KR20140043546A - Composition for skin external application comprising extract of papenfussiella kuromo - Google Patents

Abstract

A composition for external skin application comprising an extract of Papenfussiella kuromo according to the present invention is not toxic to skin cells and has abilities of inhibiting tyrosinase activity, L-DOPA oxidation, and melanin synthesis. Thereby the composition of the present invention is excellent for skin whitening and is effective for antioxidant activity, wrinkle prevention or alleviation, anti-aging, anti-skin aging for skin aging caused by ultraviolet light, and recovering damaged skin.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for external application for skin containing an extract of Chondrus follicularis,

The present invention relates to a method for producing papillomavirus kuromo extract, and more particularly, to a composition for external application for skin, which contains, as an active ingredient, an extract of fenugreek, which has antioxidant ability, anti-aging and whitening function.

The cosmetics industry is a technology-intensive, high-value-added industry in which basic science and applied technologies such as chemistry, biology, physiology, and pharmacy are applied in a technical aspect. Today, cosmetics are products that are directly used on human skin. They should have safety, efficacy and ease of use for skin. They should not only provide aesthetic functions to maintain beauty, but also protect skin, clean, moisturize, It should also have biological functions. Especially in the cosmetics industry, as the industrialization has progressed rapidly and the aging phenomenon has deepened, the importance of environment and health has been emphasized and the well - being culture has become the mega trend of modern society. Various kinds of skin diseases caused by environmental changes are increasing with the development of such advanced industries, and in the bio venture, cosmetics and biotechnology industries, researches on whitening and anti-aging using high functionality of natural materials are actively being carried out.

Particularly, recent attempts to develop products with high biological efficiency by incorporating highly functional biomaterials have been actively conducted by the cosmetics industry and bio companies. Existing cosmetic materials used in cosmetics were crude extracts based on information from plants or herbal extracts or by means of Donguibogam. On the other hand, there is an increasing need for functional cosmetic materials having superior skin function and excellent efficacy and safety by applying an extract or extraction method that has been validated on a biological basis. Especially, in Korea, marine resources are abundant on three sides, but biologic research on marine algae is insufficient and there are few cases where marine algae are applied to cosmetics.

[0002] Conventionally, as a cosmetic composition patent using a seaweed, there is Korean Patent No. 10-1040393 entitled "Cosmetic Composition Containing Seaweed Fiber". Specifically, the patent discloses a cosmetic composition will be. However, there have been no cases in which cosmetics were conventionally applied by using a hair dye.

The inventors of the present invention have made efforts to manufacture a functional skin external composition for seaweeds, and confirmed that the extract of green tea fungus has an effect of whitening, prevention of skin aging and antioxidant ability, and completed the present invention.

Accordingly, it is an object of the present invention to provide a composition for external application for skin, which contains, as an active ingredient, an extract of green tea hair having whitening, prevention of skin aging, and antioxidant ability.

In order to accomplish the above object, the present invention provides a composition for external application for skin, which comprises an extract of green foxtail as an active ingredient.

In the present invention, the extract may be a lyophilized powder of an artificial hair follicle, an organic solvent extract, an ultrasonic extract, or a hot water extract.

In the present invention, the composition may be characterized in that it is for preventing skin aging.

In the present invention, the composition may be characterized in that it is for improving skin wrinkles.

In the present invention, the composition may be characterized in that it is for restoring skin damage due to ultraviolet rays or for skin protection.

In the present invention, the composition may be characterized by being whitening.

In the present invention, the composition may be characterized as being for antioxidation.

The composition for external application for skin containing the extract of Chondrus folliculata according to the present invention is excellent in skin whitening due to inhibition of tyrosinase activity, inhibition of L-DOPA oxidation and melanin synthesis without toxicity to skin cells, It is effective for improvement, anti-aging, prevention of skin aging from ultraviolet rays, and recovery of damaged skin.

