KR20160114841A

KR20160114841A - Composition for improvement or treatment of Fat comprising Eucommiae ulmoides Oliv. Bark. extract or Adiponectin isolated therefrom as active ingredient

Info

Publication number: KR20160114841A
Application number: KR1020150041236A
Authority: KR
Inventors: 이완도
Original assignee: 이완도
Priority date: 2015-03-25
Filing date: 2015-03-25
Publication date: 2016-10-06

Abstract

The present invention relates to a composition for improving or treating obesity which comprises as an active ingredient an extract of mulberry bark extract or adiponectin isolated therefrom, wherein the mulberry extract or adiponectin, which is a main component separated from bark extract, is used as an active ingredient, Extract or lycopene separated therefrom as an active ingredient to thereby improve and treat obesity through inhibition of lipolysis and fat production in a living body.
In accordance with the present invention as described above, it is possible to provide a method for preventing and treating obesity, which is very useful for medicines and health functional foods for obesity improvement and treatment, .

Description

TECHNICAL FIELD The present invention relates to a composition for improving or treating obesity, which comprises as an active ingredient adiponectin isolated from bovine bark extract or an adiponectin isolated therefrom. Bark. extract or Adiponectin isolated therefrom as active ingredient}

The present invention relates to a composition for improving or treating obesity, which comprises, as an active ingredient, a bark extract or adiponectin isolated therefrom. More particularly, the present invention relates to a composition for improving or treating obesity, And further comprising a tomato extract or lycopene isolated therefrom as an active ingredient to thereby improve and treat obesity through inhibition of lipidation and fat production in a living body.

Eucommiae ulmoides Oliv.) Is a fairy tale-like tree with mulberry leaves and grows up to 20 meters in height. Its leaves are elliptical, pointed end, sharp edges are sharp. Eucommiae ulmoides Oliv. Bark. Is a herbal medicine which refers to the bark of Eucommiae ulmoides Oliv. (Eucommiaceae). It is used for edible and medicinal purposes.

In particular, it has been known to have a blood pressure lowering effect by treating low back pain, knee pain, cold depression and uterine bleeding. Known components include gutta-percha, alkaloid, pectin, It is known to contain components such as resins.

Tomatoes are an annual crop belonging to the family Lycopersicon esculentum Mil, originating from the South American Andes. Europe and the United States, as well as Asia and Africa are widely used around the world, and currently the country is suitable for the climate climate is cultivated nationwide. Tomatoes are known to be effective for aging tears, headache, acute hemorrhage, and inflammation. These tomatoes are rich in vitamins, organic components, free amino acids and dietary fiber, and have functions such as anti-aging, anti-degenerative disease and immunity enhancement. They also have physiological activities such as lycopene, quercetin, β-carotene, caffeic acid and phytoene Substances are reported to be contained.

Accordingly, the present inventors have attempted to improve the obesity by using the extract obtained by using the bacillus thuringiensis, the adiponectin fraction purely isolated therefrom, or the tomato extract and the lycopene isolated therefrom to utilize the lipolysis and lipogenesis inhibitory effect The present invention has been successfully completed.

Korean Patent Publication No. 10-2009-0078084

As a result, it is a main object of the present invention to provide a composition for improving or treating obesity, comprising a bivalvarial extract or adiponectin isolated therefrom as an active ingredient, and a health functional food containing the same.

Another object of the present invention is to provide a composition for improving or treating obesity and a health functional food comprising the extract or the adiponectin isolated therefrom further comprising a tomato extract or lycopene isolated therefrom as an active ingredient have.

In order to achieve the above object, the present invention provides a bamboo shoot extract having an obesity improvement and therapeutic effect, or a bamboo shoot extract to which a tomato extract is added.

In the present invention, the bamboo shoot extract is a crude bamboo shoot extract or a non-polar solvent use extract.

In the present invention, the tomato extract is a crude extract of tomato.

