KR20190090362A - A composition for imobesity containing dicaffeoylquinic acid - Google Patents

Abstract

The present invention relates to a composition for alleviating obesity containing dicaffeoylquinic acid as an active component. The composition is easier to access and has less side effects than a chemical medicine by using dicaffeoylquinic acid derived from mugwort as a natural substance and also as the active component, and thus has the activity of inhibiting toxicity and fat accumulation, glycolysis inhibitory activity, fat diet-induced obesity alleviation activity, and biochemical activity, thereby being able to be usefully utilized as a novel anti-obesity material.

Description

TECHNICAL FIELD The present invention relates to a composition for improving obesity containing dicaffeoylquinic acid as an active ingredient,

The present invention relates to a composition for improving obesity containing dicaffeoylquinic acid as an active ingredient.

Obesity is an excess accumulation of fat in the body caused by genetic cause or lifestyle habits, and causes diseases such as adult diseases and chronic degenerative diseases. In advanced countries, 2 ~ 7% of the total national health expenditure is caused by overweight and obesity have. As a result, the World Health Organization (WHO) has become a '21st Century New Infection' and the American Medical Association has defined obesity as a 'disease'. This global movement suggests that obesity is not simply a matter of beauty or physical phenomenon, but a disease that requires treatment.

Obesity is not just obesity, but it can cause various diseases and even cause mental diseases. Is a representative obesity-related disease such as type 2 diabetes, dyslipidemia, hypertension, fatty liver, gallbladder disease, coronary artery disease (angina pectoris, myocardial infarction), stroke, sleep apnea, gout, osteoarthritis, .

Koreans also have a steep increase in obesity (body mass index <30-35) and high-altitude obesity (body mass index <35). According to the '2016 Obesity White Paper' published by the National Health Insurance Corporation, the nation 's super - heavy obesity population has tripled from 9,148 in 2015 to 19,643 in 9 years. During the same period, high altitude obesity increased 2.5 times from 219,905 to 5,348,600. The obesity population increased 1.7 times from 2,331,146 to 4,060,6015. In the current trend, one out of every 17 adults in Korea will be overweight.

Therefore, in Korea, along with efforts in the medical and pharmaceutical sectors, the government is promoting the obesity campaign to promote obesity. Like medical advanced nations, Koreans are also making awareness of raising awareness of obesity and defining obesity as a disease.

The most common methods of weight control are dieting, exercise, behavior modification, and drug therapy. Diet related industries include diet foods for weight control, dietary supplements, diet drugs, , A clinic, a health center, and a clinic for obesity in hospitals.

The market for diet foods for weight loss continues to grow. Recently, interest in natural products, which are more accessible than medicines and known to have few side effects, has led to the active promotion of dietary-related health functional foods and special- to be.

On the other hand, 'health functional foods' refers to foods manufactured using raw materials or ingredients having useful functions in the human body. In recent years, weight loss has been attempted in many ways due to a social atmosphere that is biased not only in promoting health but also in appearance. In addition, the market for diet foods is getting larger as commercial diet foods are attracted attention as a way of weight control. In other words, there is an increasing interest in weight reduction using foods that are easier to access than drugs and have less side effects.

In the case of anti-obesity, classification according to the functional group is divided into digestion, appetite control, and fat metabolism control.

Digestion and appetite control are foods that inhibit the digestion of ingested foods and prevent them from being absorbed into the body. Food ingredients and ingredients that act to limit food intake by suppressing appetite are included in this subclass , Digestion and appetite control are divided into digestion control and appetite control.

Regulation of digestion is mainly related to the digestion and absorption of saccharides. In digestion fields of saccharides, digestion and absorption inhibitors are mostly related to amylase or α-glucosidase.

Appetite control is a substance that controls appetite by controlling neurobiological activity. It is largely classified into monoamines, peptides (pancreatic polypeptide family, secretin family, bombesin family and neuromedin U family, opioid peptide family, CRF family, Melanocortin, and fibroblast growth factor), cytokines (interleukins, interferons, MIP-1 and the like), and other factors (thyroid stimulating hormone releasing hormone, proopiomelanocortin, oxytocin, arginine Vasopressin, prolactin, cytosine, insulin, somatostatin, cholestystinin, enterostatin, gallanin, neurotensin, motilin, ghrelin, CART, etc.).

