KR101227737B1 - A composition comprising Ligularia stenocephala extract, fractions thereof or compounds isolated from Ligularia stenocephala extract and fractions thereof having peroxinitrite-scavenging activity - Google Patents

Abstract

The present invention relates to a composition having a peroxynitrite scavenging activity comprising a gondal extract or a fraction thereof, or a compound derived therefrom as an active ingredient, which is effective for preventing or treating obesity-related diseases and thus improving obesity-related diseases. It can be used as a pharmaceutical composition, food composition, cosmetic composition, quasi-drug composition and the like for the prevention, and treatment.

Description

A composition comprising Ligularia stenocephala extract, fractions according or compounds isolated from Ligularia stenocephala extract and fractions having having peroxinitrite- scavenging activity}

The present invention relates to a composition having peroxynitrite scavenging activity using as an active ingredient gondalbi extract or fractions thereof, or compounds derived therefrom.

Obesity is increasing due to the improvement of diet, living standards and stress all over the world, and obesity has become a social problem recently because it can develop into other diseases such as diabetes and arteriosclerosis. In order to reduce obesity, drug therapy, dietary therapy, surgical therapy, etc. are being taken, and as part of the diet, wild vegetables are used to prevent and cure diseases such as diabetes and arteriosclerosis in addition to obesity (Jeong et al. , 2006). Wild vegetables are considered to be used for the treatment of obesity and obesity-related diseases because they contain a lot of vitamins, fiber, antioxidants and the like. The inventors also found Allium victorialis (Choi et al., 2005) Pleurospermum kamtschaticum (Jung et al., 2007), and the roots of Rosa rugosa ((Park et al., 2005). ) Has reported an anti-obesity effect.

Peroxy nitrite (peroxinitrite, ONOO ^{-) -} is one of the factors that the reaction result severe damage to the cells as being generated between the nitrogen monoxide (NO) (Radi et al, the superoxide anion radical (· O _2). 1991). This induces peroxidation of proteins and lipids, results in cytotoxicity and further rapid neuronal damage (Haenen et al., 1997). In addition, peroxynitrite has been reported to cause metabolic or circulatory diseases such as atherosclerosis, diabetes, obesity, hypercholesterolemia (Korda et al., 2008; Patcher et al., 2005; Drel et al., 2007).

In Korea, Ligularia stenocephala is used as a wild vegetable belonging to the Asteraceae family, and it is also a wild vegetable eaten in parallel when eating meat. Anti-thrombotic effects of gondal have been reported, but no anti-obesity effects have been reported. Ligulariacephalins A, B, C, 5,6-dimethoxy-2-isopropylbenzofuran, euparin, (R)-(-)-hydroxytremetone, etc. (Toyoda et al., 2005) It is known that gondal leaf contains 3,4-dicaffeoydicquinic acid and 3,5-dicaffeoylquinic acid as antithrombotic components (Yoon et al., 2008). However, in addition to the 3,4-dicaffeoydicquinic acid and 3,5-dicaffeoylquinic acid, the types of caffeoylquininic acid compounds included in gondal are unknown. Antioxidant activity, anti-inflammatory activity, antimicrobial action, vasodilator action, hepatocyte protection action, and platelet aggregation inhibitory action of the caffeoylquinine acid compounds have been reported (Zhao et al., 2006).

The inventors found that while screening wild vegetables for scavenging peroxynitrite, the gondalbi methanol extract had an excellent effect of scavenging peroxynitrite and fractionating it with ether and butanol and analyzing its main active ingredient. As a result, it was confirmed that there is an anti-obesity effect on gondal methanol extract, fractions thereof, and six kinds of caffeoylquinine acid compounds which are main active ingredients thereof, and completed the present invention.

