KR20140073631A - Composition comprising extract of grifola frondosa fruit body - Google Patents

Abstract

The present invention relates to a composition comprising a Grifola frondosa fruiting body extract which contains polysaccharides of a particular molecular weight. The composition of the present invention can promote synthesis of collagen by activating elastase and inhibiting collagenase (MMP-1), thereby has an effect for improving skin elasticity and alleviating skin wrinkles.

Description

Technical Field [0001] The present invention relates to a composition comprising a fruiting body extract,

The present invention relates to a composition containing a leaf mushroom fruit body extract comprising a polysaccharide having a specific molecular weight.

Grifola frondosa is a mushroom belonging to the genus Aphyllphorales, Polyporaceae and Grifola in the taxonomic classification. It is a species of oak, broad-leaved, It is a white rot fungus which occurs in bundles around the branches and roots of dead wood, and is distributed in Korea, Japan, Europe, America and so on. Leaf mushroom is an edible but highly functional pharmacological mushroom that increases the immunity of the human body and is known to have excellent tumor suppression. Leaf mushroom has also been reported to be effective in inhibiting cancer by raising the activity of natural killer cells and macrophages involved in immunity.

The inventors of the present invention have found that the extract of fruiting body of the mushroom containing polysaccharide having a specific range of molecular weight has various effects besides anticancer, and thus completed the present invention.

Korean Patent No. 10-0438009

It is an object of the present invention to provide a composition comprising a leaf mushroom fruiting body.

In order to achieve the above object, the present invention provides a composition comprising an extract of fruiting body of mushroom fungus, wherein the extract comprises a polysaccharide having a molecular weight of 2,000 to 10,000 Da.

The composition of the present invention can activate the elastase and inhibit collagenase (MMP-1) to promote the synthesis of collagen, and can be used for anti-aging since it has the effect of improving skin elasticity and improving skin wrinkles.

The composition of the present invention also promotes the differentiation of keratinocytes and restores skin barrier function, thereby increasing the skin moisturizing power and inhibiting lipogenesis with antibacterial activity.

In addition, the composition of the present invention has an inflammation-improving effect, and it is useful because it alleviates itching and has no irritation.

In one aspect, the present invention relates to a composition comprising an extract of Fusarium oxysporum fructus, wherein the extract comprises a polysaccharide having a molecular weight of 2,000 to 10,000 Da.

In the present specification, " Grifola frondosa "is a mushroom belonging to the family Aphyllphorales, Polyporaceae, and Grifola in taxonomic classification. It is a species of oak, It is a white rot fungus which occurs in bundles around the roots and distributed in Korea, Japan, Europe, USA.

In the present specification, "fruiting body" means a collection of sporangia. Most mushrooms are composed of mycelium and fruiting bodies. Mycelia are the root parts of mushrooms, and fruiting bodies are the visible parts.

In the present specification, "extract of leaf mushroom fruiting body" is obtained by, for example, washing and drying dried leaf mushroom fruiting body or a hyssomalized leaf mushroom fruiting body obtained by extracting it in water or organic solvent, The residue and the filtrate are separated by centrifugation, and the separated filtrate is concentrated under reduced pressure to obtain an extract of fruiting body mushroom fruiting body. The organic solvent usable in the present invention may be selected from ethanol, methanol, butanol, ether, ethyl acetate, chloroform or a mixed solvent of these organic solvents and water. In view of the safety of raw materials, Of ethanol is used. After obtaining the extract by using the solvent, a liquid substance can be obtained by freezing, heating and filtering at room temperature by a conventional method known in the art. Alternatively, the solvent can be evaporated, spray dried or lyophilized But the present invention is not limited thereto, and any method generally used in the art can be used without limitation.

If the molecular weight of the polysaccharide is less than 2,000 Da, the specific structural characteristics of the polysaccharide disappear and sufficient effect is not exhibited. If the polysaccharide is more than 10,000 Da, the skin permeability falls and the sufficient effect can not be exhibited. From the above viewpoint, the molecular weight of the polysaccharide may be 2,200 to 9,800, 2,400 to 9,600, 2,600 to 9,400, 2,800 to 9,200, 3,000 to 9,000, 3,200 to 8,800, or 3,400 to 8,600 Da.

In one aspect of the present invention, the polysaccharide is selected from the group consisting of? -1,3-glucan (β-1,3-glucan) and β-1,6-glucan And includes at least one selected. The general structure of the mushroom polysaccharide is 1,6- [beta] -D-glucose in the main chain of 1,3- [beta] -D-glucose whereas the polysaccharide of the mushroom is 1,6- [beta] -D- The branched chain structure of 1,3- [beta] -D-glucose is mixed in the main chain of glucose, and the efficacy of leaf mushroom is known to be attributed to the specific structure of such leaf mushroom polysaccharide.

In the present specification, "β-glucan" is a polysaccharide containing glucose as a constituent unit, and glucose, which is a monosaccharide constituting it, is bound by β-glycosidic bonds . The structure of the monosaccharide constituting? -glucan is represented by the following formula (1).

As used herein, the term "β-1,3-glucan" means a structure in which β-glucan is bonded as 1,3-glycosidic bonds , and "β-1,6-glucan" may mean a structure in which β-glucan is bonded as 1,6-glycosidic bonds . For example, the following formula (2).

