KR101430646B1 - Extract of Chrysanthemum indicum leaves having inhibitory effects on protein tyrosine phosphatase 1B and a composition comprising the same for preventing or treating metabolic diseases - Google Patents

Abstract

The present invention provides a chrysanthemum indicum leaf extract for inhibiting protein tyrosine phosphatase 1B (PTP1B) and a composition including the same for preventing or treating metabolic diseases. According to the present invention, the chrysanthemum indicum leaf extract has PTP1B inhibitor activities and the composition including the chrysanthemum indicum leaf extract has excellent effects in preventing adipopexia, reducing blood sugar, and preventing and treating diabetes so as to be useful as an agent for preventing or treating metabolic diseases. Moreover, the composition according to the present invention is appropriate for oral administration so as to have high bio-availability when being used as an agent for preventing and treating metabolic diseases through PTP1B inhibiting functions, and is made from a natural material so that side effects including toxicity can be reduced compared to PTP1B inhibitor compounds which are produced through synthesis.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for inhibiting protein tyrosine dephosphorylase 1B, and a composition for preventing or treating metabolic diseases containing the same,

The present invention relates to a composition for inhibiting protein tyrosine dephosphorylase 1B (PTP1B) and a composition for preventing or treating a metabolic disease comprising the same, and more particularly, to a composition for preventing or treating metabolic diseases, And a composition for preventing or treating metabolic diseases such as diabetes.

In modern society, the quality of life is improved and the life span is prolonged, while the incidence of metabolic diseases is also increasing. Among metabolic diseases, obesity, diabetes, and cancer are rapidly increasing in the industrialized society, especially since the onset age is low. In particular, as the dietary life improves, the amount of calories consumed is greatly increased, resulting in serious obesity and the resulting diabetes is increasing. If diabetes persists for a long time, the blood lipid concentration increases in addition to the increase of blood sugar, leading to complications such as atherosclerosis, cardiovascular diseases such as coronary heart disease, diabetic kidney and retinal disease.

Drugs that are commonly used as diabetes medications include hypoglycemic agents to treat insulin resistance, which lacks insulin production or insulin is sufficient but insufficient to utilize sugars in the body because of low activity. The hypoglycemic agent is divided according to the mechanism of action and the action point of the drug. The sulfonylurea system mainly promotes the movement of granule cells containing insulin in the beta cells of the pancreas, and indirectly secretes insulin. Serious side effects are reported. The acetaminophen medication is a drug that moves sugar to muscle cells and prevents sugar from being produced in the liver. It is used in obese diabetic patients and has the advantage of not causing hypoglycemia, but caution should be paid to the elderly and cardiovascular disease patients. Alpha-glucosidase inhibitors inhibit enzymatic activity in the small intestine to temporarily inhibit blood sugar levels in the postprandial state, and are used for the treatment of mild diabetes because the side effects are not severe. Recently, a thiazolidinedione (TZD) -based drug that activates PPAR-gamma (peroxisome proliferator activated receptor gamma) related to the differentiation of adipocytes has also been developed. However, since such an oral hypoglycemic agent does not effectively lower blood glucose and most of known drugs have side effects such as weight gain, hypoglycemia and edema, it is urgent to develop a safer new therapeutic agent capable of restoring the normal endocrine response of the human body .

Cells use a variety of regulatory mechanisms through phosphorylation as well as protein dephosphorylation to make signaling faster and more accurate. In particular, protein tyrosine dephosphorylase 1B (PTP1B) is an important component of intracellular signal transduction and dephosphorylates several substrate proteins. Tyrosine phosphorylation is a reversible process and plays an important role in cell proliferation, cell differentiation, metabolism, migration, gene transcription, and immune response. The abnormalities of tyrosine phosphorylation are closely related to diseases such as hypertension, arthritis, diabetes, cancer and the like, which are controlled by phosphatase enzyme together with kinase. If these functions do not work properly in the human body, it will lead to disease induction and progression.

