KR101760163B1 - Composition for anti-obesity comprising extract of Sargassum serratifolium as an effective component - Google Patents

Abstract

More particularly, the present invention relates to an anti-obesity pharmaceutical composition or health functional food containing an extract of Sargasia serratifolium or an n-hexane fraction thereof as an active ingredient. to provide. Since the Sawtooth mildew extract of the present invention or the n-hexane fraction isolated therefrom is a safe substance having excellent inhibitory effect on adipocyte differentiation, fat production and fat accumulation and is free from cytotoxicity, the pharmaceutical composition for reducing body fat and treating obesity Or as an active ingredient of a health functional food.

Description

TECHNICAL FIELD The present invention relates to a composition for anti-obesity comprising an extract of Sawtooth mildew as an active ingredient,

TECHNICAL FIELD The present invention relates to a composition for anti-obesity derived from a sawtooth hat, and relates to a pharmaceutical composition or health functional food for an anti-obesity containing an extract of Sargassum serratifolium or a fraction thereof as an active ingredient.

Obesity is a disease caused by the imbalance of food intake and energy use, and can be accompanied by serious illnesses such as cancer, type 2 diabetes and arteriosclerosis (Kopelman, 2000). Recently, obesity, diabetes, dyslipidemia, and hypertension have been called metabolic syndrome. Despite the worldwide efforts to understand and respond to the causes of obesity, obesity is accelerating. The WTO warns that "Obesity is a disease that requires long-term transparency." Obesity is not simply a cosmetic issue, but a cause of various diseases, increasing the perception that it is a serious life-threatening disease. The World Health Organization (WHO) says 17 million people die each year from cardiovascular disease, an important risk factor for obesity, and warns that in 2015, 1.5 billion people worldwide, up by 50 percent, could suffer from obesity.

Obesity has been reported to be caused by complex factors such as genetic, social, and economic factors, as well as social diminution of exercise and failure to control energy homeostasis due to increased intake of high calorie food. Recently, the worldwide incidence of obesity associated with metabolic syndrome including type 2 diabetes, which is known to be caused by ingestion of high-calorie foods and deterioration of physical activity, is increasing. In 2025, the population with these metabolic syndrome doubles to about 300 million. Obesity is a result of the unbalance of energy intake and consumption of the human body, resulting in an increase in fat cell size due to fat accumulation and an increase in the number of adipocytes due to the differentiation of fibroblasts into adipocytes. The adipogenesis process of fibroblasts differentiates into adipogenesis, which involves liver X receptor α (LXRα), CCAAT enhancer-binding protein-α (C / EBPα), peroxisomeproliferators-activatedreceptor- γ (PPARγ) (Srebp-1c) are involved (Ntambi and and Kim, 2000; Rosen et al., 2000; Seo et al., 2004). These transcription factors are mostly expressed in adipose tissue and their activation when differentiating into adipocytes is the same as fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), and acetyl-CoA carboxylase- (Schultz et al., 2000), which directly enhances expression of synthetic genes and increases fat accumulation in differentiated adipocytes (Joseph et al., 2002). For this reason, in order to develop an anti-obesity agent, many studies have been conducted to find natural substances that inhibit the activity of transcription factors or genes involved in adipocyte differentiation or lipogenesis (Gonzalez-Espinosa de los Monteros et al. 2011; Niwano et al., 2009).

Currently, various methods for treating obesity have been attempted. In general, the treatment of obesity includes diet therapy, exercise therapy, appetite suppressant, diuretic, diarrhea, or drug therapy using fiber to give a feeling of fullness, behavior modification therapy to correct wrong eating habits and lifestyle, Surgery for reducing the volume of fat, and fat removal surgery such as decomposing and removing fat cells using ultrasound in plastic surgery. In addition, amphetamine, reductil, thiazolidinedione, metformin, and the like, which are associated with obesity treatment, have shown various side effects. For this reason, interest in alternative approaches including the use of medicinal plants in relation to the treatment of obesity has been increasing recently, It is necessary to develop a new preventive and remedy for obesity from natural products that have been approved for stability.