In one aspect of the present invention, the polysaccharide may be a polymer of glucose. In the present specification, the polysaccharide is obtained by linking monosaccharides of the formula (1) with 1,3-glycosidic bonds or 1,6-glycosidic bonds Or a structure comprising 1,3-glycosidic bonds and / or 1,6-glycosidic bonds. can do. For example, the following Formula 3-5 is used.

In one aspect of the invention, the extract may comprise from 0.001 to 10% by weight polysaccharide based on the total weight of the extract. If the content of the polysaccharide is less than 0.001% by weight based on the total weight of the extract, the effect of anti-aging, moisturizing, antibacterial and anti-inflammatory effects is insignificant. If the content exceeds 10% by weight, In view of the above, the composition of the present invention may contain 0.005 to 9.5% by weight, 0.01 to 9% by weight, 0.03 to 8.5% by weight, 0.05 to 8% by weight, 0.07 to 7.5% by weight, From about 0.1% to about 7%, from about 0.1% to about 6.5%, from about 0.3% to about 6%, from about 0.5% to about 5.5%, or from about 0.7% to about 5%.

In a composition according to an aspect of the present invention, the composition may be an anti-aging agent comprising an extract of a leaf mushroom fruit body as an active ingredient.

As used herein, the term " anti-aging " refers to an effect of slowing down, preventing or preventing the aging process known in the art. Specifically, it is capable of inhibiting the activity of elastase in the skin or effectively inhibiting the expression of collagenase Or to enhance collagen synthesis to promote skin elasticity and improve wrinkles, but is not limited thereto.

In one aspect of the present invention, the composition may enhance skin elasticity or improve skin wrinkles.

In a composition according to one aspect of the present invention, the composition may be a moisturizing agent containing an extract of foliar mushroom fruiting body as an active ingredient. The composition, which is an aspect of the present invention, can enhance the skin barrier function and induce the differentiation of dermal keratinocytes, thereby increasing the moisture content of the skin and making the skin moist. Therefore, the composition as one aspect of the present invention can be usefully used as a composition for preventing or improving dry skin, atopic dermatitis, contact dermatitis, psoriasis, or the like caused by incomplete epidermal differentiation.

In a composition according to one aspect of the present invention, the composition may be an antimicrobial agent containing an extract of a leaf mushroom fruiting body as an active ingredient. The composition, which is one aspect of the present invention, specifically shows excellent antibacterial activity against acne bacteria. Therefore, the composition can be effectively used in the treatment and alleviation of acne.

In one aspect of the present invention, the bacterium comprises an acne bacterium.

The present invention relates to an antiinflammatory composition comprising soybean root extract as an active ingredient from an aspect. The composition, which is an aspect of the present invention, can alleviate skin irritation and alleviate inflammation, and thus can be used for anti-inflammation. Specifically, the composition of the present invention may have an anti-allergic effect by inhibiting or inhibiting the expression of prostaglandin or interleukin, or by inhibiting or inhibiting the activity of prostaglandin or interleukin.

In one aspect of the invention, the composition may comprise from 0.001 to 10% by weight of the extract, based on the total weight of the composition.

If the content of the extract is less than 0.001% by weight based on the total weight of the composition, the anti-aging, moisturizing, antibacterial and anti-inflammatory effects are insignificant. If the content is more than 10% by weight, In view of the above, the composition of the present invention is used in an amount of 0.005 to 9.5% by weight, 0.01 to 9% by weight, 0.03 to 8.5% by weight, 0.05 to 8% by weight, 0.07 to 7.5% by weight, 7 to 10 wt%, 0.1 to 6.5 wt%, 0.3 to 6 wt%, 0.5 to 5.5 wt%, or 0.7 to 5 wt% of the extract.

In one aspect of the present invention, the composition comprises an external preparation for skin application.

In one aspect of the invention, the composition comprises a cosmetic composition.

When the composition for external application for skin according to the present invention is formulated in the form of a cosmetic composition, it may be formulated into a form of cosmetics such as a softening agent, a convergent lotion, a nutritional lotion, an eye cream, a nutrition cream, a massage cream, a cleansing cream, a cleansing foam, a cleansing water, And the formulations thereof are not particularly limited. In addition, the composition according to the present invention can also be used as a lubricant, an organic solvent, a solubilizer, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, The active ingredient may be combined with any other ingredients commonly used in cosmetics, such as ionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, And may contain adjuvants commonly used in the same cosmetic or dermatological fields. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields. In addition, the composition of the present invention may contain a skin absorption promoting substance to increase the skin improving effect.

The cosmetic composition according to the present invention may contain, in addition to the above-mentioned substances, other ingredients which can give a synergistic effect to the main effect, so long as they do not impair the main effect. The cosmetic composition according to the present invention may further comprise a moisturizing agent, an emollient agent, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cold agent or a limiting agent. The compounding amount of the above components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present invention. The amount thereof may be 0.01 to 5% by weight, specifically 0.01 to 3% by weight, have.

In one aspect of the invention, the composition comprises a health food composition.

