KR20070070311A - A composition comprising the extract of semisulospira libertine, sulfur fed duck, ulmus devidiana, pine pollen powder, acer tegmentosum maxim., bamboo salt, artemisia capillaries, platycodon grandiflorum, rhynchosiavolubilis for the prevention and treatment of obesity and hypercholesterolemia - Google Patents

Abstract

A composition for the prevention and treatment of obesity and hypercholesterolemia comprising extracts of natural products is provided to reduce the number and size of adipocytes in the tissue, and inhibit expression of genes associated with lipid synthesis including SREBP-1(sterol response element-binding protein-1), FAS(fatty acid synthase) and SCD-1(stearoyl-CoA desaturase) mRNA(messenger RNA). The composition for the prevention and treatment of obesity and hypercholesterolemia comprises the extracts of Semisulcospira libertine, sulfur fed duck, Ulmus macrocarpa Hance, pine pollen powder, Acer tegmentosum Maxim., bamboo salt, Artemisia capillaries, Platycodon Grandiflorum and black bean in a weight ratio of 20-1 : 1-10 : 5-1 : 5-1 : 5-1 : 5-1 : 5-1 : 5-1 : 2-1, wherein the composition is a pharmaceutical or health food composition; and the daily dosage of the natural product extracts is 0.0001-100 mg/kg.

Description

A composition comprising the extract of Semisulospira libertine, Sulfur Fed, which contains a complex extract consisting of mulberry, sulfur duck, root roots, pine powder, cheongsam, bamboo salt, jinjin wormwood, bellflower and black bean Duck, Ulmus devidiana, Pine Pollen Powder, Acer tegmentosum Maxim., Bamboo Salt, Artemisia capillaries, Platycodon Grandiflorum, Rhynchosiavolubilis for the prevention and treatment of obesity and hypercholesterolemia}

1 is a comparison of the tissues extracted from the hepatocytes and the HE stained adipocytes between groups,

Figure 2 compares the mRNA expression of SREBP-1 between groups,

3 compares FAS mRNA expression between groups,

Figure 4 compares the mRNA expression of SCD-1 between groups.

Recently, as the standard of living improves, the hygiene environment is improved and the dietary life is westernized, and the average life span is extended. Therefore, adult disease has emerged as the biggest medical task today, and obesity, which is a major cause of such disease, is rapidly increasing. Obesity refers to a phenomenon in which excess calories are accumulated in the form of fat in the body by ingesting excessive calories compared to calories consumed. Obesity is thought to be caused by a variety of causes, including genetic effects, environmental effects of westernized diets, and psychological effects of stress, but the exact cause and mechanisms are not clearly established. However, because obesity can act not only as a problem of obesity itself, but also as a cause of diseases such as cardiovascular disease and diabetes (Manson et al., New England J. Med., 333, pp677-685, 1995; Kopleman PG, Nature, 404 pp635-643, 2000; Must et al., JAMA , 282 , pp1523-1529, 1999). There is much interest in the treatment of obesity worldwide.

In Korea, cerebral vascular diseases such as cerebral hemorrhage and cerebral infarction due to hypertension, and coronary artery disease due to hyperlipidemia in developed countries in Gumi, have recently increased incidence of hyperlipidemia due to increased intake of animal fat in Korea. Ischemic heart and brain diseases are increasing on the basis of the Ministry of Health and Welfare, 1997 National Nutrition Survey Report , p37, 1997. Hyperlipidemia refers to abnormally increased lipids such as cholesterol and triglyceride in plasma.

Such risk factors for atherosclerosis include hyperlipidemia, hypertension, smoking, diabetes, hyperuricemia, obesity, lack of exercise, and excessive drinking (Kannel WB., Med. Clin. North. Am , 79 , pp951-956, 1995; Grodstein F., N. Engl. J. Med. , 336 , pp1769-1774, 1997), hyperlipidemia, smoking, and hypertension are known as three major risk factors in Western countries, Smoking and diabetes were the main risk factors. However, hyperlipidemia has recently emerged as a major risk factor. In Korea, the prevalence and mortality of atherosclerosis and its associated cardiovascular disease is increasing, and considering the increase in the prevalence of hyperlipidemia, one of the causes, it is urgently required to take measures for proper prevention and treatment of hyperlipidemia. As a main method for preventing cardiovascular diseases, various methods such as avoiding excessive intake of calories and limiting the amount and type of ingested fat have been proposed, and in recent years, proper intake of natural foods has become important in cardiovascular diseases. From this point of view, the active substance for reducing hyperlipidemia, which is a major cause of pulmonary disease, has been actively pursued in various fields.