Korea Patent No. 919,323 Korean Patent No. 1,326,271

Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity, which comprises an extract of Tooth mothballs as an active ingredient.

Another object of the present invention is to provide a functional health food for preventing or ameliorating obesity, which comprises an extract of Tooth mothballs as an active ingredient.

In order to achieve the object of the present invention as described above, the present invention provides a pharmaceutical composition for preventing or treating obesity comprising an extract of Tooth mothballs as an active ingredient.

In one embodiment of the present invention, the sawtooth extract may be an extract obtained by using water, an organic solvent or a mixture thereof.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane .

In one embodiment of the present invention, the Sawtooth mildew extract may have the ability to inhibit PPARγ, C / EBPα, RXRα, and SREBP-1c, which are lipogenic synthetic transfer factor genes.

The present invention also provides a functional health food for preventing or ameliorating obesity, which comprises the extract of the present invention as an active ingredient.

In one embodiment of the present invention, the sawtooth extract may be an extract obtained by using water, an organic solvent or a mixture thereof.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane .

Since the extract of Tooth mackerel or the fraction isolated therefrom according to the present invention has an effect of inhibiting the differentiation of adipocytes induced by fibroblasts and inhibiting lipid accumulation by inhibiting intracellular lipid synthesis and promotion of lipolysis, Functional food, or obesity and an adult disease caused thereby.

FIG. 1 shows the cytotoxicity of 3T3-L1 cells of the extract of the teaspoon of the alcohol extract.
Fig. 2 shows the cytotoxicity of 3T3-L1 cells of the n-hexane fraction of the toothed mats.
FIG. 3 shows the result of showing the fat content accumulated in the cells after 3 days of 3T3-L1 cells were treated with the extract of the teaspoon mine extract.
Fig. 4 shows the result of showing the fat content accumulated in the cells after treatment of 3T3-L1 cells with n-hexane fractions of sawtooth moths for 7 days.
FIG. 5 shows the results of the weight change of rats during the 6-week rearing experiment.
FIG. 6 shows the results of analysis of the expression levels of transcription factors involved in obesity by RT-PCR.

The present invention relates to a new use of a Sawtooth mildew extract which has proved to be excellent in anti-obesity activity.

In the present invention, attention has been paid to a new antimicrobial agent for preventing or treating obesity, which is a perennial giant brown algae having a diameter of 1 to 4 m. The roots are 4 ~ 5cm in diameter, conical, rubbery, and have rings. The stem is short in the circumference and divided into a number of central branches. In addition, the edges of the stem are thin, vertically rising like a midrible, with short branches at the edge, stem leaves are pointing toward the base, there are double teeth at the edge, the bubble is round and the top There are 冠葉 (冠 葉) or spiny protuberances. Especially, it is known that it grows in the southern coast of Korea and Jeju. There is no known function about the physiological activity of the sawtooth moth.

According to one embodiment of the present invention, the 3T3-L1 fibroblasts were examined for extracts of Tooth mothballs or fractions fractionated from the mothballs extract of Tooth mothballs by the present inventors. The results showed that the extracts of the serrate serotypes and the n-hexane fractions of the extracts were superior to the 3T3-L1 fibroblasts in the adipocyte differentiation process or in the inhibition of intracellular fat accumulation in differentiated adipocytes (See Figures 3 and 4), and was also found to be cytotoxic (see Figures 1 and 2). From the above results, it was found that the extract of the sawtooth moth or the n-hexane fraction thereof was used as a material effective for the treatment or prevention of obesity, which is not toxic to cells and has few side effects. The degree of anti - obesity effect was found to be proportional to the treatment concentration of saphenous moth extract.

In addition, it was confirmed that the obesity-induced model animals were superior to the control group in the treatment group treated with the extract of Tootsuna mushroom extract (see Fig. 5) And it is confirmed that the above-mentioned obesity treatment and prevention effect is exhibited. Therefore, the inventors of the present invention have confirmed that the Sawtooth mallow extract is useful as an anti-obesity agent.

