KR20110102649A - Composition comprising an extact of cirsium japonicum and the compounds isolated thereform for the treatment and preventation of obesity - Google Patents

Abstract

The present invention thistle ( Cirsium The present invention relates to a pharmaceutical composition and a dietary supplement for the treatment and prevention of obesity, comprising a japonicum extract or a compound isolated therefrom as an active ingredient, wherein the extract according to the present invention and the compound apigenin isolated therefrom are fibroblasts ( 3T3-L1 cell) inhibits the differentiation of adipocytes and inhibits the synthesis of triacylglycerol, which can be widely used to fundamentally treat and prevent obesity as well as to alleviate and improve symptoms.

Description

COMPOSITION COMPRISING AN EXTACT OF CIRSIUM JAPONICUM AND THE COMPOUNDS ISOLATED THEREFORM FOR THE TREATMENT AND PREVENTATION OF OBESITY}

The present invention thistle ( Cirsium japonicum ) extract or a composition for the treatment and prevention of obesity containing a compound separated therefrom as an active ingredient, more specifically, thistle extract having an anti-obesity effect, and the extract or a compound isolated therefrom as an active ingredient It relates to a pharmaceutical composition and health functional food for containing obesity treatment and prevention.

Obesity is a condition in which fat is excessively accumulated in the body and a high proportion of fat in the body weight.

As modern life becomes more complicated and diverse than in the past, obesity is caused by various factors such as distorted lifestyles, nutrient imbalance, lack of exercise, and excessive drinking.

In the Orient, where the frequency of obesity was not high due to the vegetarian diet, the incidence of obesity increased significantly due to the changes in diet, habits, and living environment caused by Western culture.

Obesity causes various adult diseases such as high blood pressure, heart disease, diabetes, and causes serious health problems such as increasing the morbidity to chronic diseases and shortening the life span of phosphoric acid.

There are many known causes of obesity, including genetic predisposition and environmental factors. In particular, if both parents are obese, the child is more than 50% chance of obesity, if one side obesity is about 25% obesity probability of genetic effects is very low. In addition, the reduction of energy consumption due to excessive intake of calories caused by lifestyle changes and environmental factors due to lack of exercise is also a major cause of obesity.

The methods used to treat obesity can be divided into methods of correcting lifestyles such as diet therapy, exercise therapy and behavioral therapy, and medication and surgical treatment. Correcting lifestyles is not easy and weight loss is limited and requires medication along with lifestyle correction.

The use of drugs for the treatment of obesity has been tried for a long time, but obesity treatment drugs targeting many substances that have been found to be involved in the regulation of energy homeostasis of the human body has limited clinical applications due to side effects.

There are currently two types of obesity drugs approved by the US FDA for long-term use: sibutramine (sibutramine), which inhibits reuptake of norepinephrine and serotonin, and lipolytic secretion from the pancreas and digestive system. There is an orlistat that suppresses enzymes and has an anti-obesity effect.

However, in the case of sibutramine, side effects such as increased blood pressure, insomnia, dry mouth, and dizziness are common, and it is not available to patients with cardiovascular disease and uncontrolled hypertension. Oristat has diarrhea, fat stool, and loss money. In case of less fat intake than in Koreans, such as Koreans, the effect of the drug is not clear.

Therefore, recent research and development method that can safely treat obesity using a natural-derived physiologically active substance has been actively conducted.

Conventional obesity treatments and problems by treatment Main drugs problem Thyroid Hormone (1890) Hyperthyroidism amphetamine (1930) Addictive digitalis, diuretics (1960) Death report aminorex (1970) Induced Pulmonary Hypertension fenfluramine (1990) 35% of patients on combination therapy cause heart valve abnormalities, banned in 1997 sibutramine (current) Prohibited use for patients with elevated blood pressure, insomnia, dry mouth, dizziness, cardiovascular disease orlistat (current) Diarrhea, fatty stool, faecal incontinence, not effective for Asians

Thistle ( Cirsium japonicum ) is a perennial herb belonging to the Asteraceae family, and is distributed in Korea, Japan, and northeast China. It is 50-100cm high and has white hairs and cobweb-like hairs. Root leaves remain until flowering and larger than stem leaves. Stem leaves are lanceolate oval, split like feathers, underneath the circumference, splitting the most magnetic part again, and having thorny teeth with chamfered serrate. Flowers bloom from June to August and purple to red. Gun pieces are arranged in seven to eight rows, the longer the inside. Fruits are achene and tubules are 16-19 mm long. The leaves are narrow, green, and have more thorns. Narrow leaf thistles and thorns are called thistles.

