KR102178199B1 - a composition comprising an extract of Rhus verniciflua and Eucommia ulmoides, as an active ingredient for preventing or treating obesity - Google Patents

Abstract

The present invention relates to a composition for the prevention and treatment of obesity containing a combination extract consisting of lacquer tree and chinensis as an active ingredient. Targeting the combination extract according to the present invention, it was confirmed that weight gain was excellently suppressed through an experiment to verify the efficacy of improving obesity in an obesity model due to a fat diet for the combination extract of the present invention (Experimental Example 1), and measurement of serum lipid content In the experiment, it was confirmed that the accumulation of intracellular fat and the accumulation of triglyceride and cholesterol were strongly inhibited (Experimental Example 2): In the experiment of hepatocyte tissue staining through Oil Red staining, the accumulation of intracellular fat and the accumulation of triglyceride and cholesterol It was confirmed that it strongly inhibits (Experimental Example 3); Through immuno-blotting assay, the expression of endoplasmic reticulum stress-related proteins is reduced, the expression of SREBP-1 and FAS is suppressed, the phosphorylation of mTORC1, P70S6K and 4EBP-1 is reduced, and the phosphorylation of AMPK is It was confirmed to increase (Experimental Example 4); It was confirmed that the activity of GSH, GPx, SOD, and CAT increased in a concentration-dependent manner to prevent oxidative damage to the liver due to a high fat diet by inhibiting lipid peroxide substances and maintaining the detoxification function of toxic substances in the body (Experimental Example 5). By doing so, the combination extract composed of poison ivy and chinensis of the present invention can be usefully used in the prevention or treatment of obesity.

Description

A composition comprising an extract of Rhus verniciflua and Eucommia ulmoides, as an active ingredient for preventing or treating obesity, containing a combination extract consisting of poison ivy and chinensis as an active ingredient

The present invention relates to a composition for the prevention and treatment of obesity containing a combination extract consisting of lacquer tree and chinensis as an active ingredient.

[Reference 1] Hida K, et al., Visceral adipose tissue-derived serine protease inhibitor: a unique insulin-sensitizing adipocytokine in obesity. Proc Natl Acad Sci USA. 102, pp. 10610-5, 2005

[Reference 2] Han DR, et al., Studies on morphological and chemotaxonomy and seco-triterpene glycoside component of korean Acanthopanax spp. Bull D S Pharm Sci Inst 11:1-66, 1994

[Document 3] Ho-gi Park. The method of reproducing the cuttings of gasiogapi. Journal of Medicinal Crop Science 2: 133-139, 1994

[Reference 4] Kim YH, et al., Studies on the constituents of Acanthopanax koreanum Nakai (1). Kor J Pharmacogn 16:151-154, 1985

[Ref. 5] Hahn DR, et al., A study on the chemical constituents of Acanthopanax koreanum and its pharmacobiological activities. Yakhak Hoeji 29:357-361, 1985

[Reference 6] Chung BS, et al., Studies on the constituents of Acanthopanax koreanum Nakai. Kor J Pharmacogn 17:62-66, 1986

[Document 7] Kim YH, et al. Studies on the chemical constituents of Acanthopanax koreanum. Arch Pharm Res 11:159-162, 1988

[Document 8] Kim YH, et al., Pimaradine diterpenes from Acanthopanax koreanum. J Nat Prod 51:1080-1082, 1988

[Document 9] Kim YH, et al., Diterpene glycoside form Acanthopanax koreanum . Kor J Pharmacogn 21:49-51, 1990

[Document 10] Choi HS, et al., Lupane glycosides from the leaves of Acanthopanax koreanum . Chem Pharm Bull 56:1613-1616, 2008

[Reference 11] Perry LM, et al., Medicinal Plants of East and Southeast Asia. MIT Press, Cambrige, p.41, 1980

[Document 12] Korean Patent Publication No. 10-2010-1183656

Obesity is a disease caused by the loss of energy balance in the human body due to genetic, environmental, and mental factors. According to the World Health Organization (WHO), as of 2008, 400 million adults worldwide were obese and 1.6 billion were overweight. , It is predicted that by 2015, this number will soar to 700 million and 2.3 billion, respectively, and about 23.4% of the world's population will become obese.

Increased body fat due to obesity causes complications such as diabetes, hyperlipidemia, high blood pressure, cardiovascular disease, stroke, cancer, depression, and so on, significantly lowering the quality of life and acting as a major cause of shortening lifespan. Prevention and the pursuit of healthy living' report points out obesity as a key cause of various diseases, disabilities and deaths (Hida K, et al., Visceral adipose tissue-derived serine protease inhibitor: a unique insulin-sensitizing. adipocytokine in obesity.Proc Natl Acad Sci US A. 2005;102:10610-5).

As such, obesity is steadily increasing in the number of patients due to changes in life and industrialization, and increases serious risks to health such as high blood pressure and diabetes, so the demand for obesity treatment is steadily increasing, and this trend is 5 to 10 of the total medical expenses. It is spreading rapidly in countries such as the United States, which are used in the field of obesity, but in reality there are no effective products on the market that can satisfy huge demands. As a commercial product, it is limited to appetite suppressants and liposuction inhibitors, which is not meeting expectations, and there are various side effects listed below.

Representative obesity treatments include phentermine (GSK), a psychotropic appetite suppressant, and orlistat (Roche), a fat absorption inhibitor. Phentermine is a psychotropic appetite suppressant, and if it is not effective within one month due to serious side effects of heart disease such as pulmonary arterial hypertension and arrhythmia when taken for a long period of time, it is prohibited to take it immediately, and taking it for more than 3 months is prohibited by regulations. In the case of overseas, it is known that most of the European countries except the US, Germany, Czech Republic and Denmark do not use psychotropic obesity treatment.

