KR102099147B1 - Composition for preventing, ameliorating or treating obesity comprising Heracleum moellendorffii root and Torilis japonica mixed extract as effective component - Google Patents

Abstract

The present invention relates to a composition for the prevention, improvement or treatment of obesity, which contains the mixed base of the fish roots and casualties, as an active ingredient, and the mixed base of the fish roots and casualties of the present invention compared to the sole base of the fish roots or the casualties extract. It has an excellent effect of suppressing differentiation and has a effect that the content of triglycerides is significantly reduced compared to the treatment of a single extract, so it can be usefully used as a health functional food and a therapeutic drug for the prevention and improvement of obesity.

Description

Composition for preventing, ameliorating or treating obesity comprising Heracleum moellendorffii root and Torilis japonica mixed extract as effective component

The present invention relates to a composition for preventing, improving or treating obesity, which includes an extract of fish roots and casualties mixed as an active ingredient.

Obesity refers to the case where fat is excessively accumulated in the body, and according to the national health statistics, the prevalence of obesity increased from 26.0% in 1998 to 31.4% in 2011, and is related to complications of various adult diseases. It is revealed.

In addition, obese patients have been reported to have a higher mortality rate of 2 times for cirrhosis disease, 1.6 times for cerebrovascular disease, and 1.8 times for coronary artery disease compared to those with normal weight. Currently, dietary therapy, exercise therapy, and drug therapy have been attempted as a treatment method for obesity, and among these, obesity treatment agents used as drug therapy control appetite, interfere with digestion and absorption of fat, increase energy consumption, or lipid metabolism. It was developed as a target for the regulation of, but there are various side effects. As an example, Xenical manufactured by Roche induces lipase inhibition that is separated from the pancreas and digestive system, thereby inhibiting fat absorption, but shows only 2 ~ 3% of low weight loss effect, and has side effects such as frequent diarrhea and fat stool. To accompany.

In addition, Abbott obesity treatment drug Reductil suppresses appetite by suppressing resorption of serotonin and noradrenaline, which has a weight loss effect of 5-10%, but has side effects of cardiovascular diseases such as stroke and myocardial infarction, 2010 In 2010, the European Agency for Food and Drug Safety (EMA) and the United States Food and Drug Administration (FDA) were put out of the market with prescriptions, suspensions and voluntary recall recommendations, and were discontinued in Korea by the Food and Drug Administration in October 2010.

In addition, many of the products developed as anti-obesity drugs are banned due to serious side effects. For example, aminophylline has been reported to have a wide range of side effects across the mental nervous system, circulatory system, and digestive system despite its excellent body fat-decomposing effect, and fenpruramin, dexfenfluramine, topiramay, or ephedrine are also considered to be unsuitable for obesity treatment. Sales were banned. As such, as Western medicine has shown limitations in overcoming side effects and chronic diseases of synthetic medicines, the value of natural foods and medicines with anti-obesity effects is emerging.

Meanwhile, Eosu-ri grows in mountains and fields. Stems stand upright, 70-150cm high, hollow cylinder-shaped, vertically lined, coarse hairy, and thick branches split. The leaves are alternate, 3 to 5 small leaves, and are feathers, with hairs and shorter petioles as they rise above the stem, and the lower part is wider to cover the stem. The small leaf on the end is heart-shaped and is divided into three, and the small leaf on the side is broad egg-shaped or triangular, 7-20cm long, split into 2-3 pieces, and has a sawtooth deeply dug at the edge. The flowers bloom white in July-August and run in a double inflorescence at the ends of branches and stems. In the inflorescence, 20 to 30 peduncles are divided into small peduncles, each having 25 to 30 flowers, and the flowers at the edges are larger than the flowers at the center. There are 6 petals and different sizes, and the outer petal is larger than the inner petal.

The casualty (Torilidis Fructus) is the fruit of the two-year-old casualty ( Torilis japonica (Houtt.) DC.) Of the Apiaceae (Umbelliferae). Herbal medicines made from dried fruit are called casualties, and are distributed in Korea, Japan, China, Taiwan, India, and Myanmar. It has anti-inflammatory effect, sex hormone-like action, anti-allergic action, etc. as a pharmacological effect. It is effective in itching, etc. as an astringent anti-inflammatory drug. The main components of the casualties are torilin, torilolone, and sesquies. It has been reported to contain a terpene-based compound.

