KR20180111479A - Anti-obesity functional composition comprising abeliophyllum distchum extract as an active ingredient - Google Patents

Abstract

The present invention relates an anti-obesity functional composition containing an Abeliophyllum distichum Nakai extract as an active ingredient, which shows high inhibitory activities for lipogenic genes, suppresses weight gain due to reduction of body fat and triglyceride, and exhibiting anti-obesity functions beneficial to human body. To this end, the anti-obesity functional composition contains, as the active ingredient, an extract obtained by extraction after mixing at least one or two portions selected among leaves, stems, fruits, roots, husks and seeds of Abeliophyllum distichum Nakai in the same ratio.

Description

[0001] The present invention relates to an anti-obesity functional composition comprising abscisic acid extract,

The present invention relates to an anti-obesity functional composition containing an extract of Helicobida plant which has high inhibitory activity of lipogenic gene and inhibits weight gain due to reduction of body fat and triglyceride and exhibits an anti-obesity function beneficial to human body as an active ingredient will be.

In general, obesity means a condition with excess body fat. When a person has body fat of 25% of body weight and a woman of 30% or more of body weight, clinically, when body mass index (BMI) And the present weight is defined as a case where the abnormal weight exceeds 20%.

The major causes of obesity are genetic factors, environmental factors, and energy metabolism abnormalities. Obesity can be classified into simple obesity and symptomatic obesity depending on the cause. Simple obesity is caused by overeating and lack of exercise, and symptomatic obesity is caused by endocrine, hypothalamic, genetic, frontal lobe and metabolic .

Obesity, therefore, increases the likelihood of developing diabetes and hyperlipemia, increases the risk of sexual dysfunction, arthritis, and cardiovascular disease, as well as the occurrence of cryptorchidism and cancer. can see.

Such obesity is best addressed by dietary and exercise habits, but it can also be treated with medications or health supplements. There are two types of drugs used in obesity treatment: appetite suppressants and drugs that inhibit the absorption of fat. Sibutramine and orlistat are drugs approved for long-term use.

Sibutramine is a type of appetite suppressant that can be used on a long-term basis for about one year, reducing the average 5 to 9 percent of body weight. However, due to the continuous use of medication, side effects such as headache, dry mouth (severe thirst), insomnia and constipation may occur, and blood pressure and pulse rate may increase. Therefore, when taking a long time sibutramine, blood pressure and pulse rate should be measured periodically Should be checked.

In addition, orlistat is an inhibitor of lipolytic enzymes, which prevents the digestion of fat in the body. Therefore, about 30% of the fat ingested is not extinguished and absorbed properly and is discharged to the outside of the body as it is.

Accordingly, in recognition of such a problem, Korean Patent Laid-Open Publication No. 10-2014-0058375 discloses a functional health food composition for obesity suppression containing a pear extract.

This technique extracts the active ingredient from a pupa extract or a sludge extract, and aims at weight loss and weight maintenance activity.

However, the above-mentioned technique not only has an extremely small amount of the effective ingredient extracted from the abdomen, but also has an effect that can inactivate the action mechanism of the factor which is fundamentally a cause of weight gain, for example, , The effect of being exerted through the active ingredient is weak due to the absolute lack of factors directly involved in weight loss or weight maintenance activity, that is, COX-2 and CEFP (cholesteryl ester transfer protein) Respectively.

Korean Patent Publication No. 10-2014-0058375 (Apr. Korean Patent Publication No. 10-2015-0083622 (July 20, 2015) Korean Patent No. 10-1563315 (Oct. 20, 2015)

The inventors of the present invention have made intensive studies on a composition capable of exhibiting the anti-obesity effect to the utmost without adverse effects on the human body. As a result, it has been found that a composition containing an extract of Aspergillus species as an active ingredient inhibits fat accumulation and has a high inhibitory activity Anti-obesity test, etc., and completed the present invention.

Accordingly, it is an object of the present invention to provide an anti-obesity functional composition comprising an extract of Helicobida plant as an active ingredient, which can contribute to the health improvement of the human body by promoting weight loss or weight maintenance activity.

In order to accomplish the above object, the present invention provides a method for producing a plant extract, comprising extracting an extract obtained by mixing at least one of two or more parts selected from leaf, stem, fruit, root, Characterized in that the composition further comprises one or both of a transferase or an excipient.