In the case of orlistat, it is known that the effect is less effective in Koreans with a low proportion of fat intake such as meat among the total calories intake, and side effects such as fat stool excreted without ingestion of fat, inhibition of absorption of fat-soluble vitamins, and severe liver damage have been reported. The safety question is being raised.

As obesity treatments approved by the FDA in 2012, belviq and qsymia, which are appetite suppressants, are leading the overseas market.Belviq, developed by U.S. Arena Pharmaceuticals, entered the domestic market through Ildong Pharmaceutical in the first half of 2015, but it has carcinogenic and attention deficit side effects. The controversy is still being discussed.

In addition, qsymia (phentermine + topiramate), Viverse's obesity treatment, has serious side effects such as the occurrence of deformities and an increase in heart rate.

As such, the biggest problem with obesity drugs currently used is that they have serious side effects. Although there was some effect on weight loss and maintenance, in most cases, the market was stopped due to side effects that occur during administration.

A representative case of obesity drugs discontinued because safety was not guaranteed is sibutramine (Abbott), which occupied 50% of the obesity market until 2009, and was marketed in 2010 due to serious side effects of cardiovascular diseases such as myocardial infarction, stroke, and heart disease. After stopping, he was expelled from the market.

Acomplia (Acomplia, ingredient name: rimonabant, Sanofi-Aventis), which was sold with permission in the UK in 2006 as a case of discontinuation of other developments, did not exceed the risk of depression and suicidal thoughts due to the effectiveness of obesity treatment, so the European Medicines Agency (EMA) in 2009 Merck's CB1 antagonistic obesity treatment drug test showed irritability and depression symptoms and gave up during the development stage.

As such, the use of existing obesity treatments is limited due to side effects, so the necessity and importance of obesity treatments with low side effects are emerging.

Rhus verniciflua is a deciduous tree belonging to the Anacardiaceae family, and contains ingredients such as urushiol, stellacyanin, glucuronic acid, and galactose, etc. It is known that it has a pharmacological effect of (Information Public Hyangyak Daejeon, Younglimsa, p382-384, 1998).

Eucommia ulmoides OLIV is a deciduous arboretum native to China belonging to the Eucommiaceae family.Dried root bark is referred to as Eucommia ulmoides OLIV, and its blood pressure lowering and diuretic effects are known. Ingredients are known to contain ingredients such as gupta-percha, alkaloid, pectin, jinbang, and resin (Information-exclusive Hyang-Yak Daejeon, Younglimsa, p605-606, 1998 year).

However, none of the above documents discloses or teaches the efficacy of a combination extract composed of poison ivy and chinensis as a treatment for obesity.

The present inventors have made intensive research efforts to develop substances that can effectively prevent or treat obesity and are safe for the human body, especially plant-derived substances, and confirmed that weight gain is excellently suppressed through an experiment to verify the efficacy of improving obesity in an obesity model caused by a fat diet. (Experimental Example 1), it was confirmed that the accumulation of fat in the cells and the accumulation of triglycerides and cholesterol were strongly inhibited in the serum lipid content measurement experiment (Experimental Example 2): Cells in the hepatocyte tissue staining method through Oil Red staining It was confirmed that the accumulation of fat within and the accumulation of triglycerides and cholesterol was strongly inhibited (Experimental Example 3); Through immuno-blotting assay, the expression of endoplasmic reticulum stress-related proteins is reduced, the expression of SREBP-1 and FAS is suppressed, the phosphorylation of mTORC1, P70S6K and 4EBP-1 is reduced, and the phosphorylation of AMPK is It was confirmed to increase (Experimental Example 4); It was confirmed that the activity of GSH, GPx, SOD, and CAT increased in a concentration-dependent manner to prevent oxidative damage to the liver due to a high fat diet by inhibiting lipid peroxide substances and maintaining the detoxification function of toxic substances in the body (Experimental Example 5). By doing, of the present invention By finding that the combination extract composed of poison ivy and chinensis is very effective in preventing or treating obesity, the present invention has been completed.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention and treatment of obesity, which contains a combination extract composed of poison ivy and headworm as an active ingredient.

In addition, the present invention provides a health functional food for the prevention and improvement of obesity, containing a combination extract consisting of lacquer tree and chinensis as an active ingredient.

The combination consisting of lacquer and champignons as defined herein is preferably, the weight-relative blending ratio of the individual extracts of dried lacquer and champignons is 1: 0.01 to 100 (w/w), preferably, 1: 0.5 to 50 (w/w) blended parts by weight, more preferably 1: blended parts by weight of 0.1 to 10 (w/w), even more preferably 1: blended parts by weight of 0.3 to 5 (w/w) , Most preferably 1: 1 to 3 (w/w).

The sumac as defined herein can be used as bark, dermis, wood, branch, root, branch bark, root bark, leaf, outpost, or a combination thereof, and headworm is bark, dermis, neck, branch, root, branch bark, root It is possible to use shells, leaves, outposts or combinations thereof.

The extract defined herein is water including purified water, alcohol, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, preferably water, or a mixed solvent of water and a lower alcohol having 1 to 4 carbon atoms, more preferably water Or 10 to 100% (v/v) ethanol or alcohol, even more preferably water or 20 to 80% (v/v) ethanol or extract soluble in alcohol.

A composition containing the extract of the present invention as an active ingredient contains 0.1 to 50% by weight of the extract based on the total weight of the composition.

As used herein, the term "prevention" refers to any action that suppresses or delays inflammation, allergy or asthma by administration of a composition comprising the extract.

