KR101086225B1 - Cosmetic Composition Comprising Smilax china as Active Ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition containing Smilax china L. (Liliaceae) extract as an active ingredient, characterized in that it contains 0.001 to 30.0% by weight of the extract as an active ingredient, According to the extract, the blue mule vine extract can provide various functional cosmetics as well as the effect of activating human fibroblasts, improving skin wrinkles, alleviating skin damage caused by ultraviolet rays, and alleviating skin irritation.

Blue vine, anti-aging, skin wrinkle improvement, skin irritation, cosmetics

Description

Cosmetic composition containing the extract of blue vines as an active ingredient {Cosmetic Composition Comprising Smilax china as Active Ingredient}

The present invention relates to a cosmetic composition containing the blue vine extract as an active ingredient.

Human skin undergoes various physical and chemical changes during the aging process. The causes are largely divided into intrinsic aging and photo-aging, and research on this has been actively conducted. UV rays, stress, disease conditions, environmental factors, wounds, and aging can cause free radicals to become active. If this condition is exacerbated, it destroys the antioxidant defenses in vivo and damages cells and tissues. And aging.

More specifically, lipids, proteins, polysaccharides, nucleic acids, and the like, which are major constituents of skin, are oxidized to destroy skin cells and tissues, resulting in skin aging.

In particular, the oxidation of proteins is caused by severe cuts in the skin's connective tissues such as collagen, hyaluronic acid, elastin, proteoglycan, and fibronectin, resulting in severe hyperinflammatory reactions and skin elasticity. Disrupting and worsening, DNA mutations lead to mutations, cancer, and reduced immune function.

Therefore, to protect the cell membrane by eliminating the free radicals generated by the body's metabolic process, ultraviolet radiation, or inflammatory reactions, and to protect the cell membranes, the already damaged cells must be regenerated by active metabolism to proliferate the cells. The skin can recover quickly and maintain healthy skin.

Aging involves not only these free radicals but also an enzyme called matrix metalloproteinase (MMP), a collagen degrading enzyme. In other words, the synthesis and degradation of extracellular matrix such as collagen in vivo is properly controlled, but collagen synthesis decreases as aging progresses, and the expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen, promotes the skin. Elasticity is reduced and wrinkles are formed. In addition, such degradation enzymes may be activated by ultraviolet irradiation.

Therefore, there is a need for the development of a substance capable of regulating or inhibiting MMP expression in which activation is induced in cells. Until now, most of the raw materials used as materials for cosmetics simply inhibited enzyme activity. Therefore, we tried to regulate the expression and activity of MMP expression induced by intracellular natural aging and photoaging.

In order to solve the above problems, cosmetics using natural products have recently been developed to reduce skin irritation caused by various chemicals. Natural ingredients have less side effects on the skin, and as the consumer's response to cosmetics using natural ingredients has increased recently, the development value of cosmetic ingredients is increasing.

Thus, the present inventors have studied the possibility of applying to cosmetics in a variety of natural products that have not been known, as a result, by selecting the extract from the blue vines of maple and deciduous broad-leaved arborescent and extracts from it, to inhibit the MMP-1 production, UV induction Inhibition of MMP-1 production, collagen synthesis enhancement effect, cytotoxic effect by UV irradiation, and inflammatory cytokine expression inhibition effect by UV irradiation were measured. Was found.

SUMMARY OF THE INVENTION An object of the present invention is to provide a cosmetic composition containing an extract of blue vines showing an excellent anti-aging effect such as inhibiting MMP-1 production, inhibiting MMP-1 production by UV induction, enhancing collagen synthesis, and preventing skin wrinkles. have.

In addition, an object of the present invention is to provide a cosmetic composition exhibiting the effect of inhibiting skin damage by the ultraviolet rays and the skin irritation effect when the blue vine extract extract is applied to the skin cosmetic.

It is another object of the present invention to provide a cosmetic method using the cosmetic composition containing the blue vine myrtle extract.

Therefore, the object of the present invention as described above is achieved by a skin anti-aging cosmetic composition comprising a blue vine extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for improving skin wrinkles comprising blue vine extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for inhibiting skin damage by ultraviolet rays comprising the extract of blue vines as an active ingredient.