Herein, the active ingredient is characterized in that it contains one or both components of verbascoside or polyphenol.

Meanwhile, a method for extracting an effective ingredient from the spinach includes: a) a material processing step for cutting, washing, drying, and pulverizing a desired part by collecting spinach; b) extracting a solution containing the active ingredient from the processed material of step a) through a solvent to prepare an extraction solution; c) filtering and concentrating the extract solution of step b) to prepare a concentrate; d) lyophilizing the concentrate of step c); And e) pulverizing the lyophilized concentrate of step d).

Also, the extraction solution of step b) is extracted until the concentration of the soluble solid reaches 20 - 22 ° brix, and the solvent of b) is extracted with 70% ethanol, purified water, methanol, The solvent is selected from the group consisting of glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether and hexane. Any one or two or more may be mixed in the same ratio.

The anti-obesity functional composition containing the above-described composition of the present invention as an active ingredient of the present invention has a large amount of components beneficial to human body such as polyphenol and inhibits the lipogenic gene which is a cause of obesity It is effective to reduce the body fat and promote the health of the human body.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process diagram showing a method for extracting an anti-obesity functional composition containing an extract of Helicobida plant as an active ingredient according to a preferred embodiment of the present invention.
FIG. 2 is a graph showing the results of cytotoxicity test according to the treatment concentration of the bitter gourd extract. FIG.
FIG. 3 is a photograph showing the change in lipid droplet according to the treatment concentration of the bitter gourd extract. FIG.
FIG. 4 and FIG. 5 are graphs showing the content of triglycerides according to the treatment concentration of the bitter gourd extract. FIG.
6 is a photograph showing the expression of the protein in PPARγ and C / EBPα by Western blot.
FIG. 7 is a photograph showing the expression of PPARγ and C / EBPα by RT-PCR (Real time polymerase chain reaction).
FIG. 8 is a graph showing changes in body weight according to the treatment period in the case of administration of Zygomycota leaf extract to obese rats.

The anti-obesity functional composition according to the present invention contains the extract extracted from the spruce tree as an active ingredient and can be used as a pharmaceutical or a foodstuff-acceptable transferase (precursor gene) or an excipient Or materials added for the purpose of facilitating use by increasing the amount thereof).

Herein, the active ingredient may include one or both of the components of verbascoside or polyphenol, and the verbascoside may be a compound belonging to phenylpropanoid glycoside (Wu et al., 2007; Cardinali et al., 2012) have been shown to have various biological activities such as antioxidant, anti-inflammation, antimicrobial and immunosuppressive effects, .

The method for extracting the bitterling extract according to a preferred embodiment of the present invention, as shown in FIG. 1,

a) a material processing step for cutting, washing, drying and crushing a desired part by taking a spike;

b) extracting a solution containing the active ingredient from the processed material of step a) through a solvent to prepare an extraction solution;

c) filtering and concentrating the extract solution of step b) to prepare a concentrate;

d) lyophilizing the concentrate of step c); And

and e) pulverizing the lyophilized concentrate of step d).

The extracting method for extracting Leucocandia spp. Comprising the steps as described above will be described separately for each step. First, the material is processed. In order to collect the desired part of the brackish tree, it is necessary to collect the desired part of the brackish tree by collecting the brackish wood. In order to collect the desired part of the brackish tree, the endangered wildlife, As a material for obtaining the extract of Helicobidae according to the present invention, any one or more of the leaves, stems, fruits, roots, husks and seeds formed on the spikelets that have been scraped for propagation in the organ obtained by the certificate of proliferation is cut .

The Abelophyllum distichum , Nakai) is a plant of the ash tree, and it is called "the fine line tree" which resembles the round fan of the fruit, and it is a shrub that lives only in our country. It is the only one genus in the world and one kind of natural monument. Have found that polyphenol, which is about 10 times that of green tea, has a large amount of components that act as anti-inflammatory, anti-cancer, and antioxidant.

It is most desirable to collect these species from early April to late October. This is because the functional activation of the active ingredient in the bitter gourd is most actively carried out from the beginning of April to the end of October.

As described above, the desired part of the spike is cut and taken as a material. Then, the material is washed in running water to remove foreign matter, and then water is permeated into the material to prevent contamination with harmful bacteria such as mold The dried material is dried sufficiently to remove the water completely.