In particular, PTP1B is known as an enzyme that induces type 2 diabetes by inducing insulin resistance by interfering with the signal transduction pathway of insulin by causing dephosphorylation of insulin receptor and insulin receptor substrates. PTP1B is also known to cause obesity by inhibiting leptin signal transduction by inhibiting leptin receptor and phosphorylation in Jak during signal transduction of leptin (J. Med. Chem., 2004, 47 (17), 4142 ~ 4146). In addition, in the PTP1B knock-out animal model experiment, autophosphorylation of the insulin receptor was increased, muscle cells or hepatocytes were insensitive to insulin and the insulin resistance was removed, so that type 2 diabetes was not induced, (Science, 1999, 283, 1544 to 1548), it is reported that obesity is suppressed, reproductive function is normal, and incidence of cancer is not high due to low formation, increased resistance to obesity due to overproduction of nutrition (vanadium salts) have been shown to exhibit anti-diabetic effects (J. Med. Chem., 2000, 43 (5), 995-1010). In addition, the relationship between type 2 diabetes and PTP1B is revealed, and the inhibitor thereof is expected to be developed as a therapeutic agent for diabetes (Nat. Rev. Drug. Discov, 2002, 1, 696-709). It has also been reported that PTP1B inhibitors selectively affect type 2 diabetes and obesity (JBC, 2000, 275, 10300-10307). In addition, the association of obesity and diabetes with chromosome 20q13.1, where PTP1B is present, has been reported to be genetically linked to overexpression of PTP1B variant and increased insulin levels.

As such, the biological function of PTP1B and the mechanism of inducing the disease have been revealed, which is a target of new drug development. In particular, the role of PTP1B in metabolic diseases such as diabetes, obesity, hyperlipidemia, and cancer has been elucidated and development of new drugs targeting this enzyme inhibition is progressing.

PTP1B inhibitors known so far include vanadate (sodium orthovanadate), phenylarsine oxide (PAO), dephostatin, stevastelins, Dnacin, Dysidiolide, RK682, and the like. Most of these compounds were made by adding a tyrosine phosphate-like structure to a peptide-like structure-based structure. Since many charges are present in the active site of PTP1B, these inhibitors have poor polarity due to low cell permeability, (Nat. Rev. Drug Discov., 2002, 1, 696-709), which are toxic and other side effects of most synthetic compounds and are not suitable for oral administration.

Therefore, in order to overcome the problems of these conventional PTP1B inhibitors, it has been urgently required to develop PTP1B inhibitors derived from natural materials which can be orally administered and have high stability.

Chrysanthemum indicum is a herbaceous perennial herbaceous plant. Its efficacy is well known, and it is used as medicinal herb to regulate colds, pneumonia, bronchitis, headache, gastritis, enteritis and boils, and is used as a chrysanthemum tea.

The efficacy of the leaf is not known, and most of it is abandoned.

Patent No. 10-0554343 Patent Document 10-2010-0136026 Patent No. 10-0443264

Therefore, the present inventors have continued to study the efficacy of this natural product in order to find out how to utilize the leaves of the mulberry leaves. Surprisingly, the extract of mulberry leaves has an effect of inhibiting the activity of PTP1B enzyme, Has an excellent effect in an obesity / diabetic animal model, and thus has a preventive or therapeutic effect of a metabolic disease, thus completing the present invention.

It is therefore an object of the present invention to provide a Ganoderma leaf extract having PTP1B inhibitor activity and a process for producing the same.

It is still another object of the present invention to provide a composition for preventing or treating metabolic diseases,

In order to accomplish the objects of the present invention, the present invention provides a method for producing a PTP1B inhibitor.

The method for producing a PTP1B inhibitor according to the present invention comprises

I) drying and pulverizing leaves of Chrysanthemum indicum ;

Ii) adding water to the green leaf powder and refluxing and cooling at 70 to 100 ° C for 6 to 9 hours to extract;

Iii) filtering the extract obtained in step ii);

Iv) concentrating the filtrate obtained in step iii) at a reduced pressure at 50 to 65 占 폚; And

V) lyophilizing the concentrate obtained in step iv).

