KR101059021B1 - Method for producing an inhibitory fraction of Kadipsin S derived from Snow Cordyceps sinensis and composition for improving obesity using the same as an active ingredient - Google Patents

Abstract

The present invention relates to a method for preparing a Kaepepsin S (Cathepsin S) inhibitory fraction derived from Paecilomyces tenuipes and to a composition for improving obesity using the inhibitory fraction as an active ingredient, the Kadipsin S inhibitory fraction according to the present invention Is effective in improving obesity by inhibiting the proliferation and differentiation of adipocytes.

Obesity, Paecilomyces tenuipis, Synthetic Adsorbent, Diaion HP-20, Diaion SP-850, Adipocyte Differentiation, Cathepsin S

Description

Method for production of cathepsin S inhibitory fraction from Paecilomyces tenuipes and use of flowers as antiobesity}

The present invention relates to a method for producing a Kaepepsin S (Cathepsin S) inhibitory fraction derived from Paecilomyces tenuipes and to a composition for improving obesity using the inhibitory fraction as an active ingredient, more specifically using a synthetic adsorbent resin The present invention relates to a method for producing a fraction having excellent anti-obesity effect by removing unnecessary components from the hydrothermal extract of Snow Cordyceps sinensis and selectively concentrating a component having a Kadepsin S inhibitory activity and a composition for improving obesity using the fraction as an active ingredient. .

Globally, the number of obese people is about 150 million, and in Korea, the proportion of obese people is about 30% in 2004 and the rate is increasing every year, which is recognized as a serious health problem. With the improvement of the economic level and the increasing interest in beauty, there have been various methods of treating obesity, ranging from conventional diet and exercise to drug treatment, but some of the obesity inhibitors and diet materials that are overflowing in Korea are Efficacy has not been scientifically validated. There are many ways to treat obesity, such as the cause of appetite suppression, food absorption inhibition, energy metabolism promotion, hormone regulation, and the like. However, despite various mechanisms of action, most obesity drugs have a pronounced initial effect, but gradually decrease with time and become almost ineffective after 6-8 months. In addition, when the drug is stopped, the weight is increased again, and in many cases, the drug is discontinued due to side effects. Currently, the medical community recognizes only two drugs, sibutramine and orlistat, as long-term obesity drugs. However, these drugs also raise questions about side effects.

Paecilomyces tenuipis is a kind of medicinal mushroom in which the fungus Cordyceps fungus is infected with insects and is not affected by the habitat environment and is distributed worldwide in Korea, Japan and Nepal. The chemical composition of Snow Cordyceps sinensis consists of 7.0% of water, 60.9% of crude protein, 2.4% of crude fat, and 6.4% of ash. The protein contains a total of 17 amino acids, including 8 essential amino acids that are indispensable to our body. According to a report from the Korea Food Research Institute, alanine among the amino acids in Snow Cordyceps sinensis promotes alcohol metabolism, which is effective in relieving hangover and protecting liver function, and also lowering blood cholesterol, preventing high blood pressure and stroke. In addition, glycine and tyrosine are effective in preventing dementia and Parkinson's disease. According to the Seed Industry Act, the varieties of Snow Cordyceps with this effect are registered as Silkworm Cordyceps, and the crop name is Snow Cordyceps, and is registered in the Food Code to be used as a food ingredient (July 10, 1998). Is getting.

On the other hand, the present inventors have shown through the Republic of Korea Patent Application Publication No. 10-2008-0068982 the hot water extract of Snow Cordyceps sinensis inhibits the Kadapsin S to prevent the proliferation and differentiation of fat cells, and lower the blood sugar and total cholesterol concentrations. , Has suggested a composition for improving obesity using the above-mentioned snow Cordyceps sinensis hot water extract as an active ingredient.

In addition, the present inventors have led to the present invention by developing a method for separating the substance inhibiting the activity of the Kadipsin S in the snow Cordyceps sinensis hot water extract after repeated studies.

Therefore, the present invention effectively extracts and concentrates the Kadepsin S inhibitory ingredient from Paecilomyces tenuipes , thereby producing an anti-obesity Kadapsin S inhibitor fraction that is superior in anti-obesity effects to the existing obesity inhibitors derived from It is an object of the present invention to provide a method for producing and a composition for improving obesity comprising the above inhibitory fraction as an active ingredient.