Fat metabolism control is intended for food containing ingredients and ingredients that control the metabolism of fat, and is classified as inhibiting body fat synthesis and inhibiting fat absorption.

The research on obesity and adipocyte is divided into two main parts: obesity (the mechanism of appetite regulation, the mechanism of metabolism of the body energy, the mechanism of adipocyte differentiation) and the mechanism of disease development in obesity Which is divided into two groups.

The accumulation of visceral fat is an important cause of cardiovascular disease. It has been shown that the concentration of visceral fat is increased by accumulation of visceral fat as well as the secretion of visceral fat. TNF-α secreted from adipocytes lowers insulin sensitivity, One.

Therefore, the regulation of lipid metabolism is regulated in a close mutual control relationship with the regulation of blood sugar. In the classification of anti-obesity foods having the function of inhibiting fat absorption and inhibiting the synthesis of body fat, Includes techniques in which the signaling primary messenger is involved in the regulation of lipid metabolism.

Regarding the suppression of fat absorption, Orlistat is known, and this substance is known to have a function of strongly suppressing various lipases including pancreatic lipase and carboxyester lipase. In addition, 5-campesterone is a derivative of campesterol, a kind of plant sterol (phytosterol), and is a kind of oxidized sterol. Since they are known to significantly reduce liver triglyceride (TG) and cholesterol, and are known to have an effect of hyperglycemia after meal and hyperglycemia of lipid metabolism during fasting, their anti-obesity effect It is a different feature from other obesity treatments to exert its effects, especially reducing visceral fat.

Accordingly, an object of the present invention is to provide a health functional food for preventing or ameliorating obesity containing dicaffeoylquinic acid as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity containing dicapheloquinic acid as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or ameliorating obesity containing dicaphea oil quinic acid as an active ingredient.

DISCLOSURE OF THE INVENTION In order to achieve the above object, the present inventors have succeeded in separating 3,5-diCQA (3,5-dicaffeoylquinic acid) and 4,5-diCQA (4,5-dicaffeoylquinic acid) , And 5-diCQA were tested for toxicity, inhibition of lipid accumulation, inhibition of sugar chain inhibition, activity of improving obesity induced by high-fat diet, and biochemical activity.

According to one embodiment of the present invention, there is provided an anti-obesity composition containing dicapheloquinic acid as an active ingredient.

Here, the dicafeoylquinic acid may be 3,5-dicaphelaquinic acid or 4,5-dicaphelaquinic acid.

The dicaffeoylquinic acid may be at least one of the following formulas (1) and (2).

[Chemical Formula 1]

(2)

The dicafeoylquinic acid may be one isolated from the mugwort extract or the fraction thereof.

Artemisia princeps Pampan ., Artemisia annua Linne , Artemisia apiacea HANCE , Artemisia scoparia , Artemisia keiskeana , Artemisia keiskeana , mulssuk (Artemisia selengensis TURCZ.), huinssuk (Artemisia stelleriana Bess), heat being (Artemisia iwayomogi Kitamura), hair sanssuk (Artemisia sacrorum subsp.), wormwood (Artemisia asistica Nakai), sanssuk (Artemisia montana Pampan), injinssuk (Artemisia iwayomogi Kitamura ), Artemisia capillaris Thunb ., Artemisia lavandulaefolia , Artemisia argyi , or Artemisia japonica Thunb .

In addition, the mugwort extract may be extracted with water, an alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, hexane, dichloromethane, or a mixed solvent thereof.

In one embodiment, the mugwort fraction may be obtained by fractionating the mugwort extract into a polar solvent containing ethyl acetate or acetone.

In one embodiment, the decaffeoylquinic acid may be a mixture of mugwort ethanol extract: mugwort ethyl acetate fraction in a ratio of 2: 8 to 8: 2.

According to another embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating a metabolic disease comprising the composition.

Herein, the metabolic diseases may include diabetes, hyperlipidemia, stroke, arteriosclerosis, cardiovascular disease or non-alcoholic fatty liver.

According to another embodiment of the present invention, there is provided a health functional food for preventing or ameliorating a metabolic disease comprising the composition.

According to another embodiment of the present invention, there is provided a food for preventing or improving a metabolic disease comprising the composition.