In addition, the term "treatment" used in the present invention refers to any action in which symptoms of a disease are improved or advantageously changed by administration of a composition containing the extract.

The lacquer tree extract defined herein is a method prepared by the method of extracting lacquer tree extract from which toxicity is removed while maintaining the polyphenol component by low temperature drying and low temperature extraction described in Korean Patent Application No. 10-2018-0011627 developed by the present inventor. , Specifically,

(1) The first step of crushing the ground portion of the lacquer tree, preferably the neck and the bark, to prepare a crushed product in a size of 1 to 10 mm, preferably 3 to 5 mm: (2) The crushed product is 40 to 100°C, preferably 50 A second step of first igniting urushiol in the lacquer tree by drying with hot air at a temperature of 80° C. for 1 hour to 48 hours, preferably 6 hours to 12 hours to obtain a primary ignited crushed material; (3) 1 to 10 times the weight of the crushed material, preferably 3 to 8 times by weight, more preferably 4 to 6 times by weight of purified water is added to the crushed product in which urushiol is first ignited, and then 60 to 100°C, preferably Step 3 of obtaining a secondary fired extract concentrate by extraction heating at a temperature of 70 to 90° C. for 1 to 48 hours, preferably 6 to 12 hours: (4) The second fired hot water extract concentrate is -0.01 4 to obtain an extract concentrated to 3 to 10 times, preferably 4 to 8 times by removing residual volatile substances by vacuum concentration at a pressure in the range of Mpa to -0.15 Mpa, preferably -0.08 Mpa to -0.10 Mpa step; And, (5) drying the extract concentrate to finally maintain the polyphenol component and to prepare a poison ivy extract from which toxicity is removed.

The manufacturing method of the lacquer tree extract can be prepared by effectively removing urushiol, which is toxic to humans, using a low-temperature extraction method to prepare a lacquer tree extract that is safe and contains a large amount of a polyphenol component useful for the body.

In addition, the combination extract of the present invention can be prepared as follows.

Dried lacquer is prepared by the method of extracting lacquer extract from which toxicity is removed while maintaining the polyphenol component by low-temperature drying and low-temperature extraction described in Korean Patent Application No. 10-2018-0011627, specifically, (1) The first step of crushing the ground portion of the lacquer tree, preferably the neck and the bark, to prepare a crushed product in a size of 1 to 10 mm, preferably 3 to 5 mm: (2) The crushed product is 40 to 100°C, preferably 50 A second step of first igniting urushiol in the lacquer tree by drying with hot air at a temperature of 80° C. for 1 hour to 48 hours, preferably 6 hours to 12 hours to obtain a primary ignited crushed material; (3) 1 to 10 times the weight of the crushed material, preferably 3 to 8 times by weight, more preferably 4 to 6 times by weight of purified water is added to the crushed product in which urushiol is first ignited, and then 60 to 100°C, preferably Step 3 of obtaining a secondary ignited extract concentrate by extractive heating at a temperature of 70 to 90°C for 1 hour to 48 hours, preferably 6 to 12 hours: (4) The secondary ignited hot water extract is -0.01 Mpa To -0.15Mpa, preferably -0.08Mpa to -0.10Mpa in a process of vacuum concentration to remove residual volatile substances to obtain an extract concentrated 3 to 10 times, preferably 4 to 8 times ; And, (5) a fifth step of drying the extract concentrate to finally maintain the polyphenol component and prepare a poison ivy extract from which toxicity is removed. (6) After washing and shredding the dried headworm, water containing 1 to 20 times the weight of the dried material, preferably about 4 to 8 times the volume of purified water, lower carbon atoms such as methanol, ethanol, butanol, etc. A solvent selected from alcohol, alcohol, or a mixed solvent thereof, preferably water, or a mixed solvent of water and a lower alcohol having 1 to 4 carbon atoms, more preferably water or 10 to 100% (v/v) ethanol or alcohol, Even more preferably, water or 20 to 80% (v/v) ethanol or alcohol is mixed several times and then at 30 to 150°C, preferably 60 to 120°C for 1 hour to 48 hours, preferably 8 hours to 14 hours. A sixth step of repeatedly performing the ultrasonic extraction method, hot water extraction method, room temperature extraction method, or reflux extraction method, preferably reflux extraction method for about 1 to 20 times, preferably 2 to 10 times; (7) a seventh step of obtaining a Duchung extract obtained by filtering, concentrating under reduced pressure, and drying the extract obtained in step 6; (8) The dry weight relative mixing ratio of the individual dry extract powders of the 7th step is 1: 0.01 to 100 (w/w) by weight, preferably 1: 0.5 to 50 (w/w) by weight , More preferably, 1: 0.1 to 10 (w / w) of the blended parts by weight, even more preferably 1: 0.3 to 5 (w / w) of the blending weight, most preferably 1: 1 to 3 It is possible to obtain the combined extract of the present invention through the eighth step by blending in (w/w).

A composition containing the extract of the present invention as an active ingredient contains 0.1 to 50% by weight of the extract based on the total weight of the composition.

It was confirmed that weight gain was excellently suppressed through an experiment to verify the efficacy of improving obesity in an obesity model due to a fat diet for the combination extract according to the present invention for the combination extract prepared by the above manufacturing method (Experimental Example 1), In the serum lipid content measurement experiment, it was confirmed that the accumulation of intracellular fat and the accumulation of triglycerides and cholesterol were strongly inhibited (Experimental Example 2): In the experiment of hepatocyte tissue staining through Oil Red staining, the accumulation of intracellular fat and triglycerides It was confirmed that it strongly inhibits the accumulation of cholesterol and cholesterol (Experimental Example 3); Through immuno-blotting assay, the expression of endoplasmic reticulum stress-related proteins is reduced, the expression of SREBP-1 and FAS is suppressed, the phosphorylation of mTORC1, P70S6K and 4EBP-1 is reduced, and the phosphorylation of AMPK is It was confirmed to increase (Experimental Example 4); It was confirmed that the activity of GSH, GPx, SOD, and CAT increased in a concentration-dependent manner to prevent oxidative damage to the liver due to a high fat diet by inhibiting lipid peroxide substances and maintaining the detoxification function of toxic substances in the body (Experimental Example 5). By doing so, the combination extract composed of poison ivy and headworm of the present invention can be usefully used in the prevention or treatment of obesity.