On the other hand, cholesterol is mainly produced in the liver and is present in the blood in the form of small round particles called lipoproteins, which carry cholesterol throughout the body. There are two types of lipoproteins, low density lipoprotein (LDL) and high density lipoprotein (HDL). HDL carries cholesterol from other tissues. On the other hand, LDL is produced mainly in the liver and carries cholesterol to other tissues of the human body, so a large amount of LDL accumulates a lot of cholesterol in blood vessels to promote atherosclerosis, and about 70% of blood cholesterol is present as harmful cholesterol (LDL). Kim, Kyoung-hwan, Lee Woo-joo, Pharmacology Lecture, 4th edition, p466-481, 2001).

In fact, cholesterol is an essential ingredient necessary for the normal functioning of the human body, nutrients necessary to make cells, and a component of various hormones such as corticosteroids, male hormones, and female hormones. It can help you digest fatty foods. In addition, it is a nutrient necessary for the human body, such as a component of a biological membrane and used as a starting material for hormonal synthesis. However, when too much cholesterol is accumulated in blood vessels, it is known to cause heart disease. So far, there is no preventive method other than low cholesterol diet, and taking drugs such as cholesterol-lowering drugs are effective, but their use is extremely limited because of causing side effects such as liver dysfunction due to inhibition of cholesterol synthase.

The differentiation process of adipocytes has been studied using cells such as 3T3-L1, and various transcription factors, particularly transcription factors known to be involved in localization, CAAT enhancer binding proteins (C / EBPs), It is known that PPARs (Peroxisome Proliferator Activated receptor) ADD / SREBPs (Adipocyte determination and differentiation dependent factor 1 / sterol response element binding proteins) are expressed over time and regulate the process (Bart A Jessen et al., Gene , 299, pp95-100, 2002; Darlington et al., J. Biol. Chem., 273, pp30057-30060, 1998; Brun RP et al., Curr. Opin.Cell. Biol., 8, pp826-832, 1996).

Homeostasis of fat is maintained by the balance between production and breakdown of fat. ADD1 / SREBP1 is a transcription factor that regulates fat production and regulates the synthesis and absorption of fatty acids, triglycerides, cholesterol, phospholipids (Horton JD et al., J. Clin. Invest., 109, pp 1125-1131, 2002 ). SREBPs are synthesized as an inert precursor bound to the endoplasmic reticulum membrane, and after the N terminus is cut off, it is activated and moved to the nucleus and binds to the SRE (sterol regulatory elements) of the regulatory gene promoter. Among these isomers, SREBP1c mainly regulates the synthesis of neutral lipids, while SREBP2 is known to be involved in cholesterol synthesis. Genes regulated by SREBP1c are ATP citrate lyase (ACL), acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), and stearoyl-CoA desarurase (SCD) (Osborn TF et al., J. Biol. Chem., 275, pp 32379-32382, 2000; Soazig L. L et al., J. Biol. Chem., 277, pp 35625-35634, 2002).

Current strategies for developing obesity treatments include reducing meals, suppressing caloric intake, promoting exothermic reactions, regulating energy metabolism and regulating signaling through the nervous system (Kopleman P.G., Nature, 404, pp635-643, 2000). Attempts to use obesity treatments by developing drugs to perform one or more functions for such a strategy have been continued for a long time, but it is not easy to develop drugs that have safety and efficacy at the same time.

Therefore, attempting to find a component that meets the strategy for treating obesity from natural products with proven safety may be a more efficient approach than developing therapeutics from synthetic preparations.

According to the Chinese herbal classics such as synthosis , sulfur duck, Yu-geun-pi, Yu-geun-pi, Songhwa powder, Sancheongmok, bamboo salt, Injin mugwort, bellflower, and black bean, Sesulgi ( Semisulospira libertine ), according to oriental classics such as synergism It is known to be effective in controlling ". In addition, Kim Il-hoon, Ph.D., a folk medicine scientist, said that the blue pigment in the scallop is similar to the liver pigment in humans, making it an excellent medicine for diseases of the hepatobiliary system. ).

