KR20210073055A - Composition for Anti-obesity Using a hot water Extract of Antigonon leptopus - Google Patents

Abstract

The present invention discloses an anti-obesity composition using an Antigonon leptopus extract having an activity to significantly inhibit the differentiation of 3T3-L1 mouse precursor adipocytes into adipocytes and fat accumulation. The anti-obesity composition of the present invention can be commercialized as a functional food, drug, or the like.

Description

Composition for Anti-obesity Using a hot water Extract of Antigonon leptopus

The present invention relates to a composition for anti-obesity using an antigonon leptopus hot water extract.

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, but excessive energy intake promotes the differentiation of fat cells and increases the amount of fat stored in the body, which is a direct cause of obesity. (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 lead to 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), DGAT (diacylglycerol acyltransferase), 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.

Adipocytes, 3T3-L1, 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 the increase in the 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 peppermint trees 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. .

Antigonon leptopus (Siraea prunifolia for. simpliciflora Nakai) is a medicinal plant belonging to the order Rosaceae. It is widely distributed in domestic production areas nationwide and grows well in sunny places such as mountain foot or field. In general, in traditional medicine, the root is used as a medicine, and it can be used for antipyretic, vascular tissue contraction, and branch astringent action. It can also be used for fever caused by cold, neuralgia, sore throat, sore throat (a symptom of chills and fever rising at a certain time every day), diarrhea, etc.

Nevertheless, studies on pharmacological components and uses for Antigonon Leptopus have not been conducted much, so the application use for Antigonone Leptopus is not wide.

Specifically, in Korean Patent Registration No. 10-1176526, the anti-aging effect of antigonon leptopus extract through the antioxidant effect, skin wrinkle improvement effect, skin whitening effect, and skin damage suppression effect have been known. There is no known research on it.

The present invention discloses the anti-obesity use of an antigonone leptopus extract.

An object of the present invention is to provide a composition for anti-obesity using the antigonon leptopus hot water 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 was completed by confirming that the antigonon leptopus hot water extract clearly inhibited the differentiation of 3T3-L1 mouse precursor adipocytes into adipocytes and fat accumulation.

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

As used herein, the term "antigonon leptopus extract" refers to the extraction target of antigonon leptopus leaves, stems, above-ground parts, rhizomes, roots, underground parts, outposts, or mixtures thereof with water, lower alcohols 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 It refers to an extract obtained by leaching using glycol or a mixed solvent thereof, an extract obtained by using a supercritical extraction solvent such as carbon dioxide, pentane, or a fraction obtained by fractionating the extract, and the extraction method includes the polarity of the active material, the degree of extraction, In consideration of the degree of preservation, any method such as chilling, reflux, warming, ultrasonic radiation, or supercritical extraction may be applied. In the case of the fractionated extract, a 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., hydrophobic solvent, hydrophilic solvent, or a mixed solvent thereof 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 is removed by freeze drying, vacuum drying, hot air drying, spray drying, or 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 capable of exhibiting the desired activity alone or in combination with a carrier that has no activity 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 obese or overweight.

In addition, 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 blending, 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, 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 effective amounts 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 verified for safety in the art, in order to increase and reinforce the anti-obesity effect or to enhance the convenience of taking or intake through the addition of similar activities such as fatigue improvement activity. and may further include any compound or natural extract known to have a corresponding activity. Such compounds or extracts include compounds or extracts, drugs listed in compendial documents such as pharmacopeias of each country (“Korean 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, extracts, and health functional foods that have been approved for items in accordance with the laws of each country (the "Pharmaceutical Act" in Korea) that regulate the manufacture and sale of '), compounds or extracts whose functionality is recognized are included.

