KR101802696B1 - Pharmaceutical composition for preventing or treating obesity comprising extract of pueraria lobata leaf - Google Patents

Abstract

The present invention relates to a composition containing an extract of sesame leaf as an active ingredient, wherein the sesame leaf extract of the present invention is a natural extract, which expresses an acetyl-CoA carboxylase (ACC) gene, which plays a major role in lipogenesis in the differentiation of lipoprotein somatic cells (Glycerol-3-phosphate dehydrogenase), which is involved in lipid differentiation and lipid synthesis, is inhibited and lipid accumulation is inhibited. Therefore, the composition containing the safflower extract of the present invention as an active ingredient is useful for the prevention of obesity Or as a pharmaceutical composition or a health food that can prevent or treat the disease.

Description

Pharmaceutical composition for preventing or treating obesity comprising extract of pueraria lobata leaf}

The present invention relates to a pharmaceutical composition for preventing or treating obesity containing an arrowroot leaf extract as an active ingredient.

Obesity is a metabolic disease caused by an imbalance between intake and consumption of calories, and refers to a condition in which adipose tissue is abnormally increased due to excess calories. Men are considered to be obese when their body fat is 25% of their body weight and women are over 30% of their body weight. Clinically, BMI (Body Mass Index) of 24.0 to 30.0 is defined as overweight and 3.0 or more is defined as obesity. do.

If obesity occurs and the condition persists, it acts as a cause of various diseases. Such diseases include high blood pressure, elevated blood cholesterol, diabetes, kidney disease, stroke, arteriosclerosis, fatty liver, arthritis, cancer, sleep apnea, and diabetes. have.

The causes of obesity include environmental and genetic factors such as high fat and high calorie diet, lack of exercise due to busy social environment, and endocrine abnormalities, of which 50 to 70% of obesity is attributed to environmental factors. It is known, and the rest is known to be due to genetic factors.

Obesity treatments are generally divided into three categories: appetite suppressants, stimulators of energy metabolism in the body, and inhibitors of digestive absorption. A typical obesity treatment that uses a pharmacological mechanism to suppress appetite is Reductil™ (Abbott, USA), and Exorise™ (Aco Pharma) is a representative obesity treatment that uses a pharmacological mechanism that promotes energy in the body. , France), and Xenical™ (Roche Pharmaceuticals, Switzerland) is a representative obesity treatment that uses a pharmacological mechanism that inhibits the digestion and absorption of fat.

Arrowroot is a perennial plant and has been edible as an oral crop for a long time, and has been used as a health food such as a nutrient tonic. In oriental medicine, the root is called prickly pear, and it is used as a medicinal for anti-inflammatory, antioxidant, blood pressure lowering, anti-ulcer, and liver protection. Until now, studies on the inhibition of lipid accumulation in arrowroot roots with the highest estrogen activity have been reported. , There is no research on the effectiveness of arrowhead leaves for preventing or treating obesity.

Korean Patent Registration No. 10-0645385 (2006.11.23)

The present invention is to provide a composition for preventing or treating obesity by inhibiting the differentiation of adipocytes by inhibiting the differentiation of adipocytes by providing a composition containing a natural product of arrowroot leaf extract as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating obesity containing an arrowroot leaf extract as an active ingredient.

In addition, the present invention provides a health food for preventing or improving obesity containing an arrowroot leaf extract as an active ingredient.

The present invention contains an arrowroot leaf extract as an active ingredient, and the arrowhead leaf extract provides a reagent composition for inhibiting adipocyte differentiation, characterized in that it inhibits differentiation of adipocytes into adipocytes in vitro.

In addition, the present invention provides a method for inhibiting differentiation of adipocytes in vitro, comprising the step of treating an arrowroot leaf extract on adipocytes isolated from individuals other than humans.