Apigenin, an indicator of thistle, which is widely known as an anti-inflammatory substance, was first known in 1958 as a potency related to histamine derivation from Dr. Subrt F. (Effect of apigenin on histamine liberation, Cesk Fysiol. 1958; 7 (3). (Structural characterization of a flavonoid-inducible Pseudomonas aeruginosa A-band-like O antigen of Rhizobium sp.strain) has been reported to suppress inflammation and oxidative stress and to promote carbohydrate metabolism promotor. NGR234, required for the formation of nitrogen-fixing nodules.J Bacteriol. 2005; 187 (18): 6479-87).

Apigenin is also used by many scientists to produce various cancer cells, such as breast cancer (Way, TD, Kao, MC, and Lin, JK Apigenin induces apoptosis through proteasomal degradation of HER2 / neu in HER2 / neu-overexpressing breast cancer cells). via the phosphatidylinositol-3′-kinase / Akt-dependent pathway.J. Biol.Chem. 2004: 279, 4479-489), colon cancer (Wang, W., Heideman, L., Chung, CS, Pelling, JC, Koehler , KJ, and Birt, DF Cellcycle arrest at G2 / M and growth inhibition by apigenin in human colon carcinoma cell lines.Mol.Carcinogen. 2000; 28 (2): 102-10), skin cancer (Birt, DF, Mitchell, D ., Gold, B., Pour, P., and Pinch, HC Inhibition of ultraviolet light-induced skin carcinogenesis in SKH-1 mice by apigenin, a plant flavonoid.Anticancer Res. And; 17 (1A): 85-91) , Thyroid cancer (Yin, F., Giuliano, AE, and Van Herle, AJ Growth inhibitory effects of flavonoids in human thyroid cancer cell lines.Thyroid and9; 9 (4): 369-76), leukemia (Wang, IK, Lin- Shiau, S. Y., and Lin, J. K. Induction of apoptosis by apigenin and related flavonoids through cytochrome c release and activation of caspase-9 and caspase-3 in leukaemia HL-60 cells. Eur. J. Cancer and 9; K (10): 1517-525), Pancreatic cancer (Strouch MJ, Milam BM, Melstrom LG, McGill JJ, Salabat MR, Ujiki MB, Ding XZ, Bentrem DJ The flavonoid apigenin potentiates the growth inhibitory effects of gemcitabine and abrogates gemcitabine resistance in Proliferation of human pancreatic cancer cells.Pancreas. 2009; 38 (4): 409-15) has been shown to be inhibited, and recently, it has been reported to inhibit the proliferation of prostate cancer (Shukla S., Mishra A., Fu P., MacLennan GT, Resnick MI, Gupta S. Up-regulation of insulin-like growth factor binding protein-3 by apigenin leads to growth inhibition and apoptosis of 22Rv1 xenograft in athymic nude mice.FASEB J. 2005; 19 (14 ): 2042-4).

In this study, in order to observe the cancer prevention effect of apigenin, after ingesting apigenin into mice and injecting prostate cancer cells into the body, apigenin induced self-destruction of cancer cells, which slowed down the proliferation rate and increased weight. It was found that it is not accompanied by side effects.

In addition, apigenin reduces insulin-like growth hormone-1 (IGF-1) levels, which are known to increase the risk of developing prostate cancer, breast cancer, colorectal cancer, and lung cancer, and lower insulin risk. It significantly increases the levels of hormone binding protein-2, prevents the development of prostate cancer, and inhibits the proliferation of cancer cells. In particular, there is an inhibitory effect on carcinogenic aflatoxin B1 activity, among which napigenin and apigenin are known to be most effective (Sanjeev Shukla, Anil Mishra, Pingfu Fu, Gregory T. MacLennan, Martin I. Resnick, and Sanjay). Gupta, Up-regulation of insulin-like growth factor binding protein-3 by apigenin leads to growth inhibition and apoptosis of 22Rv1 xenograft in athymic nude mice.FASEB J. 2005; 19 (14): 2042-4).

