KR101086227B1 - Cosmetic Composition Comprising the extract of Lloydia triflora Bak as Active Ingredient - Google Patents

Abstract

The present invention is Naloy dog persimmon ( Lloydia triflora Bak ) relates to a cosmetic composition containing the extract as an active ingredient, characterized in that the Nado dog persimmon extract contains 0.001 to 30.0% by weight relative to the total weight of the cosmetic composition, according to the present invention, In addition to antioxidant effects, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, melanin production inhibitory effect, inflammatory cytokine expression suppression effect by ultraviolet irradiation and skin wrinkle improvement effect can provide various functional cosmetics.

Nado dog persimmon, antioxidant, anti-aging, wrinkle, whitening, skin irritation relief, cosmetics

Description

Cosmetic composition containing Nado dog persimmon extract as an active ingredient {Cosmetic Composition Comprising the extract of Lloydia triflora Bak as Active Ingredient}

The present invention relates to a cosmetic composition containing the Nadoga persimmon 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 free radicals may be caused by aging, which intensifies the body's ability to destroy antioxidant defenses and damage 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 over-inflammatory reactions and skin elasticity. If this becomes worse, mutations in the DNA can lead to mutations, cancer, and reduced immune function.

Therefore, it is necessary to protect the cell membrane by eliminating free radicals generated by metabolic processes of the body, ultraviolet rays, and inflammatory reactions.Also, 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 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.

On the other hand, skin changes occur due to pigmentation. Factors that determine skin color include basic or partial pigmentation, such as tanning due to blemishes, freckles and ultraviolet exposure, except for race, region, gender and age differences, as well as the distribution of acne, scars and keratin. And blood circulation, stress, and health. Among the above factors, the most important factor is pigmentation.

Pigments that affect skin color include melanin, melanoids, carotene, and hemoglobin, the most important of which is melanin, the most important factor affecting the biosynthesis of melanin is the amount of ultraviolet rays and hormones in the body.

Melanin plays a major role in preventing or damaging the skin from ultraviolet rays by absorbing or scattering ultraviolet rays. There is no special maximum absorption wavelength and it absorbs light in all areas. In addition, the function of removing active oxygen species is excellent, but sometimes melanin itself generates free radicals, and other substances are reduced or oxidized by catechol or quinone in melanin structure, and melanin itself shows the properties of free radicals. Pray.

The biosynthesis of melanin is a series of oxidation processes that begins with the conversion of tyrosine, an amino acid, from the melanosomes of melanocytes to tyrosinase and conversion to dihydroxy phenylalanine. eumelanin).

This biosynthesis process is carried out in a special type of brown cells in organelles called melanosomes. Melanosomes, including melanin granules, move from the periphery of the nucleus to the dendritic end of the dendritic projections and into the cytoplasm by the action of keratinocytes. Accumulate around the nucleus.

The synthesis of melanin, the number of melanosomes, and the migration to the surrounding keratinocytes are partially affected by hormones and ultraviolet rays, and primarily by genetic influences. In addition, it is known that interferon, prostaglandin, histamine, and other metal ions such as cytokines, copper, zinc and iron, which are intracellular regulators involved in the expression of tyrosinase and the synthesis and transfer of melanin.

Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150) and some extracts inhibit tyrosinase. It has activity and is used as a whitening cosmetic, but its stability in cosmetic formulations is low, so that its use is limited due to decomposition or coloring, odor occurrence, efficacy at the biological level, unclear effect and safety problems. The above substances have been shown to have a tyrosinase inhibitory effect, but the effect is low in experiments similar to the actual biological level. Therefore, the evaluation of the inhibitory effect by melanoma cell culture similar to the biological level is required. In addition, hydroquinone is defined as a carcinogenic substance and its use is prohibited or restricted in cosmetics. Kojic acid and ascorbic acid are very unstable substances. Cosmetics containing a small amount of the above components may cause browning when stored for several weeks at room temperature.

In order to solve these various 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.

