KR20220057928A - Composition for Preventing or Treating Obesity Comprising Enzyme-Treated Euphorbia humifusa Extract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition or a food composition for anti-obesity and weight loss, containing a Euphorbia humifusa extract as an active component. The Euphorbia humifusa extract of the present invention exhibits an effect of inhibiting adipocyte differentiation and lipid accumulation, thereby being able to be usefully used for preventing and treating obesity.

Description

Enzyme-treated composition for preventing or treating obesity of silk grass extract {Composition for Preventing or Treating Obesity Comprising Enzyme-Treated Euphorbia humifusa Extract}

The present invention relates to a composition for preventing or treating obesity using a natural extract. More particularly, the present invention relates to a composition for preventing or treating obesity, which has anti-obesity and weight loss effects by inhibiting fat accumulation in adipocytes.

Obesity refers to an abnormally increased body fat due to an imbalance between energy intake and consumption. The World Health Organization (WHO) regards Asians as having a body mass index (BMI) of 23 to 25 as overweight, 25 to 30 as obese, and over 30 as highly obese.

It is known that obesity is not a disease caused by a single cause, but is caused by a complex action of several factors. The causes of obesity include genetic factors and environmental factors such as high-fat and high-calorie diet, lack of exercise due to busy social environment, and endocrine abnormalities. It is known that 50 to 70% of obesity is due to environmental factors and the rest to genetic factors.

Currently, methods for treating obesity can be divided into lifestyle modifications such as diet, exercise, and behavioral therapy, and drug or surgical treatment. Active efforts to correct lifestyle should be preceded by drug or surgical treatment, but it is not easy to correct lifestyle, and there is a limit to the weight that can be reduced only by lifestyle modification. Therefore, in many cases, drug treatment is necessary along with lifestyle modification. Many anti-obesity drugs are being developed every year for the drug treatment of obesity, but there are not many currently available anti-obesity drugs, and most are limited to chemical agents that suppress digestion or appetite. These chemical agents are known to cause mental problems in the process of controlling digestion or appetite, or often cause side effects such as diarrhea and constipation. Therefore, there is a need to develop an anti-obesity agent derived from a safe natural product that can be consumed as food.

Numerous references are referenced throughout this specification and citations thereof are indicated. The disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly describe the content of the present invention and the level of the art to which the present invention pertains.

Republic of Korea Patent Publication No. 10-1414741

The present inventors have made research efforts to develop an anti-obesity composition derived from a safe natural product that can be ingested, and as a result, the silk extract, especially the enzyme-treated silk grass extract, is safe for the human body and at the same time has a significant fat accumulation inhibitory effect. invention was completed.

Accordingly, it is an object of the present invention to provide a composition for preventing or treating obesity, including an enzyme-treated silk grass extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating obesity comprising the composition.

Another object of the present invention is to provide a food composition for preventing or improving obesity comprising the composition.

Another object of the present invention is to provide a method for preventing or treating obesity by treating a subject with a composition comprising a silk grass extract as an active ingredient, including enzyme-treated silk grass extract.

Another object of the present invention is to provide a use for the manufacture of a pharmaceutical composition for preventing or treating obesity, including an enzyme-treated silk grass extract, as an active ingredient, or a food for preventing or improving obesity.

Further objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a composition for preventing or treating obesity comprising an ethanol extract of Euphorbia humifusa as an active ingredient, wherein the extract is enzyme-treated. do.

According to another aspect of the present invention, the present invention provides a method for preventing or treating obesity, comprising administering to a subject a composition comprising an enzyme-treated ethanol extract of silkwormwood as an active ingredient.

According to another aspect of the present invention, the present invention provides a use (use) of the enzyme-treated silkworm ethanol extract for preparing a medicament for the prevention or treatment of obesity.

Silk grass ( Euphorbia humifusa ) used as an active ingredient in the composition of the present invention is an annual herb of the genus Euphorbia , which is a plant commonly grown in grass fields, yards, and roadsides. It is the largest genus of angiosperms with about 2,000 species distributed around the world, and about 20 species are known to be native to Korea. Silkweed is commonly grown on eves, roadsides, grasslands, yards, and roadsides across the country, but it is small and inconspicuous.