In one embodiment, the drying of the material is preferably dry in a shade, i.e., in a shady location, for 7 to 10 days. At this time, when the drying period of the material exceeds 10 days, the effective ingredient contained in the beech tree is discharged to the outside, so that it is difficult to achieve the object of the present invention. When the drying period is less than 7 days, It is preferable that the material is dried for 7 days to 10 days since harmful bacteria such as bacteria and the like may be inhabited and further harmful bacteria may propagate. In addition, in addition to natural drying, the material may be dried by using a drying method such as a dryer to remove water from the fine wood.

The dried material is a pretreatment step for extracting the extract of Spruce tree, and it is pulverized to a particle size of 10 mesh to 200 mesh and a moisture content of 4% to 5%.

As described above, the pulverized and processed material can be used to prepare a solution by extracting a solution containing the active ingredient through a conventional solvent extraction method.

The solvents used in the solvent extraction method include 70% ethanol, purified water, methanol, glycerin, ethyl acetate, butylene glycol, propylene glycol ), Dichloromethane, chloroform, ethyl ether, and hexane may be mixed in the same ratio.

In this extraction step, the concentration of soluble solids, minus the amount of insoluble solids in the total solids amount, is preferably extracted until the concentration is between 20 ° brix and 22 ° brix. This is because the functional components such as organic acids are present most abundantly in the above concentration range.

The extract solution is filtered through a filter (about 55 mu m) and then concentrated by a known centrifugal thin film concentrator. In one embodiment, the extraction solution may be concentrated through a centrifugal thin film concentrator rotating at 1,200 rpm at a temperature of 80 DEG C as a condition for the concentration.

As described above, the concentrate is cooled in a freezer at -30 캜 to -35 캜 for 10 to 14 hours, and then lyophilized for about 46 to 50 hours at a temperature of maximum -45 캜 , And then obtained through a publicly known pulverizer, to obtain a vinegar extract in powder form having a particle size of 80 mesh to 120 mesh.

In the meantime, the term 'extract' as used in the present specification generally means a crude extract, but in a broader sense it also includes fractions obtained by further fractionation of the extract. That is, the bitter gourd extract includes not only those obtained by using an extraction solvent but also those obtained by further applying a purification process thereto. For example, a fraction obtained by passing the above-mentioned extract through an ultrafiltration membrane having a constant molecular weight cut-off value, and a variety of additionally carried out such as separation by various chromatographs (manufactured for separation according to size, charge, hydrophobicity or affinity) The fraction obtained through the purification method is also included in the scope of the present invention.

In a further preferred embodiment of the present invention, the Herba extract obtained by the above-mentioned method may further contain any one or both of transitional enzymes or excipients, and the amount thereof is preferably 0.0001 wt% To 5.0% by weight. If it is added in an amount of less than 0.0001% by weight, it is difficult to obtain desired functionality because the effective concentration of the active ingredient is too low. If the added amount is more than 5.0% by weight, In addition, an auxiliary additive exhibiting functional activity may be further added to the composition within the above-mentioned range.

The extract may be formulated into various forms depending on the intended use and the method of use, such as a pill, a tablet, and a capsule.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples.

EXAMPLES Preparation of Samples Extracted from Leaf Tree Leaves

Leaves were collected from the spikelets cultivated in Chougok - ri, Goesan - gun, Chungbuk Province, and the collected leaves were washed and dried, and the dried leaves (200g) were cut into 5 - 6cm size. The cut leaves were plated in 10% (w / w) 70% (w / w) ethanol and extracted for 24 hours. The extracted solution was filtered with a milipore filter (55 μm), concentrated using a centrifugal thin film concentrator, and used as an experimental sample. The obtained samples were stored in a freezer maintained at a temperature of -30 DEG C until the day of the experiment.

[Experimental Example 1] Measurement of cytotoxicity against 3T3-L1 cells

Toxicity of the extracts from Leuconstorm leaves was assayed by 5- (3-carboxymethoxyphenyl) -2H-tetrazolium inner salt (MTS) assay. This is a measure of the conversion of MTS to formazan by mitochondrial dehydrogneases. 3T3-L1 cells were plated at 1 × 10 ⁴ cells / well in a 96-well plate and the samples were treated for 24, 48 and 72 hours at concentrations of 0, 25, 50, 100 and 200 μg / ml, respectively. After incubation at 37 ° C in a 5% CO ₂ incubator for 4 hours, the absorbance at 450 nm was measured using a microplate reader (Versamax, USA) Expressed as a percentage. The absorbance of each sample was corrected by comparing the absorbance of the control and the experimental group.