In order to achieve the above object, the present invention comprises the steps of: a) first filtering the hot water extract of Paecilomyces tenuipes with a filter paper and then secondary filtering with 0.45㎛ membrane using Celite 545 as a filter aid; b) adsorbing the filtrate of step a) to the synthetic adsorptive resin and then removing the inert components that are not adsorbed by 2 to 5 times the volume of the synthetic adsorptive resin; c) eluting the capsidsin S inhibitory active ingredient adsorbed on the synthetic adsorbent with dilution alcohol of 2 to 5 times the volume of the synthetic adsorbent resin from which the inactive component of step b) is removed; And d) concentration of the eluted kadipsin S inhibitory active ingredient in step c) under reduced pressure to remove alcohol components and lyophilized to powder to form a snowhopper Cordyceps sinensis derived kadepsin S inhibitory fraction. It is characterized by providing a composition for improving obesity, which contains the decayed Sap Depth derived from Snow Cordyceps sinensis prepared as an active ingredient.

Hereinafter, the present invention will be described in detail.

The method for effectively preparing the Kadipsin S inhibitory active ingredient in Snow Cordyceps sinensis is to extract snow Cordyceps sinensis with hot water and then pass it through a synthetic adsorption resin to adsorb the active ingredient, eluting the adsorbed ingredients with a suitable solvent, and then concentrating and powdering. It is made by the process of making a difference.

The manufacturing process will be described in detail for each step as follows.

a) Paecilomyces japonica (Paecilomyces tenuipes) comprising: after filtering the first hot-water extract with a filter paper using a Celite 545 filter aid to the second filter by the filtration membrane 0.45㎛

In the above step, only the missing parts of Paecilomyces tenuipes are collected and pulverized, and 20 times distilled water (L) is added per unit mass (kg), and then heated at 90 to 110 ° C. for 2 hours to obtain snow cauliflower herb hot water extract. Next, the snow Cordyceps sinensis hot water extract was first filtered through a filter paper, and then filtered through a 0.45 μm membrane using celite as a filter aid, and then powdered by freeze drying.

b) adsorbing the filtrate of step a) to the synthetic adsorptive resin and then removing the inert components which are not adsorbed by 2 to 5 times the volume of the synthetic adsorptive resin.

The step is adsorbed by passing the lyophilized powder of Cordyceps sinensis hot water extract of step a) in a suitable volume of water through a synthetic adsorption resin filled in a column, and then adsorbed with 2 to 5 times the volume of the synthetic adsorption resin. It is to remove inactive ingredients that are not.

Here, the synthetic adsorption resin uses a weight equivalent to 5 to 20 times the weight of snow Cordyceps sinensis hot water extract powder. This is because the active ingredients other than the above ranges may be too small or excessive in excess of the adsorption capacity of the resin, resulting in waste of the resin or loss of the active ingredients that have not been adsorbed.

In addition, the synthetic adsorption resin is preferably used Diaion-HP20 or Diaion-SP850, which is a highly porous synthetic adsorbent of a copolymer of styrene and divinyl benzene.

In particular, Diaion-HP20 has a large number of pores, high specific surface area and excellent adsorption capacity, and relatively large pores, making it suitable for adsorption of large molecules (MW 1,500 or more). Elution is easy. In particular, since the surface of the particles is hydrophobic (Hydrophobic), when the organic material is adsorbed to the synthetic adsorbent, hydrophobic groups in the organic molecules are adsorbed. Therefore, the organic matter (ex. Alkyl group of fatty acid) having a small degree of hydrophobic group is easily adsorbed. Elution of the adsorbed organics includes acid, alkali, polar solvent and the like.

In addition, Diaion SP850 has a very large specific surface area and uniform pore size distribution and a very small pore radius compared to Diaion HP20. Because of the large specific surface area and the small pore volume, adsorption of small molecules (<1,000mw) can be selectively performed and macromolecules can be excluded.

c) eluting the capsidsin S inhibitory active ingredient adsorbed on the synthetic adsorbent with dilution alcohol of 2 to 5 times the volume of the synthetic adsorbent resin from which the inactive component of step b) is removed.

The step is to elute the capidine S inhibitory active ingredient adsorbed on the synthetic adsorbent with 2 to 5 times the dilution alcohol of the synthetic adsorbent volume from which the inactive component is removed.