Choi, J. W., Lee, K. T., Kim, W. B., Park, K. K., Lee, J. H., Lim, S. C., Jung, H. J., Park, H. J. Effect of Allium victorialis var. platyphyllum Leaves on Triton WR-1339-induced and Poloxamer-407-induced hyperlipidemic Rats on Diet-Induced Obesity Rats, Kor. J. Pharmacogn., 36: 121-128 (2005). Choi, S. Z., Choi, S. U., and Lee, K. R., Phytochemical constituents of the Aerial parts from Solidago virga-aurea var. gigantea Miq. Arch. Pharm. Res., 27, 164-168 (2004). Costa, RM, Magalhaes, AS, Pereira, JA, Andrate, PB, and Valetao, P., Eva; uation of free radical scavenging and antihemolytic activities of quince (Cydonia oblonga) leaf: A comparative study with green tea (Camellia sinensis) , Food and Chemical Toxicology 47: 860-865 (2009). Drel, V. R., Patcher, P., Vareniuk, I., Pavlov, I., Lyzogulbov, V. V., Grovez, J. T., and Obrosova, I. G., A peroxynitrite decomposition catalyst counteracts sensory neuropathy in streptozotocin-diabetic mice. Eup. J. Pharmacol., 569, 48-58 (2007). Friedewald, W. T., Levy, R. I., and Fredrickson, D. S., Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin. Chem. 24, 499-502 (1972). Haenen, G. R., Paquay, J. B., Korthouwer, R. E., and Bast, A., Peroxynitrite scavenging by flavonoids. Biochem. Biophys. Res. Commun., 236, 591-593 (1997). Hsu, C. L., Huang, S. L., and Yen, G. C., Inhibitory effect of phenolic acids on the proliferation of 3T3-L1 preadipocytes in relation to their antioxidant activity. Agric. Food Chem., 54, 4191-4197 (2006). Jeong, S. H., Lee, K. T., Ann, H. S., Lee, J. Y., Choi, J. W., and Lim, D. G., Hypolipidemic and antioxidative effects of bangpoongtongsungsan-kamibang on diet-induced obesity rats, Kor. J. Pharmacogn., 37: 190-195 (2006). Jung, HJ, Nam, JH, Park, HJ, Lee, KT, Park, KK, Kim, WB, Choi, J. W: The MeOH extract of Pleurospermum kamtschaticum and its active component buddlejasaponin (IV) inhibits intrinsic and extrinsic hypercholesterolemia and hypercholesterolemia in the rat, Journal of Ethnopharmacology, 112, 255-261 (2007). Kim, K. H., Lee, K. H., Choi, S. U., Kim, Y. H., and Lee, K. R., Terpene and phenolic constituents of Lactuca indica L. Arch. Pharm. Res., 31, 983-988 (2008). Kobatake, Y., Saito, M., Kuroda, K., Kobayashi, S., and Innami, S. (1987) Influence of fish consumption on serum lipid and lipid peroxide concentrations in middle aged subjects. J. Japan Soc. Nutr. & Food Sci. 40: 103-107. Kooy, N. W., Royall, J. A., Ischiropoulos, H., and Beckman, J. S., Peroxynitrite-mediated oxidation of dihydrorhodamine 123. Free Radic. Biol. Med., 16, 149-156 (1994). Korda, M., Kubant, R., Patton, S., and Malinski, T., Leptin-induced endothelial dysfunction in obesity. Am. J. Physiol. Heart Circ. Physiol., 295, 1514-1521 (2008). Kwon, HC, Jung, CM, Shin, CG, Lee, JK, Choi, SU, Kim, SY, and Lee, KR, A new caffeoyl quinic acid from Aster scaber and its inhibitory activity against human immunodeficiency virus-1 (HIV- 1) integrase. Chem. Pharm. Bull., 48, 1796-1798 (2000). Marinova, E. M., Toneva, A., and Yanishilieva, N., Comparison of the antioxidative properties of caffeic and chlorogenic acids, Food Chemistry 114: 1498-1592 (2009). Noma, A. Nezu-Nakayama, K., Kita, M., Nad Okabe, H., Simultaneous determination of serum cholesterol in high- and low-density lipoproteins with use of heparin, Ca2 +, and an anion-exchange resin, Clin . Chem. 24, 1504-1508 (1978). Olmos, A., Guner, R. M., Recio, M. C., Rios, J. L., Gil-Benso, R., and Marnez, S., Interaction of dicaffeoylquinic derivatives with peroxynitrite and other reactive nitrogen species. Arch. Biochem. Biophys., 475, 66-71 (2008). Oyanagui, Y. (1984) Reevaluation of assay methods and establishment of kit for superoxide dismutase activity. Anal. Biochem. 42: 290-296. Park, HJ, Nam, JH, Jung, HJ, Lee, MS, Lee, KT, Jung, MH, Choi, JH, Inhibitory effect of euscaphic acid and tormentic acid from the roots of Rosa rugosa on high fat diet-induced obesity in the rat. Kor. J. Pharmacogn. 36, 324-331 (2005). Patcher, P., Obrosova, I. G., Mabley, J. G., and Szabo, C., Role of nitrosative stress and peroxynitrite in the pathogenesis of diabetic complications. Emerging new therapeutical strategies. Curr. Med. Chem., 12, 267-275 (2005). Radi, R., Beckman, J. S., Bush, K. M., and Freeman, B. A., Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide. J. Biol. Chem., 266, 4244-4250 (1991). Richmond, W., Use of cholesterol oxidase for assay of total and free cholesterol in serum by continuous flow analsysis. Clin. Chem. 22, 1579-1588 (1976). Toyoda, M., Yaoita, Y., and Kikuchi, M., Three new dimeric benzofuran derivatives from the roots of Ligularia stenocephala Matsum. Et Koidz. Chem. Pharm. Bull., 53, 1555-1558 (2005). Yagi, K., Lipid peroxides and human diseases, Chem. Pharm. Bull., Phys. Lipids 45: 337-351 (1987) Yoon, M. H., Cho, C. W., Lee, J. W., Kim, Y. S., An, G. H., and Lim, C. H., Antithrombotic compounds from the leaves of Ligularia stenocephala M., Nat. Prod. Sci., 14, 62-67 (2008). Zhao, Y., and Zhao, J., Advances in Caffeoylquinine Acid research. Zhonguo Zhongyao Zazhi, 31, 869-874 (2006). Ziegler, D., Nowak, H., Kempler, P., Vargha, P., and Low, P. A., Treatment of symptomatic diabetic polyneuropathy with the antioxidant alpha-lipoic acid: a metaanalysis. Diab. Med., 21, 114-121 (2004). Kim, K. H., Kim, Y. H., and Lee, K. R., Isolation of quinic acid derivatives and flavonoids from the aerial parts of Lactuca indica L. and their hepatoprotective activity in vitro. Bioorg. Med. Chem. Lett., 17, 6739-6743 (2007).

An object of the present invention is to provide a composition for the prevention or treatment of obesity-related diseases containing Ligularia stenocephala extract, fractions thereof, or compounds isolated therefrom as an active ingredient.

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of obesity-related diseases containing the composition.

An object of the present invention is to provide a food composition for the prevention or improvement of obesity-related diseases containing the composition.

An object of the present invention to provide a cosmetic composition for the prevention or improvement of obesity-related diseases containing the composition.

An object of the present invention is to provide a quasi-drug composition for the prevention or improvement of obesity-related diseases containing the composition.

An object of the present invention is 3,4-di-O-caffeoylquinic acid (3,4-DQ), 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 3,5-di O- caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ) and 3- O -caffeoylquinic acid ( It is to provide a composition for the prevention or treatment of obesity-related diseases containing at least one compound selected from the group consisting of 3-CQ or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object, the present invention is a gonadabi ( Ligularia Stenocephala ) provides a composition having a peroxynitrite scavenging activity containing an extract, a fraction thereof, or a compound separated therefrom as an active ingredient.

