KR20210129945A - Cosmetic composition for preventing skin photo-aging comprising extract of castanea crenata or fractions from thereof as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing photoaging of the skin containing Castanea crenata inner shell extract or isolated compounds thereof, scoparone and scopoletin, as active ingredients. The cosmetic composition according to the present invention has an effect of inhibiting wrinkle formation or melanin biosynthesis due to UV-induced skin damage.

Description

COSMETIC COMPOSITION FOR PREVENTING SKIN PHOTO-AGING COMPRISING EXTRACT OF CASTANEA CRENATA OR FRACTIONS FROM THEREOF AS ACTIVE INGREDIENT }

The present invention relates to a cosmetic composition for preventing photoaging of the skin containing, as an active ingredient, a Yulpi extract ( Castanea crenata ) or a mixture of scopoletin and scoparone, which are separated compounds thereof.

The skin, the largest organ of the human body, has various physiological functions such as the function of protecting various organs inside the skin from the outside, the barrier function that prevents the leakage of moisture and useful components from the inside, the function of temperature control, the function of excretion, and the function of breathing. It is a very important institution in charge. However, with age, the skin barrier function decreases as the thickness of the epidermis, dermis, and subcutaneous tissue becomes thinner, and the physiological functions of the skin are deteriorated, such as dry skin due to a rapid decrease in the moisture content of the skin, resulting in various changes and aging. .

In addition, most people's skin responds easily to sunlight during growth, but as the skin tissue gradually changes as the age increases, roughness due to sunlight, wrinkles, pigmentation spots, poor complexion, diarrhea, capillary dilatation, Black spots, purpura and easy bruising, atrophy, fibrosis, discolored areas, and, in extreme cases, precancerous and malignant tumors occur.

Ultraviolet rays are divided into UVA, UVB, and UVC regions, and only UVA and UVB act on the earth, and the UVC region is blocked by the ozone layer. UVB has the greatest effect on the skin and mainly penetrates into the epidermis and affects skin cells. UVA penetrates into the dermis of the skin and mainly affects skin cells related to collagen synthesis. These ultraviolet rays affect the skin either directly or indirectly by generating active oxygen. The effects of UV rays on skin cells are as follows. First, among skin cells, keratinocytes secrete inflammatory cytokines when irradiated with ultraviolet rays, and oxidative stress is induced, so that various types of protein kinases and phospholipases are activated in the cells. Activated these enzymes hydrolyze cell membrane lipids to release arachidonic acid. The secreted arachidonic acid is converted to prostaglandin E2 (PGE2) by the action of cyclooxygenase. Arachidonic acid and PGE2 thus formed cooperate with cytokines to cause various inflammatory reactions including redness in the skin, and ultimately accelerate skin aging.

The physiological changes of the skin due to aging can be broadly divided into three categories: first, the thinning of the epidermis, dermis, and subcutaneous tissue, which are components of the skin, and second, the lipid barrier, which functions as the skin barrier. ), the lipid composition and content change, and its function deteriorates rapidly, resulting in a decrease in the moisture content of the skin and drying of the skin, and finally, a phenomenon in which spots, freckles, pigmentation, and various skin lesions are induced. Recently, the amount of ultraviolet rays reaching the skin is gradually increasing due to environmental pollution, which causes various causes related to skin aging such as generation of free radicals, reduction of skin elasticity, pigmentation, melasma, and freckles.

In the skin, elastin fibers form cross-links with collagen fibers inside the epidermis. As we age, the elasticity of the skin rapidly decreases due to the action of the elastin-degrading enzyme, elastase, and sagging occurs. In addition, histologically, the activity of inflammatory cells increases with age, and shows a deficiency and aggregation of elastin fibers and a decrease in collagen fibers, and biochemically, the activity of elastase is remarkably reduced. Elastase is the only enzyme known so far that can degrade elastin, and inhibition of this activity or production is a way to radically reduce skin aging.