Here, it is preferable to use the diluted alcohol containing 50 to 80% by volume ethanol or 50 to 80% by volume methanol relative to 100% by volume of dilute alcohol. This is because without using 50 to 80% by volume ethanol or 50 to 80% by volume methanol can not effectively elute the active ingredients adsorbed on the resin.

d) concentrating the eluted capepsin S inhibitory active ingredient of step c) under reduced pressure to remove alcohol components and lyophilizing to powder

The step is to remove the alcohol component by concentrating the eluted Kadipsin S inhibitory active ingredient of step c) under reduced pressure and lyophilized to -70 ~ -80 ℃ to powder.

On the other hand, the composition for improving obesity containing an eyelid Cordyceps sinensis derived capidine S inhibitory fraction according to the present invention as an active ingredient can be administered orally during clinical administration and pharmaceutically acceptable according to the pharmaceutical preparation methods known in the art. Carriers and the like may be added to prepare a pharmaceutical composition. In actual clinical administration, it can be administered in a variety of oral dosage forms. When formulated, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, etc. which are commonly used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate (Calcium carbonate) carbonate), sucrose or sucrose, lactose, and gelatin. In addition to simple excipients, lubricants such as magnesium, stiarate and talc are also used. Liquid preparations for oral use include suspensions, solutions, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .

A typical daily dosage of the composition for improving obesity, containing an eyelid Cordyceps sinensis-derived capepsin S inhibitory fraction, as an active ingredient, is in the range of 1-30 mg / kg body weight, and can be administered once or in several divided doses. have. However, the actual dosage should be determined in light of several relevant factors such as the route of administration, the age, sex, weight of the patient and the severity of the patient and the dosage does not limit the scope of the invention in any aspect.

In addition, the Kadipsin S inhibitory fraction of the present invention can be expected to have little toxicity and side effects because it is extracted from the snow Cordyceps sinensis used for food or medicinal use.

In addition, the fermented milk containing the snow deciduous flower-derived Kadipsin S inhibitory fraction of the present invention as an active ingredient, the combination of freeze-dried powder and mixed fruit juice syrup of lactic acid bacteria culture medium and the Kaepsin S inhibitory fraction derived from the snow-derived C. sinensis After homogeneous at 150 bar and cooled to 10 ℃ or less to prepare a fermented milk by packaging in a container.

In addition, the beverage containing the deciduous snow Cordyceps sinensis-derived capidine S inhibition fraction of the present invention as an active ingredient, by combining the mixed juice syrup, lyophilized powder of the decapsule derived from Snow Cordyceps sinensis S. After homogeneous at 150 bar and cooled to 10 ℃ or less to prepare a functional beverage by packaging in a small packaging container such as glass bottles, plastic bottles.

In addition, the health functional food containing the snow deciduous flower-derived Kadipsin S inhibitory fraction of the present invention as an active ingredient in the eye obesity improvement effect of vitamin B1, B2, B5, B6, E and acetate esters, nicotinic acid amides, oligosaccharides and the like may be added, and other food additives may be added.

Kadipsin S is an enzyme involved in the proliferation and differentiation of adipocytes, and it is known that the inhibitory effect of obesity is inhibited when the enzyme is inhibited. Snow capsicum S-derived Kadipsin S inhibitory active fraction according to the present invention is effectively extracted and concentrated, so a small amount of high obesity inhibitory effect can be expected. Therefore, it is possible to apply to a wider range of obesity treatment products.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are not intended to limit the scope of the present invention, and ordinary changes by those skilled in the art are possible within the scope of the technical idea of the present invention.

&Lt; Example 1 >

Isolation and Purification of Kadipsin S Inhibitory Fraction from Snow Cordyceps Sinensis

1-1. Hot Water Extraction and Filtration of Snow Cordyceps Sinensis

After only collecting the dried parts of the dried snow Cordyceps sinensis pulverized completely, 20 times distilled water (L) per unit mass (kg) was mixed and heated at 90 ~ 110 ℃ for 2 hours to obtain a hot water extract. Thereafter, the hot water extract was first filtered through a filter paper (whatman No. 41), and then filtered through a filtration membrane (0.45 μm) using celite 545 as a filter aid.