According to the present invention, the arrowroot leaf extract, a natural product, inhibits the expression of the ACC (acetly-CoA carboxylase) gene, which plays a major role in adipogenesis in the differentiation process of adipocytes, and GPDH (glycerol-3), which is involved in adipogenesis and adipogenesis. -phosphate dehydrogenase) activity was confirmed to inhibit lipid accumulation.

Accordingly, the composition containing the arrowroot leaf extract of the present invention as an active ingredient may be provided as a pharmaceutical composition or health food capable of preventing or treating obesity.

Figure 1 is a result of confirming the cytotoxicity of the arrowroot leaf ethanol extract, 3T3-L1 fat progenitor cells treated with 0 to 800 μg / mL arrowroot ethanol extract for 48 hours and confirmed the cell viability MTT analysis results.
Figure 2 is a result of oil red O staining confirming the lipids accumulated during differentiation after treatment with 25 to 100 μg/mL ethanol extract of arrowroot leaves on 3T3-L1 adipocytes induced differentiation for 8 days, a is a normal non-differentiated 3T3 -L1 is the result of staining of adipocytes, b is the result of staining of lipids accumulated in the 3T3-L1 adipocytes group in which differentiation into adipocytes is induced, and c is the result of staining of 3T3-L1 adipocytes in which differentiation is induced, 25 μg/ It is the staining result of the cell group treated with mL arrowroot leaf ethanol extract, d is the staining result of the cell group treated with 50 μg/mL arrowroot leaf ethanol extract in 3T3-L1 adipocytes induced differentiation, and e is the 3T3- induced differentiation. It is the staining result of the cell group treated with ethanol extract of 100 μg/mL arrowroot leaf to L1 adipocytes.
Figure 3 is a result of confirming the content of lipids accumulated during differentiation after treatment with 25, 50, and 100 μg/mL arrowroot leaf ethanol extract for 8 days on 3T3-L1 adipocytes from which differentiation was induced, compared with the differentiated control group * **p<0.001, N is an undifferentiated normal control, and C is a differentiated control cell.
FIG. 4 is a result of treating 3T3-L1 adipocytes in which differentiation is induced with ethanol extracts of 25, 50 and 100 μg/mL arrowroot leaves for 8 days and confirming the activity of glycerol-3-phosphate dehydrogenase (GPDH) during differentiation, arrowhead ethanol Compared to the untreated cell differentiation control with the extract, **p<0.01, N is the non-differentiated normal control, and C is the cell differentiation control.
5 is a RT-PCR result confirming the expression level of ACC (acetly-CoA carboxylase) mRNA in 3T3-L1 cells treated with 50 and 100 μg/mL arrowroot leaf ethanol extract for 8 days on 3T3-L1 adipocytes. , A is the result of confirming the ACC mRNA band in the gel, and B is the result of quantifying ACC mRNA expression.

The present invention can provide a pharmaceutical composition for preventing or treating obesity containing an arrowroot leaf extract as an active ingredient.

More specifically, the arrowroot leaf extract may be extracted with water, C 1 to 4 alcohol, and a mixed solvent thereof, and more preferably, an ethanol extract, but is not limited thereto.

The arrowroot leaf extract of the present invention can inhibit differentiation into adipocytes and lipid synthesis by inhibiting the activity of glycerol-3-phosphate dehydrogenase (GPDH) and the expression of acetly-CoA carboxylase (ACC) in adipocytes.

In more detail, GPDH (glycerol-3-phosphate dehydrogenase) is an enzyme that catalyzes the ester bond between glycerol and fatty acids, and the activity of GPDH increases the degree of triglyceride synthesis in cells, so it is known as a marker for adipocyte differentiation.

According to an embodiment of the present invention, as shown in FIG. 4, when the 3T3-L1 adipocytes in which differentiation is induced were treated with ethanol extract of arrowroot leaves at concentrations of 25 and 50 μg/ml, the ethanol extract of arrowroot leaves was not treated with the differentiated cells. It was confirmed that the activities of GPDH involved in lipodifferentiation and liposynthesis were reduced by 16.8% and 17.9%, respectively.