The food may include beverages, tea, liquor, gum, candy, confectionery, and food additives.

According to the composition for improving obesity containing the dicafeoylquinic acid of the present invention as an active ingredient as described above, dicapheloquinic acid derived from mugwort, as a natural substance, is used as an effective ingredient and is easier to approach than a chemical drug and has side effects And can be usefully used as a new anti-obesity diet material because of its toxicity, fat accumulation inhibiting activity, sugar inhibiting activity, high fat diet induced obesity improving activity, and biochemical activity.

The above and other objects, advantages and novel features of the present invention will become more apparent from the following detailed description when read in conjunction with the accompanying drawings, in which: FIG. It can be understood.

FIG. 1 shows the results of cytotoxicity tests on adipocytes using MTT assay of 3,5-diCQA and 4,5-diCQA according to an embodiment of the present invention.
FIG. 2 shows the effect of 3,5-diCQA and 4,5-diCQA on lipid accumulation in adipocytes according to an embodiment of the present invention.
FIG. 3 shows the effect of 3,5-diCQA and 4,5-diCQA, CGA on the expression of adipogenesis signaling molecules in adipocytes according to an embodiment of the present invention.
FIG. 4 shows the sugar chain inhibition activity of (A) α-glucosidase and (B) α-amylase of 3,5-diCQA and 4,5-diCQA according to an embodiment of the present invention.
FIG. 5 is a graph showing the effect of the 3,5-diCQA and 4,5-diCQA on the weight gain of (A) and the weight of epididymal adipose tissue (B) in obese mice induced by high fat diet according to an embodiment of the present invention Lt; / RTI &gt;

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

The present invention relates to a composition for improving obesity containing dicaffeoylquinic acid as an active ingredient.

According to one embodiment of the present invention, it is possible to provide a composition for preventing or improving an anti-obesity agent containing, as an active ingredient, dicafeoylquinic acid derived from mugwort. Artemisia princeps Pampan ., Artemisia annua Linne , Artemisia apiacea HANCE , Artemisia scoparia , Artemisia keiskeana , Artemisia keiskeana , Artemisia keiskeana , Artemisia selengensis TURCZ.), huinssuk (Artemisia stelleriana Bess), heat being (Artemisia iwayomogi Kitamura), hair sanssuk (Artemisia sacrorum subsp.), wormwood (Artemisia asistica Nakai), sanssuk (Artemisia montana Pampan), injinssuk (Artemisia iwayomogi Kitamura) , Artemisia capillaris Thunb ., Artemisia lavandulaefolia , Artemisia argyi or Artemisia japonica Thunb . Preferably, Artemisia argyi can be used.

Artemisia argyi is a plant distributed in Korea, China and Russia. Its height is 45 ~ 120cm and its stem is straight. Leaves are opposite, thick, long lance, but fragrant. In July, a light brown flower blooms in cones. The fruit is long oval (瘦果) bear. It has mainly edited jujubes, extracts fragrant oil from the outposts, and leaves with persimmon leaves. It is mainly used as root for menstrual irregularity, uterine leiomyoma, pregnancy addiction.

In one embodiment of the present invention, the dicaffeoylquinic acid may be at least one of those represented by the following general formula (1) or (2).

[Chemical Formula 1]

(2)

Here, it is preferable that the present invention is a single dicafeoylquinic acid represented by the general formula (1).

Also, in one embodiment of the present invention, the dicafeoylquinic acid may be at least one of 3,5-dicapheloquinic acid or 4,5-dicapheloquinic acid.

In one embodiment of the present invention, dicapheloquinic acid has a fat accumulation inhibiting activity and a sugar chain inhibiting activity.

According to another embodiment of the present invention, it can be used as a pharmaceutical composition for preventing or treating obesity which contains, as an active ingredient, dicafeoylquinic acid derived from mugwort.

According to another embodiment of the present invention, it can be used as a food composition for prevention or improvement of obesity which contains, as an active ingredient, dicaphea oil quinic acid derived from mugwort.

The prevention of obesity in the present invention may be any of prevention of obesity such as digestion and appetite control, regulation of fat metabolism, inhibition of body fat synthesis, regulation of metabolism of body energy, and regulation of lipid differentiation cells.