Still another object of the present invention is achieved by a cosmetic composition for alleviating skin irritation, which comprises the extract of blue vines as an active ingredient.

Preferably, the herbaceous vine extract is characterized in that it contains 0.001 to 30.0% by weight based on the total weight of the cosmetic composition.

Also, preferably, the extract of blue vine is (a) water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, Solvent extraction using an extraction solvent selected from the group consisting of dichloromethane, hexane and mixtures thereof, (b) supercritical extraction, (c) ultrasonic extraction, or (d) fermentation from microorganisms or natural fermentation metabolism. Characterized in that extracted.

Also, preferably, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream and spray It is characterized by having any one of the formulations selected from the group consisting of.

In addition, the cosmetic composition is characterized in that it exhibits wrinkle improvement, anti-aging effect, cytotoxicity relieving effect by ultraviolet irradiation, skin damage suppression and irritation relieving effect.

Further, preferably, the cosmetic composition is effective for inhibiting MMP-1 production, inhibiting MMP-1 production by UV induction, enhancing collagen synthesis, improving skin wrinkles, reducing cytotoxicity by UV irradiation, and UV irradiation. Inhibits inflammatory cytokine expression.

In addition, another object of the present invention is achieved by a cosmetic method characterized in that the cosmetic composition is applied to human skin to prevent skin aging, improve wrinkles, inhibit skin damage caused by ultraviolet rays or to alleviate skin irritation. .

Cheongmirae vine extract according to the present invention has excellent anti-aging effects such as MMP-1 production inhibitory effect (Experimental Example 1), MMP-1 production inhibitory effect (Experimental Example 2), skin wrinkle improvement effect (Experimental Example 6) And it has the effect of relieving skin irritation caused after ultraviolet irradiation (Experimental Example 4).

Therefore, the cosmetic composition comprising the extract of the herbaceous vine extract has collagen degrading enzyme activity control effect (Experimental Examples 1 and 2), collagen synthesis enhancement effect (Experimental Example 3) skin fine wrinkles improvement effect (Experimental Example 6), cytotoxicity relieving effect (Example 4), the inflammatory cytokine expression inhibitory effect (Experimental Example 5) and the like has excellent effects.

The present invention relates to a cosmetic composition for preventing skin aging, for improving skin wrinkles, for inhibiting skin damage caused by ultraviolet rays, or for alleviating skin irritation, comprising the extract of blue vines as an active ingredient.

Smilax China (Smilax china L. (Liliaceae)) is used as an active ingredient of the cosmetic composition of the invention goes beoteo rhizome is thick and stiff as the leaves of vine plant with lily and long in the next winding. Stems grow every 2m around each bar with hook-like thorns. The leaves are alternate, round or broad oval, thick and shiny. The flowering period is May, the fruit is round, about 1cm in diameter, and ripens in red from September to October. It is distributed in Korea, Japan, China, Philippines, and Indochina, and grows at the edge of forests in the mountains. Young sprouts are eaten as herbs, and the fruit is called sweet or sour. Roots are effective for diuresis, detoxification, gusts, etc. Used for arthritis, back pain, and boils.

In the present invention, the blue mule vine extract means that it is obtained by extracting from various organs or parts (for example, leaves, flowers, roots, stems, branches, shells and seeds, etc.) of the blue mule vines, preferably the leaves, stems, branches , An extract obtained from the bark or root, more preferably an extract obtained from the leaves, branches, bark or root, and most preferably means an extract obtained from the root.

In addition, the herbaceous vine extract of the present invention may be prepared according to a conventional method known in the art, that is, using a conventional solvent, under the conditions of conventional temperature and pressure, but preferably (a) water, Anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloro Extraction is carried out using a solvent extraction method using a solvent selected from the group consisting of methane, chloroform, hexane and mixtures thereof, (b) supercritical extraction by reduced pressure and high temperature with carbon dioxide, or (c) ultrasonic extraction. Or (d) fermented and extracted by fermentation from various microorganisms or natural fermentation metabolism.