In addition, ACC (acetly-CoA carboxylase) is a gene that promotes the synthesis of long-chain fatty acids that forms malonyl-CoA, which is a substrate of FAS, regulates the rate of fatty acid synthesis, and catalyzes the first reaction step of fatty acid synthesis. According to an example, when differentiation-induced 3T3-L1 adipocytes are treated with ethanol extract at concentrations of 25 and 50 μg/ml as shown in FIG. 5, ACC genetic expression is more pronounced than differentiated cells not treated with the arrowroot ethanol extract. It could be confirmed that it was reduced by 34.6 and 33.3%, respectively.

According to another embodiment of the present invention, as a result of oil red O staining that can confirm the degree of lipid accumulation by displaying red color by reacting specifically to lipid components, 3T3-L1 in which differentiation is induced as shown in FIG. 2 It was confirmed that the fat droplets of adipocytes decreased in a concentration-dependent manner of the ethanol extract of arrowroot leaves.

From the above results, the ethanol extract of arrowroot leaves inhibits the expression of the ACC (acetly-CoA carboxylase) gene, which plays a major role in adipogenesis in the differentiation process of adipocytes, and glycerol-3-phosphate dehydrogenase (GPDH), which is involved in the differentiation of fat and fat synthesis. ) By reducing the activity, it was confirmed to inhibit lipid accumulation.

From the above results, the present invention contains an arrowroot leaf extract as an active ingredient, and the arrowhead leaf extract is a reagent composition for inhibiting adipocyte differentiation, characterized in that it inhibits differentiation of adipocytes into adipocytes in vitro. Can be provided.

In addition, the present invention can provide a method for inhibiting differentiation of adipocytes in vitro, comprising the step of treating an arrowroot leaf extract on adipocytes isolated from individuals other than humans.

The pharmaceutical composition of the present invention may be included in an amount of 0.01 to 90 parts by weight based on 100 parts by weight of the total pharmaceutical composition, and the arrowroot leaf extract is not limited thereto.

In one embodiment of the present invention, the pharmaceutical composition for preventing or treating cell damage caused by ultraviolet rays containing the arrowroot leaf extract as an active ingredient is an injection, granule, powder, tablet, pill, capsule, suppository, gel according to a conventional method. , Suspension, emulsion, drop or liquid may be used in any one of the selected formulation.

In another embodiment of the present invention, the pharmaceutical composition for preventing or treating obesity containing the arrowroot leaf extract as an active ingredient is a suitable carrier, excipient, disintegrant, sweetener, coating agent, swelling agent, lubricant, which is commonly used in the manufacture of pharmaceutical compositions. It may further include one or more additives selected from the group consisting of lubricants, flavoring agents, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders, and lubricants.

Specifically, carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil can be used, and solid preparations for oral administration include tablets, pills, powders, granules, capsules. And the like, and these solid preparations may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like in the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, and preservatives may be included. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, and the like. As the non-aqueous solvent and suspension, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like may be used. As a base material for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerogelatin, and the like may be used.

According to an embodiment of the present invention, the pharmaceutical composition is intravenous, intraarterial, intraperitoneal, intramuscular, intraarterial, intraperitoneal, intrasternal, transdermal, intranasal, inhalation, topical, rectal, oral, intraocular or intradermal It can be administered to the subject in a conventional manner via the route.

A preferred dosage of the arrowroot leaf extract may vary depending on the condition and weight of the subject, the type and degree of the disease, the form of the drug, the route and duration of administration, and may be appropriately selected by those skilled in the art. According to an embodiment of the present invention, although not limited thereto, the daily dosage may be 0.01 to 200 mg/kg, specifically 0.1 to 200 mg/kg, and more specifically 0.1 to 100 mg/kg. Administration may be administered once a day or may be divided into several doses, whereby the scope of the present invention is not limited.