Step i): drying and powdering

The germination leaves can be dried by a conventional method such as natural drying or hot air drying. As in the embodiment of the present invention, it is possible to dry for about 48 hours or more at 50 to 70 DEG C, but is not limited thereto.

Dried mutton leaves can be pulverized by conventional methods such as blinds.

Step ii): Extraction

Water is added to the green leaf powder and the mixture is refluxed and cooled at 70 ~ 100 ℃ for 6 ~ 9 hours. Water is preferably added in an amount of about 15 to 25 times that of the green leaf powder, and the extraction can be repeated twice or more as necessary.

Extraction can be performed using a Soxhlet extractor, but is not limited thereto.

Water is not limited, but preferably distilled water is used.

Step iii): Filtration

The extract obtained in step ii) is filtered to remove by-products such as wastes.

The filtration is preferably first centrifuged using a centrifuge to remove the by-products of the residue first, and then filtered with a filter paper.

Step iv): Concentration

The filtrate obtained in step iii ) is concentrated under reduced pressure at 50 to 65 ° C.

As in the embodiment of the present invention, it is possible to concentrate under reduced pressure using a rotary vacuum concentrator, but the present invention is not limited thereto.

Step v): Freeze-drying

Freeze-dry the concentrate obtained in step iv). The freeze-drying conditions are appropriately determined according to the sample and the extraction solvent.

It was confirmed by the present invention that the extract of P. falciparum had an excellent PTP1B inhibitory activity by measuring the protein chip screening using the insulin receptor as a protein substrate (see Example 2).

In another aspect, the present invention provides a composition for the prevention or treatment of metabolic diseases, including Ganoderma lucidum extract.

In the present invention, metabolic diseases include but are not limited to obesity, diabetes, hyperlipidemia, and cancer.

The composition comprising the extract of Ganoderma lucidum according to the present invention has effects such as lowering of blood glucose, increase of blood insulin concentration, suppression of body fat and inhibition of blood fatty acid in a mouse model of obesity and diabetes mellitus, and the composition is useful for the treatment of metabolic diseases such as diabetes and obesity For prevention and treatment and for suppression of fatty liver (see Examples 3 and 4).

The composition according to the present invention may further comprise a pharmaceutically acceptable carrier or excipient in addition to the extract of Ganoderma lucidum, and may be formulated into preparations for oral administration in the form of tablets, powders, granules, capsules, suspensions and emulsions have.

The effective dose of the active ingredient represented by the Gamma leaf extract may be varied depending on the patient's age, physical condition, body weight, etc., but is generally administered within the range of 10 to 500 mg / kg (body weight) / day. And may be administered once a day or several times a day within an effective daily dose range.

Since the composition according to the present invention is extracted from a natural product, Namgu Leaves, is almost toxic and is for oral administration, it is very easy to take it.

The extract of Ganoderma lucidum according to the present invention has PTP1B inhibitor activity and the composition containing Ganoderma lucidum extract has excellent effects such as blood glucose lowering, increase in blood insulin concentration, inhibition of fatty liver formation, and body fat suppression, . ≪ / RTI >

In addition, since the composition according to the present invention is suitable for oral administration, it is not only highly bioavailable when used as a preventive and therapeutic agent for metabolic diseases through PTP1B inhibitory action, but also has a natural product as its material. Therefore, compared with the PTP1B inhibitor compound prepared through synthesis, Have few side effects.

FIG. 1 is a photograph showing the inhibitory effect of PTP1B on the extract of Chrysanthemum morifolium as measured by a protein chip screening technique.
Fig. 2 is a photograph showing the lipid droplet accumulation inhibitory effect of the Ganoderma lucidum extract of the present invention by staining differentiated 3T3-L1 adipocytes with Oil Red O. Fig.
FIG. 3 is a graph showing the results of measurement of body weight change and food intake by the treatment with Matsutake leaf extract in a DIO mouse model.
FIGS. 4A and 4B are graphs showing the test results of the hypoglycemic effect and the blood insulin concentration-enhancing effect on the DIO mouse model by the treatment with the extract of Leptocorus aureus.
FIG. 5 is a graph showing the results of oral glucose tolerance test by the treatment with the extract of Leuconostoc sp.
FIGS. 6A and 6B are graphs showing changes in blood fatty acid and body fat by the treatment with Momordica fruticosa extract in a DIO mouse model. FIG.

Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples.

Example

Example 1: Preparation of cuttlefish leaf extract

The leaves of Gamguk were purchased from Yanggu residents' farming complex in Gangwon - do in July 2011. 50 g of green tea leaves were dried in a 60 ° C hot air drier for about 68 hours. The dried leaves were hand-crushed and powdered using a blender (world mix, DA280-S, Korea).

The extracts were added to the Soxhlet extractor (AP1240-2000, Joi Lab, Korea) and 20 times of distilled water was added thereto. The extracts were extracted twice by repeatedly refluxing at 80 ° C for 8 hours, followed by centrifugation using a centrifuge (4,000 rpm, 20 minutes, 4 ° C) to remove the by-products firstly, and then filtered through a filter paper (Whatman filter paper, no. 2, diameter 110 mm) to obtain a filtrate.

The filtrate was refluxed in a chiller maintained at 4 ° C while the filtrate was placed in a vessel. The filtrate was rotated in a water bath at 55 to 65 ° C, and the filtrate was centrifuged using a rotary vacuum evaporator NE, EYELA, Tokyo Fikakikai And the concentrate was freeze-dried (-85 ° C, vacuum; more than 42 hr at 5 mTorr) in a freeze dryer (FD20L, Ilshinrap, Korea).

The yield rate of the extract of the Korean lettuce leaf was 28.7%.

Example 2: Verification of PTP1B inhibitory activity using protein chip screening technique

The inhibitory activity against PTP1B was verified by protein chip screening technique.

Specifically, insulin recombinant protein (SignalChem, USA) was used as a protein substrate. Incubation was carried out at 30 DEG C for 1 hour for protein substrate phosphorylation. A small amount (1 μl) of the phosphorylated material was spotted onto coated slides (Proteogen Co., Korea) and allowed to bind overnight at 4 ° C. The next day, the cells were washed with washing buffer (PBS buffer, 5% Tween 20, pH 7.4) for 10 minutes, blocked with blocking buffer (PBS buffer, 3% BSA, pH 7.4) for 1 hour at room temperature, . As such, a slide chip prepared by coating a protein substrate, an insulin receptor, was used.

In order to observe the phosphorylation inhibition effect of protein substrate, we divided into 4 groups as follows. (0.5 μl) was used as a control, and PTP1B buffer (2 × assay buffer [20 mM HEPES, 0.2 mM EDTA, 0.2 mM EGTA, 20 mM DTT], 20 mM DTT and 10% PEG) , And 10% PEG (0.5 μl) + PTP1B (0.5 μl, 100 ng / μl) was used as a negative control and sodydimorsobanadate (Na ₃ VO ₄ , (0.5 μl, 100 μg / ml in DW) prepared in Example 1 was used as a sample group (denoted by Chry), and PTP1B (0.5 μl, final 800 μM) + PTP1B (0.5 l, 100 ng / l) was used.

Each group was treated with the protein chip prepared above, and then subjected to dephosphorylation reaction at 30 ° C for 1 hour, followed by washing in the same manner as described above. After washing, anti-phosphoinsulin receptor antibody (Catalog No. 44804G, Invitrogen, USA) was diluted to 1/100 with dilution buffer (10% BSA, 30% glycerol in 1x PBS) (Anti-rabbit IgG-Cy5, Catalog No. A10523, Invitrogen, USA) conjugated with Cy5 fluorescent material, followed by incubation at 30 ° C for 1 hour, washing in the same manner as described above, Was diluted to 1/100 with a dilution buffer and treated with 1 쨉 l of each, incubated at 30 째 C for 1 hour, washed for 10 minutes, and images were scanned using a microarray scanner system (GenePix 4300A, Molecular Devices) , And the photographs thereof are shown in Fig.