Therefore, the present inventors have studied the possibility of applying to cosmetics in various natural products, and selected the Nadogae persimmon of Liliaceae, and extracts were prepared from them, thereby enhancing skin antioxidant effect, MMP-1 production inhibitory effect, collagen synthesis As a result of measuring the effect, the inhibitory effect of melanin production, the inhibitory effect of inflammatory cytokine expression by ultraviolet irradiation, and the effect of improving skin wrinkles, it was found that the efficacy as a cosmetic product can be expected because of its excellent efficacy.

Accordingly, it is an object of the present invention to provide a cosmetic for inhibiting skin damage by anti-oxidation, anti-wrinkle, whitening, and ultraviolet irradiation containing Nadogae persimmon extract as an active ingredient.

That is, the purpose of the present invention is to contain the extract of Nadogae persimmon extract applied to skin cosmetics, skin antioxidant effect, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, melanogenesis inhibitory effect, inflammatory cytokine expression inhibitory effect by UV irradiation To provide a cosmetic composition exhibiting excellent antioxidant, anti-aging, wrinkle improvement, whitening effect and skin damage inhibiting effect.

Another object of the present invention to provide a cosmetic method using the cosmetic composition containing the Nado dog persimmon extract.

The object of the present invention as described above is achieved by the cosmetic composition for antioxidant containing Nado dog persimmon extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for improving skin wrinkles containing Nado dog persimmon extract as an active ingredient.

Another object of the present invention is achieved by a cosmetic composition for skin whitening containing Nado dog persimmon extract as an active ingredient.

Still another object of the present invention is achieved by a cosmetic composition for inhibiting skin damage by ultraviolet rays containing Nadoga persimmon extract as an active ingredient.

Still another object of the present invention is achieved by a cosmetic composition for alleviating skin irritation by ultraviolet rays containing Nadogae persimmon extract as an active ingredient.

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

In addition, preferably, the Nadogam persimmon extract 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. It is characterized in that extracted by fermentation.

Still another object of the present invention is achieved by a cosmetic method comprising applying the cosmetic composition to human skin to obtain an antioxidant effect, skin wrinkle improvement effect, skin whitening effect or skin damage suppression effect by ultraviolet rays.

Nadogae persimmon extract according to the present invention has an antioxidant effect (Experimental Example 1, 2), MMP-1 production inhibitory effect (Experimental Example 3), collagen synthesis enhancing effect (Experimental Example 4), melanin production inhibitory effect (Experimental Example 5) , Inflammatory cytokine expression suppression effect by ultraviolet irradiation (Experimental Example 6) and skin wrinkle improvement effect (Experimental Example 7).

In addition, the Nadogae persimmon extract has a whitening effect such as melanin production inhibitory effect, which is a substance causing blemishes, freckles and skin pigmentation.

Therefore, the cosmetic composition comprising the Nadogi persimmon extract extracts excellent skin damage by inhibiting the expression of inflammatory cytokines by UV irradiation along with excellent effects such as antioxidant effect, collagen degrading enzyme activity control effect, skin fine wrinkle improvement effect, anti-inflammatory, whitening, etc. It can be seen that it has an inhibitory effect and a stimulating alleviation effect.

The present invention relates to a cosmetic composition for antioxidant, for preventing skin aging, for improving skin wrinkles, for skin whitening, and for inhibiting skin damage by UV rays, which contains the extract of Nadogae persimmon as an active ingredient.

Nadogae persimmon used as an active ingredient in the cosmetic composition of the present invention ( Lloydia triflora Bak ) is a lily family plant distributed in Japan, Manchuria, Sakhalin and Kamchatka. It grows wild in Jirisan and Mudeungsan north of Jeonnam. Its habitat is mountain meadows and blooms from May to June. It is grown a lot for ornamental purposes. One leaf and one stem from the plastic stem stand straight. The stem is weak and stands straight and its branches are about 20cm in height without branching.

In the present invention, the Nadogae persimmon extract means that it is obtained by extracting from various organs or parts (eg leaves, flowers, roots, stems and seeds, etc.) of Nadogae persimmon, preferably obtained from leaves, stems, roots Extract, more preferably an extract obtained from the stem or root, most preferably an extract obtained from the root.