In the present invention, the'subject' may be a mammal including a human, but is not limited to these examples.

In addition, the present invention can provide a health food for preventing or improving obesity containing the arrowroot leaf extract as an active ingredient.

The health food may be used together with other foods or food additives in addition to the arrowroot leaf extract, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient is suitably used according to the purpose of use, such as prevention, health or therapeutic treatment. Can be determined.

The effective dose of the compound contained in the health food may be used in accordance with the effective dose of the therapeutic agent, but in the case of long-term intake for the purpose of health and hygiene or health control, it may be less than the above range, and is effective. It is clear that the ingredient can be used in an amount beyond the above range because there is no problem in terms of safety.

There is no particular limitation on the kind of health food, for example, meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, Drinks, alcoholic beverages, and vitamin complexes.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely describe the present invention to those of ordinary skill in the art.

<Reference Example> Reagents and instruments

Tannic acid, folin-ciocalteu's phenol reagent, di(ethylene glycol) [di(ethylene glycol)] reagent, DPPH(1,1-diphenyl-2-picryl hydrazyl), insulin, 3-isobutyl-1-methyl Xanthine [3-isobutyl-1-methylxanthine], dexamethasone, and oil red O were manufactured by Sigma (USA), and DMEM (Dulbecco's modified Eagle's medium), fetal bovine serum (fetal bovine) were used. Serum; FBS) and penicillin/streptomycin (P/S) were purchased and used by Lonza (USA), and other general reagents were used with special products. The sample was extracted using a rotary vacuum concentrator (R-205, B-490, Buchi, Japan). The cell line was observed using an inverted microscope (CKX41, Olympus, Japan), and the cell line was cultured using a CO ₂ incubator (MCO-15AC, Sanyo Electric, Japan).

<Example 1> Arrowroot leaf ethanol extract

A powdery sample was prepared by adding 3L of 80% ethanol to 300g of arrowroot leaves collected in Chilgok, Gyeongsangbuk-do and adding 3L of 80% ethanol to immersion for 24 hours, extracting 3 times repeatedly, filtering through filter paper, concentrating and freeze-drying. The yield was 1.2%.

<Example 2> Confirmation of cytotoxicity of ethanol extract of arrowroot leaves

1. Cell line and culture

3T3-L1 adipocytes were pre-sold in Molecular Medicine, Keimyung University College of Medicine, and were subcultured. Attached to the cell culture plate and cultured in an incubator at _{37° C. and 5% CO 2} using DMEM medium to which 10% bovine calf serum (BCS) and 1% penicillin/streptomycin (P/S) were added. .

2. MTT analysis

3T3-L1 adipocytes were dissolved in DMEM medium containing 10% BCS and 1% penicillin/streptomycin (P/S) _{and stabilized in a 37° C., 5% CO 2} incubator for 48 hours, and then used for MTT analysis. . Dispense 200 µl of stabilized 3T3-L1 adipocytes into a 96-well plate at an appropriate number of cells (1×10 ⁴ cells/well) and incubate for 24 hours in a 37°C, 5% CO ₂ incubator, and then 1 with PBS. After washing twice, the freeze-dried ethanol extract of arrowroot leaves was diluted in DMEM medium by concentration of 25-800 μg/ml, added 200 µl each, and _{cultured in a 37°C, 5% CO 2} incubator for 48 hours.

Then, wash once with PBS and add 200 µl of DMEM medium containing 0.5 mg/mL of MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] at 37°C, Incubated for 3 hours in a 5% CO _{2 incubator.} Discard the medium, add 200 µl of DMSO each, and dissolve the cells for 15 minutes in a plate stirrer, and then measure the absorbance at a wavelength of 540 nm with an ELISA reader.