FIG. 1 is a graph showing the results of cell stability test on human keratinocyte cell line of the extract of Chrysanthemum morifolium according to the present invention.
FIG. 2 is a graph showing the results of cell stability test on the human fibroblast cell line of the extract of P. falciparum according to the present invention.
FIG. 3 is a graph showing the results of a test for inhibiting tyrosinase activity of the artificial hair follicle extract according to the present invention.
FIG. 4 is a graph showing the results of L-DOPA oxidation inhibition test of the extract of P. falciparum according to the present invention.
FIG. 5 is a graph (A) and a photograph (B) showing the results of the melanin synthesis inhibitory effect test of the artificial hair follicle extract according to the present invention.
FIG. 6 is a graph showing the results of the antioxidant efficacy test of the artificial hair follicle extract according to the present invention.
FIG. 7 is a graph showing the results of testing the protective effect against UV-induced cell damage in the human keratinocyte lineage of the extract of the artificial hair follicle according to the present invention.

The present invention relates to a composition for external application for skin containing an extract of green tea hair as an active ingredient.

Papenfussiella kuromo is a brown algae distributed mainly on the south coast and the east coast. It grows in the shape of a brown thick thread, and is seaweed that grows up to about 30-50 cm attached to rocks and other seaweeds. It lives mainly in spring and early summer (February to July).

In one aspect, the present invention relates to a composition for external application for skin for improving skin comprising an extract of green beans hair follicle as an active ingredient. Specifically, the extract can be contained in an amount of 0.001 to 10% by weight based on the total weight of the composition.

For example, in the following examples, 0.001 to 0.1 wt.% Of the composition was tested, but 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10 wt. 20%, 30% or more, it has excellent antioxidant ability, whitening and wrinkle improving function.

In the present invention, the " extract " refers to an extract obtained by various methods known in the art, such as freeze-drying the powder, or cold water extraction, hot water extraction, ethanol extraction and ultrasonic extraction.

The extraction method in the present invention is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux extraction, hot water extraction, and the like. Here, in the case of hot water extraction, hot water extract is obtained by heating at 55 to 75 ° C for 6 to 8 hours in a hot water distiller. In the case of ultrasonic extraction, Sonics VC 750 can be used for 1 hour to extract and ultrasonic treatment for 3 seconds and stop for 5 seconds with the program set in Sonics VC 750 for 1 hour. In the case of cold water extraction, cold water (15-25 ° C), for example, is mixed with the dried or extracted powder of the green tea extract and extracted for 3 days to obtain a cold water extract.

Or by extraction using water, an organic solvent, or a mixed solvent thereof. The extracted liquid can be used directly or by concentrating and / or drying. When extracted with an organic solvent, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof may be used and extracted by room temperature or warming under conditions where the active ingredient of the herbal medicine is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may vary, so select an appropriate organic solvent. Particularly, in the present invention, an ethanol extract, preferably a 30-100% ethanol extract, specifically 100% ethanol extract is preferable. As described in the examples, the extract is mixed with 100% ethanol and shaken for 1 week . However, it is not necessary to shake for one week, and it is enough to extract for 5 to 8 hours.

In the present invention, the extract may be used by concentration or dilution, or distillate of the extract may be used. Specifically, in one embodiment, it may be concentrated using a reduced pressure concentrator.

In the present invention, facilitation of skin improvement includes skin protection and improvement of skin condition, prevention of skin damage from ultraviolet rays, skin whitening, prevention or improvement of skin aging and wrinkles, skin protection and alleviation of skin inflammation reaction, , Or improvement of skin barrier function, skin irritation mitigation, skin cell proliferation and regeneration ability, antioxidant ability, and collagen synthesis promoting ability.

In the present invention, the term "comprising as an active ingredient" means a composition for external application for skin, which can exhibit a skin improving effect, for example, as a composition for external application for skin, which prevents skin cell aging by inhibiting UV cell damage and inhibits tyrosinase activity Quot; means that it contains an effective amount that can exhibit antioxidant ability, whitening ability, whitening ability through L-DOPA oxidation inhibition, inhibition of melanin synthesis, and antioxidant ability.

Therefore, the composition for external application for skin according to the present invention can be applied particularly for prevention of skin aging, improvement / prevention of skin wrinkles, whitening, and antioxidation. In addition, the extract of Chondrus folliculata according to the present invention has an effect on prevention of skin damage (photoaging) due to ultraviolet rays, alleviation, and healing, so that it can be applied to prevent skin damage from ultraviolet rays and to protect skin.

In the present invention, the composition for external application for skin may be a cosmetic composition or a pharmaceutical composition.