Furthermore, the inventors of the present invention have confirmed that the extract of the present invention has an effect of inhibiting the expression of a gene related to weight loss at the level of mRNA, and the PPARγ, C / EBPα, RXRα and SREBP-1c genes (See Fig. 6).

Therefore, a composition containing an extract of Sawtooth mildew as an active ingredient can effectively prevent or treat obesity.

In the present invention, 'obesity' means that the amount of fat in the body is excessively increased. Obesity is caused by an imbalance in the synthesis and degradation of fat in adipocytes. Obesity, which is caused by excess fat accumulation, increases the risk of developing chronic degenerative diseases and increases the risk of coronary artery disease, hypertension, stroke, hyperlipemia and diabetes .

In the present invention, " prevention " means all actions that inhibit obesity or delay progression by administration of the composition of the present invention.

&Quot; Improvement " in the present invention means all the actions of improving or ameliorating symptoms of obesity by administration of the composition of the present invention.

In the present invention, " treatment " means, unless otherwise indicated, reversing, alleviating, inhibiting or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder And the term " treatment " as used herein refers to an act of treating when " treating " is defined as described above.

The term " administration " as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The term " individual " as used herein means all animals such as a human, a monkey, a dog, a goat, a pig, or a mouse having a disease in which symptoms of obesity can be improved by administering the composition of the present invention.

The term " pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, including the type of disease, severity, activity of the drug, The time of administration, the route and rate of excretion of the drug, the duration of the treatment, factors including drugs used simultaneously and other factors well known in the medical arts.

The extract of Tooth mackerel according to the present invention can be obtained by extracting and isolating from nature using an extraction and separation method known in the art, and the 'extract' defined in the present invention can be obtained by using an appropriate solvent, , And may include, for example, a sawtooth extract, a polar solvent-soluble extract, or a non-polar solvent-soluble extract.

Suitable solvents for extracting the extract from the sawtooth mackerel include, but are not limited to, water or an organic solvent. For example, purified water, methanol, ethanol, propanol, isopropanol isopropyl alcohol, isopropanol, butanol and the like, an alcohol having 1 to 4 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride ), Hexane, and cyclohexane, may be used alone or in combination. Preferably, ethanol (alcohol), n-hexane and ethyl acetate solvents can be used.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation on the production method of the sawtooth extract of the present invention, and any known method may be used.

For example, in the composition of the present invention, the first extract extracted with the hot water extraction method or the solvent extraction method can be prepared into a powder state by an additional process such as vacuum distillation, freeze drying or spray drying have. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

Therefore, in the present invention, the teaspoon mallow extract is a concept including all the extract, fraction and purified product obtained in each step of extraction, fractionation or purification, and a diluted solution, a concentrate or a dried product thereof.

The composition of the present invention comprising the above-mentioned Saw Mabs extract as an active ingredient can be used as a pharmaceutical composition or a food composition.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. 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 as an appropriate method in the field.

In one embodiment of the present invention, the teaspoon mushroom extract of the present invention may be contained in the composition at a concentration of 10 to 100 μg / ml, and the teaspoon mushroom extract of the present invention may be contained in an amount of 0.1 to 95% .

The obesity diseases in which the pharmaceutical composition of the present invention can exhibit therapeutic effects include obesity, diabetes, hyperlipidemia, fatty liver, arteriosclerosis, hypertension, cardiovascular disease, hyperinsulinemia, hyperlipidemia, hypercholesterolemia, hypertriglyceridemia But are not limited to, a metabolic syndrome in which simultaneous occurrence of a disease, a metabolic syndrome, an endothelial dysfunction, a pre-aggregation state, dyslipidemia and atherosclerotic diseases.

The composition of the present invention may also be a food composition. In particular, it has been confirmed by one embodiment that the extract of Tooth mothballs according to the present invention does not induce toxicity in cells and mice. Accordingly, the food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient as well as ordinary food compositions, in addition to containing an effective component of the Tooth moth extract.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may contain various additives such as flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and aging agents (such as cheese and chocolate), pectic acid, Salts of alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated beverages and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

The extract of Tooth mackerel, which is an effective ingredient of the present invention, is a natural substance and has almost no side effects such as chemical agents. Therefore, it can be safely used for prolonged use for the purpose of prevention or treatment of obesity.