As can be seen from FIG. 1, the inhibition rate of 93% (when the negative control was 0%) compared with the negative control group (NC) of the extract of Chukji leaf extract (Chry) showed high activity inhibition rate, and the commercial PTP1B inhibitor, (P / C), which is similar to Orbovanadate (Na ₃ VO ₄ ) positive control (P / C).

Example 3: Inhibition of fat accumulation by 3T3-L1 adipose precursor cells and Oil Red O staining method

The lipid droplet accumulation inhibitory effect of the extract of the present invention was verified by staining differentiated 3T3-L1 adipocytes with Oil Red O.

3T3-L1 adipose precursor cells (ATCC, USA) were cultured in DMEM (Dulbecco's modified Eagle's medium) medium (100 mM HEPES, 50 IU penicillin, and 50 ug streptomycin / ml, containing 10% bovine calf serum, Co., USA) at 37 ° C and 5% CO ₂ . When 3T3-L1 adipose precursor cells exhibited about 100% confluency to induce differentiation of 3T3-L1 adipose precursor cells, MDI solution (0.5 mM 1-methyl-3-iso-butyl- 1-methylxanthine, 1.0 μM desamethasone, 10 μg / ml insulin) (Sigma Co, USA).

3T3-L1 adipose precursor cells were differentiated for 3 days in a DMEM culture medium containing 10% fetal bovine serum (FBS), and a sample group (50 mu g / ml, 100 mu g / ([(+/-) - 5- [4- (1-Methylcyclohexymethoxy) -benzyl] thiazolidine-2,4-dione, C ₁₈ H ₂₃ NO ₃ S, Calbiochem Co., Germany) at a final concentration of 10 mM.

As shown above, the fat cells were treated with the MDI solution and the samples of each group, and oil red O staining (Sigma Co, USA) was performed to observe the adipocytes using the completely differentiated cells after 7 to 8 days. Specifically, after removing DMEM containing 10% FBS, the cells were washed with PBS buffer 2-3 times, fixed with 10% formalin at room temperature for 2 hours, and washed once with 60% isopropanol. The washed cells were stained with Oil Red O staining reagent (Sigma Co, USA) that specifically reacted with lipid at room temperature for 10 minutes. After dyeing, the dye was removed and washed 3-4 times with distilled water. The wells were then filled with distilled water and observed under a microscope. The resulting photographs are shown in FIG.

As can be seen from FIG. 2, it was found that the treatment of the sample group (50, 100, 200 ㎍ / ml) of the extract of the mulberry leaves inhibited lipid accumulation at each treatment concentration as compared with the positive control group.

Example 4. Verification of metabolic disease treatment effect using obesity-induced mouse model

An obesity-induced mouse model was used to demonstrate the therapeutic effect of the extract of Ganoderma lucidum extract of the present invention on metabolic diseases.

Four weeks old C57BL / 6 mice were purchased and classified into normal group and obesity / diabetic test group (7 animals). After adjusting to the animal environment for 1 week, the experimental group induced weight gain through high fat diet (HFD), D12492, 60 kcal% fat, Research Diets, New Brunswick, NJ for 12 weeks, (Diet-induced obese (DIO) model). During the experimental period, the animals were housed in an animal room set at a temperature of 22 ± 2 ° C, a relative humidity of 50 ± 5%, and a lighting time of 12 hours, and water was freed of purified water by microfiltration.

Mice in the normal group and the experimental group were administered with PBS for 3 weeks. By 19 weeks, the extracts of Momordica japonica leaf extract, the control drug (SITA for diabetes and Sibu, a treatment for obesity) ) For 3 weeks. The extracts were used as 20 mpk (mg per kilo) and 100 mpk. Citragliptin, a diabetes drug, and sibutramine, a treatment for obesity, were used as control drugs at 10 mpk and 5 mpk, respectively. The extracts and drugs were dissolved in 0.5% carboxy-methyl cellulose (CMC) solution and the route of administration was oral and daily (once / daily).

Example 4-1. Verification of anti-obesity effect by measuring weight change

The weight change of the mice was measured using an animal scales for 4 weeks at intervals of 3 days, and the results are shown in FIG.