The cosmetic composition of the present invention may contain an acceptable carrier in cosmetic preparations. Herein, "acceptable carrier in cosmetic preparation" means a compound or composition already known and used that may be included in a cosmetic preparation or a compound or composition which will be developed in the future, and which has no toxicity, instability or irritation that is acceptable to the human body upon contact with the skin. Say nothing.

The carrier may be included in the skin topical composition of the present invention in an amount of from about 1% by weight to about 99.99% by weight, preferably from about 90% by weight to about 99.99% by weight of the composition, based on the total weight thereof. However, since the ratio varies depending on the formulations described below in which the composition for external application for skin of the present invention is prepared, and its specific application site (face, neck, etc.) or its preferable application amount and the like, And should not be construed as limiting the scope of the invention.

Examples of the carrier include an alcohol, an oil, a surfactant, a fatty acid, a silicone oil, a wetting agent, a moisturizer, a viscous modifier, an emulsifier, a stabilizer, an ultraviolet scattering agent, an ultraviolet absorber, a coloring agent and a perfume. The compounds / compositions which can be used as the above-mentioned alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscous modifier, emulsifier, stabilizer, ultraviolet scattering agent, ultraviolet absorber, The person skilled in the art can select and use the appropriate substance / composition.

In an embodiment of the present invention, the composition for external application for skin according to the present invention may contain glycerin, butylene glycol, propylene glycol, polyoxyethylene hardened castor oil, ethanol, triethanolamine and the like in addition to the above- , Antioxidants, coloring agents, purified water and the like may be included as needed.

The composition for external application for skin according to the present invention can be prepared in various forms such as lotion, essence, gel, emulsion, lotion, cream (underwater type, water type, multiphase), solution, suspension ), Capsules (soft capsules, hard capsules) having a coating such as anhydrous products (oil and glycol), gel, mask, pack, powder or gelatin.

The skin of the present invention includes not only the face but also the scalp and whole body. The composition for external application for skin which can be applied to such scalp includes shampoo, rinse, treatment, hair removal agent, etc., and a body cleanser And the like can be manufactured in various forms.

The method of manufacturing the composition for external application for skin containing the foliar hair extract according to the present invention is not limited to the above-described manufacturing method, and any person skilled in the art may make modifications The skin external composition containing the extract of the artificial hair follicle according to the present invention can be prepared.

In particular, the composition for external application for skin may be prepared in the form of a general emulsified formulation and a solubilized formulation, by a known manufacturing method, in addition to the manufacturing method specifically disclosed in the present invention.

When the cosmetic composition is manufactured from a cosmetic composition, the cosmetic composition of the emulsified formulation includes nutritional lotion, cream, essence and the like. It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

In addition, suitable cosmetic formulations include, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing an oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) May be provided in the form of a follicular dispersing agent of the type, cream, skin, lotion, powder, ointment, spray or conical stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition, the composition for external application for skin of the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, As used herein means any emulsion or emulsion which is commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Adjuvants commonly used in the cosmetics or dermatological fields, such as other ingredients. And, the above ingredients can be introduced in amounts commonly used in the field of dermatology.

The composition for external application for skin according to the present invention includes functional cosmetics capable of functioning to prevent wrinkling of skin caused by collagen synthesis promoting ability and aging of skin based on aging skin cell activation ability.

Such a pharmaceutical composition may contain, in addition to the active ingredient, flaxseed extract, a "pharmaceutically acceptable carrier" which is selected from the group consisting of diluents, lubricants, binders, disintegrants, sweeteners, stabilizers and preservatives . The pharmaceutical composition may further comprise an additive. The additives may include flavoring agents, vitamins, and antioxidants. Examples of the diluent include lactose, dextrin, tapioca starch, corn starch, soybean oil, microcrystalline cellulose or mannitol as a carrier, magnesium stearate or talc as a lubricant, , And the binding agent may be polyvinylpyrrolidone or hydroxypropylcellulose. Examples of the disintegrant include carboxymethylcellulose calcium, sodium starch glycolate, polacrilin potassium, or crospovidone; examples of sweeteners include white sugar, fructose, sorbitol, or aspartame; stabilizers include sodium carboxymethylcellulose, Or xanthan gum, and the preservative may be methyl paraoxybenzoate, propyl p-hydroxybenzoate, or potassium sorbate.