In addition, the Nadogae persimmon 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, 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, Solvent extraction using a solvent selected from the group consisting of dichloromethane, chloroform, hexane and mixtures thereof (b) extraction using 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.

In the present invention, a solvent extraction method using an extraction solvent is preferably used.

In the present invention, Nadogae persimmon extract is a variety of extraction solvents, for example, water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butyl Ethylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane, and mixtures thereof, which can be obtained using at least one extraction solvent, preferably ethanol, It is obtained using 70% (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 Nadogae persimmon extract 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 Nadogae persimmon extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone the conventional purification and fermentation process. 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 according to 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 and diethyl ether can be used. The extracted sample may be vacuum filtered to recover the filtrate and then removed by a vacuum evaporator, and the extract may be obtained through a conventional extract preparation method of lyophilization.

Nadogae persimmon extract according to the present invention also includes a fermented extract, fermented extract of nado dog persimmon can be prepared as follows. After finely crushing Nadogae persimmons to 100 ~ 500 mesh, 1 ~ 50g / L is added to a conventional microbial culture, and microorganisms such as yeast strain or lactic acid bacteria are added in an 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 eliminates free radicals and free radicals, inhibits the production of MMP-1, inhibits the production of MMP-1 overproduced by ultraviolet light, enhances biosynthesis of collagen, and inhibits melanin production of melanocytes, It is possible to suppress the expression of inflammatory cytokines expressed by ultraviolet irradiation, characterized in that it comprises an effective amount of Nadogagamchae extract effective in improving wrinkles.

According to the present invention, the Nadogae persimmon extract is contained 0.001 to 30.0% by weight based on the total weight of the cosmetic composition, more preferably, the Nadogae persimmon extract contains 0.01 to 10% by weight relative to the total weight of the cosmetic composition It is characterized by. When the content of the extract is less than 0.001% by weight, no skin improvement effect is observed, and when the content of the extract is more than 30.0% by weight, the degree of improvement of the skin improvement effect against the increase of the content is insignificant, and there is a problem in the safety and stability of the formulation and economics. Neither ever

The cosmetic composition of the present invention containing the Nadogi persimmon extract has an antioxidant effect (Experimental Examples 1 and 2), MMP-1 production inhibitory effect (Experimental Example 3), collagen synthesis enhancing effect (Experimental Example 4), melanin production inhibitory effect (Experimental example 5), the inflammatory cytokine expression suppression effect (experimental example 6) by ultraviolet irradiation and the skin wrinkle improvement effect (experimental example 7) are excellent.

Therefore, the cosmetic composition of the present invention containing the Nado dog persimmon extract having such various functions has an excellent antioxidant effect, skin anti-aging effect, skin wrinkle improvement effect, skin whitening effect, skin damage relieving effect and irritation relieving effect It is characterized by having.

The components included in the cosmetic composition of the present invention may include components conventionally used in cosmetic compositions in addition to the active ingredient, such as conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, and Carrier.

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, it 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 dosage form of the present invention is a suspension, water, a liquid diluent such as ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline 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 is applied to the human skin by applying a cosmetic composition comprising the Nado dog persimmon extract of the present invention as an active ingredient, characterized in that it has an antioxidant effect, wrinkle improvement effect, skin whitening effect and skin damage inhibitory effect Provide makeup methods.

The cosmetic method of the present invention refers to all cosmetic methods using the cosmetic composition of the present invention. That is, all 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.

When the cosmetic method using the cosmetic composition containing the Nadogae persimmon extract of the present invention, the antioxidant effect, MMP-1 production inhibitory effect, collagen synthesis enhancement effect, melanin production inhibitory effect (skin whitening effect), inflammatory by ultraviolet irradiation Cytokine expression inhibitory effect (skin damage inhibitory effect) and skin wrinkle improvement 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  One: Me too  Extract manufacturer

After dipping, the roots of the dried Nadogae persimmon were refluxed three times for 5 hours with an aqueous 70% (V / V) ethanol solution, cooled, and filtered through a filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 캜 or lower. Nadogae persimmon extract was prepared by dispersing / dissolving using a 80% butylene glycol solution such that the vacuum concentrate and the freeze-dried were 1% by weight.