Preferably, in the present invention, a solvent extraction method using an extraction solvent is used.

In the present invention, the herbaceous vine extract is selected from various extracting solvents, for example, water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (eg, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene It can be obtained using one or more extraction solvents selected from the group consisting of glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, preferably ethanol, 70 It is obtained using% (v / v) ethanol aqueous solution or water, and most preferably it is obtained using 70% (v / v) ethanol aqueous solution.

On the other hand, it is apparent to those skilled in the art that the extract of blue vines of the present invention can obtain an extract having substantially the same effect using not only the aforementioned extraction solvent but also other extraction solvents.

In addition, the herbaceous vine extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract that has undergone conventional purification and fermentation. For example, decompression by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by ultrasonic extraction, separation using ultrafiltration membranes with constant molecular weight cut-off values, various chromatography (size, charge, hydrophobicity or affinity) The active fraction obtained by various purification and extraction methods additionally carried out, such as by the separation by the sex) or by fermentation products using natural or various microorganisms, is also included in the extract of the present invention.

In addition, the present invention provides a cosmetic composition, characterized in that the extract is obtained by cold condensation at room temperature, heating and filtration, further obtained by concentrating the solvent under reduced pressure or lyophilization.

The decompression by carbon dioxide and the supercritical extraction by high temperature mean supercritical fluid extraction. Generally, the supercritical fluid is a liquid having a gas when the gas reaches a critical point under high temperature and high pressure. It is gaseous, chemically similar in polarity to nonpolar solvents, and because of its properties, supercritical fluids are used for extraction of fat-soluble substances (J. Chromatogr. A. 1998; 479: 200-205).

Carbon dioxide becomes a supercritical fluid with both liquid and gaseous properties as the pressure and temperature reach a critical point through the operation of a supercritical fluid device, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the fat-soluble substance contained in the sample is extracted from the supercritical carbon dioxide.

After extracting the fat-soluble substance, the supercritical carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction container to extract components that were not extracted with pure supercritical carbon dioxide alone.

The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract the active ingredient by using a supercritical carbon dioxide or a mixed fluid in which a co-solvent is added to carbon dioxide.

The cosolvent may be used one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether.

Most of the extracted samples contain carbon dioxide. Since carbon dioxide is volatilized into air at room temperature, the extract obtained by the above method may be used as a cosmetic composition, and the cosolvent may be removed by a reduced pressure evaporator.

In addition, the ultrasonic extraction method is an extraction method using the energy generated by the ultrasonic vibration, the ultrasonic wave can destroy the insoluble solvent contained in the sample in the water-soluble solvent, the reactants located around due to the high local temperature generated Since the kinetic energy of the particles is increased, sufficient energy required for the reaction is obtained, and the effect of ultrasonic energy is induced to increase the pressure by increasing the mixing effect of the substance and the solvent contained in the sample, thereby increasing the extraction efficiency.

As the extraction solvent that can be used in the ultrasonic extraction method, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane, diethyl ether and mixtures thereof can be used. The extracted sample is vacuum filtered to recover the filtrate, and then removed by a vacuum evaporator, the extract can be obtained through a conventional extract preparation method of freeze drying.

Cheongmirae vine extract according to the present invention also includes a fermented extract, fermented extract of cheonmirae vine can be prepared as follows. Finely crush the blue mule vine about 100 ~ 500 mesh, and then add 1 ~ 50g / L of the conventional microbial culture, and add the microorganism such as yeast strain or lactic acid bacteria in the amount of 10,000 ~ 100,000 cfu / L. The culture temperature is incubated in the normal microbial culture conditions of 30 ~ 37 ℃. The pH is 5 to 7 and incubated for about 5 to 10 days under aerobic or anaerobic conditions. It can then be obtained through aging and filtration.

The cosmetic composition of the present invention inhibits the production of MMP-1, inhibits the production of MMP-1 overproduced by ultraviolet rays, enhances collagen biosynthesis, alleviates cytotoxicity by ultraviolet rays, and is expressed by inflammatory cytokines. It is characterized in that it comprises an effective amount of blue vine extract extract capable of inhibiting the expression of.