The composition according to the present invention provides various types of food additives or functional foods containing them. The composition may be formulated into fermented milk, cheese, yogurt, juice, probiotics, tablets, granules, drinks, caramels, diet bars, and the like, and may be processed into ordinary tea leaves, tea bags, And can be used in the form of various other food additives.

In one embodiment, the composition may contain other ingredients or the like that can give synergistic effects to the main effect within a range that does not impair the intended main effect of the present invention. For example, additives such as perfume, coloring agent, bactericide, antioxidant, preservative, moisturizing agent, thickening agent, inorganic salt, emulsifier and synthetic polymer substance may be further added for improvement of physical properties. In addition, it may further contain auxiliary components such as water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides and seaweed extract. The above components may be mixed and selected without difficulty by those skilled in the art depending on the purpose of formulation or use, and the amount thereof may be selected within a range that does not impair the objects and effects of the present invention. For example, the addition amount of the above components may be in the range of 0.01 to 5% by weight, more specifically 0.01 to 3% by weight, based on the total weight of the composition.

The composition according to the present invention may be in various forms such as a solution, an emulsion, a viscous mixture, a tablet, a powder, etc., and may be administered by various methods such as simple drinking, injection administration, spraying or squeezing.

In one aspect of the invention, the composition comprises a pharmaceutical composition.

When the composition according to the present invention is applied to medicines, it may be formulated into an oral or parenteral dosage form in the form of solid, semi-solid or liquid by adding an inorganic or organic carrier to the composition as an active ingredient.

Examples of the agent for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups and pellets. In addition, the parenteral administration may include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, pills, and the like. In order to formulate the active ingredient of the present invention, it can be easily formulated according to the conventional method. Surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffering agents, suspending agents and other adjuvants can be suitably used.

The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously and the like.

The effective ingredients of the pharmaceutical composition of the present invention will vary depending on the age, sex, weight, pathological condition and severity of the subject to be treated, administration route or judgment of the prescriber. Determination of the amount of application based on these factors is within the level of ordinary skill in the art and its daily dose is, for example, from 0.1 mg / kg / day to 100 mg / kg / day, more specifically from 5 mg / kg / day to 50 mg / / Day, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

[ Experimental Example  One]

1. Extraction of Polysaccharide of Leaf Mushroom

The polysaccharide was extracted from the fruiting body of mushroom fruiting body or fruiting body using the method described in Korean Patent Application No. 2001-0062648. The dried mushroom polysaccharide was prepared by pulverizing dried fruiting bodies of mushrooms and dipped in distilled water at 80 ° C to dissolve the polysaccharide component. Then, the undissolved parts and impurities were removed using a nonwoven fabric for filtration, Naturally dried or lyophilized to obtain a polysaccharide solid on a powder or a sponge.

2. Hydrolysis of polysaccharide

After extracting the extracted mushroom polysaccharide in a suitable solvent such as distilled water, sodium acetate buffer solution or the like, the solution was hydrolyzed with an acid and hydrolyzed with an enzyme.

After dissolving the leaf mushroom polysaccharide in 1N HCl at a concentration of 10% (w / v), hydrolysis was carried out at 100? For 3 hours to conduct hydrolysis with acid. The reaction mixture was neutralized to pH 7 with a 0.2N NaOH aqueous solution to terminate the hydrolysis.

In addition, the leaf mushroom polysaccharide was dissolved in 0.2 M sodium acetate buffer solution (pH 5.0) at a concentration of 1% (w / v), then the β-glucanase was added at a concentration of 20 U / ml, For a period of time to conduct hydrolysis with an enzyme. The hydrolysis was terminated by lowering the temperature.

3. Filtration of polysaccharides

The polysaccharides obtained by hydrolysis of the hydrolyzed leaves were selected from polysaccharides having a specific molecular weight by using an ultrafiltration membrane. The filtration using an ultrafiltration membrane was performed by a cross flow method.

Specifically, pressure was applied to the hydrolyzate solution of the mushroom polysaccharide to pass through an ultrafiltration membrane having a molecular weight cutoff (MWCO) of 10,000 and the filtered solution was recovered. Then, the recovered filtrate (molecular weight: 10,000 or less) was passed through an ultrafiltration membrane having a cutoff molecular weight of 2,000 by the same apparatus and method, and the filtrate passed through the membrane was discarded and the unfiltered liquid was recovered again, Only 10,000 mushroom polysaccharides were selected.

[ Reference example  One] Low molecular weight Mushroom  Production of polysaccharides

Dried leaf mushroom fruiting bodies were pulverized and prepared in powder form, and then immersed in distilled water at 80 ° C to dissolve the polysaccharide component and remove undissolved portions and impurities using a filtration nonwoven fabric. The prepared solution was naturally dried or lyophilized to obtain a polysaccharide solid on the powder.

The resulting polysaccharide solids were dissolved in 0.2 M sodium acetate buffer (pH 5.0) at a concentration of 1% (w / v). After that, beta-glucanase was added at a concentration of 20 U / ml and hydrolysis was carried out at 50? For 2 hours. Hydrolysis was terminated by lowering the temperature.