1-2. Synthetic Adsorbent Treatment with Snow Cordyceps Sinensis Extract

After filling the column with Diaion-HP20, a synthetic adsorption resin of a weight corresponding to 10 times the weight of the powder of the snow Cordyceps sinensis hot water extract of Example 1-1, and washed with alcohol and water in turn. After dissolving the snow Cordyceps sinensis hot water extract of Example 1-1 in water, it was passed through the synthetic adsorption resin Diaion-HP20 filled in the column. The column was washed with water equal to 5 times the volume of the synthetic adsorbent resin to remove inert components that were not adsorbed. The adsorbed active ingredient was eluted with 80% by volume ethanol corresponding to 5 times the volume of the synthetic adsorbent resin, and concentrated under reduced pressure to remove the alcohol component and then powdered using a freeze dryer.

Figure 1 shows the separation process of the Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis hot water extract as described above.

<Example 2>

Preparation of a pharmaceutical composition using a Kadipsin S inhibitory fraction derived from Snow Cordyceps sinensis

Hereinafter, an example of preparation of a composition for improving obesity, which contains the kadipsin S inhibitory fraction according to the present invention as an active ingredient, is not limited thereto.

Manufacture of powder

According to the preparation method of powder in the Pharmacopoeia General Formulation, it is prepared in the following component content per packet.

Kasupsin S Inhibitory Fraction from Snow Cordyceps Sinensis (Dry Powder) 300 mg

Lactose ......................................... 100 mg

Talc ............................................. 10mg

Manufacture of tablets

According to the preparation method of tablets in the Pharmacopoeia General Formulation, it is prepared in the following component contents per tablet.

Kasupsin S inhibitory fraction (dry powder) from Snow Cordyceps sinensis, 300 mg

Corn starch ........................... 100 mg

Lactose ... Mg

Magnesium Stearate ......................................... 2 mg

Manufacture of capsules

According to the preparation method of capsules in the pharmacopeia formulation, it is prepared in the following component content per capsule.

Kasupsin S inhibitory fraction (dry powder) from Snow Cordyceps sinensis ..... 300mg

Corn starch ........................... 100 mg

Lactose ... 100mg

Magnesium Stearate ......................................... 2 mg

Preparation of liquid

According to the method for preparing a liquid formulation in the Pharmacopoeia General Formulation, it is prepared in the following component content per 100 ml of liquid formulation.

Kasupsin S inhibitory fraction (dry powder) derived from Snow Cordyceps sinensis ...... 1g

Isomerized sugar ......................................... 10g

Mannitol ......................................... 5 g

Purified water ......................................

<Example 3>

Preparation of Fermented Milk Containing Kadipsin S Inhibitory Fraction from Snow Cordyceps Sinensis as an Active Ingredient

Method for producing a fermented milk consisting of a freeze-dried powder and mixed juice syrup of lactic acid bacteria culture medium and Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis of Example 1 is as follows.

The lactic acid bacteria culture medium was stirred at 95.36% of crude milk and 4.6% by weight of skim milk powder (or mixed milk powder), and the specific gravity at 15 ° C was 1.0473 to 1.0475, the titratable acidity was 0.200 to 0.220%, the pH was 6.65 to 6.70, and the brix at 20 ° C. (Brix ^o ) was mixed to 16.3 ~ 16.5%. Then, UHT heat treatment (sterilized at 135 ℃ for 2 seconds), cooled to 40 ℃, Streptococcus thermophilus bacteria and lactose (Valley laboratory, USA) each added 0.02% by weight Incubated for 6 hours, the total lactic acid bacteria in BCP medium was prepared by 1.0 × 10 ⁹ cfu / ㎖ or more, the titratable acidity is 0.89 ~ 0.91%, pH is 4.55 ~ 4.65.

Mixed fruit syrup consists of 10% by weight liquid fructose, 3% by weight white sugar, 3% by weight brown sugar, 10% by weight mixed juice concentrate (56 Brix ^o ), 0.1% by weight pectin, 0.05% by weight fresh fruit mix essence and 73.85% by weight purified water After mixing by stirring at 30 ~ 35 ℃ UHT heat treatment (sterilized for 2 seconds at 135 ℃) was prepared by cooling.