Silk grass has also been used in oriental medicine, and it is known to be effective against dysentery, diarrhea, gastritis, colitis, cough caused by a cold, bloody stool, hematemesis, uterine bleeding, hematuria, gum inflammation, herpes zoster, and sore throat. In oriental medicine, silk grass is understood to have the same efficacy by binding it together with other species of plants belonging to the genus. However, as a result of the study of the present invention, it was confirmed that the silk grass had a particularly remarkable anti-obesity effect, unlike other species of plants belonging to the genus genus.

The composition of the present invention includes an extract of silkworm as an active ingredient. The term 'extract' used while referring to silk grass in this specification includes not only the extraction result obtained by treating the silk grass with an extraction solvent, but also the silk grass processed (eg, powdered) formulated so that the silk grass itself can be administered to animals. has

When the silk grass extract used in the composition of the present invention is obtained by treating the silk grass extract with an extraction solvent, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent may be used. Suitable as polar solvents are: (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable nonpolar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkane, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- Pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF.

More preferably, the extraction solvent used in the present invention is obtained by treating silk grass with ethanol (eg, 50 wt% to 70 wt% ethanol).

As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the silk extract is not only obtained by using the above-described extraction solvent, but also includes those obtained by additionally applying a purification process here. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the silk extract of the present invention.

The silk extract used in the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying.

In a preferred embodiment of the present invention, the silk grass extract is characterized in that it is enzyme-treated.

The enzyme-treated dandelion extract exhibits an increased fat accumulation inhibitory effect as compared to a simple solvent extract of dandelion grass without enzyme treatment. The increase in the fat accumulation inhibitory effect (inhibition rate of adipocyte differentiation in adipocytes) by the enzyme treatment is at least 1%, at least 3%, at least 5%, at least 7%, at least 10%, at least 15%, at least 20%, at least 25%, or at least 30%.

The enzyme treatment is to treat the extraction result obtained by treating the silk grass raw material with an extraction solvent, or to treat the enzyme before treating the silk grass raw material with an extraction solvent (extracting solvent is applied to the result obtained by treating the silk grass raw material with an enzyme) processing) are included.

The raw material of silk grass may include above-ground parts of silk grass, leaves, stems, flowers, flower buds, seeds, sap roots, and whole plants, which may be washed and pulverized.

The enzyme may preferably be a starch degrading enzyme. The starch degrading enzyme is not necessarily limited thereto, but may be at least one selected from the group consisting of cellulase, hemicellulase, pectinase, glucosidase, glucanase, amylase, glucoamylase and isoamylase, preferably It may be one containing cellulase.

In one embodiment, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more of the treated enzyme may be cellulase and , or 100% of the treated enzyme may be cellulase.

The composition for preventing or treating obesity of the present invention acts to inhibit fat accumulation in fat cells, and through this action, various types of obesity are treated or prevented.

Obesity that can be prevented or treated by the present invention includes childhood obesity and adult obesity, and includes simple obesity (controlled obesity) and symptomatic obesity (metabolic or endocrine obesity). In addition, congenital obesity is cell proliferative obesity and acquired obesity is cell hypertrophy, abdominal obesity, visceral fat type obesity, upper body obesity, peripheral obesity, hip obesity, lower body obesity, subcutaneous fat type obesity, male type obesity and female type obesity. , but is not limited thereto.

The composition of the present invention may be prepared as a pharmaceutical composition. According to a preferred embodiment, the composition of the present invention comprises (a) a pharmaceutically effective amount of the enzyme-treated ethanol extract of the present invention; and (b) a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the efficacy or activity of the above-mentioned silkworm extract.

When the composition of the present invention is prepared as a pharmaceutical composition, the composition of the present invention includes a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil; it is not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and is preferably applied by oral administration.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. can be A typical dosage of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg/kg for adults.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by a person of ordinary skill in the art to which the present invention pertains. or it may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or an aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.