The hydrolyzed solution was passed through the ultrafiltration membrane stepwise through a cross-flow system as shown in Fig. 1 to obtain a polysaccharide having a molecular weight in the range of 2,000 to 10,000 Da. First, a polysaccharide solution having a molecular weight of 10,000 Da or less was obtained by passing through an ultrafiltration membrane having a molecular weight cut-off of 10,000 Da and then repeatedly permeated using an ultrafiltration membrane having a molecular weight cut-off of 2,000 Da. At this time, the permeated solution contained a polysaccharide having a molecular weight of 2,000 Da or less, so that the remaining portion was not permeated to obtain a solution containing a polysaccharide having a molecular weight of 2,000 to 10,000 Da.

(Content:% by weight) Component (INCI name) Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 water 78.0 78.0 78.0 79.0 78.0 78.0 1,3-butylene glycol -20.9 20.9 20.9 20.9 20.9 20.9 Leaf mushroom polysaccharide (MW 1,000) - - - - 1.0 - Leaf mushroom polysaccharide (MW 2,000) 1.0 - - - - - Leaf mushroom polysaccharide (MW 5,000) - 1.0 - - - - Leaf mushroom polysaccharide (MW 10,000) - - 1.0 - - - Leaf mushroom polysaccharide (MW 12,000) - - - - - 1.0 Phenoxyethanol 0.1 0.1 0.1 0.1 0.1 0.1 Sum 100 100 100 100 100 100

[ Test Example  1] Elastase activity inhibitory activity measurement

The inhibitory activity of the elastase activity of the mushroom polysaccharide according to the molecular weight was measured by comparing with EGCG. The elastase and substrate used were commercially available from Sigma Aldrich, USA

 EGCG 250 μM was used as a positive control, and negative control group, distilled water, was used.

The method for calculating the activity of inhibiting the activity of the elastase was as shown in the following formula (1), and the results are shown in Table 2 below.

A: absorbance at wavelength 415 nm in the absence of the test substance and addition of the enzyme

B: absorbance at a wavelength of 415 nm in the absence of the test substance and no enzyme addition

C: Absorbance at wavelength 415 nm in addition of test substance and enzyme addition

D: Absorbance at wavelength 415 nm in addition of test substance and enzyme-free addition

compound % Inhibition Untreated group 0 EGCG 65 Example 1 38 Example 2 45 Example 3 58 Comparative Example 1 6 Comparative Example 2 13 Comparative Example 3 25

The treatment of Examples 1 to 3, in which the molecular weight of the leaf mushroom polysaccharide corresponds to 2,000 to 10,000 Da, did not exhibit an inhibitory effect equivalent to that in the case where the molecular weight was large, but was comparable to Comparative Example 2 in which the molecular weight was lower than the above range And the inhibitory effect of < RTI ID = 0.0 > The mushroom polysaccharide in the above molecular weight range showed 47 ~ 80% of the effect of EGCG used as the control group.

[ Test Example  2] Collagenase ( MMP -One) Low performance

The inhibitory effect of collagenase production on the mushroom polysaccharide was measured in comparison with retinoic acid. The degree of collagenase production in the cell culture was measured using a commercially available collagenase assay device (Amersham Pharmacia, USA, Catalog #: RPN 2610).

The absorbance of a yellow 96-well plate was measured at 405 nm using a spectrophotometer and the degree of synthesis of collagenase was calculated by the following equation (2) Respectively. At this time, the reaction absorbance of the cell culture fluid collected from the non-treated group was used as a control.

compound Expression level (%) Untreated group 100 Retinoic acid 75 Example 1 73 Example 2 76 Example 3 75 Comparative Example 1 35 Comparative Example 2 45 Comparative Example 3 52

From these results, it can be confirmed that the leaf mushroom polysaccharide of the present invention has an effect of inhibiting the substrate metalloprotease (MMP-1).

[ Test Example  3] Collagen synthesis promotion effect

After human fibroblasts were cultured, the mediums containing the above Examples 1 to 3 and Comparative Examples 1 to 3 were respectively replaced. As a control group, non - treatment group was used. Relative Collagen Biosynthesis (RCB) was measured for the control and experimental groups according to the following formula (3), and the results are shown in Table 4 below.

compound Expression level (%) Untreated group 100 Example 1 125 Example 2 150 Example 3 167 Comparative Example 1 87 Comparative Example 2 90 Comparative Example 3 95

Leaf mushroom polysaccharide promotes the biosynthesis of collagen, thereby increasing the elasticity of the dermis layer, thereby preventing wrinkles of the skin and increasing skin vitality.

Compounding ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Purified water To 100 To 100 To 100 To 100 To 100 To 100 Example 1 5.00 - - - - - Example 2 - 5.00 - - - - Example 3 - - 5.00 - - - Comparative Example 1 - - - 5.00 - - Comparative Example 2 - - - - 5.00 - Comparative Example 3 - - - - - 5.00 Vegetable hydrogenated oil 1.50 1.50 1.50 1.50 1.50 1.50 Stearic acid 0.60 0.60 0.60 0.60 0.60 0.60 Glycerol stearate 1.00 1.00 1.00 1.00 1.00 1.00 Stearyl alcohol 2.00 2.00 2.00 2.00 2.00 2.00 Polyglyceryl-10pentastearate and behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 1.00 1.00 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 1.00 1.00 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 2.00 2.00 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 1.50 1.50 1.50 1.50 Caprylic / carboxy triglyceride 11.00 11.00 11.00 11.00 11.00 11.00 Cyclomedicone 6.00 6.00 6.00 6.00 6.00 6.00 Preservative, incense Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Triethanolamine 0.1 0.1 0.1 0.1 0.1 0.1

[ Test Example  4] Skin elasticity improvement efficacy confirmation

To confirm the effect of improving the skin elasticity in humans, 60 healthy women of 30 to 40 years old were divided into 6 groups of 10 persons for 6 groups of Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 3, respectively, The skin elasticity was measured using a skin elasticity meter (Cutometer SEM 575, C + K Electronic Co., Germany).

