KR101944013B1 - Antiobesitic composition comprising extract of Quercus aliena - Google Patents

Abstract

The present invention relates to an anti-obesity composition containing an alopecum oak extract, wherein the pharmaceutical composition or health functional food containing the alopecum oak extract comprises a composition for the treatment of metabolic diseases such as obesity, And can be easily used as a functional food.

Description

[0001] The present invention relates to an antiobesity composition comprising extracts of Quercus aliena,

The present invention relates to an anti-obesity composition containing an oak extract. More specifically, the present invention relates to an anti-obesity composition containing an alcohol extract of a leaf of oak, which has an effect of suppressing differentiation of adipocytes and inhibiting lipid accumulation and inhibiting oxidation reaction according to an adipocyte differentiation process.

In recent years, the population of obesity has been rapidly increasing due to improvements in living standards due to economic development, lack of exercise due to busy living environment, and excessive intake of nutrition. Obesity is a condition in which energy is excessively accumulated in the body due to unbalanced energy intake and consumption, resulting in an abnormally increased adipose tissue. The World Health Organization (WHO) considers Oriental BMI (body mass index) 23-25 as overweight, 25-30 obese, and over 30 as high obesity. The causes of obesity include environmental factors such as high fat, high calorie diet, lack of exercise due to busy social environment, endocrine abnormality, and genetic factors. Of these, about 50 to 70% of obesity is due to environmental factors And the rest is known to be caused by genetic factors. According to the statistics of the World Health Organization (WHO), the incidence of obesity is reported to increase year by year, especially in the OECD countries, the adult obesity population is increasing year by year. In Korea, the obesity population is increasing rapidly due to changes in the living environment, and the Korean adult obesity population with a body mass index (BMI) of 25 or more is increased by 40 million every year. As of 2005, 3 out of 10 were obese. According to data from the CDC, the obesity population in 1995 was 26.3% of the total population, but in 2001 it increased to 30.6% of the total population and to 31.5% of the total population in 2005, and is still showing a steady upward trend.

Quercus aliena Blume is also called jade tree, saw blade oak, and big oak tree. Origin is Korea, China, Japan and India. It grows at the foot of a mountain. Its height is 25m and its diameter is about 1m. The bark is shallowly divided like a net. Leaves are oval, 5 ~ 30cm long, 3 ~ 9cm long, with dull and pointed ends, no hairs on the top and dark green. The back side of the leaf is grayish white and the star-shaped hairs are tightly divided into 2 ~ 17 pieces. The petiole is 1 ~ 3.6cm and the edge of leaf has wavy or coarse toothed sawtooth. The flower blooms in May and is a single flower. The ear of a male flower is lanceolate and has 5 ~ 9 flowers and 6 ~ 14 stamens. The female flower has 6 horns and 2 to 4 pistil heads. It is covered with small projections of triangular shape. The fruit is nuts, 6 ~ 23mm in length, 7 ~ 16mm in diameter, elliptical, ripened in October and edible. The young leaves are used as fertilizer, and the wood is used as equipment and firewood. There are no hairs on the back of the leaves (var. Pellucida), no saw teeth (var. Acuteserrata), no saw teeth on the back of the leaves, no acneserrata for calvescens ).

The inventors of the present invention have studied various pharmacological activities possessed by such oak, and confirmed that the oakwood extract has an anti-obesity effect, thus completing the present invention.

Japanese Patent Application Laid-Open No. 1999-299451 (entitled: Water oak seed tree fruit powder having antioxidative function by lowering the concentration of cholesterol and triglyceride in blood, Applicant: IWAIZUMICO SANGYO KAIHATSU KOUSHA, Publication date: November 1999 02) Korean Unexamined Patent Publication No. 10-2011-0111568 (title of the invention: oil composition impregnated with oak component, applicant: white cast iron, published on October 12, 2011) Korean Patent No. 10-1500836 (Name of the invention: Composition of oak extract and its composition for the treatment or prevention of diabetes comprising the same as an active ingredient Applicant: Industry-academia cooperation corporation of Gyeongsang National University, Registered date: March 03, 2015)

Jin YS etc., Free radical scavenging and hepatoprotective actions of Quercus aliena acorn extract against CCl4-induced liver. Free Radic Res. 2005, 39 (12), 1351-1358.