In a paper published in a recent issue of PNAS (online edition), UC Riverside researchers also reported that apigenin, a natural substance found in vegetables or fruits, introduces p53 into the nucleus and responds to chemotherapy of cancer cells. (Cai Xin, Liu Xuan. Inhibition of Thr-55 phosphorylation restores p53 nuclear localization and sensitizes cancer cells to DNA damage, Proc Natl Acad Sci US A. 2008; 105 (44): 16958-63).

However, no disclosure or teaching has been made of the anti-obesity effect of thistle extract or apigenin isolated from thistle.

Therefore, the present inventors have continued efforts to develop obesity treatments using natural products. As a result, the extract obtained using thistle and the active fraction isolated from it are inhibited from differentiation of fibroblasts (3T3-L1 cells) into adipocytes. The present invention was completed by confirming that the synthesis of triacylglycerol was inhibited.

After all, the main object of the present invention is to provide a composition for the treatment and prevention of obesity containing thistle extract having an anti-obesity effect or apigenin isolated therefrom as an active ingredient.

In order to achieve the above object, the present invention has an anti-obesity effect thistle ( Cirsium japonicum ) extract.

In the present invention, the thistle extract is characterized in that the crude extract of the thistle or non-polar solvent soluble extract.

The milk thistle crude extract of the present invention is collected by drying thistle and powdered with a crusher, the volume of water, methanol, ethanol, butanol and the like of about 1 to 20 times, preferably about 3 to 10 times the weight of the sample As a polar solvent of C1 to C4 lower alcohol or a mixed solvent thereof, preferably a mixed solvent of water and ethanol, more preferably 60 to 100% ethanol, about 0.1 to 12 hours at 100 to 150 ° C, preferably Is obtained by using extraction methods such as stirring extraction, hot water extraction, cold extraction, warm extraction, reflux cooling extraction or ultrasonic extraction for 1 to 6 hours, preferably obtained by stirring, extraction under reduced pressure or drying under reduced pressure can do.

In addition, the milk thistle non-solvent soluble extract of the present invention is a solvent selected from the thistle crude extract obtained from hexane, chloroform, methylene chloride, dichloromethane, ethyl acetate, glycerin, propylene glycol, butylene glycol or ether, preferably dichloromethane Alternatively, ethyl acetate, more preferably, ethyl acetate may be added, followed by extraction to obtain silica gel column chromatography.

The thistle extract or the active fraction isolated therefrom inhibits the differentiation of fibroblasts (3T3-L1 cells) into adipocytes and has the effect of inhibiting the synthesis of triacylglycerol.

The present invention thus provides a new use of the thistle extract prepared by the above method or a compound isolated therefrom for the treatment and / or prevention of obesity.

Specifically, the present invention provides a pharmaceutical composition for the treatment and prevention of obesity containing the thistle extract or a compound isolated therefrom as an active ingredient.

In the present invention, the compound isolated from the thistle extract is characterized in that the apigenin (apigenin) represented by the following formula (1), the apigenin is not only an active fraction isolated from thistle chemically synthesized compounds or commercially available Anything can be used.

The pharmaceutical composition for treating and preventing obesity of the present invention comprises 0.0001 to 99.99% by weight, preferably 0.001 to 50% by weight of the thistle extract or apigenin isolated from the total weight of the composition.

In addition, the pharmaceutical compositions according to the invention may further comprise suitable carriers, excipients or diluents commonly used in the manufacture of pharmaceutical compositions.

Carriers, excipients or diluents usable in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose , Microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

In addition, the pharmaceutical compositions according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories, and sterile injections, respectively, according to a conventional method. Can be used.

When formulated, diluents or excipients such as fillers, extenders, binders, humectants, sealants, and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may be mixed with at least one excipient, for example, starch calcium carbonate, sucrose or lactose, gelatin, or the like. To prepare. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be 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. have. 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 a base material of suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

In the present invention, the pharmaceutical composition may be administered by varying the dose depending on the progress of the disease, the age, sex, physical condition, administration period, administration method, weight of the patient, diet, discharge rate, and the like of the disease. However, for the desired effect, the pharmaceutical composition of the present invention may be administered parenterally or orally in the range of 0.001 to 100 μg / kg, more preferably in the range of 0.01 to 100 μg / kg. 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.