Experimental product Skin elasticity effect Formulation Example 1 0.20 Formulation Example 2 0.31 Formulation Example 3 0.36 Comparative Formulation Example 1 0.07 Comparative Formulation Example 2 0.11 Comparative Formulation Example 3 0.13

As shown in Table 6 above, Formulation Examples 1 to 3 and Comparative Formulation Examples 2 to 3 containing the leaf mushroom polysaccharide showed more elasticity than those of Comparative Formulation 1 containing no fungicide polysaccharide . In addition, the groups to which Formulation Examples 1 to 3 were applied showed better effects than Comparative Formulation Example 2, similarly to the above results.

In addition, the results of the above Test Examples 1 to 3 show that the mushroom polysaccharide having a molecular weight of 12,000 Da, which is superior in efficacy, does not exhibit sufficient efficacy due to its reduced skin permeability when applied to human skin, It can be seen that the adjusted leaf mushroom polysaccharide can provide higher efficacy as a result of applying the formulation to the human skin because the ingredient itself has higher skin permeability even when the effect is less than that of the larger molecular weight.

Therefore, it can be confirmed that the polysaccharide of Leaf mushroom is very effective for improving skin elasticity.

[ Test Example  4] Confirmation of skin wrinkle improvement efficacy

Sixty healthy female volunteers were divided into six groups of 10 for each of the six groups of Formulation Examples 1 to 3 and Formulation Examples 1 to 3 respectively. The nutritional cream was applied to the face for one week for 12 weeks, The condition of the wrinkles was measured with a viscometer (SV600, Courage + Khazaka electronic GmbH, Germany) by taking replica. The results are shown in Table 7 below.

After 8 weeks of use
Clinical outcome R1 R2 R3 R4 R5 Formulation Example 1 0.16 0.15 0.12 0.02 0.02 Formulation Example 2 0.15 0.13 0.09 0.01 0.01 Formulation Example 3 0.14 0.13 0.08 0.01 0.01 Comparative Formulation Example 1 0.28 0.25 0.22 0.04 0.03 Comparative Formulation Example 2 0.25 0.19 0.19 0.03 0.02 Comparative Formulation Example 3 0.21 0.18 0.17 0.02 0.02

R1: Difference between the maximum value and the minimum value of the wrinkle contour line

R2: The wrinkle contour line is arbitrarily divided into 5 squares, and the average of R1 values

R3: the highest value of R1 divided by 5

R4: Average value obtained by subtracting the value of each apex and valley from the baseline of the wrinkle contour line

R5: Difference between the baseline of the wrinkle contour line minus the contour of each wrinkle

As shown in Table 7, the external preparation compositions of Formulation Examples 1 to 3 were more effective in improving skin wrinkles than the external composition compositions of Comparative Formulation Examples 1 and 2.

It can also be seen that Formulations 1 to 3 according to the present invention provide a similar or higher wrinkle-reducing effect, as compared to the case of Comparative Formulation Example 3 containing a leaf mushroom polysaccharide having a molecular weight of 12,000 Da. This can be attributed to the fact that the skin permeability of the leaf mushroom polysaccharide of 12,000 Da is lower than that of the small molecular weight leaf mushroom polysaccharide.

[ Test Example  5] Skin Moisture power  Increase effect measurement

Sixty male and female adults aged between 40 and 50 who were classified as dry skin were divided into six groups of 10 for each of Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 3, and the nutritional cream was applied twice daily for 4 weeks to the face Respectively. (24 ° C, relative humidity 40%) after 1 week, 2 weeks, and 4 weeks after application, and after 2 weeks (total 6 weeks) (Corneometer CM825, C + K Electronic Co., Germany).

Test group Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Formulation Example 1 31 33 35 28 Formulation Example 2 31 34 33 29 Formulation Example 3 32 34 35 30 Comparative Formulation Example 1 30 29 30 17 Comparative Formulation Example 2 31 30 29 20 Comparative Formulation Example 3 32 30 31 23

The results of Table 8 show that skin moisture increase rate is higher than that of Comparative Formulation Example 1 when Formulation Examples 1 to 3 and Comparative Formulation Examples 2 and 3 are applied. Compared with Comparative Formulation Example 2 having a smaller molecular weight and Comparative Formulation Example 3 having a higher molecular weight, it was found that similar effects were obtained during application, but at 6 weeks after the discontinuation of application, Formulations 1 to 3 provided better skin moisturizing effect have.