According to the present invention, the blue rice vine extract is contained 0.001 to 30.0% by weight based on the total weight of the cosmetic composition, more preferably, the blue rice vine extract is contained 0.01 to 10% by weight based on the total weight of the cosmetic composition It features. When the content of the extract of the gingiva myrtle vinegar is less than 0.001% by weight, the skin improvement effect does not appear, and when the content of the blue rice vine extract exceeds 30.0% by weight, the degree of improvement of the skin improvement effect on the increase of the content is insignificant, and there is a problem in the safety and stability of the formulation. It is not economical.

The cosmetic composition of the present invention containing the blue vine extract, MMP-1 production inhibitory effect (Experimental Example 1) and MMP-1 production inhibitory effect by ultraviolet (Experimental Example 2), collagen synthesis enhancement effect (Experimental Example 3) It is excellent in the cytotoxic mitigation effect (experimental example 4) by ultraviolet irradiation, the inflammatory cytokine expression suppression effect (experimental example 5), and the skin wrinkle improvement effect (experimental example 6) by ultraviolet irradiation.

Therefore, the cosmetic composition of the present invention containing the extracts of the blue mule vine having such various functions has an excellent skin anti-aging effect, human fibroblast activity, skin wrinkle improvement effect, skin damage relieving effect and irritation relieving effect It features.

Ingredients included in the cosmetic composition of the present invention as an active ingredient may include ingredients commonly used in cosmetic compositions in addition to the active ingredient, for example, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings Adjuvants, and carriers.

Cosmetic compositions of the present invention may be prepared in any formulation conventionally prepared in the art and include, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing , Oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like, but are not limited thereto. More specifically, the cosmetic composition of the present invention may be prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, and microcrystals are used as carrier components. Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide ether. Sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In addition, the present invention by using a cosmetic composition comprising the extract of the blue mule vine of the present invention as an active ingredient inhibits MMP-1 production, inhibits MMP-1 production by ultraviolet light, enhance collagen synthesis, alleviate cytotoxicity by ultraviolet irradiation, ultraviolet light It provides a cosmetic method characterized by suppressing inflammatory cytokine expression by irradiation and improving skin elasticity.

The cosmetic method of the present invention refers to all cosmetic methods using the cosmetic composition of the present invention. That is, all the methods known in the art using the cosmetic composition belong to the makeup method of the present invention.

The cosmetic composition of the present invention may be used alone or in duplicate, or may be used in combination with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention may be used according to a conventional method of use, and the number of times of use may vary depending on the skin condition or taste of the user.

If the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation or a surfactant-free cleansing formulation, it may be wiped off, peeled off or washed with water after application to the skin. As a specific example, the soap is liquid soap, powdered soap, solid soap and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel and cleansing pack, the surfactant-free cleansing formulation is a cleansing cream , Cleansing lotion, cleansing water and cleansing gel, but are not limited thereto.

Performing a cosmetic method using the cosmetic composition comprising the extract of the blue mule vine of the present invention, human fibroblast activity effect, skin fine wrinkles improvement effect, skin damage relieving effect and irritation relieving effect by UV irradiation, improve skin elasticity and excellent skin aging Prevention effect can be obtained.

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 describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Manufacturing example  1: Medicinal herb extract

Vine roots of finely dried blue vines were refluxed three times for 5 hours with a 70% (V / V) ethanol aqueous solution, cooled, and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 캜 or lower. Cheongmirae vine extract was prepared by dispersing / dissolving using a 50% butylene glycol solution so that the vacuum concentrate and the freeze-dried were 1% by weight.

Experimental Example  1. Of Blue Laver Vine Extract MMP -1 suppression effect

Experimental Example 1 is a substance known to have an effect of inhibiting the production of MMP-1 in order to determine whether the blue vine extract obtained in Preparation Example 1 has an effect of inhibiting the production of collagen degrading enzyme MMP-1 Phosphorus TGF-β was used as a comparative group to determine the effect of inhibiting the production of MMP-1.

