KR102371741B1 - Composition for Anti-obesity Using an Extract of Bistorta manshuriensis - Google Patents

Abstract

The present invention discloses a composition for anti-obesity using an extract of scallop tail having an activity to significantly inhibit the differentiation of 3T3-L1 mouse precursor adipocytes into adipocytes and fat accumulation.

Description

Composition for Anti-obesity Using an Extract of Bistorta manshuriensis

The present invention relates to a composition for anti-obesity using an extract of Beomtail ( Bistorta manshuriensis ).

In modern society, the obese population is increasing due to the convenient living environment according to the rapid automation, the excessive nutrition intake and the decrease in the amount of physical activity due to the increase of processed food and eating out. The World Health Organization (WHO) classifies obesity as a disease rather than a phenomenon or symptom, and reported that in 2015, the number of obese people worldwide will increase to 1.5 billion and become a serious health problem (Ann Epidemiol. 2005. 15:87-97; J Life Sci. 2013. 23:69-78). Although the cause of obesity is not clearly known, it is not caused by a single condition, but is caused by various causes such as improper diet and lifestyle, genetic factors, and decrease in physical activity (.Korean J. Food Science Nutrition 22(1) ) 110-12.2009; Korean J. Food Science Nutrition 23(5):363-367.1990).

Obesity is an increase in the number and size of fat cells due to excessive accumulation of body fat due to an imbalance between energy intake and consumption (Cell.104:531-543 2001). When the energy source is depleted after being stored, the stored fat is decomposed into free fatty acids and glycerol to be used as an energy source. (Metabolism. 30: 425-427 1981; Biochem J. 1;435(3):723-32 2011). Obesity acts as a risk factor that increases the incidence of various diseases as well as changes in body type due to the accumulation of fat in the intestines and abdomen (Korean J. Pediatr.48:126-137. 2005) When visceral fat accumulates excessively, Problems arise in metabolism, and symptoms such as abnormal secretion of hormones and abnormal secretion of cytokines occur. Due to obesity, an increase in triglyceride, LDL-cholesterol, and a decrease in HDL-cholesterol cause abnormal fat metabolism in the body, decrease insulin receptors in tissues, and also decrease insulin sensitivity, thereby inhibiting the transport of glucose into cells. It can also lead to high blood sugar and diabetes. In addition, obesity is known to be closely related to the occurrence of metabolic diseases such as hyperlipidemia, cardiovascular disease, cancer, respiratory disorders, stroke, and osteoarthritis (Med. Int. 22 385-388 1994;. Ann. Intern. Med 103:1024-1029 1985).

Physiological regulation of adipocytes can be largely divided into induction of adipocyte differentiation, fat biosynthesis, and lipid degradation. During the differentiation stage of adipocytes, expression of various transcription factors such as peroxisome proliferatoractivated receptor (PPAR), CCAAT/enhancer binding proteins (C/EBP), and sterol regulatory element binding protein (SREBP) is induced step by step in adipocytes undergoing differentiation. , regulates the expression of various adipocyte-specific genes through the interaction of each factor (Genes Dev. 14:1293-1307. 2000). Adipocytes that have completed differentiation undergo triglyceride biosynthesis using the fatty acids and glucose introduced into them. The enzymes involved in this are FAS (fatty acid synthase), ACC (acetyl-CoA carboxylase), SCD (stearoyl-CoAdesaturase), and ACS. (acyl-CoA synthetase), diacylglycerol acyltransferase (DGAT), and the like (Trends Endocrinol. Metab., 23, 56-64. 2011; Biochem. Biophys. Res. Commun., 420,805-810. 2012). The triglyceride synthesized by the action of these enzymes is used as an energy source. Lipid degradation is regulated by catecholamines and insulin according to nutritional status. Enzymes involved in lipid degradation include ATGL (adipose triglyceride lipase), HSL (hormone sensitive lipase), and MGL (monoacyl glycerol lipase). It is divided into three molecules (Trends Endocrinol. Metab., 23, 56-64. 2011; Biochem. Biophys. Res. Commun., 420,805-810. 2012). Fatty acids produced by hydrolysis are oxidized in mitochondria and consumed as energy or esterified and synthesized as triglycerides and stored in cells.