As a result, as shown in FIG. 1, when the ethanol extract of arrowhead leaves at a concentration of 100 μg/ml was treated, 3T3-L1 adipocytes showed a cell growth rate of 89.3%, whereas in the ethanol extract of arrowhead leaves at a concentration of 200 μg/ml, it was 68.8%. It appeared and showed cytotoxicity, and the maximum permissible concentration was confirmed to be 100 μg/ml.

From the above results, in all the experiments below, a concentration of 100 μg/ml or less was used.

<Example 3> Confirmation of lipid synthesis inhibitory effect of ethanol extract of arrowroot leaves

1. Cell differentiation and sample processing

In order to induce differentiation of 3T3-L1 adipocytes, the ^{cells were aliquoted into 1×10 6} cells on a 60 mm plate, and the medium was exchanged 2 days later, and the cells were cultured to 100% densely on the 4th day. DMEM medium containing 10% fetal bovine serum (FBS) and MDI solution (0.5 mM IBMX, 1 μM dexamethasone, 10/mL insulin) was treated for 2 days. At this time, in order to observe the effect of the arrowroot leaf ethanol extract on adipocyte differentiation, the arrowroot leaf ethanol extract was treated with 25, 50 and 100/mL concentrations. After 2 days of induction of differentiation, the medium was treated with arrowhead ethanol extract at each concentration, DMEM containing 10% FBS and 10/mL insulin for 3 days, and thereafter, 10% FBS and 1% penicillin once every 2 days with arrowhead ethanol extract. / The cells were differentiated for a total of 8 days by exchange with DMEM containing streptomycin (P/S).

2. Oil red O staining and lipid content check

Oil red O staining, a special dyeing method that can confirm the degree of lipid accumulation by reacting specifically to lipid components and showing red color, was used to confirm the lipid synthesis inhibitory effect of the ethanol extract of arrowroot leaves.

On the 8th day of differentiation of 3T3-L1 adipocytes differentiated by the above method, the medium was removed, the cells were washed with PBS, and then fixed with 10% formaldehyde at room temperature for 60 minutes. After removing the fixative and washing with 60% isopropanol, it was stained at room temperature using an oil red O solution that specifically reacts with lipid droplets generated in the cell.

As a result, as a result of treatment with ethanol extract of arrowroot leaves for 8 days on 3T3-L1 adipocytes induced differentiation as shown in FIG. 2 and observation at 200 magnification using an inverted microscope, undifferentiated 3T3-L1 adipocytes as shown in FIG. 2A In contrast, red color was hardly observed in FIG. 2B, whereas a large amount of red color was observed in the control adipocytes differentiated for 8 days in FIG. 2B, indicating that a large amount of lipid was accumulated. In addition, compared to the control adipocytes, in the cell group treated with the ethanol extract of arrowroot leaf 25 μg/ml (Fig. 2c), 50 μg/ml (Fig. 2d) and 100 μg/ml (Fig. 2e), the red color is less observed depending on the concentration. As a result, it was confirmed that lipid accumulation was suppressed.

In addition, the stained cells were observed under a microscope, extracted by dissolving Oil Red 0 staining using 100% isopropanol, and then the absorbance was measured at 490 nm and the lipid content in the adipocytes was expressed as a numerical value, as a percentage of the absorbance value of the control group. Indicated.

As a result, compared to the control group (C), which is 3T3-L1 adipocytes differentiated for 8 days as shown in FIG. 3, in the cell groups treated with arrowroot leaf ethanol extract 25, 50 and 100 μg/ml, concentration-dependently 60.0%, 71.1% and 73.3, respectively. % Significantly (p<0.001) the lipid content was low.

From the above results, it was confirmed that the ethanol extract of arrowroot leaves has an inhibitory effect on lipid accumulation.

3. Glycerol-3-phosphate dehydrogenase (GPDH) activity confirmation

By confirming the GPDH activity, which increases the activity specifically only in adipocytes, the differentiation of adipocytes and the degree of accumulation of triglycerides according to the ethanol extract treatment of arrowroot leaves were confirmed.