The present inventors have found that there is an excellent effect of eliminating peroxynitrite on the gondal methanol extract, fractions thereof, and analyzing the main active ingredients thereof, the gondal methanol extract, fractions thereof, and six main cafes thereof. It was confirmed that the anti-obesity effect on the oil quinic acid compounds.

As used herein, the term "peroxy nitrite (peroxinitrite, ONOO ^-) scavenging activity" is superoxide anion radical (· O ₂ ^-) to be a peroxy nitrite is produced by the reaction of the acid and nitric acid (NO) It means blocking or reducing its efficiency or removing or reducing the peroxynitrite produced. In general, hyperleptinemia develops in obesity, resulting in an increase in peroxynitrite. This nitric oxide / peroxynitrite imbalance can lead to atherosclerosis and diabetes mellitus, so the presence of peroxynitrite is an important indicator of obesity-related diseases.

According to a specific embodiment of the present invention, the present inventors confirmed the peroxynitrite scavenging effect of the gondalby extract, and more specifically, the result of the peroxynitrite scavenging effect experiment from the gondalbi extract, the IC ₅₀ value of 0.87 ± 0.33 μg / ml. This value was similar to the IC ₅₀ value of 0.89 ± 0.22 μg / ml of penicillamine, a positive control compound, indicating that the gondal methanol extract had a very strong peroxynitrite scavenging activity (FIG. 3).

In addition, the present inventors confirmed that peroxynitrite causes obesity-related diseases, and gondalby extracts and fractions have such peroxynitrite scavenging activity, thereby ultimately preventing or treating obesity-related diseases.

The obesity-related disorders include, but are not limited to, obesity, hyperlipidemia, or metabolic syndrome, which are caused by an increase in peroxynitrite resulting in a nitrogen oxide / peroxynitrite imbalance. .

According to a specific embodiment of the present invention, the present inventors administered weight (Fig. 4) and abdominal adipose tissue (epithelial cavity and epididymis when administering the gondabi methanol extract and its fraction ether fraction, butanol fraction to obese rats for anti-obesity activity measurement ) Weight (FIG. 5) significantly decreased to the level of the intestinal group, decreased total cholesterol content and LDL-cholesterol content, decreased arteriosclerosis index (AI) (Table 3), and significantly reduced antioxidant activity. (Table 4).

In the present invention, the gondalby extract is crushed after washing and drying the gondalby; And the ground gondal ratio can be obtained by extracting with a solvent selected from water, C ₁ ~ C ₄ lower alcohol or a mixed solvent thereof.

The gondalby extract comprises a solvent selected from the group consisting of water, lower alcohols of C ₁ to C ₄ and mixed solvents thereof, preferably extracted with methanol. In addition, in this invention, the said extract shall contain the extract obtained by an extraction process, the dilution or concentrate of an extract, the dried material obtained by drying an extract, or any of these modifiers or refined products. The extraction method is not particularly limited and may be extracted at room temperature or warmed under conditions where the active ingredient is not destroyed or minimized. More specifically, the method for obtaining gondal extract in the present invention is as follows. The gondalby pulverized product is prepared by using a polar solvent of C ₁ to C ₄ lower alcohols such as water, methanol, ethanol and butanol, etc., in volume of about 2 to 20 times, preferably about 3 to 5 times, dry weight or about 1 thereof. A mixed solvent having a mixing ratio of: 0.1 to 1:10 is used as the elution solvent, and the extraction temperature is 20 to 100 ° C, preferably 80 ° C, and the extraction period is about 5 hours to 10 days, preferably 5-10 Extraction is performed using extraction methods such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for a period of time. Preferably, the mixture is extracted at 1 to 5 times by reflux extraction and filtered under reduced pressure, and the filtrate is concentrated under reduced pressure at a temperature of 20 to 100 ° C., preferably at room temperature with a distillation concentrator, and soluble in water, lower alcohols or a mixed solvent thereof. One gondal ratio crude extract can be obtained.

The gondalby extract can be extracted from various organs of natural, hybrid, and variegated plants, for example from roots, stems, leaves, and flowers, as well as from plant tissue cultures, most preferably from the leaves of gondal. According to a specific embodiment of the present invention was prepared gondalby extract from the leaves of gondalby.

In the present invention, the fraction of gondalbi extract of the present invention is the step of washing and drying the gondalbi pulverized with a grinder; Extracting the pulverized gondal ratio with a solvent selected from water, a lower alcohol having ₁ to ₄ C, or a mixed solvent thereof to obtain a gondal ratio extract; And fractionating the gondalby extract into one or more solutions selected from the group consisting of ether, butanol, and water.

More specifically, the solvent fraction of the gondalby extract may be obtained by suspending the gondalby extract with water and then fractionating it with a solvent such as ether or butanol to obtain a polar solvent fraction and a nonpolar solvent fraction, respectively. In a specific aspect of the present invention, the solvent fraction is provided with an ether fraction or butanol fraction.

Method for obtaining a fraction of gondalby extract in the present invention is as follows. The obtained gondal ratio crude extract is suspended in water, and then added to a nonpolar solvent such as an ether of about 1 to 100 times, preferably about 1 to 5 times the volume of the suspension, 1 to 10 times, preferably 2 to 5 times. It can be obtained by extracting and separating the nonpolar solvent soluble layer over times. It is also possible to further carry out conventional fractionation processes (Harborne JB Phytochemical methods: A guide to modern techniques of plant analysis , 3rd Ed. p6-7, 1998). Specifically, the gondal ratio crude extract is suspended in water and then ether mixed and fractionated to obtain an ether fraction. Butanol fraction may be obtained by adding butanol to the ether fraction.