The composition of the present invention may be provided as a food composition, in which case the composition of the present invention comprises (a) a food pharmaceutically effective amount of the extract of the present invention; And (b) it may be a food composition comprising a food acceptable carrier.

When the composition of the present invention is prepared as a food composition, as an active ingredient, it includes ingredients commonly added during food production, as well as ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. may include Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents [taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, bean extract, jujube extract, licorice extract, etc. can be additionally included in addition to the silk extract of the present invention. there is.

All ingredients described in the present invention, preferably, relevant laws and regulations such as Korea, China, the United States, Europe, Japan, etc. Do not exceed the maximum use value stipulated in Korea)), etc. That is, preferably, the pharmaceutical composition, health functional food composition, drug or food according to the present invention contains the ingredients according to the present invention within the content limit allowed by the relevant laws and regulations of each country.

According to another aspect of the present invention, the present invention provides a method for preparing the composition.

The manufacturing method of the present invention is to include the steps of preparing a solvent extraction product by treating the raw material of silk grass with an ethanol solvent and treating the solvent extraction product with an enzyme; Alternatively, it may be one comprising the steps of treating an enzyme in the silk grass raw material and treating the resultant obtained by treating the enzyme with an ethanol solvent to obtain a solvent extraction result.

Since the preparation method of the present invention is a preparation method of the above-described composition, in order to avoid excessive duplication of the specification of the present invention, the common contents of the two are omitted.

The present invention relates to a pharmaceutical composition or food composition for anti-obesity and weight loss comprising an enzyme-treated silk grass extract as an active ingredient. The enzyme-treated silk grass extract of the present invention can be usefully used for the prevention and treatment of obesity by showing the effect of inhibiting adipocyte differentiation and lipid accumulation.

1 shows the results of evaluating cell viability by a method of measuring absorbance using an MTT buffer.
Figure 2 shows the results of evaluating the fat accumulation rate in adipocytes by the Oil Red O staining method after treating the silk extract at a low concentration (20 μg/mL) and a high concentration (100 μg/mL).
Figure 3 shows the results of evaluating the fat accumulation rate in adipocytes by the Oil Red O staining method after treating the silk extract at 25, 50, 100, and 200 μg/mL concentrations.
4 is a graph showing the result of FIG. 3 .
5 is a simplified view of the production process of the enzyme-treated extracts (T-1, T-2 and T-3) and Comparative Example extracts (C-1 and C-2).
Figure 6 is GM (Growth medium), DM (Differentiation medium; negative control), enzyme-treated extracts (T-1, T-2 and T-3) and Comparative Example extracts (C-1 and C-2) in the treatment of According to the fat differentiation rate (Oil Red O stained area) is shown.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art to which the present invention pertains that the scope of the present invention is not limited by these examples.

Example

I. Confirmation of Effects of Extraction Solvents

1. Preparation of silk grass extract

50 g of dried silk grass ( Euphorbia humifusa Wild., Aerial part of silk grass) was added to 10 times of water, 30%, 50%, 70% ethanol using a reflux extraction device, and the temperature was above 70℃ for 3 hours. , the extraction was repeated twice. The extract was filtered with Whatman No. 2 (150mm Φ) filter paper to remove insoluble substances, and then concentrated under reduced pressure at 50° C. with a rotary evaporator equipped with a cooling condenser. In order to completely remove the solvent of the extract concentrated under reduced pressure, purified water was added and suspended to obtain a final powder raw material using a freeze dryer, 10.36 g of hot water extract (yield 20.70%), 10.58 g of 30% ethanol extract (yield 21.15%) , 11.71 g of a 50% ethanol extract (yield 23.30%) and 11.91 g (yield of 23.81%) of a 70% ethanol extract were obtained (Table 1).

