, ** means p <0.01, which means that it is significant (ND: normal) Diet group; HF: high fat diet group; sibutramine: high fat diet + sibutramine (100 mg / kg diet); ID1216: high fat diet + yellow ethyl acetate extract (5000 mg / kg diet); ID1215: high fat diet + coral yellow Ethyl acetate extract (5000 mg / kg feed)).

  FIG. 5 shows the results of measuring blood lipid levels from an obesity-induced animal model. 5A shows blood cholesterol, FIG. 5B shows blood neutral fat, and FIG. 5C shows a comparison of blood free fatty acid concentration with high fat diet group (ND: normal diet group; HF: high fat diet group). ; Sibutramine: high fat diet + sibutramine (100mg / kg feed); ID1216: high fat diet + yellow ethyl acetate extract (5000mg / kg feed); ID1215: high fat diet + jade yellow ethyl acetate extract (5000mg / kg diet) )). FIG. 6 shows the results obtained by treating the HEK293 cell line with either yellow or yellow ethanol extract and ethyl acetate extract, and measuring the SIRT1 protein expression change by Western blot (C: negative control group; ID1216: yellow ethyl) Acetate extract; ID1215: jade yellow ethyl acetate extract; ID1216B: yellow spirit 80% ethanol extract; ID1215B: jade yellow 80% ethanol extract).

  FIG. 7 shows the results of testing the anti-obesity effect of the composition of the present invention from an obesity-inducing animal model, and measuring the body weight of mice according to time. The numerical value is the mean standard deviation value of 8 mice, * means p <0.05, ** means p <0.01, which means that it is significant (□: normal diet; ■: High fat diet; ◆: High fat diet + ID1215B (250 mg / kg); ▲: High fat diet + ID1215B (500 mg / kg; ● High fat diet + ID1215B (750 mg / kg)).

  FIG. 8 is a graph showing the results of testing the anti-obesity effect of the composition of the present invention from an obesity-inducing animal model, and measuring the weight of liver and adipose tissue. Numerical values are the mean standard deviation values of 8 mice, * means p <0.05, ** means p <0.01, which means that it is significant (ND: normal diet group; HF: high fat diet group; 250 mg / kg: high fat diet + ID1215B-250 mg / kg: 500 mg / kg high fat diet group + ID1215B-500 mg / kg; 750 mg / kg: high fat diet + ID1215B-750 mg / kg).

  FIG. 9 shows the results of a low-temperature test related to the anti-obesity effect of the composition of the present invention from an obesity induction animal model. The numerical values are the average standard deviation values of 8 mice, * means p <0.05, ** means p <0.01, which means that it is significant (■: high fat diet; ◆: High fat diet + ID1215B (250 mg / kg); ▲: High fat diet + ID1215B (500 mg / kg; ●: High fat diet + ID1215B (750 mg / kg)).

  FIG. 10 shows the results of testing metabolic syndrome-related effects from an obesity-induced animal model. FIG. 10A shows the results of measuring blood glucose concentration by oral administration of glucose. The numerical value is a standard deviation value of 8 mice, and * is p <0.05, which means that it is significant when compared with a high fat diet (□: normal diet; ■: high fat diet; ◆: High fat diet + ID1215B (250 mg / kg); ▲: High fat diet + ID1215B (500 mg / kg); ● High fat diet + ID1215B (750 mg / kg)). FIG. 10B compares the area under curves (AUC) after 8 weeks between the obese diet group and the ID1215B administration group. FIG. 11 shows the results of measuring blood lipid levels from an obesity-induced animal model. FIG. 11A shows blood cholesterol, FIG. 11B shows blood neutral fat, and FIG. 11C compares blood free fatty acid concentration with the high fat diet group. (ND: normal diet group; HF: high fat diet group; 250 mg / kg: high fat diet + ID1215B-250 mg / kg: 500 mg / kg: high fat diet + ID1215B-500 mg / kg; 750 mg / kg: high fat diet + ID1215B-750 mg /kg).