The pharmaceutical composition may be formulated into conventional pharmaceutical formulations known in the art. The pharmaceutical composition may be formulated and administered in the form of an oral administration preparation, an injection preparation, a suppository, a transdermal preparation, and a dosage form. For example, the formulations may be formulations for oral administration such as solutions, suspensions, powders, granules, tablets, capsules, pills, or expanses.

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.

Soft hair  Extract preparation

Salinity was removed by washing several times at Bogil-do coast in Jeollanam-do and 100% ethanol was immersed in 100% ethanol for one week to extract the cell contents sufficiently. 1 L was concentrated at 30 ° C for 3 hours in a vacuum concentrator And concentrated to one tenth volume. The resulting concentrated product was lyophilized, pulverized and powdered for use in the experiment.

Experimental Example 1: Cytotoxicity test of the extract of Chondrus follicle according to the present invention

(1) Cell culture

Keratinocyte (HaCaT) and fibroblast CCD986-sk were cultured in Dulbecco's Modified Eagle's Medium (DMEM), 10% fetal bovine serum (FBS), antibiotic-antimycotic (GIBCO, Cat No. 15240 -062) in a 75 Cm ² flask at 37 ° C and 5% CO ₂ . When the cells were confluent at 80% or more, subculture was performed.

 (2) Test method

  In order to determine the concentration range that does not show cytotoxicity in the cell types (keratinocyte, fibroblast) constituting the skin, the cells are cultivated, and various concentrations of the test substance are treated in the cell culture, and then the growth and proliferation or toxicity The MTT assay was used to investigate the effect of the MTT assay. This method is more reliable than MTT (3- (4,5-dimethylthiazol-2yl) -2,5 diphenyl-2H-tetrazolium bromide) which is specifically reacted with mitochondrial dehydrogenase Can be tested.

Specifically, the cells were plated in 24 wells at 1 × 10 ⁴ cells / well Or 1 × 10 ⁵ cells / well. Cells were cultured until confluence reached 50 ~ 60%. The medium was discarded, washed with PBS (phosphate buffered saline), and then a new medium containing no 10% FBS was added. After that, 50 μl of MTT solution (6.6 mg / ml, 3- (4,5-dimethylthiazol-2yl) -2,5 diphenyl-2H-tetrazolium bromide solution) is added to each well and further incubated for 3 hours. After removing the culture medium, 200 μl of dimethylsulfoxide (DMSO, Amresco, 0231-500ML) was added and the mixture was shaken for 10 minutes. Then, 100 μl of the solution was taken in 96-well plate and absorbance was measured at 540 nm with an ELISA (Enzyme-Linked Immunosorbent Assay) reader. The degree of cytotoxicity was expressed as a percentage based on the absorbance intensity of the control using pure water.

Cell stability (%) = (absorbance of test group / absorbance of control group) x100

(3) Human keratinocyte line  ( HaCaT Cell stability against)

   Cellular stability of the extracts of P. falciparum on human keratinocyte cell line HaCaT was evaluated. HaCaT, a human keratinocyte cell line, was cultured in an ATCC (American Type Culture Collection).

As a control, purified water was used. The concentration of the extracts of the females was serially diluted 2 times at a maximum concentration of 200 ug / ml, and tested at a total concentration of 5 to a concentration of 12.5 ug / ml. As a result, as shown in Fig. 1, there was no cytotoxicity up to 200 ug / ml and cell proliferation ability was about 60% (1.5 times).

(4) Human fibroblast line  ( CCD986 - SK Cell stability against)

   The cell stability of the extracts of P. falciparum on human fibroblast cell line CCD986-SK was evaluated. The human fibroblast host, CCD986-SK, was cultured in the ATCC (American Type Culture Collection).

As a control, purified water was used. The concentration of the extracts of the females was serially diluted 2 times at a maximum concentration of 200 ug / ml, and tested at a total concentration of 5 to a concentration of 12.5 ug / ml. As a result, no cytotoxicity was observed up to 200 ug / ml.

Experimental Example  2: whitening efficacy test

In order to confirm the whitening function of the soft flax extract according to the present invention, a very important role in skin melanin production was examined for tyrosinase activity and L-DOPA oxidation inhibition. Specifically, tyrosinase is a series of processes for making dopa by oxidizing tyrosine in the pigment forming organs (melanocyte) of pigment producing cells (melanocytes), which are cells producing skin pigment, and then oxidizing dopa again to form dopachrome Is known to catalyze the reaction.