Experimental Example  One: Me too  Extract NBT Measure antioxidant effect

Experimental Example 1 In order to confirm the antioxidant effect of the Nadogae persimmon extract obtained in Preparation Example 1 BHT (Butylated Hydroxytoluene), a substance known to have an antioxidant effect as a comparison group, the antioxidant activity was measured by the NBT method.

In order to measure the antioxidant effect, the active oxygen produced by xanthine and xanthine oxidase is measured by NBT method, and the test substance (nado persimmon extract, BHT) removes free radicals, that is, free radical scavenging effect. Evaluate. Free radicals are produced by xanthine and xanthine oxidase. This active oxygen reacts with Nitro Blue Tetrazolium (NBT) and the active oxygen scavenging rate (%) is measured by measuring the blue color produced by this at wavelength 560 nm.

As a measuring method

0.05M Na ₂ CO ₃ ------------------------------------ 2.4ml

2.3mM xanthine solution ---------------------------------- 0.1ml

3.3mM EDTA Solution ------------------------------------ 0.1ml

4.BSA solution ---------------------------------------- 0.1ml

5.0.72 mM NBT solution ---------------------------------- 0.1ml

6.Xanthine Oxidase Solution ----------------------------- 0.1ml

7.6 mM CuCl ₂ solution ----------------------------------- 0.1ml

① Add 1,2,3,4,5 to the vial, and add 0.1ml of sample solution to it and leave at 25 ℃ for 10 minutes.

② Add 6 and stir quickly, and start incubation for 20 minutes at 25 ℃.

③ After that, add 7 to stop the reaction and measure the absorbance St at 560nm.

④ The blank of the sample solution is measured in the same manner as in the case of measuring the absorbance St, except that distilled water is used instead of the above 6 to measure the absorbance So.

⑤ The absorbance test is performed in the same manner as in the case of measuring the absorbance St, except that distilled water is used instead of the sample solution.

⑥ The blank of the blank test was measured in the same manner as when the absorbance St was measured except that the sample solution and 6 were used instead of distilled water, and the absorbance Bo was measured.

Active oxygen scavenging rate (%) was calculated by Equation 1, the results are shown in Table 1.

St: absorbance at 560 nm after enzyme reaction of sample solution

Bt: absorbance at 560 nm after enzymatic reaction of blank test solution

So: absorbance at 560 nm before reaction without enzyme in sample solution

Bo: absorbance at 560 nm before reaction in the absence of enzyme in blank test solution

Sample Name Treatment concentration (%) Active oxygen scavenging rate (%) Nado Dog Persimmon Extract (Manufacturing Example 1) 0.01 92.2 BHT 0.01 90.2

As a result of the experiment, the active oxygen scavenging rate (antioxidant effect) was confirmed in all the samples tested at 0.01% concentration as shown in Table 1. Nadogae persimmon extract of the present invention was confirmed to have a better antioxidant effect compared to BHT at 0.01% concentration.

Experimental Example  2: Me too  Extract DPPH Measure antioxidant effect

Experimental Example 2 is to measure the antioxidant effect of the Nadogae persimmon extract obtained in Preparation Example 1, the vitamin C, a substance known to have an antioxidant effect as a comparison group by using the DPPH method (antifree radical scavenging rate ) Was measured.

The DPPH method measures the antioxidant effect by reducing power using a free group called DPPH (2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl) free radical. The free radical scavenging rate (%) is measured at a wavelength of 560 nm by comparing the degree of DPPH reduction by the test substance (nado Doga extract, vitamin C) to decrease the absorbance.

The reagent used was 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) as 61.88 mg in methanol to make 100 ml.

As a measuring method

① Add 0.15 ml of the sample solution to 0.15 ml of 0.1 mM DPPH solution in a 96-well plate, stir rapidly, and incubate for 10 minutes at 25 ° C.

(2) Then measure the absorbance St at 560nm.