GPDH activity was measured using a GPDH activity kit (Takara Bio Inc., Japan). Remove the culture medium of adipocytes treated with the differentiation medium and ethanol extract of arrowroot leaves for 8 days, collect the cells by adding an enzyme extraction buffer, and use a dilution buffer containing 2-melcaptoethanol. Was diluted.

After mixing the diluted cells in the plate containing the substrate, the change in absorbance was measured every 1 minute at 340 nm in a spectrophotometer for 10 minutes. GPDH activity was calculated by calculating the amount of enzyme to use 1 μM of the substrate for 1 minute, and the calculated value was converted to mg protein/ml. The protein content of the supernatant was measured using a Bio-red protein assay kit (BioRad, CA, USA).

As a result, it was confirmed that, compared to the control group (C), as shown in FIG. 4, the cell group treated with the ethanol extract of arrowroot leaves 25 and 50 μg/ml significantly inhibited the GPDH activity by 16.8 and 17.9%, respectively (p<0.01).

From the above results, it was confirmed that the ethanol extract of arrowroot leaves effectively inhibits adipocyte differentiation and adipogenesis.

4. ACC (acetyl-CoA carboxylase) gene expression confirmation

The effect of ethanol extract of arrowroot leaf on the expression of acetyl-CoA carboxylase (ACC) gene, which forms the substrate of FAS, malonyl-CoA, regulates the rate of fatty acid synthesis, and promotes the synthesis of long-chain fatty acids that catalyze the first reaction step in fatty acid synthesis. The effect was confirmed by reverse transcription polymerase chain reaction (RT-PCR).

Total RNA was isolated by treatment with 0.8 ml of Trizol-Reagent (Life Technologies, CA, USA) on 3T3-L1 adipocytes differentiated and treated with ethanol extract of arrowroot leaves by the above method. 160 µl of chloroform was added to the separated RNA, mixed, and centrifuged twice to separate the supernatant. After precipitating RNA using 400 µl isopropyl alcohol, washing with 70% ethanol and air drying, the RNA was dissolved in DEPC-water and stored at -80°C.

5 μg of total RNA isolated from the control group and the experimental group by the above method, respectively, was mixed with 8 μl of M-MLV RT 5×buffer, 3 μl of 10 mM dNTPs, 0.45 μl of 10,000U RNase inhibitor, and 50,000U M-MLV reverse transcriptase (Promega, USA). ) CDNA was prepared by reverse transcription using 40 µl of a solution containing 0.3 µl and 20 pmol/oligo dT (Bioneer, Korea) 3.75 µl.

Single-stranded cDNA prepared by the above method was 5×green Go Taq flexi buffer 4 μl, 10 mM dNTPs 0.4 μl, 500U Taq polymerase 0.1 μl, 25 mM MgCl ₂ (Promega, Madison, USA) 1.2 μl and ACC, FAS, GAPDH The specific forward and reverse primers of were amplified by PCR using 0.4 μl of 20 pmol/L, respectively. The primer sequence and expected size used for PCR are shown in Table 1.

GAPDH (50, 28 cycles) as an internal standard and ACC (94 30 seconds, 56 1 minute, 72 1 minute, 30 cycles) were used as the test material, and the PCR product was subjected to electrophoresis on a 1.2% agarose gel for analysis. I did.

gene primer Expected size (base pair) ACC
Forward (5'→3') TGA CCG TGG GCA CAA AGT T (SEQ ID NO: 1) 351
Reverse (5'→3') AGG AGG AAC CGC ATT TAT CGA (SEQ ID NO: 2) GAPDH
Forward (5'→3') GTA TGA CTC CAC TCA CGG CAA A (SEQ ID NO: 3) 248
Reverse (5'→3') GGT CTC GCT CCT GGA AGA TG (SEQ ID NO: 4)

As a result, it was confirmed that ACC gene expression was reduced by 34.6 and 33.3%, respectively, in the cell groups treated with 25 and 50 μg/ml arrowroot ethanol extract compared to the control 3T3-L1 adipocytes not treated with the arrowroot leaf ethanol extract as shown in FIG. 5. Could.