75% by weight of the lactic acid bacteria culture medium prepared by the above method and 0.1% by weight of the lyophilized powder of the Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis of Example 1 and 24.9% by weight of the mixed fruit juice syrup prepared by the above method was homogeneous at 150 bar After cooling to 10 ° C or less to prepare a fermented milk containing the kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis having an effect of improving obesity as an active ingredient.

<Example 4>

Beverage containing Kadipsin S inhibitory fraction derived from Snow Cordyceps sinensis as an active ingredient

Method for producing a functional beverage consisting of a lyophilized powder and mixed juice syrup of the decayed Sap derivation from Snow Cordyceps sinensis of Example 1 is as follows.

Mixed fruit syrup consists of 10% by weight liquid fructose, 3% by weight white sugar, 3% by weight brown sugar, 10% by weight mixed juice concentrate (56 Brix ^o ), 0.1% by weight pectin, 0.05% by weight fresh fruit mix essence and 73.85% by weight purified water After mixing by stirring at 30 ~ 35 ℃ UHT heat treatment (sterilized for 2 seconds at 135 ℃) was prepared by cooling.

29.9% by weight of the mixed fruit juice syrup prepared by the above method and 0.1% by weight of the lyophilized powder and 70% by weight of sterile purified water of the snowhopper Cordyceps derived Kadipsin S inhibitory fraction of Example 1 were homogenized at 150 bar and then 10 ° C. or less. After cooling to and packed in a small packaging container such as glass bottles, plastic bottles to prepare a functional beverage containing the Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis having an effect of improving obesity as an active ingredient.

Example 5

Health functional food containing Kadipsin S inhibitory fraction derived from Snow Cordyceps sinensis as an active ingredient

To 0.1% by weight of the lyophilized powder of the snowhopper Cordyceps sinensis derived Kadipsin S inhibited fraction of the first ingredient, vitamin A supplement (vitamin B1, B2, B5, B6, E and acetate ester, nicotinic acid amide) and oligosaccharides in Example 1 100 parts by weight of the lyophilized powder of the decayed S. derivation of Snow Cordyceps sinensis was added in an amount of 10 parts by weight and mixed in a high speed rotary mixer. 10% by weight of sterile purified water was added to the mixture, mixed, and molded into granules having a diameter of 1 to 2 mm. The molded granules were dried in a vacuum dryer at 40 to 50 ° C., and then passed through 12 to 14 meshes to uniformly prepare granules. The granules prepared as described above were extruded by appropriate amounts into tablets or powders or filled into hard capsules to produce hard capsule products.

<Test Example 1>

Determination of Kadipsin S Inhibitory Activity of Snow Cordyceps Sinensis Fractions

In vitro enzyme reaction was carried out using the Kadsin S enzyme and the Z-Val-Val-Arg-AMC peptide as substrates to determine the Kadipsin S inhibitory activity of the Snow Cordyceps sinensis fraction purified in Example 1 above. Was carried out. Peptides used as substrates fluoresce when decomposed by Kadipsin S, so the activity of Kadipsin S can be confirmed by measuring the amount of fluorescence after the reaction.

Enzyme reaction was carried out in the buffer solution consisting of 100mM potassium phosphate (pH 6.5), 5mM DTT, 4mM EDTA, and fractions of snow Cordyceps sinensis hot water extract purified in Example 1 above and 7ng of kadepsin S enzyme (human, recombinant from biomol), and the peptide Z-Val-Val-Arg-AMC was added as a substrate to a final concentration of 0.8 mM and mixed to a total volume of 250 μl, followed by reaction at 37 ° C. for 25 minutes. The Kadipsin S inhibitory activity was calculated by measuring the amount of fluorescence (excitation / emission = 360 nm / 460 nm) every 5 minutes during the reaction, and applying this value to the following formula.

Kadipsin S inhibitory activity (%) = [{(C-B)-(S-B)} / (C-B)]] ㅧ 100

C: Fluorescence value of positive control without sample

B: Fluorescence value of negative control without enzyme

S: Fluorescence value of the test sphere to which the sample was added

The results are shown in FIG.

As can be seen in Figure 2, the active fractions eluted with alcohol after adsorbing the Snow Cordyceps hyacinth hot water extract to the synthetic adsorption resin, the activity was very high Kadapsin S inhibitory activity, but is not adsorbed to the Snow Cordyceps S. hot water extract and synthetic adsorption resin Washed out inactive fractions were relatively very low.