3T3-L1 adipocytes were first isolated from 3T3 cells by Green and Meuth (Cell. 3(2): 127-33. 1974), and their biological properties and the ability to differentiate into adipocytes under appropriate culture conditions were revealed. It is widely used to conduct research on the differentiation process of post-adipocytes and the decomposition of accumulated fat. 3T3-L1 cells, which are preadipocytes, are differentiated into mature adipocytes by various hormones, PPAR, C/EBP, and SREBP, and accumulate intracellular triglycerides due to increased expression of related genes and related enzyme activity ( J Nutr 130: 3116S-3121S, 2000; J Nutr 130: 3122S-3126S, 2000). For functional material development research, a method of searching for materials that inhibit the differentiation process or promote lipolysis using 3T3-L1 cells as described above with the aim of inhibiting excessive accumulation of triglycerides in adipose tissue is widely used.

Existing anti-obesity drugs such as orlistat and sibutramine are known to have serious side effects such as vomiting, constipation, gastrointestinal disorders, and cardiovascular disease (Int J Obes Relat Metab Disord. Jul;25(7)). :1095-9. 2001; Obes Res. 8(6):431-7. 2000; Lancet. 6;369(9555):71-7. 2007;Front Physiol. 2014 Jun 24;5:228. 2014); Efforts to develop effective and safe substances are ongoing. It has been reported that retinol, vitamin E, vitamin U, and extracts of Japanese peppercorns have a function of inhibiting the differentiation of adipocytes (Mol Cell Biol. 16:15671575. 1996; Ann Dermatol. Feb;24(1):39- 44 2012; J Nutr. 139(1):51-7 2009; Ann Dermatol. 24(1):39-44 2012) Research on the development of safe and sustainable natural substances for anti-obesity drugs is being actively conducted. .

The present invention discloses the anti-obesity use of a pantail extract.

It is an object of the present invention to provide a composition for anti-obesity using a pantail extract.

Other objects or specific objects of the present invention will be set forth below.

As confirmed in the Examples and Experimental Examples below, the present invention has been completed by confirming that the extract of the scallop tail significantly inhibits the differentiation of 3T3-L1 mouse precursor adipocytes into adipocytes and the accumulation of fat.

Considering the experimental results of the above bar, the composition for anti-obesity of the present invention is characterized in that it contains a pantail extract as an active ingredient.

In the present specification, the term "pantail extract" refers to an extraction target of a pantail leaf, stem, above-ground part, rhizome, root, underground part, outpost, or a mixture thereof with water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.), methylene chloride , ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof means an extract obtained by leaching using , an extract obtained by using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract. , reflux, warming, ultrasonic radiation, supercritical extraction, etc. can be applied any method. In the case of the fractionated extract, the fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent has been removed by methods such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, an extract obtained by using water, an alcohol having 1 to 4 carbon atoms or a mixed solvent thereof as an extraction solvent, more preferably an extract obtained by using a mixed solvent of water and an alcohol having 1 to 4 carbon atoms as an extraction solvent do.

In addition, as used herein, the term "active ingredient" refers to a component that can exhibit a desired activity alone or can exhibit activity together with a carrier that is not active by itself.

Also, in the present specification, "anti-obesity" means a reduction in body fat, inhibition of body fat accumulation, and/or a reduction in body weight. Therefore, it is meant to include prevention of obesity and treatment of obesity, and further includes reducing body weight/body fat for beauty or health purposes (aka diet purposes), although the weight is not classified as obesity or overweight.

Also, in the present specification, the "obesity" means a state in which adipose tissue is abnormally increased, whether it is obesity due to genetic factors or obesity due to environmental factors, and according to the classification of body mass index (BMI), high It is meant to include obesity (when BMI is 30.0 or higher), obesity (when BMI is 25-30), and overweight (when BMI is 23-25).

The composition for anti-obesity of the present invention may be included in any amount (effective amount) according to the use, formulation, purpose of mixing, etc., as long as the active ingredient can exhibit anti-obesity activity, and a typical effective amount is based on the total weight of the composition. It will be determined within the range of 0.001 wt % to 20.0 wt %. As used herein, the term "effective amount" refers to the intended functional and pharmacological effects such as anti-obesity effect when the composition of the present invention is administered to a mammal, preferably a human subject to which it is applied, during the administration period as suggested by a medical professional. Refers to the amount of the active ingredient contained in the composition of the present invention. Such an effective amount can be determined empirically within the ordinary ability of one of ordinary skill in the art.