The dumplings and tomatoes may be purchased commercially or used, or harvested or cultivated in nature, but the present invention is not limited thereto.

The term "extract" as used herein refers to an active ingredient isolated from a natural product, which is prepared by an extraction process using water, an organic solvent or a mixed solvent thereof, and is prepared by extracting water, an organic solvent or a mixed solvent thereof, Includes all formulated forms. In addition, the above-mentioned extract also includes a fraction obtained by fractionating the extract obtained through the above extraction process.

The solvent of the crude extract of tomato or tomato used in the present invention may be water, an organic solvent or a mixture thereof, preferably water, anhydrous or hydrolyzate having 1 to 4 carbon atoms such as methanol, ethanol, propanol, A mixed solvent of a lower alcohol and water, a solvent such as acetone, ethyl acetate, chloroform or 1, 3-butylene glycol as an extraction solvent, stirring, hot water extraction, cold extraction, warming extraction, And the like, and then extracting the obtained extract by filtration, concentration under reduced pressure or drying, and the extraction method is not particularly limited.

In addition, the extract of the present invention for extracting the non-polar solvent is obtained by dissolving the extract obtained in the above step in a solvent selected from hexane, chloroform, methylene chloride, dichloromethane, ethyl acetate, glycerin, propylene glycol, butylene glycol or ether, Chloroform or ethyl acetate, more preferably chloroform, followed by extractive fractionation, followed by silica gel column chromatography.

Accordingly, the present invention provides a novel use of the bivalvarial extract prepared by the above method, or the bivalvarial extract supplemented with the adiponectin or the tomato extract isolated therefrom, for the amelioration and / or treatment of obesity.

Specifically, the present invention provides a pharmaceutical composition for improving and treating obesity, which comprises the bark extract or adiponectin isolated therefrom as an active ingredient.

In the present invention, the adiponectin is not only purified and purified from bovine spongiform encephalopathy but also chemically synthesized or commercially available adiponectin.

In addition, the extract has been used as a herbal medicine for a long time, and the extract of the present invention extracted therefrom also has no toxicity and side effects.

Therefore, the extract of the present invention can be used for medicines, health supplements, and the like for anti-obesity effect and is not limited thereto.

Preferably, the composition of the present invention contains 0.01 to 99.9% by weight, preferably 0.1 to 50% by weight, and more preferably 0.01 to 25% by weight of adiponectin, 0.1 to 12.5% % Is more preferable. However, the composition is not necessarily limited thereto, and may be varied depending on the condition of the body to be used, the degree of absorption in the body, the degree of disease, the form of the drug, the administration route and the period. It can be divided into several doses. The dosage is not intended to limit the scope of the invention in any way.

When the extract of the present invention is used as a medicine, it may contain only the herbal extract itself, but may further include appropriate carriers, excipients and diluents conventionally used in the production of the pharmaceutical composition, For example, powders, tablets, granules, capsules or injections, and the like.

Examples of the carrier or excipient include water, dextrin, calcium carbonate, lactose, propylene glycol, liquid, paraffin, physiological saline, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gelatin, calcium phosphate, calcium Silicates, cellulose, methylcellulose, polyvinylpyrrolidone, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals. These may be used alone or in combination of two or more. Both carriers and excipients can be used.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories, Can be used.

In the case of formulation, it is prepared using a diluent or an excipient such as a commonly used filler, a weight agent, a binder, a wetting agent, a release agent, a surfactant, etc. Solid preparations for oral administration include tablets, pills, powders, granules, capsules These solid preparations are prepared by mixing at least one excipient, for example, starch calcium carbonate, sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used. Examples of liquid formulations for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, which are commonly used simple whitening agents, various excipients such as wetting agents, sweeteners, have. 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. As a base material for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like can be used.

The pharmacologically acceptable dosage forms of the extract of the present invention or adiponectin isolated therefrom may be used in the form of their pharmaceutically acceptable salts and may be used alone or in combination with other pharmacologically active substances, Can be used.