However, none of the above-mentioned documents teaches or discloses clinical or experimental evidence that sewage is efficacious in obesity.

In addition, Seulgigi, sulfur duck, Yugeunpi, Songhwa powder, Sancheongmok, bamboo salt, Injin mugwort, bellflower and black bean, etc. have been used for a long time as an edible or herbal medicine, the extracts of the present invention extracted therefrom also have problems such as toxicity and side effects. none.

Accordingly, an object of the present invention is to suppress the weight gain and reduce the dietary efficiency, reduce the number and size of fat cells in tissues, and inhibit the expression of SREBP-1, FAS, SCD-1 mRNA, genes involved in fat synthesis To provide a composition for the prevention and treatment of obesity and hypercholesterolosis using a combination extract consisting of marshmallow, sulfur duck, yukeunpi, pine pollen, sanchomok, bamboo salt, jinjin mugwort, bellflower and black bean.

In order to achieve the above object, the present invention is a complex extract consisting of marshmallow, sulfur duck, yugeunpi, pine tree powder, mountain cheongsam, bamboo salt, jinjin mugwort, bellflower, and black bean, preferably Dahlgi 20 ~ 1: sulfur duck 1 ~ 10: 5 ~ 1: Sancheongmok 5 ~ 1: Songhwa powder 5 ~ 1: Bamboo salt 5 ~ 1: Injin mugwort 5 ~ 1: Bellflower 5 ~ 1: Black bean 2 ~ 1 The compound extract consisting of the blending weight ratio (w / w%) is an active ingredient Provided is a pharmaceutical composition for treating and preventing obesity and hypercholesterolemia.

In order to achieve the above object, the present invention is a complex extract consisting of marshmallow, sulfur duck, bamboo salt, jinjin mugwort, mountain blue, Yugeunpi, Songhwa powder, bellflower and black bean, preferably Dahlgi 20 ~ 1: sulfur duck 1 ~ 10: 5 ~ 1: Sancheongmok 5 ~ 1: Songhwa powder 5 ~ 1: Bamboo salt 3 ~ 1: Injin mugwort 3 ~ 1: Bellflower 3 ~ 1: Black bean 2 ~ 1 The compound extract consisting of the blending weight ratio (w / w%) is an active ingredient Provides health functional foods for improving and preventing obesity and hypercholesterolemia.

The extract includes an extract available in a solvent selected from water, lower alcohols having 1 to 4 carbon atoms, or a mixed solvent thereof.

Method for producing a composite extract of the present invention is as follows.

Hereinafter, the present invention will be described in detail.

The composite agent extract of the present invention, after washing the sludge, from about 1 to 10 times, preferably about 2 to 5 times water, lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof of the weight (kg) Cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction for about 1 hour to 60 hours, preferably 24 hours to 48 hours at an extraction temperature of 0 to 200 ° C., preferably 50 to 150 ° C., with a selected solvent, preferably water Extraction method, such as the first step to obtain a multi-seed extract of the present invention by filtration after repeated extraction several times by hot water extraction method;

Sulfur ducks are washed only after removing the intestines and the hair, and washed by about 1 to 5 times the weight (kg), preferably about 1 to 3 times water, lower alcohols having 1 to 4 carbon atoms, or a mixed solvent thereof. For example, cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, etc., for about 1 hour to 48 hours, preferably 20 hours to 30 hours at an extraction temperature of 0 to 200 ° C, preferably 50 to 150 ° C, with water A second step of obtaining a sulfur duck extract by extracting the extract several times, preferably by hot water extraction, and filtering so that no solids remain;

Washed root roots 10-1: Sancheongmok 10-1: Weight blending ratio of Injin mugwort 5-1 (w / w%), Preferably, root roots 5-1: Sancheongmok 5-1-1: Weight blending ratio of Injin mugwort 3-1 (w / w %) And a solvent selected from about 1 to 5 times the weight (kg), preferably about 1 to 3 times water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably 0 Extraction method such as cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction for about 1 hour to 48 hours, preferably 20 hours to 30 hours at an extraction temperature of 200 to 200 ℃, preferably 50 to 150 ℃, preferably A third step of repeatedly extracting by hot water extraction and then filtering to prevent the solids from remaining;