  FIG. 12 is a diagram comparing the difference between fat and hepatocytes by observing the liver and adipose tissue with an optical microscope 8 weeks after the obesity induction animal model. FIG. 12A is a diagram showing the difference in the size of fat cells, and FIG. 12B is a diagram showing the state of hepatocytes and the degree of fat vacuoles. (ND: normal diet group; HF: high fat diet group; 250 mg / kg: high fat diet + ID1215B-250 mg / kg: 500 mg / kg: high fat diet + ID1215B-500 mg / kg; 750 mg / kg: high fat diet + ID1215B-750 mg /kg).

  Examples of the present invention will be described in detail below.

  However, the following examples only illustrate the present invention, and the content of the present invention is not limited to the following examples.

<Example 1>
Confirmation of efficacy of SIRT1 expression enhancement extract <1-1> Comparison of SIRT1 expression enhancement level (in vitro)
In order to select the extract with the highest SIRT1 activity, yellow goose (Polygonoatum sibiricum: ID1216), yellow goose (Polygonoatum sibiricum falcatum: ID1215), Amadokororo (Polygonoatum stenophyllum: ID1214) collected from many localities in Korea Ethyl acetate extracts were compared against 4 species such as Yuri Naruko (Polygonoatum lasianthum: ID1213).

  The ethyl acetate extract was obtained by chopping each 10 g sample, immersing it in 30 ml of 100% ethanol, extracting the extracted solution repeatedly three times at room temperature for 24 hours, and filtering it with a filtration device. The methanol was removed and the methanol extract was obtained. After adding 20 ml each of ethyl acetate and water to the methanol extract obtained by the above process and mixing, the upper layer was concentrated to obtain the ethyl acetate extract.

  Each ethyl acetate extract was washed twice with ethanol to remove residual solvent, concentrated under reduced pressure, dissolved in DMSO at a constant concentration, and 0.5% (v / v) in serum-free Dulbecco's modified Eagle medium (DMEM). ) Proteins obtained after culturing HEK293 cells for 16-20 hours in medium added at a ratio were electrophoresed on 8% SDS-PAGE gel, transferred to PVDF membrane, and immunoblotted with SIRT1 and specific antibody (Cell Signaling Technology, USA) A search system to establish (immunoblotting) was established. In the control group, a DMSO-treated group containing no drug was used to select drugs whose SIRT1 expression was increased from the control group.

  As a result, as shown in FIG. 1A, SIRT1 expression-enhancing activity was confirmed for all the four ethyl acetate extracts, and the activity of yellow coral extract (ID1215) was confirmed to be the highest. Furthermore, as shown in FIG. 1B, yellow coral extract (ID1215) showed SIRT1 expression enhancing effect even at a concentration 8 times lower than that of yellow spirit (ID1216).

<Example 2>
In vivo SIRT1 expression test for ID1215 and ID1216
In order to confirm whether ID1215 and ID1216 increase SIRT1 expression in vivo, the expression pattern of SIRT1 was confirmed for each organ in C57BL6 mice.

All animal treatments were performed according to the guidelines of the Nitto Pharmaceutical (Korea) Animal Ethics Committee. 8-week-old male C57BL / 6 male mice were purchased from Oriental Bio. The experiment was conducted by keeping the breeding room temperature at 23 ° C. and the humidity at 50 ± 10%, and adjusting to the environment for one week with a period of 12 hours (day light 08: 00-20: 00). In order to facilitate absorption, fasted mice were given ID1215 and ID1216 at 500 mg / kg body weight (= 500 mpk), respectively. Five hours after administration, liver, fat and muscle were removed from the mouse. A part of the extracted tissue is rapidly frozen in liquid nitrogen and stored at −70 ° C. until use at the time of use for RNA extraction, and the remaining tissue is extracted with a protease inhibitor and Na ₃ VO for protein extraction. _The protein was quantified using a BCA quantification method in a homogenization buffer containing ₄ (10 mM Tris, pH 7.4, 150 mM NaCl, 1 mM EDTA 10% glycerol, 0.1% Triton X-100). In order to confirm the expression of SIRT1, the remaining tissue excluding the muscle was separated from the 8% SDS-PAGE gel after separating 60μg protein (100μg in the muscle), then transferred to a PVDF membrane for 10 minutes with 5% nonfat milk. Western blotting was performed with anti-SIRT1 antibody after blocking.