As shown in FIG. 3, in the control group (n = 5, the vehicle-administered group), 48 g on average, 45 g on day 3, 48 g on day 8, 47 g on day 11, 47 g on day 14, 47 g on day 18, 48 g, 26 g 49 g, and 29 g 49 g. The administration of sibutramine (Sibu), an obesity treatment, resulted in an average of 46 g daily, 44 g daily, 44 g daily, 44 g daily, g, 14 g on day 46 g, 18 g on day 47 g, 22 g on day 47 g, 26 g on day 48 g, and 29 g on day 29.

In the experimental group (n = 5) administered with 100mpk of Chrysanthemum morifolium Leaf Extract (Chry), the average was 46 g, 3 days 43 g, 8 days 44 g, 11 days 45 g, 14 days 46 g, 18 days 46 g, 22 47 g for day, 48 g for 26 days, and 48 g for 29 days. These results are similar to those of sibutramine, which is a treatment for obesity.

As a result of observing the amount of feeds consumed by animals, the amount of feed consumed was almost the same in all groups, even though weight loss was reduced in the groups treated with the extract of Leptoptera frugiperum and the group treated with obesity . From these results, it can be confirmed that the extract of Ganoderma lucidum according to the present invention has an anti-obesity effect.

Example 4-2. Verification of blood glucose lowering effect and blood insulin concentration enhancement effect

Experiment 18 One week before the autopsy, glucose was orally administered to the mice, and blood glucose levels due to the administration of the samples in each group were measured with an Accu-Check glucometer (Roche) Respectively. Serum insulin concentration was measured by enzyme-linked immunosolvent (Millipore, MO, USA). The results are shown in Figs. 4A and 4B, respectively

<Blood sugar drop>

As shown in FIG. 4A, glucose concentration was 378.3 mg / dl and 363.9 mg / dl, respectively, in the group administered with 20 mpk and 100 mpk of Chrysanthemum morifolium leaf extract (Chry) Respectively. This blood glucose reduction lasted up to 90 minutes.

<Increase in blood insulin concentration>

As shown in FIG. 4B, the amount of insulin released at 20 minutes was 7.44 ng / ml and 9.04 ng / ml in the case of the extract of Ganoderma lucidum extract, 1.4 times and 1.7 times that of the control group, respectively.

Therefore, it can be confirmed that the extract of Ganoderma lucidum according to the present invention can be used as a preventive and therapeutic agent for obesity and diabetes by demonstrating the effect of blood glucose lowering and blood insulin increase in an obesity / diabetic mouse model.

Example 4-3. Verification of biologic glucose processing ability by oral glucose tolerance test

Oral glucose tolerance test (OGTT) was performed to observe the glucose processing ability of the living body.

After fasting the day before, the normal mice (the untreated group) were treated with 10 mg and 5 mpk of contrast agents, ie, diabetic treatment, cyproglitin, and obesity treatment, respectively, and 20 mpk and 100 mpk 0.5% CMC, respectively. After 30 minutes, each mouse was administered glucose bolus (2 g / kg solved in saline; Sigma-Aldrich) and blood was collected from the tail vein at 0 minute, 20 minute, 40 minute, and 60 minute intervals The blood glucose concentration was measured with an Accu-Check glucometer (Roche) and the results are shown in FIG.

The concentration of blood glucose decreased about 4 times compared to the control (vehicle administered), and especially when compared to the positive control group, sitagliptin (Sita) Of the total blood glucose concentration.

Example  4-4. Blood fatty acid and body fat measurement Anti-obesity  Verification of effect

<Blood fatty acids>

After completion of the experiment, the animals were fasted for 24 hours and anesthetized with ether. Blood was collected from the abdominal vein and centrifuged at 3000 rpm for 10 minutes to separate the serum. The separated serum was stored in a -70 ° C freezer And thawed during the experiment. Serum concentrations of triglyceride (TRIG), HDL-cholesterol (HDL), cholesterol (CHOL) and LDL-cholesterol (LDL) were measured using a Cobas C111 automatic analyzer (Roche, Basel, Switzerland) And the results are shown in Figs. 6A and 6B.