The present invention also provides a health functional food for the amelioration of an obesity disease, which comprises an extract of Sawtooth mildew as an active ingredient.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and rings for the purpose of improving obesity diseases.

In the present invention, the term " health functional food " refers to foods manufactured and processed using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods. Or to obtain a beneficial effect in health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned "food additives" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets can be prepared by granulating a mixture obtained by mixing the active ingredient of the present invention with the excipient, binder, disintegrant and other additives in a usual manner, Or the mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule form of the health functional food can be prepared by filling a normal hard capsule with a mixture of an extract of Tooth moth basil, an active ingredient of the present invention, with an additive such as an excipient, and the soft capsule is prepared by adding an extract, And filling the mixture with a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of an effective ingredient of the present invention, which is an extract of Tooth mothballs, an excipient, a binder, a disintegrant, etc., by a conventionally known method and, if necessary, Or it may be coated with a material such as starch, talc.

The granular health functional food may be prepared by granulating a mixture of the active ingredient of the present invention and the excipient, the excipient, the binder, the disintegrant and the like in a granular form by a conventionally known method. If necessary, a flavoring agent, And the like.

Since the tooth Sargassum extract of the present invention gajigi a functional food comprising as an active ingredient has no cytotoxicity, as confirmed in the following Examples, adipocyte differentiation and lipogenesis inhibitory effect is effective for the improvement and treatment of disorders of obesity.

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

The present invention also provides a method of preventing or treating obesity diseases, which comprises administering to a mammal an extract of Sawtooth mildew.

The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

The term "therapeutically effective amount " as used herein refers to the amount of active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, The amount that induces the relief of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the treatment method of the present invention, in the case of an adult, it is preferable to administer at a dose of 0.01 mg / kg to 250 mg / kg, once to several times a day, of the extract of the present invention.

In the treatment method of the present invention, the composition comprising the extract of the present invention as an active ingredient can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, And can be administered in a conventional manner.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that the following examples are merely illustrative of the present invention and that the scope of the present invention is not limited to these examples.

≪ Example 1 >

Preparation of Sawtooth Baker's Extract and n-Hexane Soluble Fraction

The sawtooth moths used in the present invention were obtained from Sargassum serratifolium collected from Gyungjang-gun, Busan, and Tongyeong-si, Gyeongsangnam-do, and fractionated with ethanol and n-hexane. First, 2 ㎏ of powder obtained by natural drying and shredding in a dark or dark place was immersed in 8 ℓ of a fermented alcohol (95%, ethanol content, 95%, Korea) And then filtered under reduced pressure through a filter paper (Watts, USA). The filtrate thus obtained under reduced pressure was concentrated under reduced pressure using a rotary rotary evaporator at 40 ° C to remove the solvent, and the residue thus obtained was obtained as a crude extract of teaspoon. As a result, it was possible to obtain 202 g of a syrupy mother liquor extract. Then, 4 liters of a mixed solvent of 100 g of the alcohol extract and n-hexane (n-hexane), alcohol and water at a ratio of 10: 9: 1 was added and the water layer and the n-hexane layer were separated from each other. After sufficient time had passed, the mixture was separated into 2 L water layer and 2 L n-hexane layer. 2 L of n-hexane was added again to the 2 L water layer thus separated, and the process of separating 2 L of n-hexane layer was repeated 5 times. As a result, finally, a total of 10 L of n-hexane layer was concentrated under reduced pressure using a vacuum rotary condenser to remove the solvent, and then hexane fraction was obtained.