It is an object of the present invention to provide an anti-obesity composition containing an oak extract. More specifically, it is an object of the present invention to provide an anti-obesity composition containing an alcohol extract of a leaf of oak, which has an inhibitory effect on adipocyte differentiation and inhibition of fat accumulation and an oxidation reaction according to an adipocyte differentiation process .

The present invention relates to an anti-obesity composition containing Quercus aliena Blume extract.

The oakwood extract can be extracted with water, C1-C4 alcohol, or a mixed solution thereof as a solvent.

The above-mentioned oak extract can be obtained by extracting leaves.

The present invention also provides a health functional food for an anti-obesity containing an oak extract.

Hereinafter, the present invention will be described in detail.

The present invention relates to an anti-obesity composition containing an oak extract. In the present invention, it is possible to use any part such as a leaf, a branch, a twig, a stem, a bark, a woody part, a root, a fruit, etc., Leaf oak leaf extract has the most excellent anti-obesity effect than other extracts.

The almond oak extract may be extracted with water, a C1-C4 alcohol or a mixed solution thereof as a solvent. The C1-C4 alcohol may be extracted from methanol, ethanol, propanol, isopropanol, butanol and isobutanol Can be selected. The solvent may preferably be a 50 to 90% (v / v) aqueous solution of a C1 to C4 alcohol, more preferably a 50 to 90% (v / v) aqueous solution of ethanol.

The water, the C1-C4 alcohol, or the mixed solution thereof used in the preparation of the oak extract may be used in an amount of 1 to 40 times by volume (1 to 40 liters per kg), preferably 5 to 40 times The volume can be used. The extracting conditions of the oak extract may be 1 to 48 hours at 20 to 100 ° C. The above process can be repeated 1 to 4 times. However, each process is not limited thereto. As the extraction apparatus, a conventional extraction apparatus, an ultrasonic pulverization extractor, or a fractionator may be used. The thus prepared oak extract can be removed by hot air drying, vacuum drying or freeze drying. The oakwood extract can be purified by using column chromatography.

The alder oak extract can be isolated or isolated by known methods used for the separation and extraction of plant components, such as extraction with an organic solvent (alcohol, ether, acetone, etc.), partition with hexane and water, And may be fractionated or purified by a suitable combination method.

The chromatography can be carried out using silica gel column chromatography, LH-20 column chromatography, ion exchange resin chromatography, medium pressure liquid chromatography chromatography, thin layer chromatography (TLC), silica gel vacuum liquid chromatography, and high performance liquid chromatography.

In addition, the present invention provides a pharmaceutical composition for anti-obesity containing an oak extract. The oakwood extract may be added to the pharmaceutical composition of the present invention in an amount of 0.001 to 100% by weight.

The pharmaceutical compositions may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to conventional methods. Examples of carriers, excipients and diluents that can be contained in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose , Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch, calcium carbonate, sucrose, Lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The dosage of the pharmaceutical composition of the present invention will depend on the age, sex, body weight of the subject to be treated, the particular disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, humans, and the like in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Since the compound of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for preventive purposes.

In addition, the present invention provides an anti-obesity health functional food comprising an almond oak extract and a pharmaceutically acceptable food-aid additive. The oakwood extract may be added in an amount of 0.001 to 100% by weight to the health functional food of the present invention. The health functional food of the present invention includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various kinds of drinks, meat, sausage, bread, Snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gums, tea and vitamin complexes.

The present invention relates to an anti-obesity composition containing an alopecum oak extract, wherein the pharmaceutical composition or health functional food containing the alopecum oak extract comprises a composition for the treatment of metabolic diseases such as obesity, And can be easily used as a functional food. Japanese Unexamined Patent Publication No. 1999-299451, Korean Patent Publication No. 10-2011-0111568, Korean Patent No. 10-1500836 and the like have been confirmed as prior art utilizing oak, but in the present invention, leaf oak leaf extract , The concentration of extract without cytotoxicity was confirmed, and the result of presenting the content of extract having anti-obesity effect at this concentration was not confirmed.