Cell viablility (%) = 100- (A-B) / A x 100

A: Absorbance of the control group

B: absorbance of the sample treatment group

Referring to FIG. 2, after the completion of cell culture, MTS analysis was performed at a concentration of 25-200 μg / ml to confirm the cytotoxic effect of Leucillus sp. And it was confirmed that there was a small cytotoxicity from the concentration of 200 μg / ml.

[Experimental Example 2] Experimental study on anti-obesity effect of Leaf leaf extract

1. 3T3-L1 cells were obtained from the American Type Culture Collection (ATCC), Manassas, VA. The 3T3-L1 preadipocyte was cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS, Gibco Co, USA) and 1% penicillin-streptomycin (P / S, Gibco Co, USA) medium, DMEM) at 37 ° C and 5% CO ₂ /95% air.

To induce the differentiation of 3T3-L1 adipose precursor cells, the cells were divided into 1.25 × 10 ⁵ cells / well in each well. After 2 days, the medium was changed and the cells were saturated on the 3rd to 4th day. Differentiation was induced by treatment with 10% FBS and MDI solution (0.5 mM 3-iso Butyl-1-methylxanthine, 1 μm dexamethsone, 5 μg / ml insulin) (Sigma Co, USA) in the saturated state. At this time, the samples were treated at the concentrations of 25, 50, 100 and 200 μg / ml together to observe the effect of the samples on adipocyte differentiation.

After 2 days of induction of differentiation, the medium was replaced with DEME containing 10% FBS, 1% P / S and 5 μg / ml insulin, and 10% FBS and 1% P / S < / RTI >

2. Oil red O staining and TG quantitation

First, in a culture vessel containing cells to be stained, 1 ml was separately transferred for use in quantitative analysis of TG in the cell culture medium, the remaining cell culture medium was removed, and the cells were washed twice with phosphate-buffered saline (PBS). Then, in the experiment, it was fixed with 10% formalin for 60 minutes, then the fixing solution was removed, and then washed again with PBS three times. In order to stain fat in adipocyte, it was treated with filter paper filtered Oil red O solution for 60 minutes at room temperature and then washed twice with PBS. Only the lipid droplet was stained red. Observed with an electron microscope, recorded as a photograph, and expressed as the value of absorbance (OD 530 nm) (FIGS. 3 and 4). The collected cell cultures were analyzed using a LabAssay ^TM Triglyceride kit (Wako Pure Chemical Industries, Ltd., Osaka, Japan) capable of measuring the residual amount of neutral fat in the culture medium (Fig. 5).

3. Western blot analysis

The fat cells were suspended in ice-cold lysis buffer (20 mM Tris-HCl (pH 7.4), 2 쨉 m EDTA, 500 쨉 m sodium orthovanadate, 1% Triton X-100, 0.1% SDS, 10 쨉 m NaF, PMSF) and centrifuged at 13,000 g for 30 min at 4 ° C. Protein quantification was performed using supernatant. Protein quantification was measured by Bio-Rad DC protein assay (Bio-Rad Laboratories, Hercules, Calif.) using bovine serum albumin as standard. Total protein (10-50 μg) was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using 10% polyacrylamide gel and transferred to nitrocellulose membrane. Membranes were blocked with skim milk (5% dissolved in PBST (0.1% v / v Tween-20 in PBS, pH 7.2) for 1 hour and β-actin (1: 5000 dilution, Sigma, USA) (1: 5000 dilution, Cell Signaling, USA) and incubated at 4 ° C overnight. Cells were then incubated overnight at 4 ° C in a solution of mouse IgG Ab-H & L The secondary antibody was added and reacted at room temperature for one hour. Membranes were washed five times with PBST every 5 minutes for each step. The reaction results for the samples were detected using an Amersham ECL system (Amersham-Pharmacia Biotech, Arlington Heights, IL).

4. Real time polymerase chain reaction (RT-PCR) measurement

RNA was extracted from 3T3-L1 cells induced by adipocyte differentiation using Trizol (Invitrogen, Carlsbad, CA, USA). Total RNA was quantitatively analyzed using a Premix kit (Clontech, Mountain View, CA, USA) . The synthesized cDNA was PCR-amplified in 28 cycles and electrophoresed on 1.5% agarose gel. On the other hand, the PCR reaction conditions were amplification at 95 ° C for 15 minutes, 30 seconds (denaturation), 55 ° C for 30 seconds (annealing) and 72 ° C for 30 seconds.