In addition, the present invention provides a health functional food for improving and treating obesity, which contains, as an active ingredient, a tomato extract or lycopene isolated therefrom, or the adiponectin or bifidobacterium extract isolated therefrom.

The health functional food of the present invention is not limited to any particular kind and includes, for example, meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , Beverages, tea, a drink, an alcoholic beverage and a vitamin complex, and a health functional food.

According to the present invention as described above, the extract of mulberry seeds and the extract of mulberry seeds to which the tomato extract is added has excellent fat dissolving effect and lipogenesis inhibitory effect, and thus the present invention is applicable to medicines and health functional foods for the improvement and treatment of obesity Can be usefully used.

FIG. 1 is a graph showing the effect of 3T3-L1 cells on the synthesis of triacylglycerol by the concentration of the adiponectin compound isolated from the bivalvarial extract according to the present invention.

Hereinafter, the present invention will be described in detail.

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 bamboo shoot extract

(1) Preparation of Dioscorea edulis extract using hot water extraction

The dried bamboo shoots allowed to be tested were washed with distilled water and thoroughly dried. The bamboo shoots were finely pulverized by a pulverizer. The pulverized bamboo shoots were weighed 20 times in volume with 1 kg of each raw material, The mixture was cooled at room temperature, filtered through a filter paper, and then concentrated under reduced pressure using a vacuum condenser. The above-mentioned dicalcium phosphate concentrate was lyophilized to give an extract powder of dicalcium phosphate extract.

(2) Preparation of dipstick extract by organic solvent extraction

The dried bivalves were allowed to dry thoroughly with distilled water and then thoroughly pulverized by a grinder. The pulverized bivalves were extracted with 500 ml of a methanol methanol solution per kg of each raw material at 50 ° C. using an ultrasonic grinder The mixture was cooled at room temperature, filtered through a filter paper, and then concentrated under reduced pressure using a vacuum condenser. The above-mentioned dicalcium phosphate concentrate was lyophilized to give an extract powder of dicalcium phosphate extract.

Example 2. Preparation of Soluble Extracts of Non-polar Solvents

The extracts obtained in Example 1 were dissolved in distilled water and mixed again in ethyl acetate at a ratio of 1: 1 (v / v). The extracts were repeated three times for 1 hour, and the resulting diatomic ethyl acetate fraction Was decompressed, concentrated and dried to obtain an extract powder of bamboo shoot extract.

Example 3. Isolation of adiponectin from bivalve extract

The ethyl acetate extract obtained in Example 2 was subjected to silica gel flash chromatography (Silica gel 60, Merck, Germany), and the active fraction was further decompressed and dried. The active fraction was further decompressed and dried in a small amount of methanol, The solution was dissolved in a small amount of methanol at a concentration of 20 μg / ml and filtered through Millipore (0.45 nm), followed by HPLC (2487, Waters, USA) to collect 25 mg of the active substance.

At this time, 74% methanol was used as the mobile phase, the flow rate was 10 ml per minute, and the column was 20? Mm. Further, the compound was identified by a spectroscopic method (300 MHz NMR Spectroscopy DRX 300, Bruker, Germany).

Example 4. Isolation of lycopene from tomato extracts

Tomatoes purchased from Kyungdong market were washed, milled, lyophilized at -60 ℃ for 72 hours, and powdered with a mixer to prepare tomato powder.

50 g of the above tomato powder was filled in a column equipped with a glass filter, and then 1 liter of ethyl acetate was added thereto to prepare a tomato extract at 30 ° C.

The tomato extract was prepared as a concentrate by removing ethyl acetate from a vacuum concentrator (water bath temperature: 62 ° C, vacuum pressure: 200 mmHg). 1 ml of the concentrate was added to 1 liter of methanol at -10 ° C and stirred to prepare a carotenoid Carotenoids were removed and lycopene was eluted. The eluted lycopene was filtered, dried and pulverized to obtain purified lycopene powder.