(1) Tyrosinase activity inhibition test

In this test, it was confirmed whether or not the extract of P. falciparum inhibited tyrosinase activity to inhibit intracellular melanin synthesis.

30 μl of a sample of the appropriate concentration (1 mg / ml, 500 μg / ml, 250 μg / ml, 125 μg / ml, 62.5 μg / ml) was dissolved in 210 μl of 0.1 M phosphate buffer (pH 6.5), mushroom tyrosinase , Fulka Cat. No. 93898). To this solution, 40 μl of 1.5 mM tyrosine solution was added and reacted at 37 ° C for 10 to 15 minutes. The absorbance was measured at 490 nm using an ELISA reader. 0.1 M phosphate buffer (pH 6.5) was used instead of the sample solution for the analysis. Arbutin 0.1% was used as a positive control.

% inhibition of tyrosinase activity = 100-[(b-b ') / (a-a') X 100]

a: absorbance after reaction of blank sample liquid

b: absorbance after reaction of the sample liquid

a ', b': absorbance measured by replacing buffer with tyrosinase

The extracts of P. falciparum were diluted to twice the maximum concentration of 1 mg / ml and confirmed to be treated to a minimum of 62.5 ug / ml. As a result, as shown in FIG. 3, tyrosinase inhibition rate was about 42% at 0.1% (1 mg / ml) of arbutin, while about 1% of tyrosinase inhibitory effect was about 46% at 1 mg / A similar whitening effect was observed (control: 0.1 M phosphate buffer).

(2) L- DOPA  Oxidation inhibition test

The test tube was filled with 850 ul of 0.1 M phosphate buffer solution (pH 7.0) and 50 ul of the flaxseed extract diluted with the appropriate concentrations (1 mg / ml, 500 ug / ml, 250 ug / ml, 125 ug / ml, 62.5 ug / ml) and mushroom tyrosinase 1,500-2,000 U / ml, Fulka Cat. No. 93898) were added in this order and reacted at 37 ° C for 6 minutes. To this solution, 50 μl of 0.06 mM L-DOPA (L-3,4-dihydroxyphenylalanine, Sigma D9628-25G) solution was added and reacted at 37 ° C for 1 minute. After the reaction was completed, absorbance was measured at 475 nm using an ELISA reader (BioTek-MQX200R). 0.1M phosphate buffer (pH 7.0) was used instead of the sample solution, and 0.1% arbutin was used as a positive control.

DOPA oxidative activity inhibition rate (%) = 100- (reaction absorbance of each sample liquid / reaction absorbance of the blank)

As shown in FIG. 4, the L-DOPA oxidation activity was about 30% at 0.1% (1 mg / ml) of arbutin, Inhibition of L-DOPA oxidase activity by about 33% at 1 mg / ml of fusarium head extract. Therefore, it was confirmed that the extract of Rhododendron follicle has a whitening effect equivalent to that of arbutin, which is well known as a whitening effect.

(3) intracellular Melanosite  Derived melanin synthesis inhibition test

For this study, Murine Melanoma (B16F10) was incubated in 75 Cm ² flask with Dulbecco's Modified Eagle's Medium (DMEM), 10% fetal bovine serum (FBS) and Antibiotic-antimycotic (GIBCO, Cat No. 15240-062) 5% CO ₂ . When confluence of the cells reached 80%, the cells were passaged at 1 ⁵ cells / well in 6well plates, and then cultured at 5% CO ₂ and 37 ° C until the cells reached 70-80% or more. After that, the medium was removed and 0.2 mM of IBMX (3-Isobutyl-1-methylxanthine, Sigma, Cat.No:17018) and a-MSH (a-Melanocyte stimulating hormone, Sigma, Cat. No : M4135) at 10 uM to induce intracellular melanin biosynthesis. After that, the cells were replaced with fresh medium containing the extracts of brown foxtail diluted with a certain concentration (1 mg / ml, 500 ug / ml, 250 ug / ml, 125 ug / ml, 62.5 ug / ml) and further cultured for 48 hours. After removing the medium, the cells were washed with PBS (GIBCO) and treated with 0.05% trypsin-EDTA (GIBCO). The recovered cells were counted using a hematocytometer, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was removed to obtain a pellet. After drying the cell pellet at 60, 100ul of 1M sodium hydroxide solution (SAMCHUN Chemicals) containing 10% dimethylsulfoxide (DMSO, Amresco, 0231-500ML) was added to obtain intracellular melanin in 60 thermostat. The absorbance of this solution was measured at 490 nm in an ELISA reader (BioTek-MQX200R) to determine the amount of melanin per cell constant number. The degree of inhibition of melanin synthesis was expressed as a percentage based on the absorbance of the control (untreated cells) using pure water.