5. Statistical Analysis

Student t-test was performed for all test results, and the statistical significance of mean values was tested at p <0.05 level.

6. Analysis of experimental results

3 to 5, oil red O staining and TG analysis were performed to evaluate the degree of accumulation of triglyceride in the differentiation. As a result, compared with the control group cultured with MDI alone, the results were 25, 50, 100 and 200 ㎍ / ㎖, the accumulation of lipid droplet was decreased in concentration-dependent manner, and the amount of triglyceride also decreased in concentration-dependent manner, The maximum decrease of triglyceride was found at 50 ㎍ / ㎖.

In addition, as shown in FIG. 6, in order to examine the effect of PPARγ and C / EBPα, which are crucial transcription factors, on the adipocyte differentiation, . As a result, the difference between PPARγ1 and PPARγ2 was significantly inhibited, and the expression of PPARγ1 and PPARγ2, which are isoforms of PPARγ, was inhibited. PPARγ2 was expressed specifically in fat cells with PPARγ1, (42 kDa) and p30 (28 kDa), which are isoforms of C / EBPα, also increase with the differentiation of adipocytes. , And that the expression of these extracts was remarkably reduced when the extracts were treated in a concentration-dependent manner.

As shown in FIG. 7, PPARγ was decreased in a concentration-dependent manner at 25 and 50 μg / ml compared to the control, and the results of the gel electrophoresis through RT-PCR showed that the transcription factors and differentiation markers were decreased by 100 μg / Ml inhibited the expression. The expression level of C / EBPα, another transcription factor, was decreased in a concentration dependent manner at 50 μg /

[Experimental Example 3] Measurement of weight change according to the treatment period in the administration of Leucon leaf extract

After 5 weeks of age, C57BL6 (Oriental bio, male) was obtained and purified for 7 days. From 6 weeks of age, the extract was administered for 11 weeks.

The test group (60% fat calorie), product number D12492, and Research Diets (New Brunswick, New Jersey, USA) were orally administered once a day in a negative control (normal diet, 10% fat calorie) The population of each group was 10, the dose was 1000 mg / kg and 2000 mg / kg, and the negative control and the positive control were physiological saline. In addition, the test substance was dissolved in the same physiological saline solution to prepare and used daily, and the body weight was measured at intervals of one week from the day of administration.

FIG. 8 is a graph showing changes in body weight during administration of the leaf extract of Leuconostoc sp. For 11 weeks in a high fat diet-induced obese rats. As shown in FIG.

The mean body weight of the treated group (1000 mg / kg) was 4.61 g lower than that of the control group (43.56 g), the body weight of the group was decreased by 10.6%

The mean body weight of the 2000 mg / kg treated group was 5.94 g less than that of the control group (43.56 g), and the body weight of the treated group was decreased by 13.6%.

Thus, from the above results, it can be confirmed that Leuconostoc spider leaf extract has a remarkably reduced body weight and is excellent in the effect of suppressing obesity.

[Experimental Example 4] Hematological analysis in obese rats administered with Leucon leaf extract

The body weight of rats was measured weekly starting from the day of administration of Leaf leaf extract, and feed intake was measured every 2 days.

After the end of the 11-week experiment, the blood collected from the abdominal aorta of obese rats was centrifuged at 3,000 rpm for 10 minutes and then serum was taken. Blood and histological changes of obesity inhibition effects of high - fat diet supplemented with Leucocephalus leaf extract were analyzed using a biochemical automatic analyzer (7020, HITACHI) after taking serum. The results are shown in Table 1 below.