In addition, thistle extract of the present invention or the pharmaceutical dosage form of apigenin isolated therefrom may be used in the form of their pharmaceutically acceptable salts, and may also be used alone or in combination with other pharmaceutically active substances as well as in a suitable collection. Can be used.

The pharmaceutical composition of the present invention can be administered to various mammals such as rats, mice, livestock, humans, and the like. 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 is the thistle ( Cirsium japonicum ) extract or a health functional food for the improvement and prevention of obesity containing the apigenin isolated therefrom as an active ingredient.

The dietary supplement of the present invention may further include a food acceptable acceptable food supplement additive in addition to thistle extract or apigenin isolated therefrom, and may be suitably used according to a conventional method, and the mixed amount of the active ingredient may be used. It may be appropriately determined depending on the purpose, for example, prophylactic, health or therapeutic treatment.

An effective amount of thistle extract or apigenin isolated from the dietary supplement may be used according to the effective amount of the pharmaceutical composition, but for long-term intake for health and hygiene purposes or for health control purposes, It may be less than the above range, it is clear that the active ingredient can be used in an amount above the above range because there is no problem in terms of stability.

In addition, there are no particular restrictions on the type of health functional food, for example, meat, sausage, bread, chocolate, candy, snacks, confectionary, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups , Beverages, tea, drink, alcoholic beverages and vitamin complexes, health functional foods, and the like.

It may also be added to food or beverages for the purpose of improving obesity and preventing obesity. At this time, the amount of the thistle extract or apigenin isolated from the food or beverage may be added to 0.01 to 30% by weight, preferably 0.3 to 15% by weight of the total food weight, the health beverage composition based on 100ml 0.01 to 5 g, preferably 0.03 to 1 g may be added.

The health functional food of the present invention can be manufactured and processed into health functional foods in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of preventing and improving cerebral nervous system diseases.

The health functional beverage composition of the present invention is not particularly limited to other ingredients except the thistle extract or apigenin isolated therefrom as essential ingredients in the indicated ratios, and various flavors or natural carbohydrates, such as ordinary drinks, may be used. It may further include. At this time, the natural carbohydrate is, for example, monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; And conventional sugars such as polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (tauumatin, stevia extract, for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.

In addition to the above, thistle extract of the present invention or apigenin isolated therefrom is a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, such as colorants and enhancers (cheese, chocolate, etc.), pectic acid And salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, thistle extract of the present invention or apigenin isolated therefrom may contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages, and these components may be used independently or in combination. In addition, the ratio of the additive is not so important, but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of thistle extract of the present invention or apigenin isolated therefrom.

Thistle of the present invention ( Cirsium) japonicum ) extract or a compound isolated therefrom can inhibit the differentiation of fibroblasts (3T3-L1 cells) into adipocytes and inhibit the synthesis of triacylglycerol, thereby fundamentally treating and preventing obesity. It can also be widely used to relieve and ameliorate symptoms.

In addition, thistle of the present invention is an item approved as a health functional food, so thistle extract of the present invention or a compound isolated therefrom also has no problems such as toxicity and side effects, such as drugs or health functions for the treatment and prevention of obesity It can be usefully used as a food.

Figure 1 shows the HPLC profile of the compound isolated from the thistle extract according to the present invention.
Figure 2 shows the ¹ H NMR of the compound isolated from the thistle extract according to the present invention.
Figure 3 shows ¹³ C NMR of the compound isolated from thistle extract according to the present invention.
Figure 4 is a photograph showing the inhibition rate of differentiation of 3T3-L1 cells into adipocytes by concentrations of the compounds isolated from thistle extract according to the present invention (P: positive control (DEME / 10% FBS + insulin 5㎍ / ㎖); A25 : Positive control condition + apigenin 25μM; A50: positive control condition + apigenin 50μM; and A75: positive control condition + apigenin 75μM).
Figure 5 is a photograph showing the effect on triacylglycerol synthesis of 3T3-L1 cells by concentration of the compound isolated from the thistle extract according to the present invention (P: positive control (DEME / 10% FBS + insulin 5㎍ / ㎖) A25: positive control condition + 25 μM apigenin; A50: positive control condition + apigenin 50 μM; and A75: positive control condition + apigenin 75 μM).
Figure 6 is a photograph showing the inhibition rate of differentiation of 3T3-L1 cells into adipocytes by concentration in the insulin medium of the compound isolated from thistle extract according to the present invention (P: positive control (DEME / 10% FBS + insulin 5㎍ / ㎖) A25: positive control condition + apigenin 25μM; A50: positive control condition + apigenin 50μM; and A75: positive control condition + apigenin 75μM).