Melanin synthesis inhibition rate (%) = 100- (reaction absorbance of each sample liquid / reaction absorbance of negative control group)

As shown in Fig. 5, melanocyte stimulating hormone (Melanocyte stimulating hormone) was induced by 0.2XM of IBMX and 10uM of melanocyte stimulating hormone , Induced about 1.5-fold increase in melanin biosynthesis compared to the untreated control group. It was confirmed that the melanin biosynthesis was inhibited about 1.5 times when 1 mg / ml of the extract of the foxtail extract was treated.

Experimental Example  3: Soft hair  Antioxidant Test of Derivative Extracts

DPPH free radical scavenging ability was confirmed in order to confirm the antioxidative ability of the extracts of P. falciparum. This method tests the ability of 1,1-Diphenyl-2-picryhydrazyl (DPPH, Sigma D9132-1G) to eradicate free radicals on ethanol to determine the direct action of antioxidant activity. The compound 1,1-Diphenyl-2-picryhydrazyl (DPPH, Sigma D9132-1G) generates free radicals in ethanol, and a certain concentration (1 mg / ml, 500 ug / ml, 250 ug / ml, 125 ug / / ml) and the amount of free radicals was reduced to some extent.

Specifically, 0.5 ml of a 0.1 mM DPPH solution was added to 0.4 ml of ethanol, and 0.1 ml of an extract of P. falciparum diluted with constant concentrations (1 mg / ml, 500 ug / ml, 250 ug / ml, 125 ug / ml and 62.5 ug / ml) After vortexing for 10 seconds, the cells were incubated in a dark place for 30 minutes. Absorbance was measured at 517 nm using ELISA. The degree of antioxidant activity was expressed as a percentage based on the absorbance of the control group using ethanol. As a positive control, Vitamin C (AA-2G, Ascorbic Acid 2-Glucoside, DAMY chemical) Were used.

Free radical activity inhibition rate (%) = 100- (Reaction absorbance of each sample liquid / Reaction absorbance of the blank) X100

As a result, as shown in FIG. 6, it was confirmed that the antioxidant effect was about 30% as compared with the control group at 1 mg / ml.

Experimental Example  4: UV  Cell damage Protection ability  exam

The effect of UV-B-induced cytotoxicity on the recovery of cellulolytic extract of Haenafuta spp. When HaCaT, a human keratinocyte cell line, was induced by UV-B and then treated with the extract of the present invention . Specific test methods include CCD-986sk, a fibroblast cell line of human skin cells, and HaCaT cells, a human keratinocyte cell line, with Dulbecco's Modified Eagle's Medium (DMEM), 10% fetal bovine serum (FBS), and 1% Antibiotic-antimycotic ( GIBCO, Cat No. 15240-062) were incubated in a 75 Cm ² flask at 37, 5% CO ₂ . When the cells were confluenced more than 80%, they were subcultured to 1 well in ⁶ wells and cultured until the confluence reached more than 80% under cell culture conditions. After the medium was discarded, UV irradiation was performed with UV-B at 50 mJ. Then, the foxtail extract of Example 1 was treated and further cultured for 48 hours. After that, MTT solution (6.6 mg / ml, 3- (4,5-dimethylthiazol-2yl) -2,5 diphenyl-2H-tetrazolium bromide solution) was added to each well. After removing the culture medium, 200 μl of dimethylsulfoxide (DMSO, Amresco, 0231-500ML) was added and the mixture was shaken for 10 minutes. Then, 100 μl of the solution was taken in 96-well plate and absorbance was measured at 540 nm with an ELISA (Enzyme-Linked Immunosorbent Assay) reader. The degree of cytotoxicity was expressed as a percentage based on the absorbance intensity of the control using pure water.