As a prior art related to obesity, Korean Registered Patent No. 1134251 discloses a food composition for improving obesity using the casualty extract as an active ingredient, and Korean Registered Patent No. 1069418 discloses a casualty extract having glycerol acyltransferase inhibitory activity, a solvent fraction thereof Or discloses a composition comprising a compound separated therefrom, and discloses a method for preparing a diet composition and a diet agent in Chinese Patent Publication No. 104857107, but obesity containing the mixed extract of the basement and casualties of the present invention as an active ingredient It has not been disclosed for the composition for the prevention, improvement or treatment of.

The present invention has been derived by the above-mentioned needs, the present invention provides a composition for preventing, improving or treating obesity comprising an extract of fish roots and casualties as an active ingredient, and the extract of fish roots and casualties mixed with fish roots The present invention was completed by confirming that the effect of suppressing the differentiation of adipocytes is remarkable and the content of triglycerides is significantly lower than that of the underground extract or casualty extract alone.

In order to achieve the above object, the present invention provides a health functional food composition for preventing or improving obesity containing an extract of fish roots and casualties mixed as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of obesity, which contains a mixed extract of fish roots and casualties as an active ingredient.

The present invention relates to a composition for the prevention, improvement or treatment of obesity, which contains the mixed base of the fish roots and casualties, as an active ingredient, and the mixed base of the fish roots and casualties of the present invention compared to the sole base of the fish roots or the casualties extract. It has an excellent effect of suppressing differentiation and has a effect that the content of triglycerides is significantly reduced compared to the treatment of a single extract, so it can be usefully used as a health functional food and a therapeutic drug for the prevention and improvement of obesity.

Figure 1 is a result of confirming the cell viability according to the treatment of 25, 50, 75㎍ / ㎖ fish root base extract, casualty extract and fish root base + dead box mixed extract.
Figure 2 is a photograph of the results of oil red O staining of adipocytes according to the treatment of 50 μg / ml or 75 μg / ml fish root underground extract, casualty extract and fish root underground + dead mixed extract.
Figure 3 is a result of confirming the degree of suppression of fat cell differentiation according to the treatment of 25, 50, 75㎍ / ㎖ fish root underground extract, casualty extract and fish root underground + dead box mixed extract by oil red O staining method. *, **, *** were treated with 50 or 75 µg / ml of fish root underground or casualty extract, compared to 50 or 75 µg / mL of fish root underground + mixed fat extract, statistically significantly adipocytes. Differentiation was suppressed, * is p <0.05, ** is p <0.01, and *** is p <0.001.
Figure 4 is a result confirming the effect of 75 ㎍ / ㎖ fish root base extract, casualty extract and fish root base + dead box mixed extract on the accumulation of triglycerides in fat cells. ** is a statistically significantly lower content of triglyceride in the cells of the Eosu-ri basement + injured mixed extract, which is the active ingredient of the present invention, compared to the basement extract of Eosu-ri, p <0.01, and *** compared to the casualty extract. The content of triglyceride in the cells of the mixed extract of the fishery basement + boxworm, which is the active ingredient of the present invention, is statistically significantly lower, which is p <0.001.

The present invention relates to a health functional food composition for preventing or improving obesity containing an extract of fish roots and casualties mixed as an active ingredient.

The Eosu Underground and Casualty Mix extract is preferably a mixture of Eosu Underground Extract and Casualty Extract in a weight ratio of 1: 0.1 to 10, and more preferably, the Underwater Extract and Casualty Extract is mixed in a weight ratio of 1: 0.3 to 5. It is more preferably mixed in a weight ratio of 1: 0.5 to 3, most preferably in a weight ratio of 1: 1, but is not limited thereto.

The fish root extract or casualty extract may be prepared by a method including the following steps, but is not limited to this:

(1) extracting by adding an extracting solvent to the basement or casualty of fishery;

(2) filtering the extract of step (1); And

(3) Concentrating and drying the filtered extract of step (2) to prepare an extract.

In step (1), the extraction solvent is preferably selected from water, a lower alcohol of C ₁ to C ₄ or a mixture thereof, more preferably ethanol or methanol, but is not limited thereto.

In the above manufacturing method, any conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction, may be used as the extraction method. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried fishery base or casualties, more preferably 5 to 15 times, and even more preferably 10 times. The extraction temperature is preferably 20 to 100 ° C, but is not limited thereto. In addition, the extraction time is preferably 1 to 60 hours, more preferably 1 to 48 hours, but is not limited thereto. In the above method, the concentration of step (3) is preferably, but not limited to, a vacuum rotary concentrator or a vacuum rotary evaporator. In addition, drying is preferably dried under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying, and more preferably freeze drying, but is not limited thereto.

The health functional food composition using the extract from the bottom of the fishery and the casualties as an active ingredient is prepared from any one selected from beverages, pills, tablets, capsules and powders, or added to other foods or food ingredients. It can be, and can be appropriately manufactured according to conventional methods.