A fourth step of collecting the filtrate in one place and concentrating again at a temperature of 70 to 150 ° C., preferably 75 to 100 ° C. for 10 to 60 hours, preferably 40 to 50 hours;

5th step of completely drying the crushed solid content filtered in the first step, mixed at a weight blending ratio (w / w%) of Sludge 20 to 1: Bellflower 5 to 1: Black bean 2 to 1, and then completely ground to a powder ;

The powder prepared in step 5 is uniformly mixed with the concentrate of step 4, and then formulated in a ring, and then the step of step 6 is dried at 50 to 100 ℃, preferably 60 to 80 ℃.

The combination extract prepared by the method is hyperinsulinemia, obesity, insulin resistance due to the mutation of the appetite suppressing hormone leptin (Campfield, LA, Smith, FJ et al., Horm. Metab. Res ., 28 , pp619, 1996) (Meglasson, MD, Wilson, JM et al., J. Pharmacol.Exp. Ther. 266 , pp 1454, 1993; Uysal, KT, Wiesbrock. SM et al., Nature , 389 , pp 610, 1997) / 6J ob / ob mice inhibit weight gain and reduce dietary efficiency, reduce the number and size of fat cells in tissues and inhibit the expression of SREBP-1, FAS, and SCD-1 mRNA, genes involved in fat synthesis The present invention was completed by confirming the effect.

The pharmaceutical composition for the treatment of obesity containing a complex extract consisting of the marshmallow, sulfur duck, rye root, pine powder, wild blue, bamboo salt, jinjin mugwort, bellflower and black bean of the present invention is 0.1 to 50% by weight of the extract based on the total weight of the composition Included as a percentage.

Pharmaceutical compositions comprising the extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

Pharmaceutical dosage forms of the extracts of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds, as well as in any suitable collection.

Pharmaceutical compositions comprising extracts according to the invention, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterile injectable solutions, respectively, according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. Or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, Whitepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the extracts of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.

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

In addition, the present invention has the effect of inhibiting weight gain and dietary efficiency in C57BL / 6J ob / ob mice, reducing the number and size of fat cells in tissues, SREBP-1, FAS, SCD- genes involved in fat synthesis 1 For the improvement and prevention of obesity and hypercholesterolemia, which contains a complex extract consisting of Dasuggi, Sulfur Duck, Yu Geun-pi, Songhwa Powder, Sancheongmok, Bamboo Salt, Injin mugwort, Bellflower, and Black Bean, which have the effect of inhibiting the expression of mRNA, as an active ingredient Provide dietary supplements.

As defined herein, "health functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and "functional" means It means ingestion for the purpose of obtaining useful effects on health use such as nutrient control or physiological action on structure and function.

The dietary supplement for the improvement and prevention of obesity and hypercholesterolemia of the present invention comprises the extract in an amount of 0.01 to 95%, preferably 1 to 80% by weight, based on the total weight of the composition.

In addition, it is possible to manufacture and process as a health functional food in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of improving and preventing obesity and hypercholesterolemia.

For example, the health functional food in the form of tablets may be prepared as it is or granularly mixed with an excipient, a binder, a disintegrating agent or other additives in a suitable manner, and then compressed into a glidant, etc. It is made by directly compressing the functional food directly or by mixing it evenly with excipients, binders, disintegrants or other suitable additives, or by mixing the health functional foods as it is or evenly with the appropriate additives and preparing them by compression molding. Add excipients, binders or other suitable additives to the dietary supplement foods, moisten the powder evenly with a solvent, mold the wet powder into the mold at low pressure, and dry it in an appropriate manner to prepare it. In addition, the nutraceutical can be added to the health functional food in the form of tablets, if necessary, can be avoided with a suitable epidermis.

Among the health functional foods in the form of capsules, the hard capsules are usually prepared by mixing the capsules evenly with the health functional foods or excipients suitable for the health functional foods, granulated by a suitable method, or granulated with a suitable epidermal agent as it is or Soft capsules are prepared by filling them, and soft capsules are usually filled with capsules containing glycerin or sorbitol in a capsule form containing gelatin or appropriate excipients suitable for health functional foods or health functional foods. It is molded and prepared, and a coloring agent preservative etc. can be added to the said capsule base as needed.