In addition, the present inventors additionally sought to identify the active ingredient exhibiting an anti-obesity effect among the gondal extract and its fractions. The active ingredient may be separated by performing chromatography one or more times sequentially on the extract or fraction, and the type and developing solvent of the column of the chromatograph may be controlled in various ways. In a specific embodiment of the present invention, high-performance liquid chromatography (HPLC) analysis of gondal extract and fractions thereof was performed for active ingredient analysis. Lactuca Using the caffeyl quinquinate derivatives isolated from indica as standard products, the types and contents of caffeyl quinquinate derivatives in gondabi were measured, and the contents of caffeyl quinquinate derivatives in the gondabi extract and fractions were measured. As a result, by comparison with a standard foam gondalbi include 3,4-di-O-caffeoylquinic acid (3,4-DQ), 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ ), 3,5-di- O -caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ), It was confirmed that 3- O -caffeoylquinic acid (3-CQ) was contained. However, 3- O - p- coumaroyl-caffeoylquinic acid (3-pCQ) in standard foam was not detected in gondal ratio.

Therefore, in the present invention, washing and drying the gondal ratio and pulverizing with a grinder; Extracting the pulverized gondal ratio with a solvent selected from water, a lower alcohol having ₁ to ₄ C, or a mixed solvent thereof to obtain a gondal ratio extract; Fractionating the gondalby extract with one or more solutions selected from the group consisting of ether, butanol, and water to obtain ether, butanol or water fractions of gondalby; And the fraction of high-performance liquid chromatography carried out by a 3,4-di-O-caffeoylquinic acid (3,4-DQ), 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ) , 3,5-di- O -caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ) and 3 3,4-di-O-caffeoylquinic acid (3,4-DQ), 3, isolated from gondalby comprising obtaining at least one compound selected from the group consisting of O- caffeoylquinic acid (3-CQ). 5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 3,5-di- O -caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4 , 5-DQ), 5- O- caffeoylquinic acid (5-CQ), and 3- O- caffeoylquinic acid (3-CQ).

Chemical formulas of the six kinds of caffeoylquinic acid derivatives are as follows.

The six kinds of caffeoylquininic acid derivatives can be separated from natural substances, preferably the gondalbi, and the gondalbi extract can be separated from various organs of natural, hybrid, and variegated plants as mentioned above, and extracted from gondalbi leaves. Most preferred. In addition, these six caffeoylquinolinic acid derivatives may be prepared by chemical synthesis by methods known in the art.

As used herein, the term "pharmaceutically acceptable salts" refers to salts prepared by conventional methods and known to those skilled in the art. "Pharmaceutically acceptable salts" include, but are not limited to, salts derived from the following pharmacologically or physiologically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, formic acid , Benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium, alkali cometals such as magnesium, ammonium and the like.

The gondalbi extract, 4-di-O-caffeoylquinic acid (3,4-DQ) was separated fractions thereof and therefrom, 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 3 , 5-di- O -caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ) and 3- O -Caffeoylquinic acid (3-CQ) is based on ingredients obtained from natural extracts, not artificially synthesized compounds, so it is safe, toxic, and has no side effects. In addition, the composition may be used not only in humans, but also in animals such as cattle, dogs, and the like, in which obesity-related diseases may occur.

The six caffeoylquinic acid compounds contained about 67.8% in the Gondalbi methanol extract and about 94.52% in the butanol fraction, indicating a very high content (Table 2). Thus, gondalbi extracts and fractions have excellent anti-obesity activity, and their active ingredients 4-di-O-caffeoylquinic acid (3,4-DQ), 3,5-di- O -dicaffeoyl- epi- quinic acid (3,5-DeQ), 3,5-di- O -caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ) and 3- O- caffeoylquinic acid (3-CQ) can also be expected to have the same anti-obesity activity.

Therefore gondalbi extract, fractions thereof, and 4-di-O-caffeoylquinic acid (3,4-DQ), 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 3,5-di O- caffeoylquinic acid (3,5-DQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ), 3- O -caffeoylquinic acid ( 3-CQ) has excellent prophylactic and therapeutic activity against obesity-related diseases, and the present invention provides the prevention of obesity-related diseases comprising the six kinds of caffeoylquinolinic acid derivatives or pharmaceutically acceptable salts thereof as an active ingredient. Or it provides a pharmaceutical composition for treatment.

Gondalbi extract of the present invention, fractions thereof, and six kinds of caffeoylquinolinic acid derivatives isolated therefrom, or pharmaceutically acceptable salts thereof, have excellent peroxitrite (ONOO ⁻ ) scavenging activity and have weight of obese rats, Since it reduces abdominal adipose tissue and atherosclerosis index and effectively inhibits TBARS, hydroxy radical level and SOD activity level, it is effective for the prevention and treatment of obesity related diseases, and thus can be used as a pharmaceutical composition for preventing and treating obesity related diseases.

As used herein, the term "prevention" refers to any action that inhibits or delays the onset of the disease by administration of a composition comprising a gondal extract, a fraction thereof, and six caffeoylquininic acid derivatives isolated therefrom.

As used herein, the term "treatment" refers to all actions of improving or beneficially modifying the symptoms of the disease by administration of a composition comprising a gondal extract, a fraction thereof, and six kinds of caffeoylquininic acid derivatives isolated therefrom. .

The composition according to the present invention can be administered to mammals such as rats, mice, livestock, humans, etc. by various routes. All modes of administration can be expected and can be administered, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

The throughput of the pharmaceutical composition for preventing and treating obesity-related diseases used in the present invention should be a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level may refer to an individual type and severity, age, sex, It can be determined according to the type of virus infected, the activity of the drug, the sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of treatment, factors including the drug used concurrently, and other factors well known in the medical arts. Effective amounts may vary depending on the route of treatment, the use of excipients, and the possibility of use with other agents, as will be appreciated by those skilled in the art.

The pharmaceutical treatment forms of the pharmaceutical compositions for the prevention and treatment of obesity-related diseases of the present invention may be used in the form of their pharmaceutically acceptable salts, and may also be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection. Can be.