Accordingly, the present invention provides a pharmaceutical composition for preventing or treating obesity and a health functional food containing a combination extract consisting of lacquer tree and chinensis obtained by the above manufacturing method as an active ingredient.

In addition, lacquer tree and headworm are medicinal materials that have been edible or used as herbal medicines for a long time, and the extract of the present invention also has no problems such as toxicity and side effects.

The pharmaceutical composition of the present invention contains 0.1 to 50% by weight of the extract based on the total weight of the composition.

The pharmaceutical composition comprising the extract of the present invention may further include an appropriate carrier, excipient, and diluent commonly used in the preparation of pharmaceutical compositions.

Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils.

Compositions containing the extract of the present invention are formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be used.

Specifically, in the case of formulation, it may be prepared using diluents or excipients such as generally used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and these solid preparations include at least one excipient, such as starch, calcium carbonate, and sucrose. ), lactose, and gelatin may be mixed and prepared.

In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations and suppositories. As the non-aqueous solvent and suspending agent, vegetable oils such as propylene glycol, polyethylene glycol and olive oil, and injectable esters such as ethyloleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin oil, glycerol, and the like can be used.

The preferred dosage of the extract of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for a desirable effect, the extract of the present invention may be administered in an amount of (0.0001 to 100) mg/kg, preferably (0.001 to 100) mg/kg, divided once or several times a day. In the composition, the extract of the present invention may be blended in an amount of (0.0001 to 50)% by weight based on the total weight of the composition.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be expected and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dura mater and intracere broventricular injection.

In addition, the present invention provides a health functional food for the prevention and improvement of obesity, containing a combination extract consisting of lacquer tree and chinensis as an active ingredient.

"Health functional food" as defined herein refers to a food manufactured and processed using raw materials or ingredients having useful functions for the human body according to the Health Functional Food Act No.6727, and the term "functional" It means ingestion for the purpose of obtaining useful effects for health purposes such as controlling nutrients for structure and function or for physiological effects.

The health functional food of the present invention comprises 0.01 to 95% of the extract, preferably 1 to 80% by weight, based on the total weight of the composition.

In addition, health functional foods in the form of pharmaceutical dosage forms such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, or tea bags, leached teas, health drinks, etc. for the purpose of preventing or improving diseases of the present invention It can be manufactured and processed.

In addition, the present invention provides a health supplement or food additive for preventing and improving obesity, comprising a combination extract composed of lacquer tree and chinensis as an active ingredient.

In addition, the above health functional food may additionally contain food additives, and the suitability as a "food additive" shall be determined according to the General Regulations of the Food Additive Code approved by the Food and Drug Administration and general test methods, etc. It is judged according to standards and standards.

Items listed in the "Food Additives Code", for example, chemical synthetic products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as red pigment, licorice extract, crystalline cellulose, and guar gum, L- Mixed preparations, such as a sodium glutamate preparation, an alkali additive for noodles, a preservative preparation, and a tar color preparation, are mentioned.

Functional foods containing the extract of the present invention include bread, rice cakes, dried fruits, candies, chocolates, chewing gum, confectionery such as jam, ice cream products such as ice cream, ice cream, ice cream powder, milk, low-fat milk, lactose-decomposed milk , Processed milk, goat milk, fermented milk, butter milk, concentrated milk, milk cream, butter oil, natural cheese, processed cheese, milk powder, dairy products such as whey products processed meat products, processed eggs, meat products such as hamburgers, fish cake, ham , Sausages, processed fish products such as bacon, etc. Ramen, dried noodles, raw noodles, milk-tang noodles, luxury dried noodles, improved soft noodles, frozen noodles, and noodles such as pasta Fruit drinks, vegetable drinks, carbonated drinks, soy milk , Lactobacillus beverages such as yogurt, beverages such as mixed beverages Soy sauce, soybean paste, red pepper paste, Chunjang, cheonggukjang, mixed sauce, vinegar, sauces, tomato ketchup, curry, seasoning foods such as curry, margarine, shortening and pizza. It is not limited.

The health functional beverage composition of the present invention is not particularly limited to other ingredients other than containing the extract as an essential ingredient in the indicated ratio, and may contain various flavoring agents or natural carbohydrates as an additional ingredient, such as a conventional beverage. Examples of the natural carbohydrates described above include monosaccharides, (eg, glucose, fructose, etc.); Disaccharides, (eg maltose, sucrose, etc.); And polysaccharides, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. have. The ratio of the natural carbohydrate is generally about (1 to 20) g, preferably about (5 to 12) g per 100 mL of the composition of the present invention.

In addition to the above, the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and heavy weight agents (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like may be contained. In addition, the compositions of the present invention may contain natural fruit juice and flesh for the production of fruit juice beverages and vegetable beverages. These components may be used independently or in combination. The proportion of these additives is not so critical, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

In addition, the extract of the present invention may be added to food or beverage for the purpose of preventing a target disease. At this time, the amount of the purified product or compound in the food or beverage may be added in an amount of 0.01 to 15% by weight of the total food weight, and the health beverage composition is 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 mL. It can be added at a rate.