[ Test Example  6] Measurement of promoting effect of keratinocyte differentiation

In order to investigate the effect of differentiation on the differentiation of keratinocyte of leaf mushroom polysaccharide according to the molecular weight, the amount of corned envelope (CE) produced upon differentiation of keratinocytes was measured by absorbance as described below.

First, keratinocytes were isolated from the epidermis of newborns and cultured in primary culture. At this time, low calcium (0.03 mM) and high calcium (1.2 mM) treatment groups were negative control and positive control, respectively. The protein content was measured and used as a standard for assessing the degree of cell differentiation. The results are shown in Table 9 below.

Test substance The ability to differentiate in keratinocytes (%) Low calcium (0.03 mM) solution
(Negative control) 100 High calcium (1.2 mM) solution
(Positive control group) 210 Example 1 255 Example 2 263 Example 3 277 Comparative Example 1 102 Comparative Example 2 203 Comparative Example 3 213

As shown in Table 9, when Examples 1 to 3 and Comparative Examples 2 and 3 were applied, it was found that the effect of accelerating differentiation of keratinocytes was excellent. In the case of Examples 1 to 3 according to the present invention, the effect of promoting the differentiation of keratinocytes was excellent, and in particular, Example 3 is similar to that of Comparative Example 3 having a high molecular weight.

[ Test Example  7] Measurement of skin barrier function recovery effect

Ten adult male and female adult rats were treated with seven groups of Formulation Examples 4 to 6 and Comparative Formulation Examples 4 to 7, which were prepared by using the tape stripping method to damage the skin barrier, (TWEL) was measured and compared with Vapometer (Delfin, Finland) once a day for 7 days

Compounding ingredient Formulation Example 4 Formulation Example 5 Formulation Example 6 Comparative Formulation Example 4 Comparative Formulation Example 5 Comparative Formulation Example 6 Comparative Formulation Example 7 Purified water 65 65 65 65 65 65 70 Propylene glycol 30 30 30 30 30 30 30 Example 1 5 - - - - - - Example 2 - 5 - - - - - Example 3 - - 5 - - - - Comparative Example 1 - - - 5 - - - Comparative Example 2 - - - - 5 - - Comparative Example 3 - - - - - 5 -

Test group TWEL Change (%) Before processing 1 day 2 days 3 days 4 days 5 days 6 days Formulation Example 4 100 117 119 108 107 104 105 Formulation Example 5 100 119 122 117 112 109 106 Formulation Example 6 100 115 109 117 110 108 107 Comparative Formulation Example 4 100 112 113 97 73 57 46 Comparative Formulation Example 5 100 111 101 100 98 91 88 Comparative Formulation Example 6 100 113 103 101 98 95 92 Comparative Formulation Example 7 100 108 101 96 69 53 37

When Formulation Examples 4 to 6 containing the leaf mushroom polysaccharide were treated, it was confirmed that the barrier damage was recovered as the amount of transdermal water loss returned to normal rapidly.

division Formulation Example 7 Formulation Example 8 Formulation Example 9 Comparative Formulation Example 8 Comparative Formulation Example 9 Comparative Formulation Example 10 Comparative Formulation Example 11 Comparative Formulation Example 12 A Deionized Water To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 B ethanol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 PEG-60 hardened castor oil
(hydrogenated castor oil) 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4 Perfume 0.04 0.04 0.04 0.04 0.04 0.04 0.04 0.04 C Example 1 5.0 - - - - - - - Example 2 - 5.0 - - - - - - Example 3 - - 5.0 - - - - - Comparative Example 1 - - - 5.0 - - - - Comparative Example 2 - - - - 5.0 - - - Comparative Example 3 - - - - - 5.0 - - Erythromycin - - - - - - - 5.0

[ Test Example  8] Antimicrobial activity test against acne bacterium

Each of the cosmetic compositions prepared in the formulations of Formulation Examples 7 to 9 and Comparative Formulations 8 to 12 was tested for antibacterial activity against Propionibacterium acnes (ATCC 6919: medium-BHI broth), a causative strain of acne .

Item pH Propionibacterium acnes Formulation Example 7 5.7 > 50 ppm Formulation Example 8 5.7 > 50 ppm Formulation Example 9 5.7 > 50 ppm Comparative Formulation Example 8 5.7 Maximum concentration (no antimicrobial activity) Comparative Formulation Example 9 5.7 > 50 ppm Comparative Formulation Example 10 5.7 > 50 ppm Comparative Formulation Example 11 5.7 Maximum concentration (no antimicrobial activity) Comparative Formulation Example 12 5.7 > 100ppm

As the ppm concentration in MIC was smaller, the concentration of ppm was small in Formulations 7 to 9 and Comparative Formulations 9 to 10, and in particular, in Formulations 7 to 9 The concentration of ppm was remarkably smaller than that of Comparative Formulation Example 12 using erythromycin, which is a known acne remedy, and it can be confirmed that the composition containing the leaf mushroom polysaccharide has much better antibacterial activity against the test bacteria.

[ Test Example  9] Lipid synthesis Lipogenesis ) Inhibition test

3T3-L1 cells, a fibroblast cell line of mice, were cultured.