The pharmaceutical composition for preventing and treating obesity-related diseases of the present invention may be prepared in pharmaceutical formulations using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. have. In the preparation of the formulations, it is preferred that the active ingredient is mixed with or diluted with the carrier, or enclosed in a carrier in the form of a container.

Therefore, the pharmaceutical composition for the prevention and treatment of obesity-related diseases of the present invention, powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions according to conventional methods It may be used in the form of a formulation, and may further include suitable carriers, excipients and diluents commonly used in the preparation of the composition.

For example, carriers, excipients and diluents which may be included in the pharmaceutical compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose in the compound. Or lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Examples of the liquid preparation for oral administration include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweetening agents, have.

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In addition, the present invention provides a food composition for preventing and improving obesity-related diseases, including gondal extract, fractions thereof, and six kinds of caffeoylquinine acid derivatives or pharmaceutically acceptable salts thereof as an active ingredient.

The food composition of the present invention includes the form of pills, powders, granules, acupuncture, tablets, capsules or liquids, and the like to which the composition of the present invention can be added, for example, various foods, for example, Drinks, gums, teas, vitamin complexes, and dietary supplements.

As an essential ingredient that may be included in the food composition of the present invention, there are no particular limitations on other ingredients other than the composition including the gondalby extract, fractions thereof, and six kinds of caffeoylquininic acid derivatives. Supplementary additives or natural carbohydrates and the like may be included as additional ingredients.

In addition, as mentioned above, the food supplement additive may further include food supplement additives, including food supplement additives conventional in the art, such as flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like. .

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. .

In addition to the above, the food composition of the present invention can be used as a flavoring agent such as a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. Others may contain pulp for the production of natural fruit juices and fruit juice drinks and vegetable drinks. These components can be used independently or in combination.

In addition, the food composition of the present invention is any one of meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, gum, ice cream, soup, beverage, tea, functional water, drink, alcoholic beverage and vitamin complex It may be in the form of.

In addition, the present invention relates to a cosmetic composition comprising a gondalbi extract, a fraction thereof and a composition comprising six kinds of caffeoylquinolinic acid derivatives as an active ingredient. The cosmetic composition according to the present invention may be formulated into a lotion, gel, water-soluble powder, fat-soluble powder, water-soluble liquid, cream or essence by adding a pH adjusting agent, flavoring agent, emulsifier, preservative, etc. as necessary in the usual cosmetic preparation method. have.

In addition, the present invention relates to a quasi-drug composition comprising a composition comprising gondalbi extract, fractions thereof, and six kinds of caffeoylquinolinic acid derivatives as active ingredients.

That is, the composition of the present invention may be added to the quasi-drug composition for the purpose of preventing or ameliorating obesity-related diseases. When the gondalbee extract of the present invention, its fractions and six kinds of caffeoylquininic acid derivatives are used as quasi-drug additives, the extracts, fractions and six kinds of caffeoylquinic acid derivatives can be added as they are or used together with other quasi-drugs or quasi-drug components. Can be suitably used according to conventional methods. The mixed amount of the active ingredient may be suitably determined depending on the purpose of use (prevention, health or therapeutic treatment). Preferably, the quasi-drug composition may be used for the preparation of antiseptic cleaners, shower foams, gagrins, wet tissues, detergent soaps, hand washes, humidifier fillers, masks, ointments, coating agents or filter fillers.

The present invention is a composition having a peroxynitrite scavenging activity using a gondalby extract or a fraction thereof, or a compound derived therefrom as an active ingredient is effective in preventing or treating obesity-related diseases, thereby improving, preventing, and treating obesity-related diseases. For pharmaceutical compositions, food compositions, cosmetic compositions, and quasi-drug compositions and the like can be used.

1 shows standard compounds 1-7 used for quantitative analysis.
FIG. 2 shows HPLC chromatograms of seven standard compounds, gondal methanol extracts, and fractions thereof (ether and butanol fractions).
Figure 3 shows a comparison of the peroxynitrite scavenging effect of L- penicillamine and gondal methanol extract.
Figure 4 shows the effect of gondal methanol extract and its butanol fraction on the body weight of rats fed a high fat diet and rats fed a normal diet at 5th week (W5) and 6th week (W6).
FIG. 5 shows the effect of gondal methanol extract and its butanol fraction on the abdominal fat tissue (relaparotomy and epididymis) of rats fed a high fat diet and rats fed a normal diet.

Hereinafter, preferred embodiments of the present invention will be presented to assist in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

Example  1. Experimental method and experimental material

plant

In spring 2009, L. stenocephala (Asteraceae) leaves were collected from Gangwon-do, South Korea. The leaves were air dried at room temperature and powdered. This plant is It was identified by Won-Bae Kim (Highland Agriculture Research Institute, Rural Development Administration, Pyeongchang, Gangwon-do, Korea).

Extraction and fractionation

Extracts of leaves of L. stenocephala (2 kg), air-dried three times using methanol (10 L) at 80 ° C. for 7 hours, were refluxed. The extract was concentrated using a rotary evaporator under reduced pressure to make viscous body (342 g). The extract (330 g) was suspended in 800 mL water and partitioned three times with 800 mL ether. The remaining aqueous layer was then partitioned three times with 800 mL butanol. The ether and butanol soluble portions were then concentrated and dried with a freeze-dryer to give ether and butanol powder solid fractions (108 g and 119 g, respectively).