Circular functional foods are usually mixed with excipients, binders, disintegrants, etc., and then molded into a spherical form in a suitable manner, and the coating is carried out with white sugar or other suitable coating agent, or starch, talc or You can also be greeted with a suitable substance.

The health functional food in the form of granules is usually made by adding the excipients, binders, disintegrating agents, etc. into the health functional foods as it is or evenly mixing them, and then granulating them in a proper way to make the particles as even as possible. , Mating agent, etc. can be added. For the health functional food in the form of granules, No. 12 (1680 μm), No. 14 (1410 μm) and No. 45 (350 μm) are used for the next particle size test. It should be less than 5.0% and pass through No.45 to less than 15.0% of the total amount.

The definitions of the excipients, binders, disintegrants, lubricants, copulation agents, flavoring agents, etc. of the present invention are those described in the literature known in the art and include those having the same or similar functions. , Korean College of Pharmacy, 5th Edition, p33-48, 1989).

Hereinafter, the present invention will be described in detail by the following Examples and Experimental Examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Experimental Examples.

Example 1 Preparation of a Combination Extract

1-1. Sample Preparation

15kg of live marshmallows, 1 sulfur duck fed with more than 1 year of sulfur, 2kg of yugeunpi (knee bark bark), 9kg of refined purple salt (bamboo salt), 2kg of wild cheongsam, 2kg of songhwa powder, 1kg of jinjin mugwort, Gilkgyeong) 1kg, black bean 0.6kg is prepared.

1-2. Preparation of Concentrate

After washing the marshmallows cleanly, add three times the amount of purified water and make it for 36 hours.The sulfur duck is washed after removing only the intestines and the hair and ironed with twice the amount of purified water for 24 hours to filter out the solids. 2 kg of root roots, 1 kg of Injin mugwort, and 2 kg of Sancheongmok were put together, and 2 times the amount of purified water was added and filtered for 36 hours. The filtrates were collected in one place, and then concentrated again for 48 hours while maintaining the temperature of 75 ° C. to 90 ° C.

1-3. Manufacture of powder

In Example 1-2, 4.5 kg of the filtered sage is picked out and completely dried. Dried marshmallow, bellflower, and black bean were ground to a powder.

1-4. Preparation of Vitam-S

The powder prepared in Example 1-3 is uniformly mixed with the concentrated solid of Example 1-2, formulated in a ring and hot-air dried at 73 ° C. The pill was dissolved in distilled water, homogenized with a stirrer, and used as a sample of the present invention (hereinafter referred to as Vitam-S; BS).

Reference Example 1. Preparation of Laboratory Animals

The mouse used in this study was purchased from Orient Co., Ltd., 6 weeks old C57BL / 6J ob / ob mice of about 20-25g. Mice were kept in a plastic rat cage and raised in an animal room under conditions of 23 ± 2 ° C, 50 ± 10% relative humidity, and light / dark cycle (12 / 12hr), and water and feed were freely consumed. The experimental animals were used after adapting to the animal room environment for one week before starting the experiment. Mice were divided into four groups of eight animals as follows. The control group was treated with distilled water (Con), and the sample group was treated with metformin at a concentration of 250 mg / kg (BS250) and 500 mg / kg (BS500), and metformin 300 mg / kg (MT300) for 8 weeks. During oral administration.

Reference Example 2. Data Analysis and Statistics Processing

 All experimental results are expressed as mean ± SE. The statistical significance was treated with Student's t-test compared to the Ob / ob control group.

Experimental Example 1. Measurement of weight change and dietary efficiency

To test the effect of Vitam-S on obesity, weight gain, dietary intake and dietary efficiency were compared among the experimental groups. Pre-test body weights were reared for one week to adapt to the animal room environment, weighed immediately prior to dosing, and weighed for the first time after the experiment and weighed exactly the same eight weeks later. Dietary efficiency was calculated using the following equation. Each mouse gained eight weeks of body weight divided by eight weeks of diet.