③ The blank of the sample solution is measured in the same manner as when the absorbance St is measured except that methanol is used instead of the 0.1 mM DPPH solution to measure the absorbance So.

④ The blank test is performed in the same manner as in the case of measuring the absorbance St, except that distilled water is used instead of the sample solution to measure the absorbance Bt.

⑤ Blank in the blank test was measured in the same manner as when the absorbance St was measured except that methanol was used instead of the 0.1 mM DPPH solution and distilled water was used instead of the sample solution.

The result of free radical scavenging rate (%) was calculated by Equation 2, and the results are shown in Table 2 below.

St: absorbance at 560 nm after free radical scavenging of sample solution

Bt: absorbance at 560 nm after free radical scavenging of blank test solution

So: Absorbance at 560 nm before reaction without free radicals in sample solution

Bo: absorbance at 560 nm before reaction without free radicals in blank test solution

Sample Name Treatment concentration (%) Free radical scavenging rate (%) Nado Dog Persimmon Extract (Manufacturing Example 1) 0.05 88.3 vit-C 0.05 93.5

As shown in Table 2, the Nadogae persimmon extract according to the present invention has a superior antioxidant effect than the antioxidant effect of vitamin C, a substance known to have a free radical scavenging rate (antioxidant effect) at a concentration of 0.05%. Confirmed.

Experimental Example  3. Me too  Extract MMP -1 suppression effect

Experimental Example 3 is known to have the effect of inhibiting the production of MMP-1 in order to determine whether the Nadogi persimmon extract obtained in Preparation Example 1 has an effect of inhibiting the production of collagen degrading enzyme MMP-1. TGF-β (100 ng / ml, Roche, US) was used as a comparative group to measure 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 incubation for 24 hours in the serum-free DMEM medium to the Nadogi persimmon extract of Preparation Example 1 was added to a final concentration of 200 ㎍ / ㎖ and added for 48 hours in a serum-free DMEM medium not containing Nadoga persimmon extract as a control Incubated with

After incubation, the supernatant of each well was 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 3 below. Inhibition rate was calculated and the results are shown in Table 3.

Test substance % Inhibition of MMP-1 production Nado dog persimmon extract (production example 1) 55.7 TGF-β 53.5

As shown in the results of Table 3, the Nadogae persimmon extract of the present invention exhibited an inhibition rate of 55.7% when the inhibition rate of MMP-1 production was measured at a concentration of 200 µg / ml, and the inhibition rate of TGF-β was 53.5%. Indicated. In view of the difference between the concentration of TGF-β (100 ng / ml) and the concentration of Nadogae persimmon extract of the present invention (200 ㎍ / ml), which are known to have an inhibitory effect on MMP-1 production, several kinds of substances were mixed. Indicative of the MMP-1 inhibition rate as described above at the extract level shows that the Nadogaechaechae extract of the present invention has an inhibitory effect on MMP-1 production.

Experimental Example  4. Me too  Enhancement of Collagen Synthesis by 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 Nadogae persimmon extract was further incubated for 48 hours in serum-free DMEM medium added with a final concentration of 100 ㎍ / ㎖ and serum-free DMEM medium containing no Nadogae persimmon 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 Nadogae persimmon extract was 0.01% (w / v). The experiment was conducted. Collagen biosynthesis increase rate was calculated according to the following equation (4), the results are shown in Table 4.

Test substance Collagen biosynthesis growth rate (%) Nado dog persimmon extract (production example 1) 71.4 TGF-β 69.6

Table 4 shows the results obtained by measuring the collagen synthesis enhancement effect of the culture medium added Nadogi persimmon extract and TGF-β using fibroblasts. According to Table 4, the Nadogae persimmon extract of the present invention showed a biosynthesis rate of 71.4% when the collagen biosynthesis increase rate was measured at a concentration of 0.01%, and TGF-β, a substance known to enhance collagen synthesis, was 69.6%. It was found that the biosynthesis rate of. Even considering the difference between the concentration of TGF-β (10 ng / ml) and the concentration of Nadogae persimmon extract of the present invention (0.01% (w / v)), which are known to have collagen synthesis-promoting effects, several kinds of substances are mixed. The collagen biosynthesis rate of the above degree at the level of the extracted extract shows that the Nadogae persimmon extract of the present invention has a collagen synthesis enhancing effect.