In the present invention, the decaffeoylquinic acid may be prepared by extracting mugwort with an extraction solvent or extracting with an extraction solvent, followed by fractionating the extract with a fraction solvent. In the present invention, it is not limited to an extraction solvent, but water, a C 1 to C 4 alcohol, a polar solvent such as ethyl acetate or acetone, a nonpolar solvent such as hexane or dichloromethane, or a mixed solvent thereof may be used. More preferably, ethanol can be used. In one embodiment of the present invention, the extract is extracted with 80% ethanol as the solvent to prepare a mugwort extract containing dicafeoylquinic acid, and dicapheloquinic acid can be fractionated from mugwort extract using ethyl acetate.

The term "extract " used in the present invention means a preparation which is obtained by squeezing mugwort with an appropriate leaching solution and concentrating by evaporating the leaching solution, and is not limited thereto. The extract, diluted solution or concentrate of the extract, A dried product obtained by drying the extract, a controlled preparation thereof, or a purified product thereof. The dicafeoylquinic acid can be prepared from wormwood using common extraction, isolation and purification methods known in the art. The extraction method may be, but not limited to, hot water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction.

In order to prevent or improve the obesity of dicaphea oil quinic acid, it is possible to use the yellow mugwort extract or fraction alone or a mixture thereof. Preferably, the yellow mugwort extract and the yellow mugwort fraction obtained by extracting or fractionating yellow mugwort with the respective solvents can be mixed to improve the effect of preventing or improving obesity.

In the present invention, the term "prevention" means any action that inhibits or delays the development of obesity and its complications by administration of the pharmaceutical composition according to the present invention.

As used herein, the term "treatment" refers to any action that alters the suspicion of obesity and the symptoms of an onset subject by administration of the pharmaceutical composition. Those skilled in the art will be able to ascertain the precise criteria of the disease for which the composition of the present invention is effective by referring to the data presented by the Korean Medical Association, will be.

In the present invention, anti-obesity is a complication due to obesity and means metabolic diseases.

Preferably, the metabolic disease can be provided as a composition for the prevention and treatment of metabolic diseases including diabetes, hyperlipidemia, stroke, arteriosclerosis, cardiovascular disease or non-alcoholic fatty liver.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects, Factors well known in the art and other medical disciplines, including health status, type of obesity, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and rate of release, duration of treatment, &Lt; / RTI &gt; The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art.

In the present invention, the pharmaceutical composition may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, liquid preparations, creams, lotions, pastes or solid percutaneous topical preparations, suppositories, Sterile injectable solutions, and sprays. &Lt; RTI ID = 0.0 &gt; [0040] &lt; / RTI &gt;

The therapeutically effective amount of the pharmaceutical composition of the present invention may vary depending on a variety of factors, such as the method of administration, the site of administration, the condition of the patient, and the like. Therefore, when used in the human body, the dosage should be determined in consideration of safety and efficacy. It is also possible to estimate the amount used in humans from the effective amount determined through animal experiments. Such considerations in determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman ' s Pharmacological Basis of Therapeutics, 10th ed. (2001), Pergamon Press; And E.W. Martin ed., Remington ' s Pharmaceutical Sciences, 18th ed. (1990), Mack Publishing Co.

In the present invention, the pharmaceutical composition may contain one or more active ingredients showing the same or similar functions by adding dicapheylquinic acid isolated from wormwood to a pharmaceutically acceptable salt.

The pharmaceutical compositions of the present invention may also include carriers, diluents, excipients, or a combination of two or more thereof, which are commonly used in biological agents. The pharmaceutically acceptable carrier is not particularly limited as long as the composition is suitable for in vivo delivery, for example, Merck Index, 13th ed., Merck & Inc. A buffered saline solution, a buffer solution, a dextrose solution, a maltodextrin solution, glycerol, ethanol, and one or more of these components may be mixed and used, and if necessary, an antioxidant, a buffer, Conventional additives may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into main dosage forms such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules or tablets. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science (Mack Publishing Company, Easton PA, 18th, 1990) in a suitable manner in the art.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, Wherein the starch is selected from the group consisting of lactose, mannitol, sugar, arabic gum, pregelatinized starch, cornstarch, powdered cellulose, hydroxypropyl cellulose, opaques, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, Calcium, white sugar, dextrose, sorbitol and talc may be used. The pharmaceutically acceptable excipient according to the present invention is preferably, but not limited to, 0.1 part by weight to 90 parts by weight of the dicaphelaquinone acid fraction.