Fibroblasts (Korean Cell Line Bank, South Korea), which are human normal skin cells, were seeded in 48-well microplates (Nunc, Denmark) at 1 × 10 ⁶ cells per well, and treated with DMEM medium (Sigma, United States) and 37 ° C. After culturing for 24 hours at 24 hours in a serum-free DMEM medium to which the final extract of blue vine extract of Preparation Example 1 was added at a final concentration of 200 ㎍ / ㎖ and a serum-free DMEM medium containing no blue vine extract as a control for 48 hours It was.

After incubation, the supernatants of each well were collected and the amount of ng / ml newly synthesized MMP-1 was measured using an MMP-1 assay kit (Amersham, USA), and MMP-1 production was performed according to Equation 1 below. The percent inhibition was calculated and the results are shown in Table 1.

At this time, TGF-β (100 ng / ml, Roche, United States), a substance known to have an effect of inhibiting MMP-1 production, was used as a comparison group.

Test substance % Inhibition of MMP-1 production Cheongmirae Vine Extract (Manufacturing Example 1) 68.7 TGF-β 23.5

As shown in the results of Table 1, the blue vine extract of the present invention showed an excellent inhibition rate (%) of MMP-1 production almost three times higher than TGF-β.

Experimental Example  2. Induction of Ultraviolet Rays by Extracts MMP -1 suppression effect

In general, MMP-1 is known to increase its amount by ultraviolet light. Therefore, the experiment was conducted to see if the blue vine extract of the present invention inhibits the amount of MMP-1 that is increased by ultraviolet light. MMP-1 production inhibition effect measurement by UV induction was carried out in the same manner as in Experiment 1 except that a certain amount of UVB treatment prior to sample treatment. Cheongmirae vine extract used in this experiment was obtained from Preparation Example 1, but is not limited thereto. Inhibition rate of MMP-1 production of the herbaceous vine extract was calculated according to Equation 1, and the results are shown in Table 2 below. At this time, TGF-β (100 ng / ml, Roche, United States), a substance known to have an inhibitory effect on the production of MMP-1, was used as a comparison group, and the extracts of blue vines were tested at a concentration of 0.01% (w / v). .

Test substance % Inhibition of MMP-1 production Cheongmirae Vine Extract (Manufacturing Example 1) 65.3 TGF-β 32.5

Table 2 shows the results of measuring the amount of collagen degrading enzyme MMP-1 secreted in the culture medium to which the blue vine extract and TGF-β culture medium were added after UVB irradiation to the cultured fibroblasts. Inhibition rate (%) of the production of MMP-1 at 0.01% (w / v) concentration of the extract of the present invention is about 2 times better than TGF-β, a substance known to inhibit MMP-1 production by UV rays. The effect was shown.

Experimental Example  3. Collagen Synthesis Enhancement Effect of Extracts

Fibroblasts, human normal epithelial cells, were seeded in 48-well microplates at 1 × 10 ⁶ cells per well and incubated in DMEM medium for 24 hours. The herbaceous vine extract was further incubated for 48 hours in serum-free DMEM medium added with a final concentration of 100 ㎍ / ㎖ and serum-free DMEM medium not containing the herbaceous vine extract as a control. After incubation, the supernatant of each well was collected and measured in ng / ml by measuring the amount of procollagen (procollagen) type I C-peptide (PICP) using a collagen kit (Takara, Japan). Measured. In the comparison group, TGF-β (100 ng / ml, Roche, United States), a substance known to have an effect of enhancing collagen synthesis, was used as a comparison group, and the extracts of blue vines were 0.01% (w / v). Was carried out. Collagen biosynthesis increase rate (%) was calculated according to the following equation (2), the results are shown in Table 3.

Test substance Collagen Biosynthesis Growth Rate (%) Cheongmirae Vine Extract (Manufacturing Example 1) 87.4 TGF-β 69.6

According to Table 3, it can be seen that the collagen biosynthesis increase rate (%) of the extract of the herbaceous vine extract of the present invention is superior to that of TGF-β, which is known to have an effect of improving collagen synthesis.