For example, according to the “Health Functional Food Act” in Korea, Garcinia cambogia bark extract, conjugated linolenic acid (free fatty acid), conjugated linolenic acid (triglyceride), green tea extract, and chitosan, which have been recognized as functional as 'body fat reduction', are health functional foods. Lactobacillus gasseri BNR17 (Lactobacillus gasseri BNR17), L-carnitine tartrate, green mate extract, green coffee bean extract, sesame leaf extract, soybean germ extract, etc. , lactoferrin (milk refined protein), lemon balm extract mixed powder, mate hot water extract, seaweed, etc. complex extract (xantigen), fermented vinegar pomegranate complex, puer's tea extract, seomoktae peptide complex, vegetable oil diglyceride, Wild mango seed extract, medium-chain fatty acid (MCFA)-containing oil, Coleus forskohlii extract, chitooligosaccharide, finger root extract powder, hibiscus, etc., and L-glutamic acid-derived GABA-containing powder recognized for its 'blood pressure control'functionality; Katsuobushi oligopeptide, nattobacterial culture powder, samoktae peptide complex, salmon peptide, olive leaf extract, sardine peptide, casein hydrolyzate, coenzyme Q10, grape seed enzyme-decomposed extract powder, Haitai oligopeptide, etc., 'blood triglyceride DHA-concentrated oils and fats, globin hydrolyzate, indigestible maltodextrin, bamboo leaf extract, vegetable oil diglyceride, refined sardine fish oil, refined squid oil, etc., which have been recognized for their ‘improvement’ functionality, and L-arabinose whose functionality has been recognized for ‘glycemic control’ , nopal extract, cinnamon extract powder, guava leaf extract, indigestible maltodextrin, freeze-dried silkworm powder, hemp spirit extract, banaba leaf extract, upper leaf extract, etc., fermented amino acid complex recognized for its 'fatigue improvement' functionalities These compounds or extracts include fruit extract, rhododendron extract, etc., L-theanine, asyaganda extract, hydrolyzate milk protein, and leaf extract recognized as 'anti-stress'. something to do.

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, bread, confectionery, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be prepared.

In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to Korea's "Health Functional Food Act", or confectionery, beans, tea, and beverages according to each food type according to the Food Ordinance of Korea's Food Sanitation Act (Ministry of Food and Drug Safety Notification "Food Standards and Specifications") , food for special use, 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 added to and mixed with or infiltrated into food in manufacturing, processing or preserving food. Since they are consumed daily and for a long period of time with food, their safety must be ensured. 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. 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 fragrance, 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 one, 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, dandelion leaves, bamboo leaves, cinnamon, chrysanthemum leaves, and jasmine. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo biloba, etc. 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.

Calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used as preservatives, and acacia gum, carboxymethylcellulose, xanthan gum, and pectin can be used as emulsifiers. 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. 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 the thickening agent, a suspending agent, a settling agent, a gel-forming agent, a bulking agent, and the like can be used.

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

Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. 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 type of product.

In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the national food regulations or food additives regulations.

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

The pharmaceutical composition of the present invention may be prepared as an oral dosage form or a 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 the 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 pharmacopeias 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, together with a suitable carrier according to methods known in the art. 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, its magnesium and calcium salts, polyethylene glycol, and the like. , agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, and the like.

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 a nasal inhalant, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, the carrier is Witepsol ( 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, patient's severity, and administration route. 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 the antigonon leptopus hot water 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 an antigonone leptopus 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 antigonone leptopus extract

After adding 95% ethanol by weight of 10 times to the dry powder of Antigonon leptopus (extraction site, outpost), leaching at room temperature for 24 hours, and filtering with Whatman paper No. 2, the filtrate was reduced pressure Concentrated and freeze-dried to prepare a solid antinogon leptopus extract.

< Experimental example > Obesity improvement activity experiment

Mouse precursor adipocytes 3T3-L1 (ATCC CL-173) were cultured in 10% BCS DMEM medium at 37° C. and 5% CO ₂ conditions. 3T3-L1 precursor adipocytes were aliquoted in a 24-well plate at ^{a cell number of 5×10 4} /well, and then maintained at 100% confluency for 2 days. 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. Thereafter, the culture medium was replaced with 10% FBS DMEM culture medium every 2 days for 4 days. During adipocyte differentiation induction, 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 3 repeated experiments for each concentration.

The results are shown in FIG. 1 , and referring to FIG. 1 , it can be seen that the antigonone leptopus extract clearly has the activity of inhibiting the differentiation of preadipocytes into adipocytes and the activity of inhibiting the accumulation of fat.

Claims (4)

Anti-obesity food composition comprising an antigonon leptopus extract as an active ingredient.
According to claim 1,
The anti-obesity food composition, characterized in that the antigonon leptopus extract is obtained by extraction with water, ethanol, or a mixed solvent thereof.
A pharmaceutical composition for anti-obesity comprising an antigonon leptopus extract as an active ingredient.
4. The method of claim 3,
The antigonon leptopus extract is an anti-obesity pharmaceutical composition, characterized in that it is obtained by extracting the leaves of antigonon leptopus and extracting it with water, ethanol, or a mixed solvent thereof.

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