As shown in FIG. 6A, when LDL was administered 20 mpk of Chrysanthemum morifolium extract, the vehicle-treated control (46.9 mg / dl) and the positive control (citagliptin: 49.43 mg / dl; sibutramine: 52.84 mg / dl).

As shown in FIG. 6B, CHOL decreased to a level similar to that of the patients treated with Citigliptin (231.89 mg / dl), which is a therapeutic agent for diabetes mellitus, compared to those treated with sibutramine (259.9 mg / dl) Respectively. In particular, in the case of TRIG, when the extract was administered, the extract was reduced to a level similar to that of the positive control.

From these results, it can be seen that the extract of Ganoderma lucidum extract has an anti-obesity effect.

<Body fat>

Mice were sacrificed, and inguinal fat, epididymal fat, and liver fat were extracted and washed with physiological saline, and the water was removed with a filter paper, and the weight was measured. The results are shown in Table 1.

weight Groin Epididymis Liver fat Total fat Lean Mean 31.00 1.92 1.33 4.22 3.25 S.D. 1.90 1.39 0.94 0.17 2.31 Vehicle Mean 49.10 2.57 5.92 5.18 8.50 S.D. 3.10 0.53 0.65 0.85 1.04 Sita
10 mpk Mean 48.90 2.82 5.52 4.65 8.34 S.D. 3.70 0.14 1.12 0.71 1.24 Sibu
5 mpk Mean 48.60 2.54 5.17 5.52 7.71 S.D. 1.80 0.65 0.38 0.88 0.53 Chry
20 mpk Mean 49 2.80 5.42 4.18 8.23 S.D. 3.11 0.67 0.73 0.67 1.24 Chry
100 mpk Mean 48.10 2.79 5.84 4.32 8.63 S.D. 3.30 0.55 0.90 0.95 1.08

As shown in Table 1, epididymal fat was reduced by 8.45% and 1.4%, and liver fat by 19.31% and 16.61%, respectively, compared with the control group (vehicle-treated group) Especially, liver fat decreased more than control group.

From these results, it can be seen that the extract of Ganoderma lucidum extract has an anti-obesity effect.

Claims (12)

I) drying and pulverizing the transplanting leaves;
Ii) adding water to the green leaf powder and refluxing and cooling at 70 to 100 ° C for 6 to 9 hours to extract;
Iii) filtering the extract obtained in step ii);
Iv) concentrating the filtrate obtained in step iii) at a reduced pressure at 50 to 65 占 폚; And
And (v) lyophilizing the concentrate obtained in step (iv), wherein the protein tyrosine dephosphorylase 1B (PTP1B) inhibitor is used as an active ingredient. A composition for preventing or treating diabetes and obesity comprising the protein tyrosine dephosphorylase 1B (PTP1B) inhibitor prepared according to the production method of claim 1 as an active ingredient. The composition for preventing or treating diabetes and obesity according to claim 2, wherein the composition is effective for lowering blood glucose and increasing insulin concentration in blood. The composition for preventing or treating diabetes and obesity according to claim 2, wherein the composition is effective for inhibiting the blood concentration of fatty acid. The composition for preventing or treating diabetes and obesity according to claim 2, wherein the composition is effective for inhibiting fatty liver. A composition for prevention or treatment of diabetes and obesity comprising an extract of watery mildew (water) as an active ingredient. delete The composition for preventing or treating diabetes and obesity according to claim 6, wherein the diabetes is type 2 diabetes. The composition for preventing or treating diabetes and obesity according to claim 6, wherein the composition further comprises a pharmaceutically acceptable carrier or excipient. The composition for preventing or treating diabetes and obesity according to claim 6, wherein said composition is for oral administration. [Claim 11] The composition for preventing or treating diabetes and obesity according to claim 10, wherein the composition is formulated into any one selected from the group consisting of tablets, powders, granules, capsules, suspensions and emulsions. delete

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