HPLC  Instruments, Reagents, and Basic Compounds for Analysis

Varian HPLC system (Walnut Creek, CA, USA) was used to analyze the caffeoylquinic acid contained in the methanol extract of L. stenocephala and its fractions (ether and butanol). The Varian HPLC system includes a Prostar 210 solvent delivery module, a Prostar 325 UV-Vis detector and a 20 mL sample loop (Rheodyne, Rohnert Park, CA, USA). Shiseido (Chuoku, Tokyo, Japan) The compounds of the assay samples were isolated on Capcell Pak C18 column (5 mL, 250 mm × 4.6 mm ID). All solvents used for analysis were HPLC grade obtained from JT Baker (Phillipsburg, NJ, USA). Lactuca Seven standard caffeyl quinquinate isolated from indica L. The one provided by Kang Ro Lee (College of Pharmacy, SungKyun Kwan University, Suwon, Korea) was used. The seven caffeoylquinolinic acids are: 3,4-di- O -cafeoylquinic acid (3,4-DQ), 3,5-Di- O- dicaffeoyl- epi- quinic acid (3,5-DeQ ), 3,5-di- O -cafeoylquinolinic acid (3,5-DQ), 4,5-di- O -cafeoylquinic acid (4,5-DQ), 5- O -caoylquinic acid (5-CQ), 3- O - cafe five days quinic acid (3-CQ), 3- O - p -coumaroyl- cafe five days quinic acid (3-pCQ). The structures of the seven standard compounds are shown in FIG. 1.

Cafe Oil Quinine Acid  For analysis HPLC  Condition

For HPLC analysis, test samples (methanol extract, ether and butanol fractions) and seven standard compounds were dissolved in 80% methanol and filtered through a 0.50 mm sringe filter and the filter liquid was filtered into the (20 mL) analyzer. Injected. The fluidized bed was a 0.05% phosphoric acid mixture in water (solvent A) and methanol (solvent B). The gradient system is as follows: 0-10 minutes, 60% A: 40% B; 10-20 minutes, 50% A: 50% B; 20-30 minutes, 40% A: 60% B; 30-35 min, 60% A: 40% B. Chromatography was performed at 1.00 mL min ^{- 1} flow rate. The UV detector was fixed at 246 nm. The extract was analyzed by HPLC after the R2 value of the regression equations was 0.990 or more.

Peroxynitrite -Clearing activity analysis

The modified Kooy's method was used to assess peroxynitrite (ONOO ⁻ ) scavenging activity, which involves monitoring fluorescent rhodamine 123, which was rapidly produced from non-fluorescent DHR 123 in the presence of ONOO ⁻ ( Kooy et al., 1994). That is, rhodamine buffer (pH 7.4) is 50 mM sodium phosphate dibasic, (50 mM sodium phosphate dibasic), 50 mM sodium phosphate monobasic, 90 mM sodium chloride, 5 mM potassium chloride (potassium) chloride), and 100 μM DTPA. The final DHR 123 concentration was 5 μΜ. In this assay the buffer was prepared before use and placed on ice. Plant extracts were dissolved in 10% DMSO (fc 5 μg / mL). Background and final fluorescence intensities were measured and dissolved in 0.3 N sodium hydroxide for 5 minutes after treatment with or with ONOO ⁻ (10 μM). The fluorescence intensity of oxidized DHR 123 was measured using a microplate fluorescence reader FL 500 (Bio-Tek Instruments Inc., Winooski, VT, USA) at excitation and divergence frequencies of 480 and 530 nm, respectively. Peroxynitrite-erase activity values were calculated as the final fluorescence intensity minus background fluorescence intensity via DHR 123 oxidation detection.

Experimental animals and treatments

Four weeks old, 140 ± 10 g Sprague Dawley (SD) white rats were purchased from Hyochang Science Co., Ltd., and then healthy animals were used for the experiment after one week of quarantine and purified breeding. Breeding of this test was carried out in the laboratory animal breeding room of Kyungsung University Pharmacy set up with temperature 22 ± 3 ℃, relative humidity 50 ± 10%, lighting time 12 hours (07: 00 ~ 19: 00). Feed was freely ingested for drinking water, which was a solid feed for animals (Central Experimental Animal, Seoul). 24 hours before the experiment time only watering and fasting. At this time, the experimental animals were treated within a predetermined time (10: 00-12: 00 am) in consideration of the daily variation of enzyme activity. The experiment is certified by the Hard University Animal Care and Use Committee. All procedures were performed in accordance with the Guidelines for the Management and Use of Laboratory Animals, published by the National Institutes of Health. The gondal MeOH extract, the Et ₂ O fraction and the BuOH fraction were administered orally with oral jonde at 100, 200 mg / kg for 2 weeks using 4% tween 80 dissolved in saline.

Induction of obesity and hyperlipidemia

The high fat dietary composition is as listed in Table 1. High fat diets were ingested for 6 weeks to induce obesity in experimental animals. The antiobesity effect was observed by oral administration of the sample for the last 2 weeks out of 6 weeks.

¹⁾ Cellulose: Sigma Co. LTD., USA

²⁾ Mineral mixture (g / kg diet) based on Rogers and Haper (1965) type, including: calcium phosphate dibasic 500.0, sodium chloride 74.0, potassium citrate monohydrate 220.0, potassium sulfate 52.0, magnesium oxide 24.0, magnesium carbonate 3.5, ferric citrate 6.0, zinc carbonate 1.6, cupuric carbonate 0.3, potassium iodate 0.01, chromium potassium sulfate 0.55, sucrose, finely powered make 1,000

³⁾ Vitamin mixture (g / kg diet): thiamine HCl 0.6, biotin 0.02, riboflavin 0.6, cyanocobalamine 0.001, pyridoxine HCl 0.7, retinyl acetate 0.8, nicotinic acid 3.0, DL-tocopherol 3.8, Ca-pantothenate 1.6, 7-dehydrocholesterol 0.0025 , folic acid 0.2, methionine 0.005, sucrose, finely powered make 1,000

Serum preparation

Blood was collected from the abdominal aorta by gently anesthetizing the experimental animals with CO ₂ , and the collected blood was left for 30 minutes and centrifuged at 3,000 rpm for 10 minutes to separate serum, and the content of lipid protein and hydroxyl radicals. And superoxite dismutase activity measurement.