2. Preparation and Culture of Preadipocytes (3T3-L1)

Mouse fibroblasts, 3T3-L1 preadipocytes, are the most frequently used cell line for adipocyte differentiation tests, and were used after receiving from ATCC (American Type Culture Collection, USA). 3T3-L1 preadipocytes were cultured in an incubator maintained at 5% CO ₂ , 37° C. using DEME growth medium containing 10% calf serum and 1% penicillin-streptomycin solution. In order to eliminate the phenomenon of overdensity in the proliferation of cell numbers, the growth medium was exchanged every 48 hours to maintain an appropriate number of cells.

3. Cytotoxicity and viability assays

The MTT assay was performed to confirm the appropriate concentration without cytotoxicity and toxicity of the silk grass extract. 3T3-L1 cells were inoculated in a 96-well plate at a concentration of 2x10 ⁴ cells/well, and cultured until 100% confluent by changing the culture medium once every 2 days. The cultured cells were treated with 200, 500, and 1000 μg/mL concentrations of extracts for each ethanol ratio, respectively, and cultured at 37° C., 5% CO ₂ for 8 days. After incubation, absorbance was measured using MTT buffer. As a normal control group, the experimental procedure was repeated three times using a group (Vehicle) in which cells were treated with only a medium. Cell viability was calculated by Equation 1 below. The results are shown in FIG. 1 .

[Equation 1]

Cell viability (%) = (absorbance of sample treatment group / absorbance of control group) x 100

As confirmed in FIG. 1 , the hot water extract of silkworm and the extract by ethanol ratio maintained a cell viability of 80% or more at a concentration of 200 to 1000 μg/mL.

4. Confirmation of the inhibitory effect on the differentiation of adipocytes of the silk extract

The fat accumulation rate was checked to determine whether the non-dandelion extract had the effect of inhibiting the differentiation of 3T3-L1 cells into adipocytes. To confirm this, stabilized 3T3-L1 cells were aliquoted in a 48 well plate at a density of 2x10 ⁵ cells/well, cultured, and maintained for 2 more days when the 100% confluent time point reached. 3T3-L1 cells were cultured in DMEM medium containing 10% fetal bovine serum containing MDI and 1% penicillin and differentiated into adipocytes for 2 days. In the first adipocyte differentiation test, during the adipocyte differentiation process, the culture medium was treated with silk extract at low concentration (20 μg/mL) and high concentration (100 μg/mL), and in the second test, 25, 50, 100, 200 μg It was treated in the culture medium for each /mL concentration. Thereafter, the degree of adipocyte differentiation was observed on the 8th day, when differentiation was completed. As a normal control group, only the medium was treated (Vehicle), and as a positive control group, Garcinia cambogia was treated.

In order to observe the degree of differentiation of adipocytes, Oil Red O staining that specifically responds to Lipid droplets was performed. Specifically, the medium was removed from the cells induced by adipocyte differentiation as described above and treated with 10% formalin at room temperature for 1 hour and fixed. It was then washed with 60% isopropanol, and each well was completely dried. After treatment with Oil Red O dye for 1 hour, it was washed twice with 60% isopropanol and once with sterile distilled water. For elution of the bound Oil Red O, it was treated with 100% isopropanol for 10 minutes, and the absorbance of the eluate was measured at 520 nm.

The fat accumulation rate in adipocytes was calculated by Equation 2 below, and the results are shown in Table 2 and FIGS. 2 to 4 .

[Equation 2]

Fat accumulation rate (%) = (absorbance of sample treatment group / absorbance of control group) x 100

1S: silk grass hot water extract, 3S: silk grass 50% ethanol extract, 4S: silk grass 70% ethanol extract

As shown in Table 2 and FIGS. 2 to 4, it was confirmed that the concentration-dependent inhibition of accumulation of fat in adipocytes was confirmed by the treatment with the extract, which is an active ingredient of the present invention, in a fat cell. In particular, it was found that the 50% (3S) and 70% (4S) ethanol extracts of silk grass of the present invention more effectively inhibited the differentiation of pre-adipocytes into adipocytes, and thus the prevention or treatment effect of obesity was significantly superior.