In the process of manufacturing the health functional food, the content of the extract according to the present invention added to food including beverages may be appropriately added or subtracted as needed.

For the combined extract of the present invention, it was confirmed that weight gain was excellently suppressed through an experiment to verify the effect of improving obesity in an obesity model due to a fat diet (Experimental Example 1), and accumulation and neutralization of intracellular fat in a serum lipid content measurement experiment. It was confirmed that it strongly inhibits the accumulation of fat and cholesterol (Experimental Example 2): In the experiment of hepatocyte tissue staining through Oil Red staining, it was confirmed that the accumulation of intracellular fat and the accumulation of triglycerides and cholesterol were strongly inhibited ( Experimental Example 3); Through immuno-blotting assay, the expression of endoplasmic reticulum stress-related proteins is reduced, the expression of SREBP-1 and FAS is suppressed, the phosphorylation of mTORC1, P70S6K and 4EBP-1 is reduced, and the phosphorylation of AMPK is It was confirmed to increase (Experimental Example 4); It was confirmed that the activity of GSH, GPx, SOD, and CAT increased in a concentration-dependent manner to prevent oxidative damage to the liver due to a high fat diet by inhibiting lipid peroxide substances and maintaining the detoxification function of toxic substances in the body (Experimental Example 5). By doing so, the combination extract composed of poison ivy and chinensis of the present invention can be usefully used in the prevention or treatment of obesity.

1 is a diagram showing changes in body weight, dietary intake and fasting glucose levels for HFD-induced liver damage (experimental results are expressed as mean±SD obtained from three individual experiments):
Figure 2 is a diagram showing the changes in triglyceride, total cholesterol and LDL-cholesterol levels in liver (A) and serum (B) (experimental results are expressed as mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):
3 is a representative diagram confirming the fat accumulation of hepatocyte fragments through Oil Red staining. (Experimental results are expressed as mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P < 0.05 vs. HFD):
Figure 4 is a diagram showing the effect on the HFD-induced ER stress response ((A) immunoblotting using antibodies against anti-CHOP, p-PERK, PERK, p-eIF2α, eIF2α, and β-actin Quantitative analysis of antibody expression was performed using a given loaded control group, and the experimental results are expressed as mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):
5 is a diagram showing the effect on HFD-induced lipid biosynthesis ((A) immunoblotting was performed using antibodies against anti-SREBP-1, FAS, and β-actin, and quantitative analysis of antibody expression It was carried out using a given loaded control group, and the experimental results are expressed as mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):
6 is a diagram showing the effect on HFD-induced mTORC1 activity ((A) immunoblotting is anti-p-mTOR, mTOR, p-p70S6K, p70S6K, p-4EBP-1, and 4-EBP It was performed using an antibody against -1, and the quantitative analysis of antibody expression was performed using a given loaded control, and the experimental results are expressed as mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):
7 is a diagram showing the effect on HFD-induced AMPK activation ((A) immunoblotting was performed using antibodies against anti-p-AMPK and AMPK, and quantitative analysis of antibody expression was performed at a given loading Was performed using a control group, and the experimental results are expressed as the mean±SD obtained from three individual experiments, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):
Figure 8 is a diagram showing the effect on antioxidant activity (GSH (A), GPx (B), SOD (C), and CAT (D) activities were performed in HFD-induced rats, the experimental results are three individual experiments Denoted as mean±SD, ^# P <0.05 vs. Con, ^* P <0.05 vs. HFD):

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Reference Example 1: Preparation of lacquer tree extract by low temperature drying and low temperature extraction (Korean Patent Application No. 10-2018-0011627 Manufacturing Method)

The washed and dried lacquer top part was crushed into 3~5mm size (Rhus verniciflua, Imsil Cheese & Food Research Institute, Imsil-gun, Jeollabuk-do). Thereafter, hot air drying was performed at 50 to 80° C. for 6 to 12 hours, and urushiol was firstly ignited. Thereafter, 4-6 times (weight ratio) purified water was added to the dried lacquer chips, and hot water extraction was performed at 70-80°C for an average of 8 hours to prepare a secondary ignition extract concentrate. Thereafter, the extracted concentrate was concentrated under reduced pressure in the range of -0.08Mpa to -0.110Mpa to remove residual volatile substances to prepare an extract concentrated about 6 times. Then, the extract concentrate was lyophilized to obtain a powdery poison ivy extract (hereinafter referred to as ILF-RW).

Reference Example 2: Preparation of chinensis extract

After washing and drying the bark (Eucommia ulmoides, Imsil Cheese & Food Research Institute, located in Imsil-gun, Jeollabuk-do), about 8 times the amount of purified water is added to extract hot water at 85~90℃ for an average of 8 hours. The filtered extract was concentrated under reduced pressure and freeze-dried to obtain a Duchung extract. (Referred to as ILF-EW).

Example 1: Preparation of mixed extract

The lacquer tree extract and chinensis extract were completely mixed at a ratio of 1:1 (w/w) by weight to prepare a lacquer tree chinensis extract mixture (hereinafter referred to as “ILP-RE”), which was used as a sample of the following experimental example.