Experimental Example  5: Me too  Inhibitory Effects of Extracts on Melanogenesis in B16F1 Melanogenic Cells

Experimental Example 5 is to determine the whitening effect by looking at the degree of inhibition of melanin production on B16F1 melanin forming cells in order to confirm the whitening effect of the Nadogae persimmon extract obtained in Preparation Example 1. B16F1 melanin forming cells used in Experimental Example 5 are cell strains derived from mice, and cells that secrete a black pigment called melanin.

Samples were treated during the artificial culture of these cells to evaluate the degree of reduction of melanin black pigment. B16F1 melanin forming cells used in the present experiment was used by receiving from the American Type Culture Collection (ATCC).

The melanin biosynthesis inhibitory effect of B16F1 melanin forming cells was measured as follows.

B16F1 melanin forming cells were divided into 6 well plates at a concentration of 2 X 10 ⁶ per well, and the cells were attached and incubated for 72 hours by treating the Nadogae persimmon extract according to Preparation Example 1 at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, and the cells were counted and centrifuged to recover the cells. Quantification of intracellular melanin was performed with a slight modification of the method of Rotan: Cancer Res., 40: 3345-3350, 1980. The cell pellet was washed once with PBS, and then 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added to vortex for 5 minutes to disrupt the cells. Melt the extracted melanin by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and then measured the absorbance of melanin at 405 nm with a microplate reader. Melanin production inhibition rate (%) was measured. The melanin production inhibition rate (%) of B16F1 melanin forming cells was calculated by the following Equation 5, the results are shown in Table 5. At this time, hydroquinone and arbutin, which are known to have melanin production inhibitory effects, were used as a comparison group.

A: Melanin amount in wells without sample

B: Melanin amount in the wells to which the sample was added

Sample Name Treatment concentration (%) Melanin production inhibition rate (%) Nado dog persimmon extract (production example 1) 0.1 61 Hydroquinone 0.1 65 Arbutin 0.1 42

As can be seen from the results of Table 5, it can be seen that the Nadogae persimmon extract of the present invention significantly inhibits melanin production in B16F1 melanin forming cells. In the case of Nadogae persimmon extract, the melanin production inhibition rate was 61% at 0.1% concentration, and melanin production inhibition was lower than that of hydroquinone when compared to arbutin and hydroquinone, substances known to have a whitening effect at the same concentration. Although it showed an effect, it showed an excellent melanin production inhibitory effect than arbutin. Therefore, it can be seen that the cosmetic composition comprising the extract of Nado dog persimmon of the present invention has an excellent effect on inhibiting melanin production.

Experimental Example  6: Me too  Inhibitory Effects of Extracts on Inflammatory Cytokine Expression by Ultraviolet Irradiation

Experimental Example 6 is to evaluate the inhibitory effect of the inflammatory cytokine expression expressed by UV irradiation of the Nadogae persimmon extract obtained in Preparation Example 1, 24-fibroblasts (Fibroblast) isolated from human epidermal tissue 5 x 10 ⁴ pieces were placed in a well 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 processing the Nadogae persimmon 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 Nadogae persimmon extract. In comparison group, 0.1% of indomethacin, an inhibitor of inflammatory cytokine expression, was used. The amount of IL-1α was quantified using an enzyme immunoassay (Enzyme-linked Immunosorbent Assay), and the expression inhibition rate (%) of inflammatory cytokines (IL-1α) was calculated by Equation 6 below. Shown in

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 Nado dog persimmon extract 0.1% 55.3 0.2% 62.3 Indomethacin 0.1% 53.2

According to Table 6, Nadogae persimmon extract inhibits the expression (production) of inflammatory cytokine IL-1α by UV irradiation at 5 %% at 0.1% concentration, and 62.3% IL-1α when treated at 0.2% concentration. It was found that the expression of was suppressed. In particular, it was found that the Nadogae persimmon extract according to the present invention showed an excellent inflammatory cytokine expression rate than indomethacin, an inflammatory cytokine expression inhibitor at the same concentration. Therefore, it can be seen that the Nadogae persimmon extract of the present invention effectively protects the inflammation caused by ultraviolet irradiation even at low concentrations.