Body weight and weight measurement of abdominal adipose tissue

Body weight change was measured every week from the start of the experiment, the weight of the adipose tissue was calculated by taking the fat in the abdominal cavity and peri testicles on the last day of the sample input.

Determination of Total Cholesterol Content

Total cholesterol content in serum was measured using an AM 202-K assay kit (Asan Pharm. Co., Korea) prepared by an enzyme method such as Richmond (1976). In ice-cold phase, 3 ml of solution (containing 20.5 U / L cholesterol esterase, 10.7 U / L cholesterol oxidase, 1.81 g / L sodium hydroxide, 13.6 g / L potassium phosphate monobasic, 1.88 g / L phenol) are mixed with 20 μl of serum. Incubate at 37 ° C. for 5 minutes. To determine the total cholesterol content, the absorbance was measured at a wavelength of 550 nm against the reagent blank. Blood content was calculated according to a standard calibration curve.

High density lipoprotein - cholesterol  ( HDL -C) determination of content

Experiments were carried out using kit reagents (AM 203-K, Asan) prepared by the enzyme method of Noma et al. (Noma et al., 1978). 0.2 ml of sedimentation reagent (containing 0.1% dextran and magnesium chloride 0.1M) was added to 20 µl of serum, and the mixture was mixed well and allowed to stand at room temperature. 0.1 ml of the supernatant was mixed well with 3.0 ml of enzyme solution and incubated at 37 ° C. for 5 minutes. The absorbance was measured at a wavelength of 500 nm using the reagent blank as a control. The content was expressed in mg / dl according to the standard calibration curve.

low density lipoprotein  ( LDL ) - cholesterol  Content measurement

Low density lipoprotein-cholesterol (LDL-C) content was calculated by the following equation according to Fridewald et al. (Friedewald et al., 1972). LDL-C = [Total Cholesterol Amount-(HDL-C + Triglyceride / 5)]

Blood TBARS  Determination of concentration

Yagi et al. Serum lipid peroxide content was quantified by measuring serum TBARS levels according to the method of (1987) et al. 4.0 ml of 1 / 12N H ₂ SO ₄ was added to 20 µl of serum, mixed with 0.5 ml of phosphotungstic acid, and left for 5 minutes, followed by centrifugation, taking only the precipitated serum protein and again 1 / 12N H ₂ SO _4. 2.0 ml and 10 ml of 10% phosphotungstic acid were added and centrifuged. Take the precipitate and add 4.0 ml of distilled water and 1.0 ml of thiobarbituric acid-acetic acid (1: 1), react at 60 ° C for 60 minutes, cool at room temperature, add 5.0 ml of n-BuOH, and centrifuge at 3000 rpm for 15 minutes. The resulting red n-BuOH was taken and absorbance was measured using a spectrofluorometer (Ex: 515 nm, Em: 553 nm).

Blood Hydroxyl radical  Concentration measurement

Kobatake et al. Serum hydroxy radical levels were measured according to the method of (1987) and the like. In other words, 34.8 μl serum was added to 333.3 μl including 0.54 M NaCl, 0.1 M potassium phosphate buffer (pH 7.4), 10 mM NaN ₃ , 7 mM deoxyribose, 5 mM ferrous ammonium sulfate, and distilled water and mixed well in vortex at 37 ° C. It was left for 15 minutes. 67 ml of this mixture was taken and mixed with 75 ml of 8.1% sodium dodecyl sulfate, 67 ml of 20% acetic acid and distilled water. Then, 222 ml of 1.2% thiobarbituric acid was added thereto, followed by heating in a water bath (100 ° C.) for 30 minutes, cooling at room temperature, and centrifugation at 700 × g for 5 minutes, using a spectrophotometer to absorb absorbance at a wavelength of 532 nm. The amount of hydroxyl radical (nmole / mg protein) was determined by standard calibration curve.

Blood Superoxide dismutase ( SOD ) Active measurement

Blood SOD activity is described by Oyanagui et al. It was measured using the method described in (1984). It was quantified according to the method of Mirsa and Fridovich. Vortex in laboratory tube containing 100 ml serum, 500 ml distilled water, 200 ml Reagent A (3 mM hydroxylamine / 3 mM hypoxanthine), 200 ml Reagent B {7.5 mU / ml xanthine oxidase (XOD), 0.1 mM EDTA-2Na} Allow to stand for 30 minutes after generating. 2 ml Reagent C (sulfanilic acid 300 mg / N-1-naphthyl-ethylenediamine 5.0 mg of 16.7% acetic acid) was added to the reaction mixture and allowed to vortex and left at room temperature for 20 minutes. To determine the level of SOD activity in serum, the UV-VIS absorbance of the reaction mixture at 550 nm was measured and quantified according to standard calibration curves.

Example  2. Experimental results

Peroxinitrite -Erasing effect and Of cafe oil quinic acid  Component analysis

As shown in FIG. 3, the gondal methanol extract effectively inhibited peroxynitrite formation in a concentration-dependent manner. The level of inhibition was similar to that of the positive control L-penicillamine. In the experiment of peroxynitrite scavenging effect from the extracts of Korean Asteraceae, gondal ratio showed the most remarkable activity, and the IC ₅₀ value was 0.87 ± 0.33 μg / ml. This value was similar to the IC ₅₀ value of 0.89 ± 0.22 μg / ml of penicillamine, a positive control compound, and it was evaluated that the gondal methanol extract had a very strong peroxynitrite scavenging effect.