Dietary efficiency = [weight gain (g / 8 weeks)] / [food intake (g / 8 weeks)]

As a result, as shown in Table 1, the weight gain of the control group increased by 9.37 ± 2.16g for 8 weeks, while the Vitam-S administration group increased 5.89 ± 2.79g and 6.01 ± 1.65g at 250 and 500 mg / kg doses. Compared with the control group, it showed 37%, 36% of weight increase activity. In the metformin-administered group, the positive control drug, the body weight increased by 6.56 ± 1.23g during the same period, which inhibited the 30% weight gain compared to the control group. In addition, as a result of comparing the weight gain rate according to the diet intake for 8 weeks between groups, the group treated with Vitam-S 250, 500 mg / kg and metformin showed 25%, 23%, and 17% reduction in dietary efficiency, respectively. Indicated. The decrease in dietary efficiency compared to the control group means that the weight gain is smaller than the total food intake for 8 weeks. It was confirmed that the dietary efficiency of the BS250 administration group was reduced by 25% compared to the control group, showing the greatest activity, and thus it was useful as a pharmaceutical composition for preventing and treating obesity.

Body weight gain, dietary intake, and dietary efficiency of BS and metformin-treated groups Experimental group Body weight (g) Weight gain (g) Dietary Intake (g / 8 weeks / mouse) Dietary Efficiency (x 10 ^-3) Before experiment After the experiment Control 47.05 ± 2.44 57.37 ± 3.05 9.37 ± 2.16 244.16 ± 2.81 38.38 BS250 46.49 ± 3.39 52.39 ± 3.43 * 5.89 ± 2.79 * 204.99 ± 3.34 * 28.73 BS500 46.92 ± 3.04 52.93 ± 2.89 6.01 ± 1.65 ** 204.93 ± 2.86 * 29.33 MT300 46.87 ± 2.20 53.11 ± 3.23 6.56 ± 1.23 * 205.38 ± 2.21 ** 31.94 Each represented by mean ± SD (n = 8). * P <0.05, ** P <0.01 vs. Con.

Experimental Example 2. Weight and histological observation of liver and epididymis

2-1. Measurement of weight change in liver and epididymis

After 8 weeks of experiment, the weights of the livers and the epididymal fat pads extracted from ether-analyzed C57BL / 6J ob / ob mice were compared. The BS250, BS500, and MT300-administered groups showed a decrease in weight between 24%, 20%, and 20%, respectively, compared to the control group, but there was no statistically significant difference. Liver weight to body weight tended to decrease in the three groups compared to the control group, but no significant difference was found. On the other hand, the epididymis showed a significant decrease of 19% in the BS250 group compared with the control group.

Experimental group Liver weight (g) Liver weight (g) / weight (g) Weight of epididymis (g) Control 3.29 ± 1.36 5.18 ± 0.45 2.69 ± 0.40 BS250 2.50 ± 0.28 5.05 ± 0.56  2.17 ± 0.20 * BS500 2.63 ± 0.33 5.07 ± 0.57 2.60 ± 0.75 MT300 2.64 ± 0.39 5.07 ± 0.54 2.52 ± 0.32 Values are expressed as mean ± SD (n = 8) * P <0.05 vs. Con.

2-2. Histology of the liver and epididymis

Tissues extracted from C57BL / 6J ob / ob mice were fixed using 10% neutral buffered formalin. After the dehydration and embedding process to produce a paraffin block and a tubular section of 5 ㎛ thickness to prepare a paraffin removed by xylene (xylene), 100%, 95%, 90%, 80%, 70% alcohol Hydrated. The staining method was dehydrated with Hematoxylin-Eosin staining, sutured with Canadian balsam, and observed with an optical microscope (BX-50, Olympus, Japan).

As a result of comparing the size of the adipocytes of the epididymis between groups, the area of the control group was 4,186 ± 221.4mm ² , while that of the group treated with Vitam-S 250 mg / kg was 3,427 ± 118.0 mm ² , which was 18% larger than the control group. There was a significant difference decreasing. In addition, it was shown in Figure 1 by comparing the HE-stained tissues and tissues of the tissues between the groups. In the Vitam-S group, the tissues were smaller than the control group, and the number of lipid droplets in the hepatocytes was significantly reduced. From this result, it was confirmed that Vitam-S has improved obesity and hypercholesterolemia.