Cell Stability (%) = (absorbance of test group / absorbance of control group) x100

As shown in FIG. 7, when the cells were treated with UV-B, the cells were damaged by about 40% as compared with the untreated control. In addition, the treatment of the extracts of P. falciparum from 1 mg / ml to 62.5 ug / ml restored UV damage at the most concentrations. In the 1 mg / ml treatment group, the cell damage was completely recovered, Cell proliferative ability was about 15%.

Preparation of cosmetic composition

Cosmetics of emulsified form such as nutritional lotion, cream, essence, etc. and cosmetics of solubilized form such as softened longevity were prepared with the cosmetic containing the extract of the present invention as an active ingredient.

Production Example 2-1: Lotion

According to the following formulation, it was prepared according to the conventional lotion preparation method.

Raw material name Weight% (w / w) glycerin 5.0 Dipropylene glycol 3.0 Hyaluronic acid 0.5 Polyoxyethylene hardened castor oil 0.1 Polyethylene oleyl ethyl 0.1 ethanol 5.0 antiseptic 0.15 Beauty Suitable amount flash Suitable amount Yeondu Hair Extract 2.0 Purified water to 100

Production Example 2-2: Essence

According to the following prescription, it was prepared according to a conventional method for producing essence.

Raw material name Weight% (w / w) Cetostearyl alcohol 1.0 Self emulsifying monostearic acid 1.0 Wax 0.5 Squalane 5.0 Isocetyl octanoate 3.0 Dimethylsiloxane 0.3 Sorbitan monostearate 0.5 Polyethylene glycol monostearate 8.0 glycerin 4.0 Propylene glycol 0.2 Carboxy polymer 0.22 Triethanolamine 0.25 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Yeondu Hair Extract 7.0 Purified water to 100

Manufacturing example  2-3: Lotion

According to the following formulation, it was prepared according to a conventional lotion preparation method.

Raw material name Weight% (w / w) Cetostearyl alcohol 0.8 Self emulsifying monostearic acid 1.0 Wax 0.5 Stearic acid 0.5 Liquid paraffin 7.0 Squalane 5.0 Macadamia oil 3.0 Isocetyl octanoate 2.0 Dimethylsiloxane 0.3 Sorbitan monostearate 0.5 Polyethylene glycol monostearate 1.2 glycerin 4.0 Propylene glycol 4.0 Betaine 4.0 Carboxy polymer 0.12 Triethanolamine 0.15 antiseptic 0.25 Spices Suitable amount flash Suitable amount Yeondu Hair Extract 5.0 Purified water to 100

Manufacturing example  2-4: Cream

According to the following formulation, it was prepared according to a conventional cream production method.

Raw material name Weight% (w / w) Cetostearyl alcohol 3.0 Self emulsifying monostearic acid 1.5 Chin type monostearate 1.5 Wax 0.5 Liquid paraffin 8.0 Squalane 7.0 Isocetyl octanoate 4.0 Refined jojoba oil 4.0 Dimethylsiloxane 0.3 Sorbitan monostearate 1.0 Polyethylene glycol monostearate 1.2 glycerin 6.0 Propylene glycol 4.0 Betaine 4.0 Xanthan gum 0.06 Triethanolamine 0.10 antiseptic 0.25 Spices Suitable amount flash Suitable amount Yeondu Hair Extract 7.0 Purified water to 100

Manufacturing example  2-5: Gel

According to the following prescription, it was prepared according to a conventional gel preparation method.

Raw material name Weight% (w / w) glycerin 4.0 Propylene glycol 4.0 ethanol 10 Polyoxyethylene hardened castor oil 0.1 Carboxy polymer 0.30 Triethanolamine 0.30 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Yeondu Hair Extract 1.0 Purified water to 100

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (6)

An external composition for skin containing the extract of Papenfussiella kuromo as an active ingredient. [Claim 7] The composition for external application for skin according to claim 1, wherein the extract is an ethanol extract, a hot water extract, or an ultrasound extract. The composition for applying the external of the skin according to claim 1, wherein the composition is for preventing skin aging. The external preparation composition for skin according to claim 1, wherein the composition is for improving skin wrinkles. The composition for applying the external of the skin according to claim 1, wherein the composition is for whitening. According to claim 1, wherein the composition is an external composition for skin, characterized in that for antioxidant.

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