Fig. 1 shows the result of cytotoxicity of the oakwood extract.
Fig. 2 shows the result of inhibition of adipocyte differentiation by the oak extract of Goa.
FIG. 3 shows the results of measurement of ROS scavenging activity of Ganoderma lucidum extract in adipocyte differentiation.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

≪ Example 1: Preparation of goose oak extract >

The oak extract was prepared by dipping 100 g of leaf in 1000 ml of 70% (v / v) ethanol solution and then extracting at room temperature (25 ℃) for 24 hours. The extraction procedure was repeated three times, and the obtained extracts were all combined. The extract was filtered with a filter paper (Whatman, No.3, Maidstone, Kent, UK), concentrated under reduced pressure using a rotary vacuum concentrator (EYELA, N-3000, Tokyo, Japan) and then lyophilized (Ilshinbiobase Co., Ltd.). , Ltd, Yangju, Korea).

Example 2. Identification of anti-obesity activity of gooseberry extract

Example 2-1. Preparation of sample

The oak extract obtained in Example 1 used in the experiment was diluted with 100% DMSO (dimethyl sulfoxide) to a final concentration of 25 μg / ml, 50 μg / ml and 100 μg / ml in a cell culture condition Respectively.

Example 2-2. 3T3-L1 cell culture

3T3-L1 cells were purchased from ATCC and cultured in high-glucose DMEM (Dulbecco's modified Eagle's medium) in the presence of 10% BS (bovine serum) and 1% penicillin / streptomycin (P / S) at 37 ° C in a 5% CO ₂ incubator . When the 3T3-L1 cells in the preadipocyte state are filled to 100% in the cell culture dish, a high concentration of 10% FBS (fetal bovine serum) and 1% P / S are added for differentiation into adipocytes Glucose DMEM was used and 0.5 mM Isobutylmethylxanthine (IBMX), 1 μM Dexamethasone (DEX) and 1 μg / ㎖ insulin were added for 48 hours. Cells were cultured and tested in a 96-well plate and a 24-well plate by dividing cells according to the purpose of the experiment.

Examples 2-3. Evaluation of cytotoxicity using XTT assays

The cytotoxicity of 3T3-L1 cells was assessed using an XTT assay kit (2-methoxy-4-nitro-5-sulfophenyl) -2H-tetrazolium-5-carboxanilide innersalt assay kit Respectively.

XTT and PMS were added to the cells to increase the bioreduction by adding PMS (phenazine methosulfate), which serves as an Election coupling agent to XTT, and then the Tetrazolium ring of XTT by mitochondrial dehydrogenase of living cells was decomposed to form Formazan crystal Formazan crystals are dissolved in aqueous solution and become yellow. This yellow was measured by ELISA (VersaMax ELISA Microplate Reader, Molecular Devices, Sunnyvale, USA) and used for cytotoxicity evaluation.

For this purpose, 3T3-L1 preadipocyte cells were plated in 96-well plates at a concentration of 1 × 10 ^{6 on the} day before the experiment and cultured for 24 hours. The concentration of oak extract was 25 ㎍ / ㎖, 50 ㎍ / ㎖ and 100 ㎍ / ㎖, respectively, and cultured in a cell incubator for 24 hours. Then, 1 ㎖ of XTT and 20 ㎕ of PMS reagent were mixed, and 20% of the media volume was added. After reacting for 4 hours in a cell incubator, absorbance at 450 nm and 690 nm was measured using a spectrophotometer The results are shown in Fig.

As shown in FIG. 1, the effect of the oak extract on the survival rate of 3T3-L1 cells was examined by XTT assay at a concentration of 25 μg / ml, 50 μg / ml and 100 μg / The extract shows no cytotoxicity up to 100 [mu] g / ml.