  As a result, as shown in FIG. 2, the expression of SIRT1 protein was increased in a similar manner from both fat and muscle in ID1216 and ID1215.

<Example 3>
Anti-obesity efficacy test for ID1215 and ID1216
Through the SIRT1 expression test, both ID1215 and ID1216 were expected to show anti-obesity effects. To compare the anti-obesity efficacy against ID1215 and ID1216, which had high SIRT1 activity enhancement efficacy in in vitro experiments, a DIO model test was conducted.

<3-1> Production of obesity-inducing mouse model with high-fat diet The test was conducted for 12 weeks while feeding 8-week-old C57BL6 mice with a diet containing ID1215 and ID1216 added to a high-fat diet containing 40% fat.

In a positive control group for anti-obesity efficacy in a high fat diet, sibutramine was mixed at 0.01%, ID1215 and id1216 were mixed at 0.5%, and kept refrigerated until use. The components of the feed are as shown in Table 1.

  Animal preparation proceeded as in Example 2. Mice freely fed a high fat diet for 12 weeks. Test group is general treatment group (ND) not containing beef tallow, non-drug-treated high fat diet group (HF), ID1215 and ID1216 groups containing 5000 mg per kg of high fat diet feed (ID1215, ID1216), high fat Ten animals were tested, divided into groups containing 100 mg sibutramine per kg diet.

<3-2> Measurement of body weight change The body weight was measured once a week. After the experiment, all groups of mice collected plasma, dissected and weighed liver and abdominal fat. Some liver tissues and adipose tissues were observed by tissue staining.

  As a result, as shown in FIG. 3, the obesity feed administration group to which ID1216 and ID1215 were added respectively decreased in body weight after 12 weeks compared with the high fat diet group by 8.6% and 25.9%, respectively, and the positive control group sibutramine was administered. The group had an obesity-suppressing effect at the beginning, but it has ceased after 8 weeks. Furthermore, as shown in Table 2 and FIG. 4, the drug-administered group had an abdominal adipose tissue amount of 3.2% compared to the high-fat diet group compared to the high-fat diet group at the time of necropsy after 12 weeks. 41.9%, and sibutramine administration group decreased 3.2%. The liver mass decreased by 6.2% for ID1216 and 31.2% for ID1215 compared to the fat diet group, and there was no change in liver mass in the sibutramine administration group. As a result of histological analysis of liver tissue, it was confirmed that fat that was densely distributed over a wide range from the liver tissue of the high-fat diet group was locally and mildly alleviated in the ID1215 administration group (results are shown in the figure). Not shown).

<3-3> Blood test After the experiment was completed, blood was collected from each group of mice to separate plasma. Cholesterol was analyzed using an automatic dry chemical analyzer (A25 analyzer, Biosystems), neutral fat was measured using Triglyceride assay Kit (Cayman), and free fatty acid was measured using NEFA (Wako) according to the manufacturer's guidelines. .

As a result, as shown in FIG. 5 and Table 3, as an index related to metabolic syndrome, the total blood cholesterol level was decreased by 10.6% in the ID1216 administration group and 27.8% in the ID1215 administration group compared to the high fat diet group, and neutral fat and free It was confirmed that fatty acids decreased by 14.9% and 15.4% in the ID1216 administration group, and decreased by 27.0% and 38.2% in the ID1215 administration group, respectively. In the case of ID1216, the effect of suppressing body weight gain and the decrease in the amount of fat and liver tissue are weak, but metabolic syndrome-related effects are relatively high. ID1215 improves obesity and all metabolic syndrome diseases compared to ID1216 Expected to be highly effective.