Experimental Example  4: Cytotoxicity-Reducing Effect of Ultraviolet Irradiation of Cheongmirae Vine Extract

Experimental Example 4 was carried out to evaluate the mitigating effect of cytotoxicity by ultraviolet irradiation of the extract of blue vines obtained in Preparation Example 1.

Fibroblasts (fibroblasts) were placed in 24-well test plates 1 X 10 ⁵ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with UV 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B15W, Sankyo Dennki, Japan), after which PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. It was incubated for 24 hours after the blue vine extract treatment. After 24 hours, the medium was removed, and 500 μl of cell culture medium and 60 μl of MTT solution (2.5 mg / ml) were added to each well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04N) was added thereto. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, followed by measurement of 565 nm absorbance in a microplate reader. Cell viability (%) was measured by Equation 3 below, and cytotoxicity relaxation rate (%) by UV was calculated by Equation 4.

Bo: 565 nm absorbance of wells that only developed cell culture media

Bt: 565 nm absorbance of the wells that developed the colorless reaction wells

St: 565 nm absorbance of the wells that reacted the sample treated wells

Bo: cell viability of wells without UV irradiation and no sample treatment

Bt: cell viability of wells that were not irradiated with UV light

St: Cell survival rate of wells treated with UV

Sample Name Treatment concentration Cytotoxic alleviation rate (%) Blue Laver Vine Extract 0.1% 23.5 0.2% 34.7

According to Table 4, the cytotoxic mitigation rate of the extract of blue vines was 23.5% at 0.1% and 34.7% at 0.2%, showing that the cytotoxicity mitigation rate increased with increasing treatment concentration. The extract was found to effectively protect against cytotoxicity caused by UV light. Through the above experimental results, it could be seen that the extract of the present invention, the blue vine extract effectively protects cell damage caused by ultraviolet rays even at low concentrations.

Experimental Example  5: Inhibitory Effect of Cheongmirae Vine Extract on Inflammatory Cytokine Expression by Ultraviolet Irradiation

Experimental Example 5 is to evaluate the inhibitory effect of the inflammatory cytokine expression expressed by UV irradiation of the extract of blue vines obtained in Preparation Example 1, 24-well fibroblasts isolated from human epidermal tissue 5 x 10 ⁴ pieces were put on the test plate and attached for 24 hours.

Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with 10 mJ / cm 2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then PBS was removed and FBS was not added to the cell culture medium (DMEM). Medium) 350 μl was added.

After treatment with the blue vine extract to be evaluated here was incubated for 5 hours. 150 µl of the culture supernatant was taken to quantify IL-1α to determine the inhibitory effect of inflammatory cytokine expression of the extract of Blue Crab. The amount of IL-1α was quantified using an enzyme immunoassay (Enzyme-linked Immunosorbent Assay), and the percent inhibition of expression of inflammatory cytokines (IL-1α) was calculated by Equation 5 below.

Bo: IL-1α production in wells without UV irradiation

Bt: IL-1α production in wells that were not irradiated with UV light

St: IL-1α production amount of wells that were irradiated with UV light

Sample Name Treatment concentration (%) Inflammatory cytokines
% Inhibition of expression Blue Laver Vine Extract 0.1% 57.3 0.2% 68.7

According to Table 5, it can be seen that the extract of blue vine extract inhibits the production of inflammatory cytokine IL-1α by UV rays at 5 %% at 0.1% concentration and 68.7% at 0.2% concentration. It was found that the production of IL-1α was inhibited in proportion to the concentration. Therefore, it was found that the extract of blue vines of the present invention effectively prevents inflammation caused by ultraviolet rays even at low concentrations.

Hereinafter, based on the results of the above experimental examples, to present a cosmetic composition containing the blue vine extract as an active ingredient, but the formulation of the present invention is not necessarily limited thereto.