Examples 2-4. Inhibition of adipocyte differentiation and evaluation of fat accumulation inhibition by oil red O staining

In order to measure the amount of fat accumulation in 3T3-L1 cells by oil red o staining, 3T3-L1 cells were cultured in 24 well plates for 8 days. After removing, 0.5 ml of a 10% formalin solution was added and left at room temperature for 5 minutes. 3T3-L1 cells differentiated with the same amount of formalin solution were left at room temperature for more than 1 hour, formalin was removed, and cells were washed with 0.5 ml of 60% isopropyl alcohol solution to completely dry the cells. The fully dried cells were stained with a pre-prepared working solution (Oil Red O: DDW = 6: 4), and the cells were washed 4 times with distilled water It was completely dried. The oily red O bound to the lipid components accumulated in the cells was eluted with a 100% isopropyl alcohol solution and absorbance was measured at 490 nm using a spectrophotometer. At this time, the degree of differentiation of 3T3-L1 cells was compared and analyzed using 5 mM NAC (N-acetyl-L-cysteine) as a positive control. The results are shown in FIG.

2, the amount of fat accumulation was significantly decreased in a concentration-dependent manner in the oak extract of Goa, and the oak extract was effective in inhibiting the differentiation of adipocytes and suppressing the accumulation of fat (see Fig. 2 con - extract without treatment).

On the other hand, since the oak-derived composition conventionally used for medicinal purposes is generally used as an acorn which is a fruit derived from oak, the extract is prepared from acorn oak, When the leaf extract showed a fat accumulation amount of about 75% as shown in FIG. 2, the acorn extract showed 95% or more fat accumulation and the leaf extract had better fat accumulation inhibitory effect than the acorn extract .

Examples 2-5. Evaluation of ROS abatement efficacy using NBT assay

To determine the amount of reactive oxygen species (ROS) produced by the differentiation process of adipocytes using the Nitroblue tetrazolium assay, 24-well plates were seeded with 25, 50, 100 μg / mL) to induce differentiation. By using this after 3T3-L1 After removing the culture medium of the cells with sterile PBS (phosphate-buffered saline, pH 7.4) 2 times wash, 0.2% (w / v) NBT was added to 0.2 ㎖ (nitroblue tetrazolium) solution, CO ₂ And incubated for 90 minutes in the incubator. After the staining, the plate was rinsed twice with distilled water and dried. DMSO (the ROS accumulated in the cells reacted with the NBT solution to produce dark blue formazan, which was eluted from the cells to measure the absorbance) and 1 N potassium hydroxide : 3 (v: v). The same amount of distilled water was added thereto, and the absorbance at 570 nm was measured using a spectrophotometer. The results are shown in FIG.

Referring to FIG. 3, the ROS content of the oak extract-treated adipocytes was significantly decreased in a concentration-dependent manner (the con-extract-treated group of FIG. 3).

≪ Formulation Example 1 >

200 g of the oak extract of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. To this mixture was added a 10% gelatin solution, which was pulverized and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of talc and 5 g of magnesium stearate was made into tablets.

<Formulation Example 2: Food Preparation>

Formulation Example 2-1. Manufacture of cooking seasonings

The oak extract of the present invention was added to the cooking seasoning at 1 wt% to prepare a cooking sauce for health promotion.

Formulation Example 2-2. Manufacture of flour food products

The oak extract of the present invention was added to wheat flour in an amount of 0.1% by weight, and bread, cake, cookies, crackers and noodles were prepared using the mixture to prepare a food for health promotion.

Preparation Example 2-3. Manufacture of soups and gravies

The health promotion soup and the juice were prepared by adding the oak extract of the present invention to the soup and juice at 0.1 wt%.

Formulation Example 2-4. Manufacture of dairy products

The oakwood extract of the present invention was added to milk in an amount of 0.1 wt%, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 2-5. Vegetable juice manufacturing

0.5 g of the oak extract of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare health promotion vegetable juice.

Formulation Example 2-6. Manufacture of fruit juice

Healthy fruit juice was prepared by adding 0.1 g of the oak extract of the present invention to apple juice or 1,000 ml of grape juice.

Claims (6)

A composition for anti-obesity characterized by containing a leaf extract of Quercus aliena Blume, which is extracted with water, a C1-C4 alcohol or a mixed solution thereof as a solvent, to inhibit differentiation of adipocytes. delete delete The present invention relates to a quercus aliena Blume leaf having an anti-obesity health function comprising an extract of a leaf oak extracted with water, a C1-C4 alcohol or a mixed solution thereof as a solvent, food. delete delete

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