When the methanol extract was analyzed by HPLC chromatogram, all six kinds of caffeoylquinic acid were detected. Gondalabi was 3,5-di-O-caffeoylquinic acid (9.6 min), 4,5-di-O-caffeoylquinic acid (17.1 min), 3,4-di-O-caffeoylquinic acid (2.7 min), 5-O It was found for the first time that -caffeoylquinic acid (4.5 min), 3-O-caffeoylquinic acid (8.9 min) and 3-Op-coumaroyl-caffeoylquinic acids (2.9 min). The values in parentheses are the retention time of the compound. However, it was observed that it did not contain 3-O- p -coumaroylquinic acid used as a standard compound. The total content of caffeoylquinic acid in the plant material was 11.60% on average, and 3,5-di-O-caffeoylquinic acid among the six components contained 25.85% in methanol extract and 41.32% in butanol fraction. In the butanol fraction, six caffeoylquinic acids accounted for 94.52%. As a result, the content of caffeoylquinic acid compound was found to be very high in gondal ratio. HPLC chromatograms of the standard compound and gondal methanol extract are shown in FIG. 2 and the contents of the individual compounds, the total content in the plant and the total content in the extract are shown in Table 2.

will have ^a value indicated by mean ± SD values of three duplicate experiments ,, ^b ND: Not Detected Bar

obesity Rat  Changes in body weight and abdominal adipose tissue

To test the anti-obesity effect of this extract As shown in Fig. 1, a high fat diet was orally administered to rats for 6 weeks to induce obesity. Gondalbi methanol extract, ether fraction and butanol fraction were orally administered for the last 2 weeks to test the inhibitory effect on obesity. As a survey item, weight change (FIG. 4) and a change in weight of adipose tissue (FIG. 5) were measured.

Methanol extract, ether fraction and butanol fraction were all active, but butanol fraction showed the most significant obesity inhibitory effect. Butanol fraction significantly reduced body weight and abdominal adipose tissue weight to normal group level. From the above experimental results, it can be predicted that the gondal methanol extract can be developed as a very effective extract having an anti-obesity effect. Butanol fractions contain the highest amount of caffeoylquinic acid (94.52%).

Check the weight of abdominal adipose tissue 5 is shown. The weight changes of the celiac adipose tissue and epididymal adipose tissue were significantly reduced in the methanol, ether and butanol fractions. In particular, the 200 mg / kg butanol fraction showed the most remarkable effect in reducing the weight of posterior cavity and epididymal adipose tissue by 53.4% and 45.8%, respectively.

Obese Rat in  Change in Atherosclerosis Index

Serum total cholesterol, HDL-cholesterol, and LDL-cholesterol of the rats were examined. Table 3 shows the results. Gondalbi methanol extract, ether extract and butanol extract all significantly reduced total cholesterol and LDL-cholesterol but did not significantly change serum HDL-cholesterol (Table 3).

In the atherosclerosis risk index, the normal rats showed 0.42 ± 0.09 and the control group increased 1.81 ± 0.13. All of the administered groups were shown to be beneficial for atherosclerosis by reducing the risk of atherosclerosis. In particular, the butanol fraction administration group exhibited the greatest atherosclerotic risk index of 1.22 ± 0.11 (Table 3).

The analytical procedure is described in the experimental method. Values are the average ± S.D. of the six experiments. Values according to the same letter do not differ significantly (p <0.05).

Atherosclerosis Index (AI) = (total cholesterol-HDL cholesterol) / HDL cholesterol

Obese Rat in TBARS Effect on

Levels of TBARS, hydroxy radicals and SOD activity were significantly increased in the control group compared to the normal group (Table 4). This suggests an increase in reactive oxygen species (ROS), lipid peroxidation and oxidative stress. Methanol extract and all fractions inhibited increased TBARS, hydroxy radical levels and increased SOD activity levels. In particular, this inhibitory effect was most remarkable in the butaneol 200 mg / kg administration group.

The analytical procedure is described in the experimental method. Values are the average ± S.D. of the six experiments. Values according to the same letter do not differ significantly (p <0.05).

SOD 1 Unit: defined as the amount of enzyme that inhibits adrenochrome formation rate by 50%

Claims (22)

delete delete delete delete delete delete delete delete delete delete delete delete delete delete delete A first step of obtaining gondal extract by extracting gondal with methanol; And
From the extract gondalbi 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid 3,5-di- O- dicaffeoyl- epi from gondalby, comprising a second step of separating one or more compounds selected from the group consisting of (5-CQ) and 3- O- caffeoylquinic acid (3-CQ) -quinic acid (3,5-DeQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ), and 3- O -caffeoylquinic acid (3- CQ) at least one compound separation method selected from the group consisting of.
A first step of obtaining gondal extract by extracting gondal with methanol;
A second step of dividing the gondalby extract with butanol to obtain a fraction of gondalby; And
From the fraction 3,5-di- O -dicaffeoyl- epi -quinic acid (3,5-DeQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid ( 5-CQ) and 3- O -caffeoylquinic acid (3-CQ ), 3,5-di- O from gondalbi comprising a third step of separating the at least one compound selected from the group consisting of -dicaffeoyl- epi - quinic acid (3,5-DeQ), 4,5-di-O-caffeoylquinic acid (4,5-DQ), 5- O -caffeoylquinic acid (5-CQ) and 3- O -caffeoylquinic acid (3-CQ At least one compound selected from the group consisting of: A pharmaceutical composition for the treatment of obesity, which contains methanol extract of Ligularia stenocephala as an active ingredient.
Gonalbi (Ligularia stenocephala) Food composition for the prevention or improvement of obesity containing methanol extract as an active ingredient.
A pharmaceutical composition for treating obesity, comprising butanol fraction or ether fraction of methanol extract of Ligularia stenocephala as an active ingredient.
Food composition for the prevention or improvement of obesity containing butanol fraction or ether fraction of methanol extract of Ligularia stenocephala as an active ingredient.
A first step of obtaining gondal extract by extracting gondal with methanol;
A second step of fractionating the gondalby extract with ether to obtain a fraction of gondalby; And
Method of separating the fractions from 5- O -caffeoylquinic acid (5-CQ ) 5- O -caffeoylquinic acid (5-CQ), from gondalbi comprising a third step of separating.

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