In addition to the active ingredient, the anti-obesity composition of the present invention has already been proven in the art for safety and in order to enhance the convenience of taking or ingestion through the addition of similar activities such as blood pressure control activity or increase/reinforcement of the anti-obesity effect. It may further include any compound or natural extract known to have the corresponding activity. Such compounds or extracts include compounds or extracts, drugs listed in compendial documents such as pharmacopeias of each country (“Korea Pharmacopoeia” in Korea) and health functional food regulations of each country (“Health Functional Food Standards and Specifications” announced by the Ministry of Food and Drug Safety in Korea). Each country's laws governing the manufacture and sale of compounds or extracts and health functional foods that have been approved for items in accordance with the laws of each country (the "Pharmaceuticals Act" in Korea) governing the manufacture and sale of '), compounds or extracts whose functionality is recognized are included.

For example, garcinia cambogia bark extract, conjugated linolenic acid (free fatty acid), conjugated linolenic acid (triglyceride), green tea extract, chitosan, Lactobacillus gasseri, whose functionality has been recognized as 'body fat reduction' according to the Korean 「Health Functional Food Act」 BNR17 ( Lactobacillus gasseri BNR17), L-carnitine tartrate, green mate extract, green coffee bean extract, sesame leaf extract, soybean germ extract, etc., mixed powder of aloe vera extract powder, lactoferrin (milk refined protein), lemon balm extract , Mate hot water extract, seaweed complex extract (xantijen), fermented vinegar pomegranate complex, Puer’s tea extract, Seomoktae (snow bean) peptide complex, vegetable oil diglyceride, wild mango seed extract, medium-chain fatty acid (MCFA)-containing oil, Complex extracts such as Coleus forskohlii extract, chitooligosaccharide, finger root extract powder, hibiscus, etc., L-glutamic acid-derived GABA-containing powder, Katsuobushi oligopeptide, Nattobacterium culture powder, Seomoktae ( Peptide complex, salmon peptide, olive leaf extract, sardine peptide, casein hydrolyzate, coenzyme Q10, grape seed enzyme-decomposed extract powder, Haitai oligopeptide, etc. Decomposition product, indigestible maltodextrin, bamboo leaf extract, vegetable oil diglyceride, refined sardine refined fish oil, refined squid oil, etc., L-arabinose, nopal extract, cinnamon extract powder, guava leaf extract, whose functionality has been recognized as 'glycemic control', Indigestible maltodextrin, freeze-dried silkworm powder, hemp spirit extract, Banaba leaf extract, upper leaf extract, etc., fermented amino acid complex whose functionality has been recognized as 'relieving fatigue', safflower extract, rhododendron extract, etc., as 'anti-stress' L-theanine, ashwagandha extract, milk protein hydrolyzate, and leaf extract, etc., which have been recognized for their functionality, may correspond to these compounds or extracts.

In a specific embodiment, the composition of the present invention may be identified as a food composition.

The food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, breads, sweets, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be manufactured.

In addition, the food composition of the present invention may have any product classification as long as it conforms to the enforcement laws at the time of manufacturing and distribution in legal and functional classification. For example, it is a health functional food according to the Health Functional Food Act of Korea, or confectionery, beans, tea, and beverages according to each food type according to the Food Ordinance of the Korea Food Sanitation Act (“Food Standards and Specifications” announced by the Ministry of Food and Drug Safety). , special purpose food, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives can be generally understood as substances that are mixed or infiltrated with food in manufacturing, processing, or preserving food. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Ordinance in accordance with the laws of each country that regulates the manufacture and distribution of food (“Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Specifications” notified by the Ministry of Food and Drug Safety), food additives are classified into chemically synthetic products, natural additives, and mixed preparations in terms of ingredients. It is classified into an agent, a preservative, an emulsifier, an acidulant, and a thickener.

The sweetener is used to impart appropriate sweetness to food, and both natural and synthetic ones may be used in the food composition of the present invention. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents are used for the purpose of improving taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural product, the purpose of nutritional enhancement in addition to flavor may be concurrently used. The natural flavoring agent may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or obtained from green tea leaves, horseradish leaves, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. In some cases, a synthetic flavoring agent may be used, and the synthetic flavoring agent may include esters, alcohols, aldehydes, terpenes, and the like.

As a preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, and pectin can be used. acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like may be used as acidulants. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste. As a thickening agent, a suspending agent, a settling agent, a gel-forming agent, a bulking agent, etc. can be used.