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Example 1 Preparation of Thistle Crude Extract

Gathered in the wild, thistle Cirsium japonicum ) was pulverized and milled to powder, and then 5 times 70% ethanol to the weight was added to extract the refluxed hot water three times at 80 ° C for 3 hours. 15 g was obtained.

Example 2. Preparation of Thistle Nonpolar Solvent Soluble Extract

The crude ethanol extract obtained in Example 1 was dissolved in distilled water, ethyl acetate was mixed at a ratio of 1: 1 (v / v), and the distribution was repeated three times. The ethyl acetate extract was concentrated under reduced pressure, concentrated and dried. 1.63 g of ethyl acetate extract was obtained.

Example 3 Isolation of Apigenin from Thistle Extract

Silica gel flash chromatography (Silica gel 60, Merck, Germany) was performed on the ethyl acetate extract obtained in Example 2, and the active fractions were decompressed and dried again, and then dissolved in a small amount of methanol at a concentration of 20 µg / ml. The mixture was filtered through Millipore (0.45 nm), followed by HPLC (2487, Waters, USA) to fractionate 25 mg of the active substance.

At this time, HPLC used 74% methanol as the mobile phase, the flow rate was 10 ml per minute, and the column used Odyss 20 × 250 mm. In addition, the aliquoted compound was identified by spectroscopic method (300 MHz NMR Spectrscopy DRX 300, Bruker, Germany).

1 to 3 show HPLC profiles, ¹ H and ¹³ C NMR spectra of active materials according to the invention. From the above results, it was confirmed that the active material according to the present invention was apigenin (aligenin) represented by Chemical Formula 1.

[Formula 1]

Experimental Example 1. Confirm the effect of inhibiting obesity

The fibroblasts (3T3-L1 cells) were cultured in a culture dish of 10 cm in diameter using a culture medium for animal cell growth (see Table 2 below) at 37 ° C. in a 5% CO ₂ incubator. Incubate with fresh medium every three days.

DMEM FBS Sodium
bicarbonate Insulin Dex Mix Propagation 13.4 g / ℓ 10% 3.7 g / ℓ - - - Eruption induction
Badge-1 13.4 g / ℓ 10% 3.7 g / ℓ 5 μg / ml 1 μM 0.5 mM Eruption induction
Badge-2 13.4 g / ℓ 10% 3.7 g / ℓ 5 μg / ml

However, DMEM is Dulbecco's modified Eagle's medium, FBS is Fetal Bovine serum, Dex is Dexamethasone and Mix is 1-methyl-3-isobutylxanthine.

The aliquots were washed twice with phosphate buffered saline solution and desorbed with 0.25% trypsin-EDTA before the cells were in the confluent stage (numerically grown to fill cells throughout the culture dish). Dividing into 6-well plate at a ratio of 1: 4, adding differentiation-inducing medium (Mix) and then differentiation-inducing medium (medium-1: 0-48 hours; medium-2 every 9 hours for 9 days) : 48 hours later) and thistle extracts obtained in Examples 1 to 3 or compounds isolated therefrom were added at concentrations of 25, 50, and 75 μM, respectively.

Adipose cells were stained with an oil-red O reagent, which stains red, and confirmed by phase contrast microscopy (ㅧ 200).

In addition, to confirm the effect of the thistle extract of the present invention or a compound isolated therefrom on the synthesis of triacylglycerol (TG) of 3T3-L1 cells, cells cultured in each experimental group were desorbed with Tris-EDTA buffer. Next, the cells were pulverized with an ultrasonic grinder. Prepare a calibration curve of the standard solution according to the instructions written in the BCA Protein Assay Kit (thermo Sci., USA), dilute the desorption material obtained from the cells in a kit reagent, and warm at 37 ° C for 10 minutes. Within 60 minutes, the absorbance was measured at a wavelength of 550 nm using the reagent blind as a control, and the amount of triacylglycerol per protein (µg / mg protein) was measured according to the formula.