From the above results, it was confirmed that the ethanol extract of arrowroot leaves reduced the expression of ACC genes induced in the early stage of adipogenesis, thereby inhibiting adipogenesis in 3T3-L1 adipocytes.

On the other hand, the arrowroot leaf extract according to the present invention can be formulated in various forms depending on the purpose. Hereinafter, some formulation methods containing the arrowroot leaf extract according to the present invention as an active ingredient are exemplified, and the present invention is not limited thereto.

<Formulation Example 1> Formulation example of pharmaceutical composition

<Formulation Example 1-1> Preparation of injection

Arrowhead leaves ethanol extract 10 mg, sodium metabisulfite 3.0 mg, methylparaben 0.8 mg, propylparaben 0.1 mg, and an appropriate amount of sterile distilled water for injection were mixed and prepared so that the final volume became 2 ml by a conventional method, and then 2 ml dose Filled into ampoules and sterilized to prepare injections.

<Formulation Example 2-1> Preparation of tablets

Tablets were prepared by mixing 10 mg of arrowroot leaf ethanol extract, 100 mg of lactose, 100 mg of starch, and an appropriate amount of magnesium stearate, followed by tableting according to a conventional tablet preparation method.

<Formulation Example 3-1> Preparation of capsules

10 mg of arrowroot leaf ethanol extract, 50 mg of lactose, 50 mg of starch, 2 mg of talc, and an appropriate amount of magnesium stearate were mixed and filled into gelatin capsules according to a conventional capsule preparation method to prepare a capsule.

<Formulation Example 2> Preparation of health food

Arrowroot leaf ethanol extract 0.5 mg, vitamin mixture appropriate amount (vitamin A acetate 70 μg, vitamin E 1.0 mg, vitamin B 1 0.13 mg, vitamin B 2 0.15 mg, vitamin B 6 0.5 mg, vitamin B 12 0.2 μg, vitamin C 10 mg) , Biotin 10 µg, nicotinic acid amide 1.7 mg, folic acid 50 µg, calcium pantothenate 0.5 mg) and an appropriate amount of an inorganic mixture (ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, monobasic potassium phosphate 15 mg, second 55 mg of calcium phosphate, 90 mg of potassium citrate, 100 mg of calcium carbonate, and 24.8 mg of magnesium chloride) were mixed, and then granules were prepared and health food was prepared according to a conventional method.

As described above, a specific part of the present invention has been described in detail, and for those of ordinary skill in the art, it is obvious that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

<110> INDUSTRY ACADEMIC COOPERATION FOUNDATION KEIMYUNG UNIVERSITY <120> Pharmaceutical composition for preventing or treating obesity comprising extract of pueraria lobata leaf <130> ADP-2015-0088 <160> 4 <170> KopatentIn 2.0 <210> 1 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> ACC forward <400> 1 tgaccgtggg cacaaagtt 19 <210> 2 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> ACC reverse <400> 2 aggaggaacc gcatttatcg a 21 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> GAPDH forward <400> 3 gtatgactcc actcacggca aa 22 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH reverse <400> 4 ggtctcgctc ctggaagatg 20

Claims (5)

The present invention relates to a reagent composition for inhibiting GPDH activity, which comprises an extract of perilla leaves as an active ingredient and inhibits glycerol-3-phosphate dehydrogenase (GPDH) activity in vitro. delete [Claim 3] The reagent composition according to claim 1, wherein the safflower extract inhibits ACC (acetyl-CoA carboxylase) expression. [2] The reagent composition according to claim 1, wherein the reagent composition comprises 0.01 to 90 parts by weight of a sesame leaf extract per 100 parts by weight of the total reagent composition. delete

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