Experimental Example 1 Analysis of lipidation effect

(1) MTT analysis

Fibroblast cultures of the present invention were tested for cytotoxicity by MTT (final concentration 0.4 μg / ml) after 48 hours of culture.

As a result, it was confirmed that the cytotoxicity analysis showed no cytotoxicity to fibroblasts in any sample at various concentrations.

(2) Confirmation of fat dissolving effect -

Fibroblasts (3T3-L1 cells) were cultured from ATCC and differentiated from mast cells. The culture medium for animal cell proliferation was cultured in a 10% FBS, 37 ° C, 5% CO ₂ incubator. The medium was incubated at 2 to 3 days intervals with a fresh medium.

The cells were washed twice with a phosphate buffered saline solution before the cells became confluent (cells were grown to the full size of the culture dish) and incubated with 0.25% trypsin-EDTA (1: 4 to 0: 48 hours; medium - 1: 4: 1: 4) was added to a 6-well plate at the ratio of 1: 2: after 48 hours), the extracts of the bivalves obtained in Examples 1 to 3 or the adiponectin compounds isolated therefrom were added at a concentration of 10, 40 and 80 μM, respectively.

In order to confirm the effect of the extract of the present invention or the adiponectin compound isolated therefrom on the synthesis of triacylglycerol (TG) in fibroblast (3T3-L1 cell), the cells cultured in each experimental group were treated with Tris After desorption with EDTA buffer, the cells were pulverized with an ultrasonic mill. A calibration curve of the standard solution was prepared according to the instructions given in the BCA Protein Assay Kit (BCA ™ Protein Assay Kit, Thermo Sci., USA), and the desorbate obtained from the cells was diluted with a certain amount of the kit reagent and heated at 37 ° C. for 10 minutes , The absorbance was measured at a wavelength of 490 nm as a control of the blinded reagent within 60 minutes, and the amount (쨉 g / mg protein) of triacylglycerol per intracellular protein was measured and quantified according to a calculation formula.

As a result, it was confirmed that the sample having a high content of the adiponectin compound according to the present invention was effective for lipolysis, and it was confirmed that the synthesis of triacylglycerol in the fibroblast (3T3-L1 cell) was also inhibited in the insulin medium .

(2) Confirmation of lipolytic effect - Insecticide extract + Tomato extract

Twenty male Sprague-Dawley (SD) male rats aged 4 weeks were purchased and used. They were randomized complete block design after a week of high-fat diet, normal diet (N diet), high fat diet (HFD), Eucommiae ulmoides Oliv Diet ; ED; 10% Eucommiae ulmoides Oliv and Tomato Diet (ETD)) were divided into 5 groups according to the treatments. [Temperature: 22 ℃, humidity: 50% For 4 weeks.

The diets used in the experiment were based on AIN-93G (Reeves 1993) and are shown in Table 1. The extracts (1.5%) and tomato extract (5%) were injected at a fixed time of 4 weeks at a constant daily rate of 3 mL per kg of body weight based on the amount of rat ingested. All experimental diets were refrigerated during the rearing period. The body weight was measured at a constant time every day, and the body weight before the start of the experiment was subtracted from the body weight before the start of the experiment.

HFD (kcal%) ND (kcal%) Protein 20 20 Carbohydrate 34 63 Energy from Fat (%) 58 16 Casein 20 20 Sucrose 10 10 Dextrose 5 13 Cellulose 5 5 Soybean oil 4 7 Lard (80%) 36.5 0 Vitamin Mix One One Mineral Mix 3.5 3.5 L-Cystine 0.5 0.5

The daily weight gain of the rats tended to be higher in the high fat diet group (HFD) than in the normal diet group (ND), and obesity induction due to high fat diet could be observed. As a result, as shown in Table 2, the body weight tended to decrease slightly in the group administered with the bark extract and in the group administered with the bark extract and the tomato extract, the body weight tended to be further decreased.