  In summary of the efficacy test results related to obesity and metabolic syndrome for ID1215 and ID1216 in the above examples, all of these compositions were determined to be non-toxic when administered at 500 mg / kg for a long period of time, It has been found that it not only exhibits an anti-obesity effect equivalent to or better than sibutramine, which is currently on the market as a therapeutic agent for obesity, but also improves various labels associated with obesity and metabolic syndrome.

<Example 4>
Establishing the optimum extraction conditions for the extract When using an organic solvent such as ethyl acetate, the separation and purification process is complicated or impossible to use due to residual organic solvent when used for food or the human body. Since a case occurred, an extraction method using ethanol (alcohol) that could be used was established. Therefore, when the mass of the active ingredient content of the present invention in the extract was high and the extraction amount was large, the optimum extraction conditions were tested for the purpose of making the process easy and reducing the production unit cost.

  The method for obtaining the ethyl acetate extract is the same as that of Example 1. After chopping 10% of the sample, immerse it in 30 ml of 100% methanol and extract it three times for 24 hours at room temperature. Filter the extract with a filter, remove the methanol with a vacuum, and extract the methanol extract. Obtained. To the methanol extract obtained by the above process, 20 ml each of ethyl acetate and water were added and mixed with the methanol extract, and the upper layer was concentrated to obtain the ethyl acetate extract.

  Therefore, in order to simplify the extraction process, extraction was performed as follows using 80% ethanol. A 10 g sample cut into 30 ml of 80% ethanol and extracted three times for 24 hours, and the extracted solution is filtered with a filtration device. The solvent is removed with a decompression device and 80% ethanol extract is obtained. Was obtained. The 80% ethanol extract showed an extract of 17% relative to the weight of the sample, and the extraction rate was 85 times higher than that of the ethyl acetate extract 0.2%.

<Example 5>
SIRT1 expression enhancement test of ethanol extract of ID1215 and ID1216

  Each extract extracted in Example 4 was dissolved in DMSO at a constant concentration, and the possibility of SIRT1 activity was tested by the method of Example 1.

  In order to select a solvent-specific extract with the highest SIRT1 activity, we compared the 80% ethanol extract and ethyl acetate extract of yellow japonica and yellow spirit of the present invention collected from many localities in Korea. In the experiment, the extract was treated with HEK 293 cells according to concentration as in Example 1, and then the increased expression of SIRT1 was confirmed by Western blot.

  As a result, as shown in FIG. 6, yellow yellow 80% ethanol extract (ID1216B) and yellow yellow spirit 80% ethanol extract (ID1215B) are yellow yellow ethyl acetate extract (ID1216) and yellow yellow spirit ethyl acetate extract (ID1215). It showed similar SIRT1 expression enhancement effect.

  In addition to the advantages in terms of stability and economics of the extract, it was administered to mice at a dose of 250, 500, 750 mg / kg against 80% ethanol extract (ID1215B), which is highly effective in enhancing intracellular SIRT1 expression. However, the anti-obesity effect was verified.

<Example 6>
Anti-obesity efficacy test against ID1215B

<6-1> Production of obesity-inducing mouse model with high-fat diet The test was conducted once a day at 250,500,750mg / kg with ID1215B dissolved in PBS while feeding a diet containing 40% fat to an 8-week-old C57BL6 mouse. Changes in body weight were measured by the method of oral administration for 8 weeks, and after 8 weeks, changes in various obesity and related indices were compared with those of a general diet and a group of obese diet mice.

  The test group is a general feed treatment group (ND) that does not contain beef tallow, a non-drug-treated high-fat diet group (HF), and a group that receives 250 mg / kg ID1215B once a day while giving a high-fat diet (ID1215B-250 mg / kg), 500 mg / kg administration group (ID1215B-500 mg / kg), and 750 mg / kg administration group (ID1215B-750 mg / kg), 8 mice were tested. ND and HF were administered PBS once a day for the same test results as the ID1215B administration group.