The composition of the present invention may be administered orally or non-orally (for example, intravenously, transdermally, intraperitoneally or topically) or orally administered in accordance with a desired method, and the dose may be appropriately determined depending on the patient's body weight, The range varies depending on diet, time of administration, method of administration, excretion rate, and severity of the disease.

Examples of the liquid preparation for oral administration of the composition of the present invention include suspensions, solutions, emulsions, syrups, and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, Etc. may be included together. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.

The composition of the present invention may include a health functional food. The term "health functional food " as used in the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and rings using raw materials and components having useful functions in the human body. Here, 'functional' refers to the structure and function of the human body to obtain nutritional effects and obtain useful effects for health use such as physiological action. The health functional food of the present invention can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and components which are usually added in the conventional technical fields. In addition, the formulations of the above health functional foods may also be manufactured without limitations as long as they are acceptable as health functional foods. The composition for food of the present invention can be manufactured in various forms, and unlike general pharmaceuticals, it has the advantage that there is no side effect that may occur when a drug is used for a long period of time, and is excellent in portability, Can be ingested as an adjunct to improve the effects of other obesity improvement and treatment.

There is no limitation on the kind of health food to which the composition of the present invention can be used. In addition, it may be prepared by mixing the appropriate auxiliary ingredients, which may be contained in health functional foods, with known additives, depending on the selection of the person skilled in the art, including the dicapheloquinic acid derived from mugwort of the present invention. Examples of foods that can be added include dairy products, such as meat, sausage, bread, chocolates, candies, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Vitamin complex, and the like, and can be prepared by adding to the juice, tea, jelly, and juice prepared from the extract of the present invention as a main component.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the embodiments described below are examples for embodying the contents of the present invention, and thus the present invention is not limited thereto.

&Lt; Example 1 >

1-1 Materials Preparation

The 3T3-L1 adipocyte precursor cells used in the present invention were purchased from Korean Cell Line Bank (KCLB, Seoul). The 3T3-L1 adipocyte precursor cells used in the present invention were 3,5-dicaffeyolquinic acid (3,5-diCQA) acid (4,5-diCQA) was purchased from Chengdu Biopurify Phytochemicals Ltd. (Chengdu, China). (MTT) assay kit was purchased from Sigma Chemical Co. (St. Louis, Mo., USA) as a reagent, and 3T3-dTh- Dulbecco's modified Eagle's medium and fetal bovine serum were purchased from Gibco BRL Co. (Grajd Island, NY, USA), and penicillin, streptomycin, Sodium bicarbonate and HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).

Preparation of 1-2 dicafoyloquinic acid

De café five days quinone acid (dicaffeyolquinic acid) in the present invention was used for the extracts and fractions prepared from the Yellow mugwort (Artemisia argyi). The ethanol extracts of freeze - dried Yellow mugwort were extracted with ethanol (EtOH) and the yellow mugwort ethyl acetate fractions were extracted with ethyl acetate (EtOAc). Ethanol extract of Yellow sea mugwort and Ethyl acetate fraction of Yellow sea mugwort were found to contain different contents of 3,5-dicaffeoylquinic acid (3,5-diCQA) and 4,5-dicaffeoylquinic acid (4,5-diCQA). The yellow mugwort extract and fractions prepared in this Example were concentrated and stored at -20 ° C.

1-3 Preparation of Mixture of Yellow Sea Mugwort Extract

In this Example, a mixture of ethanol extract of Mugwort mugwort and ethyl acetate fraction of Mugwort mugwort was prepared.

Ethanol extract (EtOH): Yellow mugwort ethyl acetate fraction (EtOAc) was mixed with 2: 8, 4: 6, 6: 4 and 8: 2, respectively.