The food composition of the present invention may contain, in addition to the food additives as described above, bioactive substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as food additives.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Minerals include calcium preparations such as calcium citrate, magnesium stearate Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.

In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the product type.

In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Ordinance of each country or the Food Additives Code.

The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention may be prepared as an oral dosage form or parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of a combination of two or more routes is a case in which two or more formulations of drugs according to the route of administration are combined. For example, one drug is first administered by an intravenous route and the other drug is secondarily administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopoeia of each country including the "Korea Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, according to methods known in the art together with a suitable carrier It can be prepared in a formulation such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, grease Serol etc. are mentioned. In the case of formulation activity, an appropriate binder, lubricant, disintegrant, colorant, diluent, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch ferrist, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and oleic acid as lubricant. sodium, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, magnesium salts and calcium salts thereof, polyethylene glycol, etc., and the disintegrant is starch, methyl cellulose , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine.

When the pharmaceutical composition of the present invention is prepared for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or isotonic solution such as 5% dextrose may be used. . When formulated for transdermal administration, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pasta agents, liniment agents, air rolls, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like can be used.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.

A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and the route of administration. It can be in the range /kg. Administration may be performed once or divided into several times a day. Such dosages should not be construed as limiting the scope of the invention in any respect.

As described above, according to the present invention, it is possible to provide a composition for anti-obesity using a pantail extract. The composition for anti-obesity of the present invention may be commercialized as a functional food, drug, or the like.

1 is a photomicrograph of 3T3-L1, a mouse precursor cell, treated with a pantail extract by concentration, and then stained with Oil Red O reagent and the result of quantifying the amount of fat in the cells.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

< Example > Preparation of pantail extract

After adding 10 times the weight of 70% ethanol to the dry powder of the beomtail (extraction part, whole plant), leaching at room temperature for 24 hours, filtering with Whatman paper filter paper No. 2, and then concentrating the filtrate under reduced pressure and freezing It was dried to prepare a solid pantail extract.

< Experimental example > Obesity improvement activity test

Mouse progenitor adipocytes, 3T3-L1 (ATCC CL-173), were cultured at 37° C. and 5% CO ₂ in 10% BCS DMEM medium. 3T3-L1 precursor adipocytes were seeded in a 24-well plate with a cell number of 5×10 ⁴ /well, and then maintained for 2 more days when 100% confluency reached. Preadipocytes were differentiated into adipocytes with 10% FBS DMEM medium containing MDI (0.5 mM 3-isobutyl-1-methylxanthine (Calbiochem 410957), 1 uM dexamethasone (Calbiochem 265005), 1 ug/ml insulin (Sigma I9278)). was induced for 2 days, and after 48 hours of incubation, it was cultured for 2 days with 10% FBS DMEM containing 1 ug/ml insulin. After that, every 2 days, the culture medium was replaced with 10% FBS DMEM for 4 days. During the induction of adipocyte differentiation, the extracts of Examples were treated in each culture medium at a concentration of 50 and 100 μg/ml, and the degree of adipocyte differentiation was observed on the 8th day, when differentiation was completed. The degree of adipocyte differentiation was primarily confirmed through a microscope through Oil Red O staining, and the degree of adipocyte staining was measured by measuring the amount of differentiated fat in the cells using an ELISA Reader (SPECTRAmax 340PC, Molecular Devices, USA) at 510 nm absorbance. did. The results were calculated through three repeated experiments for each concentration.

The results are shown in FIG. 1 . Referring to FIG. 1 , it can be seen that the pantail extract clearly has the activity of inhibiting the differentiation of preadipocytes into adipocytes and the activity of inhibiting the accumulation of fat (IC ₅₀ =82.9 μg/mL).

Claims (4)

A food composition for anti-obesity comprising the water and ethanol mixed solvent extract of Beomtail ( Bistorta manshuriensis ) as an active ingredient.
The method of claim 1,
The extract is a food composition for anti-obesity, characterized in that the 70% ethanol extract of the pantail whole plant.
A pharmaceutical composition for anti-obesity comprising an extract of a mixed solvent of water and ethanol of Beomtail ( Bistorta manshuriensis ) as an active ingredient.
4. The method of claim 3,
The extract is a pharmaceutical composition for anti-obesity, characterized in that it is a 70% ethanol extract of the pantail whole plant.

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