<6-2> Measurement of body weight change The body weight was measured once a week. After the experiment, all groups of mice collected plasma and weighed liver and abdominal fat. Some liver tissue and adipose tissue were observed by tissue staining.

  As a result, as shown in FIG. 7, in the ID1215B administration group, the body weight after 8 weeks decreased by 17.0, 15.4, and 17.0% in the 250,500,750 mg / kg administration group as compared with the high fat diet group, respectively. Furthermore, as shown in Table 4, FIG. 8 and FIG. 12, the amount of abdominal adipose tissue in the drug-administered group decreased by 10% or more from all ID1215B-administered groups at the time of autopsy after 8 weeks compared to the high-fat diet group. However, the size of adipocytes also decreased (see FIG. 12A). Liver mass decreased by 18.6, 14.0, and 24.4% in the 250, 500, and 750 mg / kg groups compared to the high-fat diet group, respectively. As a result of histological analysis of liver tissue, it was confirmed that fat that had been distributed extensively and densely from the liver tissue of the high fat diet group was locally alleviated in the ID1215B administration group (see Fig. 12B). .

<6-3> Low temperature test A low temperature test was performed to confirm the cause of weight loss. Mice were measured and recorded in the rectum every 8 hours for 8 hours in a cold space at 4 ° C., which was compared to the initial body temperature.

  As a result, as shown in FIG. 9, the ID1215B-administered group significantly overcome the decrease in body temperature that appears in the high-fat diet group, and the weight gain suppression effect of ID1215B administration is the result of increased heat dissipation through active metabolism in the body. It turns out that it is.

<6-4> Glucose tolerance measurement for insulin resistance To measure glucose tolerance due to insulin resistance, blood was collected from the tail of a mouse fasted for 16 hours or more (0 minutes), and glucose was measured. After injection at 2 g / kg, blood was collected at 10, 20, 30, 60, and 120 minutes, and blood glucose was measured and recorded.

  As a result, as shown in FIG. 10, the area under the curve (AUC, Area under curves) of the group administered ID1215B at 250,500,750 mg / kg dose was 14236 *, 8387 *, 7789 *, respectively, compared to the high fat diet group Reduced glucose tolerance by 31.6, 59.7 and 62.6%, respectively (* is p <0.05).

<6-5> Blood test After the experiment was completed, blood was collected from each group of mice to separate plasma. Toxicity-related indicators and cholesterol were analyzed with an automatic dry chemical analyzer, and neutral fat and free fatty acid were measured using Triglyceride assay Kit (Cayman) and NEFA (Wako), respectively, according to the manufacturer's guidelines.

  As a result, as shown in Table 5 and FIG. 11, there was no significant difference in cholesterol in blood as an indicator for metabolic syndrome, but neutral fat was 30.1% in the ID1215B administration group compared to the high fat diet group, respectively. It decreased by 42.5 and 47.2%, showing concentration dependence, and free fatty acids also decreased by 7.1, 7.1, and 14.3%, respectively. Therefore, ID1215B is expected not only to suppress weight gain but also to have a high effect of improving various metabolic syndrome diseases.

  Furthermore, as shown in Table 6, the ID1215B administration group is concerned about the abnormalities of hepatotoxicity-related indices (ALT, AST, etc.) seen in the obesity induction group to a level similar to that of the normal diet group, Indications related to nephrotoxicity (creatine, BUN, etc.) did not show significant changes compared to the normal diet group.

<6-6> Optical Microscopic Observation of Liver and Adipose Tissue After the animal experiment was completed, optical microscope observation was performed after H & E (hematoxylin and eosin) staining in order to confirm changes in the liver and adipose tissue.

  The excised liver and adipose tissue were cut into a certain size, fixed in 10% formalin by the usual method, washed with water, and a block was prepared through the process of embedding parabin. The prepared block was sliced with a thickness of 4 μm, and hematoxylin and eosin staining were performed to observe general changes.