Further, Paecilomyces japonica hot-water extract, then the inactive fractions and the enzyme reaction by treating the active fraction for each concentration to obtain IC ₅₀ values are shown in Table 1 below.

As can be seen in Table 1, it was found that the IC ₅₀ value for the Kadipsin S was reduced to about one-fifth through the fractionation process using the synthetic adsorbent resin of the Snow Cordyceps sinensis extract.

Therefore, the present invention was confirmed to be effective in selectively purifying fractions that effectively inhibit the activity of the Kadipsin S by separating the snow Cordyceps sinensis hot water extract using a synthetic adsorption resin.

<Test Example 2>

Obesity Inhibition of Kadipsin S Inhibitory Fraction from Snow Cordyceps Sinensis

2-1 Measurement of weight gain inhibitory effect

In order to determine the obesity inhibitory effect of the Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis according to the present invention, animal experiments using a high fat diet were conducted. Experimental animals were used after adapting C57BL / 6 mice to a three-week-old male with a high-fat diet for one week. Eight rats of all experimental groups were placed completely and the experimental groups were five groups. The composition ratio of the feed administered to each experimental group is shown in Table 2 below. The high-fat diet was D12451 (45% fat, 5.252 cal / g) from Research diets, and the powders of snow Cordyceps sinensis (Hyper Cordyceps sinensis hot water extract, inactive fraction, and active fractions) added to the feed of each experimental group were lyophilized and powdered. After the conversion, 0.2 ~ 1% (w / w) of the high fat diet was mixed well with the high fat diet.

Here, the experimental group ingested the active fraction (kadipsin S inhibitory fraction derived from Snow Cordyceps sinensis), which was eluted with alcohol after adsorbing the high fat diet and Snow Cordyceps Sinensis hot water extract on the synthetic adsorptive resin, was used as the The experimental group ingested 20,000 'high fat diet group', the experimental group ingested high fat diet and 1% snow Cordyceps sinensis extract, the '1% hot water extract group', and the high fat diet and 0.2% Snow Cordyceps hot water extract. The experimental group ingested was the '0.2% hot water extract group' and the non-adsorbed fraction group was the experimental group that consumed the inactive fraction washed out without being adsorbed by the high fat diet and the synthetic adsorbent.

For each experimental group, the experimental diet was administered for a total of four weeks, and body weight and feed intake were measured at two intervals per week.

The results are shown in Figure 3 and Table 3.

As can be seen in Figure 3 and Table 3, the 0.2% hot water extract group and the non-adsorption fraction group showed a weight gain similar to that of the high-fat diet group, showing almost no weight gain inhibitory effect, whereas the Kadipsin S inhibitor group Compared to the high-fat diet group, weight gain was significantly suppressed, and the weight gain was also suppressed in the 1% hot water extract group. By the way, when comparing the weight gain of the Kadipsin S inhibitor group and the weight gain of the 1% hot water extract group, it can be confirmed that the weight gain of the Kadipsin S inhibitor group is 1/5 of the weight gain of the 1% hot water extract group. From these results, it was found that the weight-inhibition effect of the active fraction eluted with alcohol after adsorbing the Cordyceps sinensis hot water extract to the synthetic adsorption resin was more than five times higher than the weight-inhibition effect of the Snow Cordyceps sinensis hot water extract.

2-2 Measurement of Changes in Adipocyte Size

In addition, in order to compare the size of the adipocytes of each of the experimental groups of 2-1, the animals were sacrificed and then opened, abdominal fat was extracted, immediately fixed in 10% formalin solution, and the tissues were sectioned to perform Hematoxylin & Eosin staining. Adipose cells of each experimental group were observed by light microscopy.

The results are shown in FIG.

As can be seen in Figure 4, in the 0.2% hot water extract group and the non-adsorption fraction group, as in the weight change results of Test Example 2-1, the size of fat cells did not decrease, as in the high-fat diet group, while Kadsin S Inhibition group and 1% hot water extract group showed much smaller fat cell size than high-fat diet group, indicating that weight-inhibiting effect was due to fat cell proliferation and differentiation inhibition.