II. Check the effect of enzyme treatment

1. Preparation of enzyme-treated silk grass ethanol extract

The effect of enzyme treatment was confirmed using two types of enzymes. Specifically, 20 g of dried silk grass purchased from China (Euphorbia humifusa Wild.) was crushed, 3% enzyme and 210 mL of water were added to the raw material, and enzyme treatment was performed at 50 to 60° C. for 3 hours. 490 mL of 70% ethanol was added to the obtained treated product, extracted twice at 70° C. for 3 hours, and concentrated under reduced pressure at 50° C. with a rotary evaporator equipped with a cooling condenser to completely remove the solvent. Purified water was added to the concentrate under reduced pressure to suspend, and then freeze-dried using a freeze dryer to obtain the final enzyme-treated silk grass extract (T-1 and T-2).

In addition, as a comparative example, a 70% ethanol extract of silk grass was prepared in the same manner as described above, except that the enzyme was not treated. That is, 700 mL of 70% ethanol was added to 20 g of pulverized silk grass, extracted twice at 70° C. for 3 hours, and then concentrated under reduced pressure and freeze-dried to obtain a final 70% ethanol silk grass extract (C-1).

The types of enzymes used, the amount and yield of the silk extract recovered are shown in Table 3 below, and the enzyme-treated ethanol extract (T-1 and T-2) and the ethanol extract (C-1) manufacturing process as a comparative example are simplified in FIG. shown to be

enzyme Rohament CL
Laminex BG2 Enzyme treatment
doesn't exist T-1 T-2 C-1 Recovery (g) 6.16 6.34 6.46 transference number(%) 30.8 31.7 32.3

Rohament CL (Vision Biochem Co., Ltd.): Cellulase

Laminex BG2 (Vision Biochem Co., Ltd.): Cellulase

2. Preparation of enzyme-treated silkworm water extract

After extracting 1 kg of silk grass with 15-20 L of water at 100° C., twice for 3 hours, it was concentrated to 20Brix using a rotary evaporator to prepare a water extract concentrate of silkworm. 20 g of enzyme (Laminex BG2) was treated with 100 g of the prepared concentrate, stirred at 55° C. for 1 hour, left still, and then inactivated at 100° C. for 1 hour to obtain a freeze-dried product through freeze-drying (T-3).

In addition, as a comparative example, except that the enzyme was not treated, a water extract of silkworm was prepared in the same manner as in the preparation method of T-3 (C-2). That is, 15-20 L of water was added to 1 kg of silk grass, extracted twice at 100° C. for 3 hours, and then concentrated under reduced pressure and freeze-dried to obtain a final silk grass water extract.

The type of enzyme used, the amount and yield of the silk extract recovered are shown in Table 4 below, and the preparation process of the enzyme-treated water extract (T-3) and the water extract (C-2) as a comparative example is briefly shown in FIG. 5 .

enzyme Laminex BG2 No enzymatic treatment T-3 C-2 Recovery (g) 103 274.2 transference number(%) 103 27.4

3. Confirmation of the inhibitory effect of enzyme-treated silk grass extract on adipocyte differentiation

(1) Preparation and culture of preadipocytes (3T3-L1)

Mouse fibroblasts, 3T3-L1 preadipocytes, are a widely used cell line for the study of adipocyte metabolism, and were purchased from ATCC (American Type Culture Collection, USA). 3T3-L1 preadipocytes were cultured in an incubator maintained at 5% CO2 and 37°C using DMEM culture medium containing 10% calf serum and 1% penicillin-streptomycin solution. In order to solve the over-density phenomenon of cell proliferation, subculture was performed every 48 hours to maintain an appropriate number of cells.

(2) Confirmation of the inhibitory effect on the differentiation of adipocytes of the enzyme-treated silk grass extract

In order to check whether the enzyme-treated silk grass extract has the effect of inhibiting the differentiation of 3T3-L1 cells into adipocytes, the fat accumulation rate of adipocytes was checked. Stabilized 3T3-L1 cells were seeded in a 24 well plate at a density of 1x10 ⁵ cells/well, and after 48 hours, differentiation medium (0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 μM Dexamethasone, 10 μg/mL Insulin) ), and adipocyte differentiation was induced while the Insulin medium was replaced a total of 4 times at 2-day intervals. During the differentiation process, the enzyme-treated silk grass extract was mixed with the differentiation medium at a concentration of 100 μg/mL and treated. Thereafter, the degree of adipocyte differentiation was observed on day 14, when differentiation was completed.