Formulation example  1. Flexible Cosmetics

Formulation examples of the flexible longevity was prepared as shown in Table 6 in accordance with a conventional method.

ingredient Content (unit: weight%) Blue Laver Vine Extract
glycerin
1.3-butylene glycol
PEG 1500
Allantoin
DL-panthenol
E.T.A.-2NA
Benzophenone-9
Sodium hyaluronate
ethanol
Octicdodeceth-16
Polysorbate 20
Preservative, fragrance, coloring
Distilled water 3.0
5.0
3.0
1.0
0.1
0.3
0.02
0.04
5.0
10.0
0.2
0.1
a very small amount
Balance Sum 100

Formulation example  2. Converging Cosmetics

Formulation examples of convergent longevity was prepared as shown in Table 7 in accordance with a conventional method.

ingredient Content (unit: weight%) Blue Marrow Vine Extract
glycerin
1.3-butylene glycol
Allantoin
DL-panthenol
E.T.A.-2NA
Benzophenone-9
Sodium hyaluronate
ethanol
Polysorbate 20
Witch hazel extract
Citric acid
Preservative, fragrance, coloring
Distilled water 1.5
2.0
2.0
0.2
0.2
0.02
0.04
3.0
15.0
0.3
2.0
a very small amount
a very small amount
Balance Sum 100

Formulation example  3. Nutritional Cosmetics

Formulation example of nutrient cosmetics was prepared as shown in Table 8 in accordance with a conventional method.

ingredient Content (unit: weight%) Blue Laver Vine Extract
Stearyl alcohol
lanolin
Polysorbate 60
Sorbitan stearate
Cured Vegetable Oil
Mineral oil
Squalane
Trioctanoine
Dimethicone
Tocopherol Acetate
Carboxy Vinyl Polymer
glycerin
1.3-butylene glycol
Sodium hyaluronate
Tri ethanolamine
Preservative, fragrance, coloring
Distilled water 1.5
1.5
1.5
1.3
0.5
1.0
5.0
3.0
2.0
0.8
0.5
0.12
5.0
3.0
5.0
0.12
a very small amount
Balance Sum 100

Formulation example  4. Nutrition Cream

Formulation of nutrition cream was prepared according to the conventional method as shown in Table 9.

Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation Example 2 ingredient Content (unit: weight%) Blue Laver Vine Extract
Retinol
Lipophilic monostearate
Cetearyl Alcohol
Stearic acid
Beeswax
Polysorbate 60
Sorbitan stearate
Cured Vegetable Oil
Squalane
Mineral oil
Trioctanoine
Dimethicone
Sodium magnesium silicate
glycerin
Betaine
Triethanolamine
Sodium hyaluronate
Preservative, fragrance, coloring
Purified water 3.0
-
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance -
-
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance -
3000 IU
2.0
2.2
1.5
1.0
1.5
0.6
1.0
3.0
5.0
5.0
1.0
0.1
5.0
3.0
1.0
4.0
a very small amount
Balance Sum 100 100 100

Formulation example  5. Massage Cream

Formulation example of the massage cream was prepared according to the conventional method as shown in Table 10 below.

ingredient Content (unit: weight%) Blue Laver Vine Extract
Lipophilic monostearic acid glycerin
Stearyl alcohol
Stearic acid
Polysorbate 60
Sorbitan stearate
Isostearyl Isosterate
Squalane
Mineral oil
Dimethicone
Hydroxyethyl Cellulose
glycerin
Triethanolamine
Preservative, fragrance, coloring
Distilled water 3.0
1.5
1.5
1.0
1.5
0.6
5.0
5.0
35.0
0.5
0.12
6.0
0.7
a very small amount
Balance Sum 100

Formulation example  6. Essence

Formulation examples of the essence was prepared according to the conventional method as shown in Table 11.

ingredient Content (unit: weight%) Blue Laver Vine Extract
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
E.T.A.-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxy Vinyl Polymer
Triethanolamine
Octyldodecanol
Octyldodeceth -16
ethanol
Preservative, fragrance, coloring
Distilled water 5.0
10.0
5.0
2.0
0.1
0.3
0.02
0.04
0.1
8.0
0.2
0.18
0.3
0.4
6.0
a very small amount
Balance Sum 100

Formulation example  7. Pack

Formulation examples of the pack was prepared according to the conventional method as shown in Table 12 below.