Experimental Example 1. Obesity improvement efficacy verification in obesity model due to high fat diet

1-1. Experiment method

Seven-week-old male SD rats (250 g) were purchased from Orient, and after being acclimated to normal solid feed for one week in an animal breeding facility environment, 8 rats were placed per experimental group, and 2 rats were separated and reared per cage. In order to induce obesity, an obesity induction experiment was conducted using general powdered food and high fat powdered food (60% high fat diet, calorie composition: crude fat (60.9%), protein 23.6%; 58Y1, Orient Bio) as a powder feed for experimental animals (Orient). Used for The experimental group consisted of a normal diet group (NCD), a poison lacquerworm extract mixture administration group (ILF-RE), a high fat diet group (HFD), a high fat diet group and a poison lacquer chinensis extract mixture administration group (HFD+ILF-RE) 40, 80 and 120 mg/kg. Divided (refer to Table 1), a high-fat diet was consumed for 8 weeks, and diet and water were freely ingested. The animal breeding room maintained a temperature of 20±2℃ and a relative humidity of 50±10%, and was controlled at intervals of 12 hours in a light and dark cycle.All animal experiments were conducted by the Institutional Animal Care and Use Committee (IACUC, cuh- IACUC-2018-9) was approved and conducted in compliance with the ethical standards for animal testing. (Table 1)

Group setting denomination Set Treatment group Control(NCD) Normal diet 8 of them HFD High fat diet. 8 of them HFD + ILF-RE40mg/kg ILF-RE 40mg/kg 8 of them HFD + ILF-RE80mg/kg ILF-RE 80mg/kg 8 of them HFD + ILF-RE120mg/kg ILF-RE 120mg/kg 8 of them ILF-RE120mg/kg ILF-RE 120mg/kg 8 of them

1-2. Weight measurement

The body weight and organ weight were measured once a week for 8 weeks, and the weight of the organ was measured after the liver was removed from the experimental animal whose abdomen was cut, washed with physiological saline, and water was removed with filter paper.

1-3. Fasting blood glucose measurement

Blood glucose levels were measured by ACCU-CHEK Active (Roche Diagnostics, Mannheim, Germany) after blood collection from the tail vein before sacrifice after maintaining fasting for more than 14 hours before measurement.

1-4. Changes in body weight, dietary intake and fasting blood sugar

Changes in body weight and dietary intake as shown in Table 2 and FIG. 1 showed that there was an increase in body weight due to a high fat diet during the experiment, but in the group treated with the high fat diet and the poison ivy chinensis extract mixture at the same time, the weight gain was 50.8%. It was confirmed that it was suppressed. (See Table 2 and Fig. 1)

Specifically, as a result of measuring the body weight of the experimental animals included in each group, 8 weeks later, compared to the animal group that consumed only high fat feed (576.2±41.23g, the animal group administered the example sample (oral, 80 mg/kg) And 120mg/kg) showed the weight gain inhibitory effect (515.2±15.62g, 519.2±26.5g) (p<0.05), and it was confirmed that the weight gain inhibitory effect of the example samples was excellent (see Table 3. )

Change in weight Test group/test period 0 wk 1 wk 2 wk 3 wk 4 wk 5 wk 6 wk Control(NCD) Mean 282.62 339.87 385.5 409. 438 456 460 S.D. 6.85 9.59 7.63 9.1 10.17 15.75 14.2 HFD Mean 278.37 351.87 416.75 464.12 500.37 534.37 576.2 S.D. 8.84 19.14 24.68 29.68 34.12 38.72 41.23 HFD + ILF-RE40mg/kg Mean 281.37 351 413. 456.37 493.12 526 542.3 S.D. 6.1 11.54 13.09 16.8 18.95 21.47 15.2 HFD + ILF-RE80mg/kg Mean 275.62 344.6 400.25 439.37 471.87 507.5 515.2 S.D. 6.55 14.18 20.85 23.03 29.36 33.21 15.62 HFD + ILF-RE120mg/kg Mean 281.87 351.62 408.37 444.12 481.25 511.87 519.2 S.D. 10.30 22.05 29.16 33.25 38.00 43.20 26.5 ILF-RE120mg/kg Mean 281.75 339.37 376.37 405.87 429.5 447.12 454.23 S.D. 8.16 15.4 18.41 20.07 24.76 27.67 24.23 ^# p<0.05 versus control, *p<0.05, compared between HFD group

Changes in fasting blood sugar Test group/analysis method Fasting blood sugar (mg/dl) Control(NCD) Mean 84 S.D. 5.19 HFD Mean 87.12 S.D. 7.32 HFD + ILF-RE40mg/kg Mean 97.12 S.D. 5.27 HFD + ILF-RE80mg/kg Mean 94.37 S.D. 11.56 HFD + ILF-RE120mg/kg Mean 95.5 S.D. 6.6 ILF-RE120mg/kg Mean 91.37 S.D. 9.6

In addition, as a result of measuring blood sugar on an empty stomach to determine the presence or absence of metabolic abnormalities due to a high-fat diet, it was confirmed that there was no difference in fasting blood sugar in the group treated with a high-fat diet, a high-fat diet, and a poison ivy chinensis extract. (See Table 3 and Fig. 1)

Experimental Example 2. Serum lipid content measurement experiment

2-1. Experimental process

After breeding was completed, the experimental animal of Experimental Example 1 was fasted for 14 hours, anesthetized with ketamine (8001,d Yuhan Corporation), and blood was collected from liver tissue and abdominal artery.

The collected blood was allowed to stand at room temperature for 30 minutes and then centrifuged at 3000 rpm for 10 minutes with a centrifuge (5430, Eppendrof) to obtain and analyze serum.

Asancet triglyceride measurement reagent (AM-157S-K, Asan Pharmaceutical Co., Ltd.) and cholesterol measurement reagent (AM 202-K, Asan Pharmaceutical Co., Ltd.) were used to measure liver tissue and blood. Liver tissue and serum were placed in a reagent for measuring triglyceride and cholesterol, and reacted at 37°C. Each sample was quantified and transferred to a 94 well plate for measurement using an ELISA device.

2-2. Experiment result

In the high-fat diet group, the accumulation of triglycerides and cholesterol in liver tissues and the secretion of triglycerides and cholesterol in serum were increased (see Fig. 2 and Table 4-Table 5).