<Experimental Example 1>

In vitro anti-obesity effect of n-hexane-soluble fractions,

&Lt; 1-1 > Treatment of teaspoon mine extract and n-hexane soluble fraction

The mouse-derived fibroblast 3T3-L1 obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) was inoculated into DMEM medium supplemented with 10% bovine serum (BS), 100 units / ml penicillin and 100 μg / ml streptomycin And cultured at 37 ° C in a 5% CO 2 environment. 3T3-L1 cells cultured as described above were dispensed into a 96-well plate at a concentration of 1.5 × 10 ³ / well, and then samples of different concentrations (0, 1.0, 2.5, 5.0 and 10 μg / And the DMEM medium containing the ethanol extract of nidulans and the n-hexane fraction obtained in Example 1) for 24 hours.

&Lt; 1-2 > Spearman extract and n- Hexane Of the soluble fractions  Cytotoxic effect

As in Experimental Example 1-1, 100 μl of the medium was taken from the 3T3-L1 cell culture treated with the Sawtooth mulberry extract or the hexane fraction, and each well was dispensed into a 96-well plate together with 5 μl of the MTS solution and reacted for 1 hour , And the absorbance was measured at 490 nm using a microplate reader. The measurements were determined as the mean value of three replicate experiments, and the relative cell viability (% of control) was calculated by comparing the determined values with the control group.

As a result, as shown in FIG. 1 and FIG. 2, it was confirmed that the extracts of Tephalan myrtle extract and hexane fraction did not show cytotoxicity up to a concentration of 10 μg / ml, and the cytotoxicity of the extracts of Tephalan myrtle extract and hexane fraction (See Figs. 1 and 2).

&Lt; 1-3 > Treatment of Sawtooth mung bean extract and n-hexane-soluble fraction for inhibiting adipocyte differentiation and lipogenesis inhibition

3T3-L1 cells cultured as described in Experimental Example 1-1 were dispensed at a concentration of 2.5 × 10 ⁴ cells / well in a 24-well plate and then cultured for another 2 days in a confluent state . Cells were further cultured for 4 days in DMEM medium supplemented with fetal bovine serum (FBS), MDI, and ethanol extracts of different concentrations (0, 1.0, 2.5, 5.0, 10 ㎍ / And cultured for a total of 7 days.

<1-4> Cytotoxic effect of extracts of teaspoon mushroom extract inhibiting adipocyte differentiation and adipogenesis

To confirm the lipogenesis inhibitory effect of the n-hexane fraction obtained in Example 1 on the differentiation of adipocytes, it was examined by the method described in Lee et al. (Plant Food and Human Nutrition 2014 Vol. 69. 137-141) Experiments were carried out according to the method for analyzing the specific intracellular fat accumulation. Three days after the treatment of 3T3-L1 cell differentiation with different concentrations of MDI (0, 1.0, 2.5, 5.0, 10 ㎍ / ㎖), the medium containing MDI was treated with 10% FBS, 170 nM insulin and alcohol extracts of different concentrations were treated two times at 2-day intervals. Cells were washed with cold PBS on day 8 and collected. The harvested cells are disrupted with an ultrasonic disrupter (Branson Ultrasonics, Danbury, Conn.) And centrifuged at 14,000 rpm for 10 minutes to obtain supernatant. Triglycerides in the supernatant were analyzed by the GPO-trinder kit (Sigma-Aldrich, St Louis, MO, USA) according to the instructions.

As a result, as shown in Fig. 3, the extracts of Tephriton japonica showed a concentration-dependent inhibition of 3T3-L1 lipogenesis within a concentration range of 0-10 / / ml, (Fig. 3). As shown in Fig.

&Lt; 1-5 > Hexane Of the soluble fractions  Inhibition of adipocyte differentiation and fat production

3T3-L1 cells cultured as described in Experimental Example 1-1 were dispensed at a concentration of 2.5 × 10 ⁴ cells / well in a 24-well plate, and then cultured for another 2 days when the cells were saturated. FBS and MDI were added to the DMEM medium, and the cells were cultured for 3 days. Then, the fraction of n-hexane with FBS, insulin and different concentrations (0, 1.0, 2.5, 5.0, 10 ㎍ / On day 8, the cells were washed with cold PBS and collected. The harvested cells are disrupted with an ultrasonic disrupter (Branson Ultrasonics, Danbury, Conn.) And centrifuged at 14,000 rpm for 10 minutes to obtain supernatant. Triglycerides in the supernatant were analyzed by the GPO-trinder kit (Sigma-Aldrich, St Louis, MO, USA) according to the instructions.