From these results, it was confirmed that adiponectin of the bivalvar extract and lycopene of the tomato extract act to increase the anti-obesity effect by interaction.

ND HFD ED ETD Initial Body weight (g) 115.3 ± 11.1 118.3 ± 5.2 118.1 ± 5.2 115.7 ± 3.8 Final Body weight (g) 270.0 + - 23.1 300.0 ± 21.5 281.3 ± 18.2 273.8 ± 17.7 Weight gain (g / day) 5.5 ± 4.7 6.4 ± 3.1 5.8 ± 2.9 5.6 ± 2.7

Experimental Example 2 Statistical analysis

To confirm the reproducibility of the experimental results, each experiment was performed twice or three times, and the analysis of the experimental results was performed by one-way ANOVA using the Bonferroni method using sigma-plot.

Hereinafter, formulation examples of the composition containing the extract of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.

Formulation Example 1. Manufacture of tablets

Adiponectin 10 mg

Lactose 100 mg

100 mg of corn starch

2 mg of magnesium stearate

The above components are mixed and tablets are prepared by tableting according to a conventional method for producing tablets.

Formulation Example 2. Manufacture of Powder

20 mg

Lactose 100 mg

10 mg of talc

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

Formulation Example 3. Preparation of capsules

10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

0.2 mg of magnesium stearate

The above components are mixed according to a conventional preparation method of capsules, and filled in gelatin capsules to prepare capsules.

Formulation Example 4. Manufacture of health food

1000 mg adiponectin

300 mg lycopene

Vitamin mixture quantity

Vitamin A acetate 70 쨉 l

Vitamin E 1.0 mg

0.13 mg of vitamin B ₁

0.15 mg of vitamin B ₂

0.5 mg of vitamin B ₆

0.2 g of vitamin B ₁₂

Vitamin C 10 mg

10 μg of biotin

Nicotinic acid amide

50 엽 of folic acid

Calcium pantothenate 0.5 mg

Mineral mixture quantity

1.75 mg ferrous sulfate

0.82 mg of zinc oxide

Magnesium carbonate 25.3 mg

15 mg of potassium phosphate monobasic

Secondary calcium phosphate 55 mg

90 mg of potassium citrate

Calcium carbonate 100 mg

24.8 mg of magnesium chloride

Although the composition ratio of the vitamin and mineral mixture is relatively mixed with a suitable ingredient for health food, the compounding ratio may be arbitrarily modified. The ingredients may be mixed according to a conventional method for producing healthy food, Granules can be prepared and used in the manufacture of a health food composition according to conventional methods.

While the present invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. It will be appreciated by those skilled in the art that numerous changes and modifications may be made without departing from the invention. Accordingly, all such appropriate modifications and changes, and equivalents thereof, should be regarded as within the scope of the present invention.

Claims

A composition for treating or ameliorating obesity, comprising as an active ingredient adiponectin isolated from bovine bark extract or adiponectin isolated therefrom.

The method according to claim 1,
The composition for improving or treating obesity according to claim 1, wherein the extract is 0.1 to 50 wt%, and the adiponectin is 0.1 to 12.5 wt%.

The method according to claim 1,
The extract is preferably a water-soluble or water-insoluble alcohol having 1 to 4 carbon atoms such as water, methanol, ethanol, propanol and butanol, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, chloroform or 1,3- The composition for improving or treating obesity is characterized in that it is extracted using glycol.

The method according to claim 1,
Wherein said extract is extracted with hexane, chloroform, methylene chloride, dichloromethane, ethyl acetate, glycerin, propylene glycol, butylene glycol or ether.

2. The composition of claim 1,
A composition for improving or treating obesity, which further comprises a tomato extract or lycopene isolated therefrom as an active ingredient.

4. A health functional food for improving or treating obesity comprising the extract of bivalves of claim 1 or adiponectin isolated therefrom as an active ingredient.

[7] The health functional food according to claim 6,
A tomato extract or lycopene isolated therefrom as an effective ingredient.