Experimental Example 1: Inhibition of toxicity and fat accumulation

1-1 Inhibitory effect on toxicity and fat accumulation

The differentiation of 3T3-L1 preadipocyte into adipocyte was carried out with 0.5 mM 3-isobutyl-1-methylxanthine (Sigma, USA), 1 μM dexamethasone (Sigma, USA), 10 μg / USA) was treated with 10% FBS-DMEM (MDI medium). 2 × 10 ⁵ cells were plated on a 24-well tissue culture plate and replaced with medium containing 10% FBS for 2 days. After 2 days, the medium was changed to a medium containing MDI, and the samples were treated at different concentrations and treated with 4-times insulin, 3,5-diCQA and 4,5-diCQA, respectively, at intervals of 2 days. Finally, 3,5-diCQA and 4,5-diCQA were treated and after 2 days, the degree of adipocyte differentiation and the inhibition of differentiation by the sample were observed at a magnification of 200 times using a phase contrast microscope, The inhibition of differentiation and TG production inhibition were analyzed by Oil Red O staining. The cells were washed with 1 × phosphate buffered saline (PBS), fixed with 10% formalin, treated with Oil Red O staining solution, and stained for 30 minutes. After staining, the stained fat was extracted with 100% isopropanol and measured at 500 nm using a multi-plate reader.

FIG. 1 shows the results of cytotoxicity tests on adipocytes using MTT assay of 3,5-diCQA and 4,5-diCQA according to an embodiment of the present invention. (A) is a cytotoxicity test result of 3,5-diCQA, and (B) is a cytotoxicity test result of 4,5-diCQA. FIG. 2 shows the effect of 3,5-diCQA and 4,5-diCQA on lipid accumulation in adipocytes according to an embodiment of the present invention. (A) is the result of 3,5-diCQA and 4,5-diCQA in 3T3-L1 adipocytes when stained with Oil Red O, and (B) is the visualization of (A). As a result of the MTT assay, it was confirmed that the cytotoxicity of 3,5-diCQA and 4,5-diCQA, which were analyzed as main substances of mugwort, did not exist at 100 uM, and ORO staining was used to confirm the fat accumulation inhibitory activity. When fat accumulation of 3T3-L1 adipose precursor cells was elicited, up to 150% of fat accumulation occurred. However, when 4,5-diCQA and 3,5-diCQA were treated, fat accumulation decreased to 140% and 134%, respectively These results suggest that 4,5-diCQA is superior to 3,5-diCQA in inhibiting lipid accumulation.

1-2 Identification of Protein Expression Changes Associated with Fat Accumulation

3 × 10 ⁵ cells were seeded in a 6-well tissue culture plate and 3,5-diCQA, 4,5-diCQA and a positive control CGA (chlorogenic acid) After cell lysate treatment, the protein concentration was determined by Bradford assay, and electrophoresis was performed using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), blotted onto polyvinylidene difluoride (PVDF) membrane, . The results were then analyzed with the iBright ™ CL1000 Imaging System (Thermo Fisher, Waltham, MA).

FIG. 3 shows the effect of 3,5-diCQA and 4,5-diCQA, CGA on the expression of adipogenesis signaling molecules in adipocytes according to an embodiment of the present invention. (A) shows the change of p-JNK protein causing cell death, (B) shows the change of p-Akt protein which suppresses the expression of p-AMPK, and (C) And the change of p-AMPK in which fat accumulation is suppressed. As a result, the expression of p-JNK protein in the 3,5-diCQA and 4,5-diCQA of the present invention was significantly lower than that of CGA (chlorogenic acid), which is a positive control group, And the expression of p-AMPK protein, In addition, 4,5-diCQA in the 3,5-diCQA and 4,5-diCQA of the present invention was found to increase lipid accumulation-inhibiting protein expression compared with 3,5-diCQA, and the expression of p- JNK protein expression was also decreased.

1-3 inhibition activity of α-amylase and α-glucosidase

The inhibition of α-amylase activity was measured by dissolving α-amylase (porcine pancreatic α-amylase, sigma) solution in sample solution and 0.02 M sodium phophate buffer (pH 6.9 with 0.006 M sodium chloride) And the absorbance was measured at 540 nm using a microplate reader.

α-Glucosidase activity was measured by dissolving ρ-nitrophenol-α-D-glucopyranoside (PNPG, Sigma) in 0.1 M phospate buffer (pH 6.9) The absorbance was measured at 405 nm using a microplate reader.