As a result, as shown in Fig. 4, the n-hexane fraction of the toothed mackerel showed a concentration-dependent inhibition of the fat production of 3T3-L1 within a concentration range of 0-10 ㎍ / ml, The hexane fraction was found to be excellent in anti-obesity because it had an effect of inhibiting adipocyte differentiation and adipocyte formation (see FIG. 4).

< Experimental Example  2>

In-Bibo extract of alcoholic beverages in vivo ) Check your weight loss effect

In order to examine the in vivo weight gain inhibition effect of the extract of the Teaspoon mugwort extract obtained in Example 1, the specific method described in the literature of Seo et al. (Korean Society of Food Science and Nutrition, 2010 39 (11), 1604-1610) In-vivo experiments were performed.

Sixty adult C57BL / 6 male mice (Samtaco, Korea) at 7 weeks of age were acclimated for one week in a breeding room of environment at a temperature of 22 ± 2 ° C., humidity of 55 ± 5% (HF), high-fat diet (0.1%), and a low-fat diet (0.1%) were fed a control diet containing 12 normal rats, a normal diet (0.25% SSE), which was administered with 0.25% Sawtooth milt seed extract, and the third (0.1% SSE) treated with high-fat diet and 0.5% Sawtooth moth extract And 0.5% SSE, respectively. The composition of normal diet and high fat diet is shown in Table 1. The body weights of the mice were measured immediately before initiation of doses of the extracts of the teaspoonful and once a week after administration. The diets used in the experiments were prepared by modifying Rodent Diet with 60% kcal fat (Research Diets, Inc.).

Dietary composition of feed

As a result, as shown in FIG. 5, the body weight gain was significantly decreased in the first to third experimental groups as compared with the positive control group (HF). In particular, after 6 weeks, the body weight gain of the first experimental group added with the extract of Tephritidae was similar to that of the normal group, and the weight of the second experimental group and the third experimental group was lower than that of the experimental group 5).

In addition, the fat tissue of the experimental animals was significantly reduced by the extract of the teaspoon mushroom (see Table 2). In addition, GOT and GPT were decreased significantly by the addition of the extracts of the extracts of the teaspoon of 6 weeks, and the toxic effects of the extracts of the teaspoon were not observed See Table 3).

<Table 2> Changes in adipose tissue in experimental animals

<Table 3> Biochemical indicators of laboratory animals

From the above results, it was found that the extracts of Tephritidae were effective in inhibiting weight gain due to high fat diet in in vivo, and thus, And it was confirmed that it was excellent.

<Experimental Example 3>

In vivo inhibition of weight loss-induced gene expression of extracts from the teaspoon myrtle extract

The expression levels of PPARγ, C / EBPα, RXRα and SREBP-1c, which are the transcription factors involved in fat synthesis in adipose tissue, were determined by real-time RT-PCR Respectively. First, 10% RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) was added to the adipose tissue and the mixture was ground with a micro homogenizer and centrifuged at 14,000 rpm The supernatant was obtained. The same amount of TRISol solution was added to the supernatant to isolate mRNA. CDNA was prepared using RNA as a template using reverse transcriptase and random primers. cDNA quantification was performed using QuantiTect SYBR Green reagents kit (Qiagen) using DNA Engine Opticon 2 Continuous Flourescence Detection System (Applied Biosystems). The sequences of the primers used in the experiments are shown in Table 4.

Table 4: Sequence of primers used in real-time RT-PCR

As a result, as shown in Fig. 6, the expression factors involved in lipogenesis were significantly decreased in the first to third experimental groups as compared with the positive control (HF).

From the above results, it was found that the extracts of Teapot mushroom extract inhibited the expression of genes involved in lipid synthesis in vivo, thereby inhibiting weight gain. Thus, And it was confirmed that there was an excellent effect in the present invention.

Examples of formulations for the composition of the present invention are illustrated below.