Hereinafter, based on the results of the above experimental examples, to present a cosmetic composition containing Nadogae persimmon 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 7 in accordance with a conventional method.

ingredient Content (unit: weight%) Nado dog persimmon 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 8 in accordance with a conventional method.

ingredient Content (unit: weight%) Nado dog persimmon 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 examples of nutrient lotion was prepared as shown in Table 9 according to a conventional method.

ingredient Content (unit: weight%) Nado dog persimmon 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 as shown in Table 10 in accordance with a conventional method.

Formulation Example 4 Comparative Formulation Example 1 Comparative Formulation Example 2 ingredient Content (unit: weight%) Nado dog persimmon 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 as shown in Table 11 in accordance with a conventional method.

ingredient Content (unit: weight%) Nado dog persimmon 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 12 below.

ingredient Content (unit: weight%) Nado dog persimmon 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 13.

ingredient Content (unit: weight%) Nado dog persimmon 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  7. Me too  Effect of Extracts on Skin Wrinkles

Clinical trials were carried out on the skin wrinkle improvement effect of the cosmetic containing Nado dog persimmon extract of the present invention. Clinical trials were evaluated through the actual use test of each cosmetic.

The experiment was compared to the nutritional cream of Formulation Example 4 shown in Table 10 containing the Nadogae persimmon extract and the cream substituted Nado dog persimmon extract with purified water Comparative Formulation Example 1, a retinol which is a substance known to have an anti-wrinkle effect As Comparative Formulation Example 2, the effect of improving skin wrinkles due to the use of cosmetics was identified and compared to humans.

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 10. 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 shown in Table 14 below.

[Table 14]

Wrinkle Improvement Effect of Nutritious Cream Containing Nado Dog Persimmon Extract (Gross Eye Evaluation)

Wrinkle improvement Effective rate (%) Great slightly none Formulation example 4 (nado dog persimmon extract) 24 4 2 93.3 Comparative Formulation Example 1 (purified water) 3 4 23 23.3 Comparative Formulation Example 2 (Retinol) 22 4  4 86.6

According to the experimental results, the cream of Formulation Example 4 containing the Nadogae persimmon extract according to the present invention showed a wrinkle improvement effect more than 3.4 times compared to the cream of Comparative Formulation Example 1 not containing the Nadogae persimmon extract. 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.

Claims (8)

Antioxidant cosmetic composition containing Nado dog persimmon extract as an active ingredient. Cosmetic composition for improving skin wrinkles containing Nado dog persimmon extract as an active ingredient. Cosmetic composition for skin whitening containing Nado dog persimmon extract as an active ingredient. Cosmetic composition for inhibiting skin damage by ultraviolet rays containing Nado dog persimmon extract as an active ingredient. Cosmetic composition for skin irritation relief containing Nado dog persimmon extract as an active ingredient. The cosmetic composition according to any one of claims 1 to 5, wherein the Nadogae persimmon extract is contained in an amount of 0.001 to 30.0 wt% based on the total weight of the cosmetic composition. The method according to any one of claims 1 to 5, wherein the Nadogam persimmon extract is (a) water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, Solvent extraction using an extraction solvent selected from the group consisting of chloroform, butyl acetate, diethyl ether, dichloromethane, hexane and mixtures thereof, (b) supercritical extraction, (c) ultrasonic extraction or (d) fermentation from microorganisms B. A cosmetic composition, characterized in that extracted using fermented and extracted by using natural fermentation metabolism. A cosmetic composition according to any one of claims 1 to 5, wherein the cosmetic composition is applied to human skin to obtain an antioxidant effect, a skin wrinkle improvement effect, a skin whitening effect or a skin damage suppression or irritation relief effect by ultraviolet rays. Way.