As a result, it was confirmed that the apigenin according to the present invention can differentiate 3T3-L1 cells into adipocytes (see FIG. 4), and also inhibit the synthesis of triacylglycerol of 3T3-L1 cells in insulin medium. (See FIG. 5).

In addition, as shown in Figure 6, it was found that the apigenin according to the present invention inhibits the expression of proteins that induce differentiation of 3T3-L1 cells into adipocytes.

Examples of the preparation of a pharmaceutical composition comprising the thistle extract of the present invention or a compound isolated therefrom are illustrated as follows, but the present invention is not intended to be limited thereto but is intended to be described in detail.

Formulation Example 1 Preparation of Tablet

Apigenin 10mg

Lactose 100mg

Starch 100mg

2 mg magnesium stearate

The tablets were prepared by mixing the above components and tableting according to a conventional method for producing tablets.

Formulation Example 2 Preparation of Powder

Thistle extract 20mg

Lactose 100mg

Talc 10mg

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

Formulation Example 3 Preparation of Capsule

Thistle extract 10mg

3mg of crystalline cellulose

Lactose 14.8mg

0.2 mg magnesium stearate

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

Formulation Example 4 Preparation of Injection

Apigenin 10mg

Mannitol 180mg

Sterile distilled water for injection 2974 mg

pH adjuster 26 mg

(2 ml) per 1 ampoule according to the usual injection preparation method.

Formulation Example 5 Preparation of Liquid

Thistle extract 20mg

10 g of isomerized sugar

Mannitol 5g

Purified water

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, And sterilized to prepare a liquid preparation.

Formulation example  6. Manufacture of health food

Apigenin 1000mg

Vitamin mixture proper amount

70 μl of Vitamin A Acetate

Vitamin E 1.0mg

Vitamin B ₁ 0.13 mg

Vitamin B ₂ 0.15mg

Vitamin B ₆ 0.5mg

0.2 μg of vitamin B ₁₂

Vitamin C 10mg

10 μg biotin

Nicotinamide 1.7mg

Folate 50 ㎍

Calcium Pantothenate 0.5mg

Mineral mixture

Ferrous Sulfate 1.75mg

Zinc Oxide 0.82mg

Magnesium Carbonate 25.3mg

15 mg potassium monophosphate

Dicalcium Phosphate 55mg

Potassium Citrate 90mg

Calcium Carbonate 100mg

Magnesium chloride 24.8mg

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 Health Beverage

Thistle extract 1000mg

Citric Acid 1000mg

Oligosaccharide 100g

Plum concentrate 2g

1 g of taurine

900 ml of purified water

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

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

As described above, specific portions of the contents of the present invention have been described in detail, and for those skilled in the art, these specific techniques are merely preferred embodiments, and the scope of the present invention is not limited thereto. Will be obvious. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

Thistle with anti-obesity effect ( Cirsium japonicum ) extract. The method of claim 1,
The extract is thistle extract, characterized in that the crude extract or non-polar solvent soluble extract. Pharmaceutical composition for the treatment and prevention of obesity containing thistle extract as an active ingredient. The method of claim 3, wherein
The extract is a pharmaceutical composition, characterized in that the crude extract or non-polar solvent soluble extract. The method of claim 3, wherein
The extract is characterized in that the fibroblasts (3T3-L1 cells) to inhibit the differentiation of adipocytes, inhibit the synthesis of triacylglycerol (triacylglycerol) to have an anti-obesity effect. A pharmaceutical composition for treating and preventing obesity containing apigenin represented by Formula 1 as an active ingredient.
[Formula 1]

The method of claim 6,
The apigenin is thistle ( Cirsium japonicum ) pharmaceutical composition, characterized in that isolated from. The method of claim 6,
The apigenin inhibits the differentiation of fibroblasts (3T3-L1 cells) into adipocytes, inhibits the synthesis of triacylglycerol, and has a anti-obesity effect. Health functional food for the improvement and prevention of obesity containing thistle extract or apigenin isolated from it as an active ingredient. The method of claim 9,
The health functional food is a health functional food, characterized in that the powder, granules, tablets, capsules or beverage form.

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