FIG. 4 shows the sugar chain inhibition activity of (A) α-glucosidase and (B) α-amylase of 3,5-diCQA and 4,5-diCQA according to an embodiment of the present invention. 3,5-diCQA and 4,5-diCQA α-amylase and glucosidase Compared with acarbose, the inhibitory activity was lower than that of acarbose. However, it was confirmed that 4,5-diCQA showed a higher sugar inhibition activity than 3,5-diCQA. These results demonstrate the usefulness of digestion and absorption inhibitors in the field of digestion of saccharides.

<Experimental Example 2> Improvement effect of 3,5-diCQA and 4,5-diCQA on obesity

2-1 Improvement of obesity induced by highland diet

4-week-old C57BL / 6 male mice were purchased from an experimental animal supplier (Samtako Bio Korea, Korea) and adapted for 7 days. All animals were kept at a constant temperature (24 ± 1 ° C) and humidity (55%), and 12 hours of light and 12 hours of shading were supplied with sufficient amounts of drinking water and feed. All groups except the normal diet were fed a high fat diet with a fat content of 60% for 16 weeks after induction of obesity. The subjects were divided into 3 groups of 7 rats per each group. To examine the anti-obesity effect of the sample extracts from 17 to 21 weeks, the high-fat diet and the sample were orally administrated. Body weight and dietary intake were measured once every 7 days while observing animal conditions during the experiment.

FIG. 5 is a graph showing the effect of the 3,5-diCQA and 4,5-diCQA on the weight gain of (A) and the weight of epididymal adipose tissue (B) in obese mice induced by high fat diet according to an embodiment of the present invention Lt; / RTI &gt; The effects of 3,5-diCQA and 4,5-diCQA on obesity-induced rats in a 16-week high-fat diet resulted in some suppression of body weight gain in the 4-week dietary group However, the effect was insufficient. However, the accumulation of visceral fat was found to significantly reduce visceral fat in the group taking 3,5-diCQA and 4,5-diCQA fractions.

2-2 Biochemical analysis through serum analysis

After the oral administration of the experimental diet, the test animals were anesthetized with ethyl ether and blood was collected from the heart. The collected blood was placed in a 37 ° C water bath and the serum was centrifuged at 15,000 rpm for 15 minutes at 4 ° C as an analytical sample Biochemical analysis. In the experiment 2-2, biochemical tests were conducted using an automatic hematology analyzer (DRI-CHEM 4000i, FUGIFILM, Tokyo, Japan) for serum lipid, total cholesterol, HDL-cholesterol concentration, BUN, CRE, GLU, GOT and GPT LDL-cholesterol was calculated according to the method of Friedewald et al., Total cholesterol- (triglyceride + HDL-cholesterol) / 5 and VLDL-cholesterol was calculated as total cholesterol- (HDL-cholesterol + LDL-cholesterol) Respectively.

The following Table 1 shows the results of the experiments for the determination of 4 glutamic oxaloacetic transaminase (GOT), glutamine pyruvic transaminase (GPT), blood urea nitrogen (BUN), creatine (CRE), lactate dehydrogenase (LDH), total cholesterol (TCHO), triglyceride , 5-diCQA, and high-density lipoprotein cholesterol and total cholesterol ratio (HTR) and low-density lipoprotein cholesterol (LDLC).

As shown in Table 1, in the serum analysis results, it was confirmed that the levels of GOP and GTP, which are confirmed as indicators of liver damage and fatty liver, are decreased in the group ingested with the 4,5-diCQA fraction of the above example. It was also found that the amount of total cholesterol (TCHO), triglyceride (TG) and LDLC decreased. In other words, 4,5-diCQA seems to be very helpful in improving liver damage, fatty liver formation and dyslipidemia caused by obesity.

<Experimental Example 3> Improvement of obesity effect of mixture of Mugwort extract and fraction

The mixture of the ethyl acetate fraction of mugwort yellow mugwort and the ethanol extract of mugwort mugwort, which was confirmed in Example 1, was prepared and the mixing ratio having the optimal obesity improving effect was confirmed.

Analysis of the material of the EtOAc fraction of the waxy mugwort of Example 1 from the UPLC Q-TOF / MS system revealed that 3,5-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and dicaffeoylquinic acid acid glucoside was identified as the main substance (Fig. 6).