ingredient Content (unit: weight%) Blue Marrow Vine Extract
Polyvinyl alcohol
Cellulose gum
glycerin
Fiji 1500
Cyclodextrin
DL-Panthenol
Allantoin
Monomethylammonium Glycyric Acid
Nicotinamide
ethanol
Fiji 40 Cured Castor Oil
Incense, pigment, preservative
Distilled water 3.0
15.0
0.15
3.0
2.0
0.15
0.4
0.1
0.3
0.5
6.0
0.3
a very small amount
Balance Sum 100

Experimental Example  6. Effect of Cheongmirae Vine Extract on Skin Wrinkle

A clinical experiment was conducted on the skin wrinkle improvement effect of the cosmetic composition containing the extract of Blue vines of the present invention. Clinical trials were evaluated through the actual use test of each cosmetic.

The experiment was compared to the formulation of Example 1 containing the medicinal cream of the extract shown in Table 9 and the cream replacing the bluish vine extract with purified water in Comparative Example 1, compared to the retinol which is a substance known to have an anti-wrinkle effect As Formulation Example 2, the effect of improving skin wrinkles caused by the use of cosmetics in humans was identified and compared.

The cosmetics of Formulation Example 4 and Comparative Formulation Example 1 used in this comparative evaluation experiment are in the form of a cream, the composition of which is shown in Table 9. For 60 women (20-35 years old), the experimental group was divided into three groups, each of 20 groups of Formulation Example 4 experimental group, Comparative Formulation Example 1 treatment group, and Comparative Formulation Example 2 treatment group. After months, the wrinkle improvement effect was visually evaluated to confirm the degree of wrinkle improvement.

After completion of the experiment, the skin wrinkle improvement effect was evaluated by visual observation by an experienced doctor before and after using the product for three months. The experimental results are as shown in Table 13 and FIG.

[Table 13]

Wrinkle improvement Effective rate (%) Great slightly none Cream of Formulation Example 4 24 4 2 85.0 Comparative Formulation Example 1 (purified water) 3 4 23 25.0 Comparative Formulation Example 2 (Retinol) 22 4  4 86.6

 According to the experimental results of Table 13 and FIG. 1, the present invention (the cream of Formulation Example 4) containing the extract of blue vine extract is about 3.4 times higher anti-wrinkle effect than the cream of Comparative Formulation Example 1 containing no extract of blue vine extract Showed. It was found that the wrinkle improvement effect was superior to that of the cream of Comparative Formulation Example 2 containing retinol, a substance known to have an anti wrinkle effect. Moreover, the skin irritation could not be observed in the skin of the subjects who apply the cosmetic composition of the present invention to the skin.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that such a specific technology is only a preferred embodiment, and the scope of the present invention is not limited thereto. Therefore, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

1 is a view showing the skin wrinkle improvement effect according to an embodiment of the present invention.

Claims (8)

Skin anti-aging cosmetic composition comprising the extract as an active ingredient. A cosmetic composition for improving skin wrinkles comprising the extract of blue vines as an active ingredient. Cosmetic composition for inhibiting skin damage by ultraviolet rays comprising the extract of blue vines as an active ingredient. Cosmetic composition for alleviating skin irritation comprising the extract of blue vines as an active ingredient. The cosmetic composition according to any one of claims 1 to 4, wherein the blue vine extract is contained in an amount of 0.001 to 30.0% by weight based on the total weight of the cosmetic composition. The method according to any one of claims 1 to 4, wherein the blue vine extract is (a) water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform Solvent extraction using an extraction solvent selected from the group consisting of butyl acetate, diethyl ether, dichloromethane, hexane and mixtures thereof, (b) supercritical extraction, (c) ultrasonic extraction or (d) fermentation from microorganisms, Cosmetic composition, characterized in that extracted by fermentation using natural fermentation metabolism. The cosmetic composition according to any one of claims 1 to 4, wherein the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion emulsion, wax foundation Cosmetic composition, characterized in that it has any one formulation selected from the group consisting of a pack, massage cream and spray. A cosmetic method according to claim 1, wherein the cosmetic composition of any one of claims 1 to 4 is applied to human skin to prevent skin aging, improve wrinkles, inhibit skin damage by ultraviolet rays, or alleviate skin irritation.

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