In order to observe the degree of differentiation of adipocytes, Oil Red O staining, which reacts specifically to Lipid droplets, was performed. Specifically, the medium was removed from the cells induced by adipocyte differentiation as described above, washed twice with phosphate buffered saline, and fixed by treatment with 4% formalin at room temperature for 1 hour. After washing once with phosphate buffered saline, washed with 60% isopropanol, each well was completely dried. After treatment with Oil Red O dye for 30 minutes, it was washed 4 times with sterile distilled water. After drying each well, adipocytes were observed under a microscope. For the elution of the dyed Oil Red O, it was treated with 100% isopropanol for 10 minutes, and the absorbance of the eluate was measured at 520 nm.

The fat differentiation rate (Oil Red O stained area) in adipocytes was calculated by Equation 1 below, and the results are shown in Tables 5 and 6 below.

[Equation 1]

Fat accumulation rate (%) = (absorbance of sample treatment group / absorbance of control group) x 100

As can be seen in Table 5, compared to the negative control group, it was confirmed that the rate of accumulation of fat in adipocytes was significantly inhibited by the treatment with the active ingredient of the present invention, Silkworm extract, except for T-3. Therefore, it was found that the silk grass extract of the present invention inhibits the differentiation of pre-adipocytes into adipocytes, and thus is effective in preventing or treating obesity.

On the other hand, in the case of the water extract of silkworm, the fat accumulation inhibitory effect was reduced in T-3 treated with the starch enzyme than in Comparative Example (C-2) that was not treated with the enzyme, whereas in the case of the ethanol extract of silkworm, the enzyme treatment was not performed. It was confirmed that the fat accumulation inhibitory effect was increased in T-1 and T-2 treated with starch than in Comparative Example (C-1).

Claims (12)

A composition for preventing or treating obesity comprising an Euphorbia humifusa ethanol extract as an active ingredient, wherein the extract is enzyme-treated. The composition for preventing or treating obesity according to claim 1, wherein the enzyme is a starch degrading enzyme. The method of claim 1, wherein the enzyme is at least one selected from the group consisting of cellulase, hemicellulase, pectinase, glucosidase, glucanase, amylase, glucoamylase and isoamylase. composition. The composition for preventing or treating obesity according to claim 1, wherein the enzyme comprises cellulase. [Claim 2] The composition for preventing or treating obesity according to claim 1, wherein the ethanol extract of silk grass is 50% to 70% by weight of the ethanol extract. [Claim 2] The composition for preventing or treating obesity according to claim 1, wherein the ethanol extract of silk grasses inhibits fat accumulation in adipocytes. The composition for preventing or treating obesity according to claim 1, wherein the ethanol extract of silk grass is a freeze-dried powder. The method according to claim 1, wherein the extract is a result obtained by treating a starch degrading enzyme on an extraction result obtained by treating a silk grass raw material with an ethanol solvent, or a result obtained by treating a silk grass raw material with an ethanol solvent. A composition for preventing or treating obesity, characterized in that it is an extraction result obtained by (a) any one of claims 1 to 7, and (b) a pharmaceutical composition for preventing or treating obesity comprising a pharmaceutically acceptable carrier. (a) any one of claims 1 to 7, and (b) a food composition for preventing or improving obesity comprising a food pharmaceutically acceptable carrier. Silk grass ( Euphorbia humifusa ) processing the enzyme in the raw material; and
Treating the resultant obtained by treating the enzyme with an ethanol solvent to obtain a solvent extraction result
A method for preparing a composition according to claim 1 comprising a. Silk grass ( Euphorbia humifusa ) preparing a solvent extraction result by treating the raw material with an ethanol solvent; and
Treating the solvent extraction result with an enzyme
A method for preparing a composition according to claim 1 comprising a.

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