Fat analysis in liver tissue Test group/analysis method Triglycerides (mg/g) Total cholesterol (mg/g) LDL-cholesterol (mg/g) Control(NCD) Mean 54.36 49.06 60.0 S.D. 5.8 7.4 5.4 HFD Mean 124.45 197.62 115 S.D. 4.6 4.7 4.5 HFD + ILF-RE40mg/kg Mean 103.41 162.42 94 S.D. 4.6 5.4 4.8 HFD + ILF-RE80mg/kg Mean 93.64 133.14 84 S.D. 7.4 5.1 6.1 HFD + ILF-RE120mg/kg Mean 88.97 100 80 S.D. 6.2 7.6 4.9 ILF-RE120mg/kg Mean 51.85 51.20 56 S.D. 7.7 11.09 4.9 ^# p<0.05 versus control, *p<0.05, compared between HFD group

Analysis of fat in plasma Test group/analysis method Triglycerides (mg/g) Total cholesterol (mg/g) LDL-cholesterol (mg/g) Control(NCD) Mean 70.87 46.86 57.0 S.D. 6.5 7.92 6.8 HFD Mean 154 179.86 97 S.D. 7.1 6.7 8.1 HFD + ILF-RE40mg/kg Mean 142 154.50 95.00 S.D. 6.9 8.1 7.2 HFD + ILF-RE80mg/kg Mean 137 123.18 84 S.D. 4.9 6.4 5.90 HFD + ILF-RE120mg/kg Mean 115 90.54 74.00 S.D. 8.1 7.6 6.1 ILF-RE120mg/kg Mean 77.04 46.80 53 S.D. 6.1 8.69 5.9 ^# p<0.05 versus control, *p<0.05, compared between HFD group

Experimental Example 3. Adipose tissue accumulation confirmation experiment

3-1. Triglyceride accumulation confirmation experiment

To check the accumulation of triglycerides in hepatocytes, after fixing with formalin for 20 minutes, washing twice with PBS (Phosphate Buffer Saline), staining with oil red O stain (O9755, Sigma) for 30 minutes, and then using an inverted microscope ( Carl Zeiss, Germany) observed the accumulation of triglycerides in the cells.

3-2. Tissue immunostaining

After fixing the tissue with formalin, paraffin formalin and formalin fixation are performed by diluting the anti-4-HNE antibody (ab46545,abcam), the primary related protein for the protein to be analyzed, to a concentration of 1/500 after deparaffinization and rehydration. Treated on a slide glass and reacted at room temperature for 1 hour. After washing with TBS (Tris-Buffer saline) solution three times, repeating three times with TBS, washing again, adding Streptavidin-peroxidase-containing solution (P039701-2, Dako) and reacting for 15 minutes, followed by TBS ( Tris-Buffer saline) was repeated three times and washed again. Newly prepared substrate solution (3mg of 3-amino-9-ethylcarbazole in 10mL of sodium acetate buffer, pH 4.9, 500 μL of dimethylformamide and 0.03% hydrogen peroxide (K3461, Dako) was added, and color development was observed for 5-30 minutes, Color development was controlled using distilled water.

The nuclei were stained with a dyeing reagent (Mayer'hematoxylin:), washed three times with distilled water for 5 minutes, mounted with a culture solution (Cytomation aqueous mounting medium), air-dried, and observed with an optical microscope (Carl Zeiss, Germany) DHE (Dihydroethidium, Germany). ;D11347, molecular probe) staining is performed by diluting DHE (100 mg/mL) in DHE (100 mg/mL) 1:1500 for 10 minutes at 37° C. and then washing the slide twice with PBS under a fluorescence microscope (Carl Zeiss, Germany). ) Was observed.

3-3. Experiment result

Fat accumulation in hepatocytes was confirmed through Oil Red staining. It was confirmed that the group administered with the high-fat diet and the extract mixture of poison ivy chinensis strongly inhibited the accumulation of intracellular fat and the accumulation of triglycerides and cholesterol (Fig. 3).

Experimental Example 4. Immunoblotting experiment

4-1. Immuno-blotting assay

The hepatocyte homogenate was separated by SDS-PAGE and transferred to a PVDF membrane. After blocking with 5% skim milk, CHOP, p-eIF2α, p-mTOR, mTOR, p-P70S6K, P70S6K, p-4EBP-1, 4EBP-1, p-AMPK, or AMPK (Cell Signaling Technologies, Inc. , Danvers, MA, USA) and p-PERK, PERK, eIF2α, SREBP-1, FAS, or b-actin (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA).

4-2. Suppression of endoplasmic reticulum stress in liver tissue

As a result of confirming the expression of endoplasmic reticulum stress due to a high fat diet, in the high fat diet group, the expression of endoplasmic reticulum stresses P-PERK, p-eIF2α, and CHOP increased, but in the group administered simultaneously with a high fat diet and lacquerworm extract mixture, endoplasmic reticulum stress It was confirmed that the expression of related proteins was decreased. (Fig. 4)

4-3. Inhibition of fat synthesis in liver tissue

As shown in FIG. 5, the expression of SREBP-1 and FAS, which are related proteins related to fat synthesis, increases in the high-fat diet alone group, but SREBP-1 in the group administered with the high-fat diet and the lacquerworm extract mixture in a concentration-dependent manner. , It was confirmed that the expression of FAS is suppressed. This confirmed that the synthesis of fat due to the high-fat diet was protected by the lacquer chinensis extract mixture (Fig. 5).