  The groups treated with ID1215B at all concentrations did not turn yellowish brown like the high fat diet group, but showed a bright red color similar to that of the general diet group, and the extent and extent of vacuolar degeneration were also clearly reduced. As a result of adipose tissue analysis, it was confirmed that the size of adipocytes was small in the ID1215B administration group of all doses compared to the high fat diet group (see FIG. 12).

<Example 7>
Single and repeated dose toxicity studies
In order to confirm the possibility of induction of toxicity when ID1215B was orally administered, a single toxicity and 4-week repeated dose toxicity test was conducted.

  Single toxicity is 6 weeks old male ICR mice each as a group, and ID1215B is orally administered at 1.000-5.000mg / kg body weight orally for 12 weeks. The abnormal findings were observed through autopsy after death. No animals were moribund in the ID1215B administration group at all concentrations, and abnormal organ findings were not found, and it was judged that there was no toxicity (results not shown).

  Repeated dose toxicity consists of 6 male 6-week-old ICR mice in one group, and ID1215B was orally administered at 1.000 to 3.000 mg / kg body weight once a day for 28 days. General symptoms and viability were observed, and experimental animals were necropsied to observe the findings of organ abnormalities. None of the animals moribund from the ID1215B administration group at all concentrations, and no findings of organ abnormalities were found. As shown in Table 7, indicators related to liver toxicity such as ALT and AST and kidneys such as creatine and BUN There were no significant changes in all indicators related to toxicity.

  ID1215B also confirmed single dose (1,000 to 3,000 mg / kg body weight) and repeated dose toxicity (500 to 1,500 mg / kg body weight) for 7 days by the above method, but survival rate at all concentrations, abnormalities at autopsy There were no significant differences from the vehicle in the findings, blood biochemical indices related to hepatotoxicity and renal toxicity (results not shown).

  In summary of the efficacy and toxicity test group results associated with obesity and metabolic syndrome for ID1215B in the above examples, the composition is administered in a single high dose of up to 5.000 mg / kg and repeated at a dose of 3.000 mg / kg. In addition to being not toxic, it was found that anti-obesity efficacy and various labels related to metabolic syndrome were improved without toxicity even when administered at a maximum dose of 750 mg / kg for a long period of time.

  The present invention relates to a pharmaceutical or food composition for the prevention and treatment of obesity, obesity complications or metabolic syndrome, and a preparation for the prevention and treatment of obesity, obesity complications or metabolic syndrome, which contains blue jade or yellow spirit extract as an active ingredient. The present invention relates to a novel use of yellow jade or yellow extract and a method for treating obesity, obesity complications or metabolic syndrome using the extract, wherein the yellow jade or yellow extract contains intracellular SIRT1 protein. Because it has the effect of maintaining a high level and reducing body weight, abdominal fat and glucose tolerance, it should be used as an active ingredient in pharmaceutical compositions or food compositions for the prevention and treatment of obesity, obesity complications or metabolic syndrome Can do.

Claims (5)

  A pharmaceutical composition for the prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome, comprising Polygonatum falcatum extract or Polygonatum sibiricum extract.   A food composition for prevention and amelioration of one or more diseases selected from the group consisting of obesity, obesity complications, and metabolic syndrome, containing a yellow yellow extract or yellow extract.   The extract is selected from the group consisting of water, ethanol, alcohol, methanol, propanol, isopropanol, butanol, acetone, ether, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane, petroleum ether, diethyl ether, and benzene. 3. The composition according to item 1 or 2, which is the above solvent extract.   Use of yellow or yellow extract for producing a preparation for the prevention and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome.   Prophylaxis and treatment of one or more diseases selected from the group consisting of obesity, obesity complications and metabolic syndrome, characterized by administering an effective amount of jade yellow extract or yellow extract to an individual in need thereof Method.

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