In addition, the ethanol extract (EtOH) and the ethyl acetate fraction (EtOAc) of the yellow mugwort were mixed in the ratio of 2: 8, 4: 6, 6: 4 and 8: 2 respectively and 3T3- Accumulation inhibitory effect was confirmed (Fig. 7). Experimental results showed that the mixture of EtOAc: EtOH (8: 2) at a concentration of 100 μg / mL, an undetectable concentration, showed a significantly better fat accumulation inhibitory activity than the other mixtures.

<Experimental Example 4> Inhibitory activity on sugar degradation of a mixture of mugwort extract and fraction

As a result of measurement of α-amylase and α-glucosidase inhibitory activity, the mixture of EtOAc: EtOH (8: 2) showed a superior sugar-degradation inhibitory activity when compared with other mixtures (Fig. 8). Specifically, in the α-amylase inhibitory activity experiment, the EtOAc: EtOH (8: 2) mixture showed significantly better α-amylase inhibitory activity than the other mixtures. The mixture of EtOAc and EtOH (8: 2) showed the most inhibitory effect on the α-glucosidase inhibitory activity. As a positive control, Acarbose (250 μg / mL), which is an alpha-glucosidase inhibitor, α-glucosidase inhibitory activity was confirmed.

As a result, it was confirmed that the (8: 2) mixture of ethyl acetate fraction and ethanol extract of yellow mugwort increased the obesity improvement effect significantly.

As described above, the 3,5-diCQA and 4,5-diCQA of the present invention are confirmed to have anti-obesity functionalities. This is a natural substance that is easier to access than chemical drugs and has less side effects, so it is useful as a new anti-obesity and metabolic disease treatment and improvement material.

As described above, the composition for improving obesity containing dicafeoylquinic acid as an active ingredient according to the present invention has been described in detail. However, those skilled in the art of the present invention, It will be apparent to those skilled in the art that various modifications and additions may be made without departing from the scope of the present invention. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. The scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents are included in the scope of the present invention .

Claims (12)

An anti-obesity composition comprising deca-coke oil quinic acid as an active ingredient. The method according to claim 1,
Wherein said dicafeoylquinic acid is 3,5-dicapheylquinic acid or 4,5-dicaphelaquinic acid. The method according to claim 1,
Wherein the dicaffeoylquinic acid is at least one of the following Formula 1 or Formula 2:
[Chemical Formula 1]

(2)

The method according to claim 1,
Wherein the decaffeoylquinic acid is separated from the mugwort extract or the fraction thereof. 5. The method of claim 4,
The mugwort is sajabal mugwort, tufts mugwort (Artemisia princeps Pampan.), Robin wormwood (Artemisia annua Linne), tarragon (Artemisia apiacea HANCE), bissuk (Artemisia scoparia), clear daessuk (gaejebissuk, Artemisia keiskeana), mulssuk (Artemisia selengensis TURCZ.), huinssuk (Artemisia stelleriana Bess), heat being (Artemisia iwayomogi Kitamura), hair sanssuk (Artemisia sacrorum subsp.), wormwood (Artemisia asistica Nakai), sanssuk (Artemisia montana Pampan), injinssuk (Artemisia iwayomogi Kitamura), sacheolssuk Wherein the composition is selected from the group consisting of Artemisia capillaris Thunb ., Artemisia lavandulaefolia , Artemisia argyi , Artemisia japonica Thunb . 5. The method of claim 4,
Wherein the mugwort extract is extracted with water, an alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, hexane, dichloromethane or a mixed solvent thereof. 5. The method of claim 4,
Wherein the mugwort fraction is obtained by fractionating the mugwort extract with a polar solvent containing ethyl acetate or acetone. The method according to claim 1,
Wherein the decaffeoylquinic acid is a mixture of mugwort ethanol extract: mugwort ethyl acetate fraction in a ratio of 2: 8 to 8: 2. 9. A pharmaceutical composition for the prevention or treatment of metabolic diseases comprising the composition of any one of claims 1 to 8. 10. The method of claim 9,
Wherein the metabolic disease includes diabetes, hyperlipidemia, stroke, arteriosclerosis, cardiovascular disease, or nonalcoholic fatty liver. A health functional food for preventing or improving a metabolic disease comprising the composition of any one of claims 1 to 8. A food for preventing or improving a metabolic disease comprising the composition of any one of claims 1 to 8.

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