In the above, 3T3-L1 adipocytes were used for staining (S4136, sigma-aldrich). The control group was prepared by using a test medium or a test medium containing no comparative substance. Lt; / RTI > The degree of inhibition of fat accumulation was graded by dividing the degree of staining by +++, +, +, -, which means that the degree of staining is greater toward +++. The results are shown in Table 14 below.

sample % Inhibition Control group +++ Comparative group + Example 1 - Example 2 - Example 3 - Comparative Example 1 ++ Comparative Example 2 + Comparative Example 3 -

As shown in Table 14, the amount of fat accumulated in the adipocytes was small and the lipid synthesis inhibition effect was low. In Examples 1 to 3, the lipid mushroom polysaccharides according to the present invention, It is shown that there is a superior lipid synthesis inhibitory effect than caffeine which is an inhibitory substance.

[ Test Example  10] Acne improvement and Sebum secretion  Test for reduction and irritation

Eighty individuals with acne were divided into eight groups each by 10 persons, and the subjects corresponding to each group were allowed to use the cosmetic composition prepared in Formulation Examples 7 to 9 and Comparative Formulation Examples 8 to 12 for one month. Acne improvement scale is from 1 point to 5 points, 1 point is 'No', 3 points are 'Normal' and 5 points are 'Very agree'. The experimental results are shown in Table 15 below with an average score of 10 persons.

Inflammatory acne improvement Acne-bloating period Recurrence of acne Decrease sebaceous glands Presence or absence of stimulation Formulation Example 7 2.8 4 days radish 2.3 0/10 Formulation Example 8 3.8 3 days radish 2.4 0/10 Formulation Example 9 3.7 3 days radish 2.2 0/10 Comparative Formulation Example 8 1.7 11th U 1.7 0/10 Comparative Formulation Example 9 2.5 5 days U 2.0 1/10 Comparative Formulation Example 10 2.7 7 days radish 2.1 0/10 Comparative Formulation Example 11 1.5 13th U 2.0 0/10 Comparative Formulation Example 12 4.2 2 days radish 4.1 9/10

As shown in Table 15, in the case of using formulations 7 to 9 containing the mushroom polysaccharide and comparative formulations 9 to 10, it was found that the acne did not recur when compared with the comparative formulations 8 and 11, Overall, there is an excellent effect on the improvement of acne, and in particular, in the case of Formulations 7 to 9, the excellent acne improvement effect is shown.

On the other hand, Comparative Formulation Example 12 containing the antimicrobial activity standard shows the effect of improving acne but it is not suitable for long-term use due to strong skin irritation in use. However, the composition according to the present invention is not stimulated, It is also suitable for use.

[ Test Example  11] Inflammation improvement effect

1. Inhibitory effect of prostaglandins

The anti - inflammatory effect was evaluated by the inhibitory effect of prostaglandin production. The effect of macrophyllous polysaccharide on the macrophages was measured.

At this time, the inhibitory activity on the production of prostaglandin of the mushroom polysaccharide was determined by setting the difference of the prostaglandins produced in the LPS-treated group and the untreated group to 100%, and by treating the LPS and the sample together in a reduced proportion of the prostaglandin The results were compared with those of the control group, and the results (inhibitory effect on the production of prostaglandins) are shown in Table 16 below.

Blank 100% Control group (aspirin treatment group) 25.0% Example 1 23.4% Example 2 22.8% Example 3 24.1% Comparative Example 1 0.3% Comparative Example 2 15.7% Comparative Example 3 19.1%

As shown in Table 16 above, the treatment with Examples 1 to 3 and Comparative Examples 2 to 3 containing the leaf mushroom polysaccharide showed a high inhibitory effect on the production of prostaglandin. Especially, when Examples 1 to 3 were treated with aspirin As in the control group, the effect of inhibiting the production of prostaglandin is very high.

Thus, it can be seen that the leaf mushroom polysaccharide of the present invention can provide excellent inflammation improving effect.

Also, it can be seen that the polysaccharide of Leaf mushroom can prevent the skin trouble by improving the expression of prostaglandin, which is a skin inflammation factor.

2. IL-8 production inhibitory effect

One day before the experiment, normal human skin keratinocyte (NHEK, supplier: Lonza) was cultured.

After culturing in a 5% CO ₂ incubator at 37 ° C for 24 hours, the culture was taken and ELISA for interleukin-8 (IL-8) was performed. The results are shown in Table 17 below. The ELISA used the experimental method of BD science.

division IL-8 secretion (pg / ml) Untreated Control (Control) 935.12 PGSA (10 [mu] g / ml) 4812.60 PGSA (50 占 퐂 / ml) 5895.08 PGSA (50 占 퐂 / ml) + LPS (1 占 퐂 / ml) 6814.91 Example 1 1131.23 Example 2 998.78 Example 3 1007.44 Comparative Example 1 1454.13 Comparative Example 2 1223.76 Comparative Example 3 1234.29

In Table 17, it was shown that the leaf mushroom polysaccharide reduced and inhibited the secretion of IL-8, which was increased by PGSA and LPS, and that IL-8 secretion was remarkably enhanced in the case of Example 1 to 3, .

Therefore, it can be seen that the composition for external application for skin of the present invention can provide a superior anti-inflammatory effect by significantly reducing the secretion of IL-8 increased by PGSA and LPS.