Sex Group / Dose
(Mg / kg) GOT
(U / L) GPT
(U / L) T-Bili
(Mg / dl) T-Chol
(Mg / dl) TG
(Mg / dl) HDL
(Mg / dl) LDL
(Mg / dl) Male G2
Saline Mean 101.78 65.58 0.03 201.97 43.98 100.60 30.28 S.D. 28.30 36.24 0.02 26.41 15.01 8.73 3.46 N 10 10 10 10 10 10 10 G3
1,000 Mean 83.02 31.12 ^* 0.05 217.05 28.55 ^* 103.48 25.20 ^* S.D. 18.02 18.49 0.02 21.20 3.91 3.74 2.75 N 10 10 10 10 10 10 10 G4
2,000 Mean 75.98 22.53 ^* 0.07 ^* 206.78 27.98 ^* 98.94 23.60 ^* S.D. 13.96 5.81 0.01 20.59 5.59 5.52 2.89 N 10 10 10 10 10 10 10

Significantly different from the control by Student's t-test: ^* p < 0.05

As shown in Table 1, in the control group fed only with high fat diets, the triglyceride level was 43 mg / dl compared with 28.55 mg / dl and 27.98 mg / dl, respectively, , And the blood triglyceride levels decreased by 33.6% and 34.9%, respectively.

In contrast, in the group fed with high fat diets, low-density lipoprotein (LDL) was 30.28 mg / dl, compared with 25.2 mg / dl and 23.6 mg / dl in the groups treated with high- And low-density lipoprotein levels decreased by 16.8% and 23.6%, respectively.

Therefore, the above results show that the extract of Leucumber Leaf has a remarkable effect of significantly reducing blood triglyceride and low density protein in blood.

[Experimental Example 5] Inhibitory effect on the accumulation of fat and body fat in obese rats due to ingestion of Leucocephalus leaf extract

The body weight of rats was measured weekly starting from the day of administration of Leaf leaf extract, and feed intake was measured every 2 days.

After 11 weeks of experimentation, blood samples from obese rats were sacrificed, and liver, subcutaneous fat, visceral fat, and testicular fat were extracted to examine the fatty liver, testicular fat, and subcutaneous fat , Visceral fat weight were measured and analyzed. The results are shown in Table 2 below.

Sex Group / Dose
(Mg / kg) Liver
(g) Perirenal
fat
(g) Epididymal
fat
(g) Subcutaneous
fat
(g) Male G2
Saline Mean 1.721 1.038 2.224 2.573 S.D. 0.388 0.241 0.380 0.913 N 10 10 10 10 G3
1,000 Mean 1.399 0.974 2.210 2.377 S.D. 0.326 0.176 0.515 0.810 N 10 10 10 10 G4
2,000 Mean 1.271 ^* 0.987 2.291 2.197 S.D. 0.124 0.154 0.307 0.572 N 10 10 10 10

Significantly different from the control by Student's t-test: ^* p < 0.05

The liver weights of 1000 mg / kg and 2000 mg / kg treated group were 1.399 g and 1.271 g, respectively, which were 18% and 26.4%, respectively, compared with 1.721 g of the control group, Respectively,

The mean subcutaneous fat weight was 2.573g in the group fed high fat diets, while the mean weight was 2.377g and 2.197g in the group treated with Leucocephalus leaf extract, which showed a maximum decrease of 14.6%.

Thus, from the above results, it can be deduced that Leucillife leaf extract significantly reduces body fat and thus has an excellent effect of inhibiting obesity.

Claims (7)

Wherein the extract contains an extract from the bracken tree as an active ingredient. The method according to claim 1,
Wherein the extract is obtained by mixing at least one of two or more parts selected from leaf, stem, fruit, roots, husks and seeds of the bracken tree in the same ratio. The method according to claim 1,
Wherein the composition further comprises one or both of a transferase and an excipient. The method according to claim 1,
The method for extracting the active ingredient from the above-
a) a material processing step for cutting, washing, drying and crushing a desired part by taking a spike;
b) extracting a solution containing the active ingredient from the processed material of step a) through a solvent to prepare an extraction solution;
c) filtering and concentrating the extract solution of step b) to prepare a concentrate;
d) lyophilizing the concentrate of step c); And
and e) pulverizing the lyophilized concentrate of step d). &lt; RTI ID = 0.0 &gt; 8. &lt; / RTI &gt; 5. The method of claim 4,
Wherein the extract solution of step b) is extracted until the concentration of soluble solids reaches 20 - 22 ° brix. 5. The method of claim 4,
The solvent of step b)
It is recommended to use 70% ethanol, purified water, methanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, chloroform ), Ethyl ether, and hexane are mixed in equal proportions. The anti-obesity composition according to claim 1,
The method according to claim 1,
Wherein the active ingredient contains one or both of the components of verbascoside or polyphenol as an active ingredient.

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