Examples of foods to which a health functional food composition using an extract of fish roots and casualties mixed as an active ingredient of the present invention can be added is meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums , Dairy products including ice cream, various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and may include all health functional foods in a general sense.

The health functional food composition is a variety of nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alkylic acid and salts thereof, organic acids, protective colloids It may contain a thickening agent, pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like. In addition, it may contain flesh for the preparation of natural fruit juices and vegetable drinks. These ingredients can be used independently or in combination.

The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional components. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, synthetic sweeteners such as saccharin and aspartame can be used.

In addition, the present invention relates to a pharmaceutical composition for the prevention or treatment of obesity containing an extract of fish roots and casualties mixed as an active ingredient.

The pharmaceutical composition of the present invention may further include a carrier, an excipient, or a diluent, in addition to the fish root base and casualty mixed extract, but is not limited thereto.

The pharmaceutical composition of the present invention may be various formulations, oral or parenteral. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc.These solid preparations include at least one excipient in one or more compounds such as starch, calcium carbonate, sucrose or lactose ( lactose) and gelatin. In addition, lubricants such as magnesium stearate, talc and the like are used in addition to simple excipients. Liquid preparations for oral administration include suspending agents, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used as diluents, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, can be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. As non-aqueous solvents and suspension solvents, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, twin tween 61, cacao butter, laurin butter, and glycero gelatin may be used.

The composition of the present invention may be administered orally or parenterally, and when parenterally administered, it is preferable to select an injection method for external application to the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine epidural or cerebrovascular. It is not done.

The composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, and activity of the drug , Sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs and other factors well known in the medical field. 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 a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, 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.

The dosage of the composition of the present invention can be used in a variety of ranges depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate and disease severity.

Hereinafter, the present invention will be described in more detail using examples. It is apparent to those skilled in the art that these examples are only intended to illustrate the present invention in more detail and that the scope of the present invention is not limited by them.

One. Fishery  Preparation of underground extract

Underwater fish ( Heracleum ) belonging to Umbelliferae moellendorffii Hance , sale number: 005-066) was used by pre-sale from the plant extract bank, Korea Research Institute of Bioscience and Biotechnology (KRIBB, Daejeon, Korea). The dried and shredded fishery basement was extracted with 99.9% methanol for 48 hours at room temperature. The extract from the bottom of the fishery was filtered and concentrated under reduced pressure with a reduced pressure concentrator. Thereafter, the fish root extract was dissolved in DMSO and stored at -20 ° C so as to have a concentration of 100 mg / ml.

2. Casualties ( Torilis  japonica ) Preparation of extract

Casualties were cultivated on farms (producers: Udongjin) located in Yeongcheon-si, Gyeongsangbuk-do, and casualties collected in August-September were purchased from Goodherb, a herbalist. After the casualties were pulverized using a sample grinder, 70% (v / v) ethanol at an 8-fold capacity per g of powder was completely immersed, and then extracted at room temperature for 72 hours. After the casualty extract was filtered, it was concentrated under reduced pressure in a heating bath at 40 ° C using a reduced pressure concentrator (EYELA N-1200A, SB1200, A-1000S, CCA1111). The casualty extract was dissolved in DMSO to store a concentration of 100 mg / ml and stored at -20 ° C.

3. Cell culture

3T3-L1 adipose progenitor cells were cultured using DMEM (Dulbeco's Modified Eagle's Medium) medium containing 10% calf serum at 37 ° C and 5% CO2. At this time, the cell density was periodically passaged so as not to exceed 70%.

4. MTT Assay  Cytotoxicity analysis

Before performing the anti-obesity activity assay, an MTT assay was performed to determine the effect of the sample on cell viability and to determine the concentration of the sample that does not induce cytotoxicity.

After dispensing 1 × 10 ⁵ 3T3-L1 cells to a 24-well culture plate and attaching them for 24 hours, treatment with the extract from the bottom of the fishery and the casualty extract at concentrations (25, 50, 75 μg / ml), respectively, and cultured for 48 hours Did. After treating the fish root extract and casualty extract, 20 μl of MTT solution (5 mg / ml) was added and reacted for 3 hours, and after discarding all the cultures on the plate, 150 μl of DMSO was added to the wells.

Thereafter, the plate was allowed to stand at room temperature for 5 minutes while shaking, so that the formazan was completely decomposed and absorbance was measured at 595 nm.