[ Test Example  12] Itching relief assessment

The day before the experiment, keratinocytes (cell line name: HaCaT source: ATCC) were cultured.

The difference between the minimum value and the maximum value obtained by measuring the flex of 80 seconds after the treatment of Examples 1 to 3 and Comparative Examples 1 to 3 and 2 U / ml trypsin or 5 μM PAR-2 active peptide (SLIGKV) And the inhibition rate (%) of intracellular calcium influx of calcium ions is compared with the difference between the minimum value and the maximum value in the treatment of 2 U / ml trypsin or 5 μM PAR-2 active peptide (SLIGKV) Respectively.

Treating material Inhibition rate of intracellular influx of calcium ions (%) Trypsin (2 U / ml) PAR-2 active peptide (5 [mu] M) Example 1 34% 40% Example 2 41% 43% Example 3 37% 42% Comparative Example 1 0% 2% Comparative Example 2 23% 27% Comparative Example 3 25% 31%

As can be seen from Table 18, the inflow of intracellular calcium ions by trypsin or PAR-2 activating peptide (SLIGKV) was decreased with the treatment of leaf mushroom polysaccharide. In particular, 3 treatment significantly reduced the intracellular calcium ion influx. Accordingly, the composition for external application for skin containing the foliar mushroom polysaccharide of the present invention can provide an excellent antinociceptive effect by effectively inhibiting the itching-inducing PAR-2 activity.

Hereinafter, a formulation example of the composition according to the present invention will be described. However, the pharmaceutical composition and the cosmetic composition can be applied to various formulations, which are not intended to limit the present invention but merely to illustrate the present invention.

[Formulation Example 1] Soft capsule

300 mg of mushroom fruiting body containing polysaccharides of 2,000 to 10,000 Da molecular weight, 2 mg of palm oil, 8 mg of palm kernel oil, 4 mg of yellow beans and 6 mg of lecithin were mixed and 400 mg / A capsule was prepared.

[Formulation Example 2] Tablets

300 mg of mushroom fruiting body containing polysaccharides of 2,000 to 10,000 Da molecular weight, 100 mg of glucose, 50 mg of red ginseng extract, 96 mg of starch and 4 mg of magnesium stearate were mixed and 40 mg of 30% ethanol was added to form granules , Dried at 60 ° C and tableted using a tablet machine.

[Formulation Example 3] Granules

300 mg of mushroom fruiting body containing polysaccharides of 2,000 to 10,000 Da molecular weight, 100 mg of glucose, 50 mg of red ginseng extract and 600 mg of starch were mixed and 100 mg of 30% ethanol was added to form granules, followed by drying at 60 ° C The granules were then formed and filled into bubbles. The final weight of the contents was 1 g.

[Formulation Example 1] Softening lotion (skin lotion)

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 0.1 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Phage-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 2] Nutritional lotion (Milk lotion)

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 0.5 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Wax 4.0 Polysorbate 60 1.5 Caprylic / capric triglyceride 5.0 Squalane 5.0 Sorbitacecequioleate 1.5 Liquid paraffin 0.5 Cetearyl alcohol 1.0 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 3] Nourishing cream

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 1.0 glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Squalane 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Triethanolamine 0.1 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 4] Massage cream

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 1.5 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Wax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 Vaseline 3.0 paraffin 1.5 Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 5] Pack

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 0.1 glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Nonyl phenyl ether 0.4 Polysorbate 60 1.2 Ethanol preservative 6.0 Good Preservative, coloring, fragrance Suitable amount Purified water Balance

[Formulation Example 6] ointment

Compounding ingredient Content (% by weight) Mushroom fruiting bodies containing polysaccharides having a molecular weight of 2,000 to 10,000 Da 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / capric triglyceride 3.0 Squalane 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.0 Wax 4.0 Preservative, coloring, fragrance Suitable amount Purified water Balance

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (14)

A composition comprising an extract of fruiting body mushroom fruiting body,
Wherein the extract comprises a polysaccharide having a molecular weight of 2,000 to 10,000 Da.
The method according to claim 1,
Wherein the polysaccharide is at least one selected from the group consisting of? -1,3-glucan and? -1,6-glucan.
The method according to claim 1,
Wherein the extract comprises from 0.5 to 20% by weight polysaccharide based on the total weight of the extract.
The method according to claim 1,
Wherein the composition is for anti-aging.
5. The method of claim 4,
Wherein the composition enhances skin elasticity or improves skin wrinkles.
The method according to claim 1,
Wherein the composition is for moisturizing.
The method according to claim 1,
Wherein the composition is for antibacterial.
8. The method of claim 7,
Wherein the bacterium is an acne bacterium.
The method according to claim 1,
Wherein said composition is anti-inflammatory.
The method according to claim 1,
Wherein said composition is for the treatment of obesity.
The method according to claim 1,
Wherein the composition comprises from 0.5 to 20% by weight of the extract, based on the total weight of the composition.
12. The method according to any one of claims 1 to 11,
Wherein the composition is a cosmetic composition.
12. The method according to any one of claims 1 to 11,
Wherein the composition is a health food composition.
12. The method according to any one of claims 1 to 11,
Wherein the composition is a pharmaceutical composition.