Experimental Example 3. Expression of fat synthesis related genes

3-1. RNA Isolation and RT-PCR

RNA was isolated from hepatocytes to compare mRNA expression of SREBP-1 and its related genes, FAS and SCD-1, in liver tissue. Total RNA from the liver was isolated using guanidine thiocyanate-water saturated phenol / chloroform and acidic sodium acetate (Chomczynski, P. and Sacchi, N., Anal. , 162 , pp 156, 1987). The total RNA amount was measured by absorbance at 260 nm and 280 nm using a UV spectrophotometer. 1 μg of total RNA was reverse transcribed into cDNA using Moloney murine leukemia virus transcriptase and oligo dT (15) (Promega, USA). Primers for each enzyme are shown in Table 3 and SEQ ID NOs: 1-8 below.

SREBP-1 sense primer 5'-GCGCTACCGGTCTTCTATCA-3 ', anti-sense 5'-TGCTGCCAAAAGACAAGGG-3 ' FAS sense primer 5'-GATCCTGGAACGAGAACAC-3 ' anti-sense 5'-AGACTGTGGAACACGGTGGT-3 ' SCD-1 sense primer 5'-CGAGGGTTGGTTGTTGATCTGT-3 ' anti-sense 5'-ATAGCACTGTTGGCCCTGGA-3 ' GAPDH sense primer 5'-CAACTTTGGCATTGTGGAAGG-3 ', anti-sense 5'-ATGGAAATTGTGAGGGAGATGC-3 '

PCR buffer conditions were mixed with 20 mM Tris-HCl (pH 8.4), 50 mM KCl, 1.5 mM MgCl ₂ , 0.5 mM dNTP, 2.5 units Taq DNA polymerase (TaKaRa, medicals, Japan), 0.05 μg c DNA, 0.5 μM primer It was. PCR conditions were denatured at 94 ℃, bound at 57 ℃, elongated at 72 ℃, PCR products were electrophoresed using a 1% agarose gel. Density is measured in the GS-700 image densitometer and is shown in FIGS.

Figure 2 is a comparison of the mRNA expression of SREBP-1 between the groups compared to the control group BS250, BS500, MT300 administration group showed the expression of 61%, 46%, 106%, respectively, dose-dependent 39% in the Vitam-S administration group , 54% expression inhibitory activity. Figure 3 is a comparison of the expression of FAS mRNA in the BS250, BS500, MT300 group compared to the control group was 49%, 40%, 70% expression, respectively, dose-dependent 51%, 60% of the Vitam-S administration group It showed expression inhibitory activity. In the metformin-treated group, 30% FAS mRNA expression was also suppressed compared to the control group, which is consistent with the results of Lin et al., Which confirmed the expression of FAS protein by metformin administration in ob / ob mice by immunochemistry (Matthews, DR, Hosker, JP et al., Diabetologia, 28 , pp 412, 1985). Figure 4 is a comparison of the mRNA expression of SCD-1 between the groups showed 84%, 50%, 90% expression compared to the control, respectively, showing 16%, 50%, 10% expression inhibition pattern. Vitam-S administration group also showed a dose-dependent inhibitory effect on the expression of SCD-1 mRNA, obesity the extract of the complex consisting of sesame, sulfur duck, rye root, pine pollen, wild blue, bamboo salt, jinjin mugwort, bellflower and black bean of the present invention And it was confirmed that it can be usefully used as a composition for the treatment and prevention of hypercholesterolemia.

The combination extract of the present invention may be administered in the following formulations, and the following formulation examples are merely to illustrate the present invention, thereby not limiting the contents of the present invention.

Formulation Example 1 Preparation of Powder

Combination Extract 300 mg

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Combination Extract 300 mg

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Combination Extract 300 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium Stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

       Combination Extract 300 mg

Mannitol 180 mg

Sterile distilled water for injection 2974 mg

Na ₂ HPO _{4 12} H ₂ O 26 mg

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Combination Extract 300 mg

10 g of isomerized sugar

5 g of mannitol

Purified water

According to the conventional method of preparing a liquid solution, each component is added to the purified water to dissolve, the lemon flavor is added appropriately, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by the addition of purified water, and then filled into a brown bottle. The solution is prepared by sterilization.