<Test Example 3>

Cordycepin Analysis of Kadipsin S Inhibitory Fraction from Snow Cordyceps Sinensis

3-1 Cordycepin Content Analysis

According to Republic of Korea Patent No. 10-0799116 cordycepin (cordycepin) is shown to play a role in inhibiting the differentiation of fat cells. Therefore, in order to confirm whether the weight increase inhibitory effect of the snow Cordyceps sinensis-derived Kadipsin S inhibitory fraction according to the present invention is derived from cordycepin, the Codyselin Strain extract from Snow Cordyceps sinensis and the Kadsin S inhibitory fraction derived from Snow Cordyceps sinensis were used by HPLC. Sepine content was analyzed.

HPLC analysis was performed using UV detector (280nm), column was Agilent Eclipse XDB-C18 (4.6 × 250mm, 5㎛), mobile phase was Deionized Water: Methanol: Formic acid (85: 14: 1). 5 is shown.

As can be seen in Figure 5, while a small amount of cordycepin was detected in the snow Cordyceps sinensis hot water extract, it was found that cordycepin was not detected in the Kadipsin S inhibitory fraction.

3-2 Determination of Cordycepin Inhibition of Kadipsin S

In addition, in order to confirm whether the weight increase inhibitory effect of the snow Cordyceps sinensis-derived capepsin S inhibitory fraction according to the present invention is derived from cordycepin, whether or not cordycepin was inhibited by capepsin S was tested in the same manner as in Test Example 1 above. The results are shown in FIG.

As can be seen in Figure 6, it was found that cordycepin does not have a capidine S inhibitory activity.

Through the above Experimental Example 3, it was confirmed that the snow Cordyceps sinensis hot-water extract according to the present invention has a weight gain inhibitory activity by other kadepsin S inhibitors other than cordycepin.

1 is a view showing a process for preparing a decay S-derived fraction derived from Snow Cordyceps sinensis.

Figure 2 is a graph showing the Kadipsin S inhibitory activity of the fractions isolated from Snow Cordyceps.

Figure 3 is a graph showing the average weight change of the four weeks of each experimental group in the animal experiment.

Figure 4 is a photograph showing the results of observation of the adipose tissue sections of each experimental group by optical microscope after staining with Hematoxylin & Eosin (200 times).

FIG. 5 is an HPLC analysis chromatogram of Kadipsin S inhibitor.

Figure 6 is a graph showing the results of experiments on the capidine S inhibitory activity of cordycepin.

Claims (9)

a) first filtering the hot water extract of Paecilomyces tenuipes with a filter paper and then secondary filtering with 0.45㎛ membrane using Celite 545 as a filter aid; b) adsorbing the filtrate of step a) to the synthetic adsorptive resin and then removing the inert components that are not adsorbed with water of 2 to 5 times the volume of the synthetic adsorptive resin; c) eluting the capsidsin S inhibitory active ingredient adsorbed on the synthetic adsorbent with dilution alcohol of 2 to 5 times the volume of the synthetic adsorbent resin from which the inactive component of step b) is removed; And d) the method for producing a decay-derived Kaepdsin S inhibited fraction, characterized in that it comprises the step of depressurizing the eluted Kadipsin S inhibitory active ingredient of step c) to remove the alcohol component and lyophilized to powder. The method of claim 1, The method of claim 1, wherein the weight of the synthetic adsorbent resin is 5 to 20 times the weight of the powdered snow Cordyceps sinensis extract powder. The method of claim 1, The synthetic adsorptive resin is Diaion-HP20 or Diaion-SP850, characterized in that the method of producing a snow deciduous Cordyceps sinensis derived Kadipsin S inhibitory fraction. The method of claim 1, The dilution alcohol is a method for producing a snow cabbage leaf-derived Kadipsin S inhibition fraction, characterized in that the ethanol or methanol content of 50 to 80% by volume with respect to 100% by volume of dilute alcohol. Kaepsin S inhibitory fraction derived from Snow Cordyceps sinensis, which is prepared by the method of claim 1. The composition for improving obesity, characterized in that containing the Kadipsin S inhibitory fraction of claim 5 as an active ingredient. Fermented milk, characterized in that containing the Kadipsin S inhibitory fraction of claim 5 as an active ingredient. The beverage characterized by containing the kadipsin S inhibitory fraction of claim 5 as an active ingredient. Health functional food, characterized in that containing the Kadipsin S inhibitory fraction of claim 5 as an active ingredient.

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