&Lt; Formulation Example 1 > Preparation of pharmaceutical preparation

The pharmaceutical preparation comprising the teaspoon extract of the present invention, or its n-hexane fraction, is prepared as follows.

<1-1> Preparation of powder

Sawtooth mung bean extract or n-hexane fraction 2 g

Lactose 1g

The above components are mixed and packed in airtight bags to prepare powders.

<2-2> Preparation of tablets

100 mg of naphtha extract or n-hexane fraction

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

&Lt; 1-3 > Preparation of capsules

100 mg of naphtha extract or n-hexane fraction

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, they are filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.

&Lt; 1-4 >

Sawtooth extract or n-hexane fraction 1 g

1.5 g of milk

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they are prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of naphtha mellifera extract or n-hexane fraction

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol is added and dried at 60 캜 to form granules, which are then filled in a capsule.

< Formulation example  2> Manufacturing of food

Foods containing the sawtooth extract of the present invention, or its n-hexane fraction, are prepared as follows.

<2-1> Production of flour food

0.5 to 5.0 parts by weight of the Sawtooth mulberry extract or n-hexane fraction of the present invention is added to wheat flour, and bread, cake, cookies, crackers and noodles are prepared by using the mixture to prepare a health improving food.

<2-2> Production of dairy products

5 to 10% by weight of the sawtooth extract or n-hexane fraction of the present invention is added to milk, and the milk is used to produce various dairy products such as butter and ice cream.

<2-3> Manufacturing of Sunshine

Brown rice, barley, glutinous rice, and yulmu are dried by a known method and dried, and then the mixture is ground to a powder having a particle size of 70 mesh. Black beans, black sesame, and perilla are steamed and dried by a known method, and then powdered with a powder having a particle size of 70 mesh by a grinder. The tofu mushroom extract or n-hexane fraction of the present invention is concentrated under reduced pressure in a vacuum concentrator, dried by spraying and dried in a hot-air drier, and pulverized to a size of 70 mesh with a pulverizer to obtain a dried powder. The above-prepared grains, seeds and sawtooth extracts or dried powders of n-hexane fractions are prepared in the following proportions.

Cereals (30% by weight of brown rice, 15% by weight of yulmu, 20% by weight of barley)

Seeds (7% by weight perilla, 8% by weight black bean, 7% by weight black sesame)

Dried powder (3% by weight) of naphtha extract or n-hexane fraction,

Manure (0.5% by weight), and

Rhizome (0.5% by weight).

< Formulation example  3> Manufacturing of beverage

<3-1> Health drink  Produce

1000 mg of naphtha extract or n-hexane fraction

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 2 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 ° C for about 1 hour. The resulting solution was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, It is used in the production of the health beverage composition of the invention.

The composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment. However, the blending ratio may be arbitrarily varied depending on the regional and national preference such as the demand level, the demanding country, and the intended use.

<3-2> Vegetable juice  Produce

Wheat germ extract or 5 g of n-hexane fraction is added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

<3-3> Of fruit juice  Produce

Healthy fruit juice is prepared by adding 1 g of Sawtooth mung bean extract or n-hexane fraction to 1,000 ml of apple or grape juice.

Claims (7)

A pharmaceutical composition for preventing or treating obesity comprising an extract of Sargassum serratifolium as an active ingredient. The method according to claim 1,
The pharmaceutical composition for preventing or treating obesity is characterized in that the sawtooth moth extract is extracted with water, an organic solvent or a mixture thereof. 3. The method of claim 2,
Wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane. Composition. The method according to claim 1,
The pharmaceutical composition for preventing or treating obesity is characterized in that the Sawtooth mildew extract has the ability to inhibit PPARγ, C / EBPα, RXRα and SREBP-1c, which are lipogenic synthesis transfer factor genes. A food composition for preventing or ameliorating obesity, comprising an extract of Tooth mackerel as an active ingredient. 6. The method of claim 5,
The food composition for preventing or improving obesity is characterized in that the sawtooth moth extract is extracted with water, an organic solvent or a mixture thereof. The method according to claim 6,
Wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane .

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