Formulation Example 6 Preparation of Healthy Food

1000 mg of pills in combination extract

Vitamin mixture proper amount

70 μg of Vitamin A Acetate

Vitamin E 1.0 mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15 mg

Vitamin B ₆ 0.5 mg

0.2 μg of vitamin B ₁₂

Vitamin C 10 mg

10 μg biotin

Nicotinic Acid 1.7 mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dibasic calcium phosphate 55 mg

Potassium Citrate 90 mg

Calcium Carbonate 100 mg

Magnesium chloride 24.8 mg

Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 7 Preparation of Healthy Drink

       Combination Extract 1000 mg

Citric acid 1000 mg

100 g oligosaccharides

Plum concentrate 2 g

1 g of taurine

Add 900 ml of purified water

After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed, sterilized and refrigerated It is used to prepare a healthy beverage composition of the present invention.

Although the composition ratio is mixed with a component suitable for a favorite beverage in a preferred embodiment, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.

The pharmaceutical composition for the prevention and treatment of obesity, containing the extract of the complex consisting of the marshmallow, sulfur duck, rye root, pine powder, calendula, bamboo salt, jinjin mugwort, bellflower, and black bean of the present invention is to suppress the weight gain in C57BL / 6J ob / ob mice and It is effective in reducing the number and size of fat cells in tissues and inhibiting the expression of SREBP-1, FAS, and SCD-1 mRNA, which are involved in fat synthesis, to treat obesity and hypercholesterolemia. It can be usefully used as a preventive pharmaceutical composition and health functional food.

<110> S & B CO., LTD. <120> A composition comprising the extract of Semisulosira libertine,          Sulfur fed Duck, Ulmus devidiana, Pine Pollen Powder, Acer          tegmentosum Maxim., Bamboo salt, Artemaisia capillaries,          Platycodon Grandiflorum, Rhynchsiavolubilis for the prevention          and treatment of obesity and hypercholesterolemia <130> DIF / 2005-07-0003 / HJ <160> 8 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SREBP-1 sense primer <400> 1 gcgctaccgg tcttctatca 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> SREBP-1 anti-sense primer <400> 2 tgctgccaaa agacaaggg 19 <210> 3 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> FAS sense primer <400> 3 gatcctggaa cgagaacac 19 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> FAS anti-sense primer <400> 4 agactgtgga acacggtggt 20 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> SCD-1 sense primer <400> 5 cgagggttgg ttgttgatct gt 22 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> SCD-1 anti-sense primer <400> 6 atagcactgt tggccctgga 20 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> GAPDH sense primer <400> 7 caactttggc attgtggaag g 21 <210> 8 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> GAPDH anti-sense primer <400> 8 atggaaattg tgagggagat gc 22

Claims (4)

A pharmaceutical composition for the treatment and prevention of obesity and hypercholesterolemia, containing as an active ingredient extracts consisting of Dasulgi, Sulfur Duck, Yugeunpi, Songhwa Powder, Sancheongmok, Bamboo Salt, Injin mugwort, Bellflower, and Black Bean. The method according to claim 1, marshmallow 20 ~ 1: sulfur duck 1 ~ 10: yugeunpi 5 ~ 1: mountain cheongmok 5 ~ 1: pine flower powder 5 ~ 1: bamboo salt 5 ~ 1: jinjin mugwort 5 ~ 1: bellflower 5 ~ 1: black bean 2 A pharmaceutical composition for the treatment and prevention of obesity and hypercholesterolemia, comprising as an active ingredient an extract of a combination consisting of ~ 1 blending weight ratio (w / w%). Health functional food for improving and preventing obesity and hypercholesterolemia, which contains complex extracts consisting of Dasulgi, Sulfur Duck, Bamboo Salt, Injin mugwort, Sancheongmok, Yugeunpi, Songhwa Powder, Bellflower, and Black Bean as active ingredients. The method according to claim 3, sledulgi 20 ~ 1: sulfur duck 1 ~ 10: yugeunpi 5 ~ 1: mountain cheongmok 5 ~ 1: pine flower powder 5 ~ 1: bamboo salt 3 ~ 1: jinjin mugwort 3 ~ 1: bellflower 3 ~ 1: black bean 2 A dietary supplement for improving and preventing obesity and hypercholesterolemia, comprising as an active ingredient an extract of a combination consisting of ~ 1 blending weight ratio (w / w%).

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