4-4. Regulation of mTORC1 activity through ER stress induced by a high fat diet

mTOR is an essential signaling pathway that detects intracellular nutritional status. The activity of mTOR1 due to a high fat diet phosphorylates downstream P70S6K and 4EBP-1 (Hida K, et al., Visceral adipose tissue-derived serine protease inhibitor: a unique insulin-sensitizing adipocytokine in obesity.Proc Natl Acad Sci USA. 102, pp. 10610-5, 2005).

As shown in Figure 6, in the high fat diet group, the phosphorylation of mTORC1 and the lower signal transducers P70S6K and 4EBP-1 increased, but the phosphorylation of mTORC1, P70S6K and 4EBP-1 in the group administered simultaneously with the high fat diet group and the poison ivy chinensis extract mixture It was confirmed that is decreased.

This means that the activity of mTORC1 is related to endoplasmic reticulum stress and intracellular fat accumulation due to a high fat diet, and the sumac cephalophyllum extract mixture regulates the activity of mTORC1 to inhibit endoplasmic reticulum stress and intracellular fat accumulation (Fig. 6).

4-5. Regulation of AMPK activation

AMP-activated protein kinase (AMPK) is a type of serine/threonine kinase that is activated when ATP is deficient and increases the production of intracellular energy. Activated AMPK reduces the concentration of malonyl-CoA by inhibiting ACC activity in cells. Malonyl-CoA is a substance that inhibits carnitine palmitoyltransferase-1 (CPT-1), a rate limiting enzyme in the introduction of fatty acids into the mitochondria. When the concentration of malonyl-CoA decreases through AMPK activation, CPT-1 is produced. Inhibition of the reaction is reduced, resulting in increased fatty acid oxidation. (2).

As shown in FIG. 7, in order to find out whether the lacquerworm extract mixture activates AMPK in liver tissue, phosphorylation of AMPK decreases in the high-fat diet group, but in the group to which the high-fat diet and the lacquerworm extract mixture were dose-dependently administered simultaneously. It was confirmed that phosphorylation of AMPK was increased. (Fig. 7)

Experimental Example 5. Antioxidant Effect Experiment

5-1. Measurement of superoxide production in tissue

In order to measure the superoxide produced in liver tissue, an experiment was performed as follows using dihydroethidium (DHE) by applying the method described in the literature (3).

After adding 10 μM dihydroethidium (Dihydroethidium; D11347, molecular probe) to the tissue and staining for 30 minutes at 37°C, the slide was washed twice with PBS (Phosphate Buffer Saline), and then tissue using a microscope (Carl Zeiss, Germany). My superoxide production was measured.

5-2. Experiment result

As shown in FIG. 8, GSH, GPx, SOD and CAT activities were observed in order to find out whether the increase in oxidative stress caused by the high fat diet protects liver toxicity through antioxidant activity caused by the poison ivy chinensis extract mixture.

In the high-fat diet group, GSH, GPx, SOD and CAT activities were significantly decreased, but in the group administered with the high-fat diet and Succulent chinensis extract mixture, GSH, GPx, SOD, and CAT activities were increased in a concentration-dependent manner.

Therefore, this study demonstrates that the poison lacquerworm extract mixture has antioxidant activity in liver toxicity induced by a high fat diet, thus inhibiting lipid peroxides and maintaining the detoxification function of toxic substances in the body, thereby protecting the liver from oxidative damage caused by a high fat diet. Confirmed. (Fig. 8)

5-3. Statistical analysis

Values presented are expressed as mean ± SD. Statistical significance of the differences between treatment groups was determined by one-way ANOVA using Graph Pad Prism 4.0 (San Diego, CA).

Hereinafter, examples of the preparation of the composition containing the herbal extract of the present invention will be described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

ILF-RE 20 mg

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablet

ILF-RE 10 mg

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3. Preparation of Capsule

ILF-RE 10 mg

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

ILF-RE 10 mg

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na2HPO4,12H2O 26 mg

It is prepared with the above ingredients per ampoule (2 ml) according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

ILF-RE 20 mg

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid preparation, each ingredient is added to the purified water to dissolve it, added lemon scent, mixed with the above ingredients, and then adjusted to 100 ml by adding purified water, then filled in a brown bottle and sterilized to prepare a solution. do.

Formulation Example 6. Manufacture of health food

ILF-RE 1000 mg

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

Nicotinic acid amide 1.7 mg

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

Ferrous sulfate 1.75 mg

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

Potassium citrate 90 mg

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for health food, but it may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. , To prepare granules, and can be used to prepare a health food composition according to a conventional method.

Formulation Example 7. Preparation of healthy beverage

ILF-RE 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 ml by adding purified water

After mixing the above ingredients according to a conventional health drink manufacturing method, stirring and heating at 85° C. for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 liter container, sealed and sterilized, and then stored in a refrigerator. It is used in the manufacture of health beverage composition.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the purpose of use.

Claims (10)

A pharmaceutical composition for the prevention or treatment of obesity, which contains a combination extract consisting of a combination weight of 1: 0.01 to 100 (w/w) of individual extracts of the above-ground part of lacquer tree and the bark of lacquer. delete delete The pharmaceutical composition according to claim 1, wherein the extract is an extract soluble in water including purified water, alcohol, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Health functional food for the prevention or improvement of obesity containing a combination extract consisting of a weight-relative mixing ratio of the individual extracts of the above-ground part of lacquer tree and the bark of lacquer: 1: 0.01 to 100 (w/w) as an active ingredient. delete delete The health functional food according to claim 5, wherein the extract is an extract soluble in water including purified water, alcohol, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. A health supplement for preventing or improving obesity, containing a combination extract consisting of a combination weight of 1: 0.01 to 100 (w/w) by weight of the individual extracts of the above-ground part of the lacquer tree and the bark of lacquer. delete

Images