5. Cell differentiation

3T3-L1 cells were dispensed into 24 well culture plates so that there were 1 × 10 ⁵ cells and cultured for 3 days, followed by differentiation with MDI medium. MDI medium was treated with 0.5 mM IBMX, 1 μM dexamethasone, and 1 μg / ml insulin in 10% FBS / DMEM. This time point was set as day 0 of differentiation, and after 2 days of differentiation, 1 µg / ml of insulin medium was replaced, and after 2 days of incubation, it was maintained for 8 days with 10% FBS medium. Underwater extracts and casualty extracts were added on day 0 and day 2 of differentiation and treated for 4 days. Thereafter, the cells were incubated with 10% FBS medium for 4 days.

6. Oil Red  O (Oil-Red-O) staining

After 8 days of differentiation, the degree of adipocyte differentiation and the degree of inhibition of differentiation by extracts were analyzed through Oil Red O staining. After completion of the adipocyte differentiation and substance treatment (25, 50, 75 µg / ml, respectively), the cells were washed with 1 × phosphate buffered saline (PBS), then added with 10% formalin and fixed at room temperature for 1 hour. After removing formalin and washing the wells with 60% isopropanol, the isopropanol was completely blown off from the hood. Filtered oil red O solution was added to the dried well, left for 10 minutes, and then washed with distilled water to confirm adipocyte differentiation by microscopy. The stained well was dissolved by adding 100% isopropanol and absorbance was measured at 500 nm.

7.TG ( Triglyceride ) Assay

The amount of TG in adipocytes was measured using a commercially available TG assay kit (Triglyceride Assay Kit-Quantification, ab65336). On the 8th day of differentiation, 3T3-L1 cells were washed with cold phosphate-buffered saline (PBS), then crushed with 5% NP-40 solution, and the total amount of TG in the crushed solution was determined by enzymatic method according to the protocol enclosed with the kit. .

Example  1. Eosu Underground and Casualty Extract 3 T3 - L1  Effects of fat progenitor cells on cell viability

 The effect of the basement and casualty extracts on the cell viability of 3T3-L1 adipocyte progenitor cells was analyzed using MTT assay.

As a result, as shown in FIG. 1, even if treated with a concentration of 25 to 75 µg / ml, both the fish root underground extract, the casualty extract, and the fish root underground + dead box mixed extract did not cause cytotoxicity.

Example  2. Eosu Underground Extract, Casualty Extract and Eosu Underground + Injured Mixed Extract induced by MDI 3 T3 - L1  Effects of fat progenitor cells on fat production

In order to confirm the anti-obesity activity of Eosu Underground Extract, Casualty Extract, and Eosu Underground + Injured Mixed Extract, the effect on fat production of 3T3-L1 fat progenitor cells induced in differentiation with MDI was confirmed, and the results of Example 1 above. Based on the analysis of the fat generation inhibitory ability of the base extract, the casualties extract and the base extract + boxworm mixed extract was carried out at a concentration of 25, 50, 75㎍ / ㎖.

2 and 3, compared to the case where the fish root base or casualty extract was treated alone, in the case of the fish root base + dead box mixed extract of the present invention, adipocyte differentiation inhibitory activity was high, in particular, 75 When treated at a concentration of ㎍ / ㎖, it was confirmed that the adipocyte differentiation inhibitory activity of the adipocyte basement + wounded mixed extract showed a significant suppression of adipocyte differentiation activity compared to the treatment of the single extract.

Example  3. Effects of the Underwater Extract, Casualty Extract, and Underwater + Dead Box Blend Extract on Triglyceride (TG) Content in Fat Cells

Quantitative analysis was performed using the TG ELISA kit to determine how the fish root extract, casualty extract, and fish root extract + dead box mixed extract affect the accumulation of triglycerides in fat cells.

As a result, as shown in FIG. 4, when the 75 μg / ml fish root extract and casualty extract were each treated, the difference was not significantly compared to the control, but 75 μg mixed in a weight ratio of 1: 1 It was confirmed that the amount of TG was significantly decreased in the mixed extract of the fishery basement + injured box.

Claims (6)

A health functional food composition for preventing or improving obesity, containing an extract of fish roots and casualties mixed as an active ingredient in a weight ratio of 1: 0.1 to 10. delete [6] The composition of claim 1, wherein the fish root extract and casualty extract are extracted using C ₁ -C ₄ lower alcohol, water, or a mixture thereof as a solvent. A pharmaceutical composition for the prevention or treatment of obesity, containing an extract of fish roots and casualties mixed as an active ingredient in a weight ratio of 1: 0.1 to 10. delete [5] The pharmaceutical composition for preventing or treating obesity according to claim 4, further comprising a carrier, an excipient, or a diluent in addition to the extract from the basement and the casualties.

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