A person's skin color is largely determined by the amount of melanin, hemoglobin, and carotene, among which melanin plays the most important role. Melanin not only determines a person's skin color, but also acts as a UV absorber and a free radical scavenger. When it is produced in the skin, pigmentation occurs within the skin, causing darkening of the skin color, spots, and freckles.

The melanin synthesis process is as follows: First, in melanocytes, using tyrosine as a substrate, an enzyme called tyrosinase produces dopaquinone (DOPA quinone), and from dopaquinone through an autoxidation reaction and an enzymatic reaction, a copolymer melanin is produced. Melanin produced in this way is transferred to keratinocytes through melanosomes and comes out to the surface of the skin through a 28-day keratinization process in keratinocytes. However, in this process, an excess of melanin is produced by a factor that promotes melanin production, and if melanin is not completely eliminated by keratinization, a pigmentation phenomenon appears. Therefore, in order to prevent this pigmentation phenomenon, it is possible by controlling some processes in the melanin production process.

Korean Patent Registration No. 10-1040507 discloses a cosmetic composition containing an extract of Magnolia magnolia, which has an anti-photoaging effect.

In addition, Korean Patent No. 10-0308178 discloses a cosmetic composition containing yulpi extract and having an effect of improving skin wrinkles.

The present inventors confirmed the various skin physiological activities of natural plants and their isolated compounds to effectively improve the photoaging phenomenon, and as a result, among natural plant extracts, a mixture of yulpi ( Castaneacrenata ) extract and its isolated compounds, scofarone and scopoletin, is effective for skin aging. By confirming that it effectively prevents, the present invention was completed.

The above object is achieved by a cosmetic composition for preventing photoaging of the skin containing, as an active ingredient, a mixture of yulpi ( Castanea crenata ) extract or an isolated compound thereof, scoparone and scopoletin.

Preferably, the mixture of scoparone and scopoletin is characterized in that it is mixed in a weight ratio of 1-10:10-1.

Also preferably, the prevention of skin photoaging includes inhibiting wrinkle production or inhibiting melanin biosynthesis according to skin damage caused by UV.

Also preferably, the cosmetic composition may contain yulpi extract or a mixture of scofarone and scopoletin, which are isolated compounds thereof, in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition.

Also preferably, the yulpi extract may be extracted with an extraction solvent selected from the group consisting of water, anhydrous or hydrated lower alcohol having 1-4 carbon atoms, acetone, ethyl acetate and butyl acetate.

Also preferably, the cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. It may have a formulation selected from

The present invention promotes collagen biosynthesis and inhibits MMP-1 activity, inhibits elastase enzyme activity, and tyrosina It is characterized in that it inhibits enzyme activity, inhibits melanin synthesis, and promotes cell proliferation.

All technical terms used in the present invention, unless otherwise defined, have the following definitions and have the meanings as commonly understood by one of ordinary skill in the art in the relevant field of the present invention. In addition, although preferred methods and samples are described herein, similar or equivalent ones are also included in the scope of the present invention.

The term "about" refers to 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, means an amount, level, value, number, frequency, percentage, dimension, size, amount, weight or length varying by 4, 3, 2 or 1%.

Throughout this specification, unless the context requires otherwise, the terms "comprises" and "comprising" include the steps or elements presented, or groups of steps or elements, but any other step or element, or It is to be understood that a step or group of elements is not excluded.

The present invention relates to a cosmetic composition for preventing photoaging of skin containing a mixture of yulpi ( Castanea crenata ) extract or an isolated compound thereof, scoparone and scopoletin, as an active ingredient.

Yulpi (Castanea crenata) used in the present invention is an inner bark obtained from a chestnut fruit of the Oak family, and has been used as a treatment for ailments such as nausea, leprosy, and rhinitis under the name of yulbu since ancient times.

Yulpi extract of the present invention can be prepared according to a conventional method known in the art, that is, under conditions of conventional temperature and pressure, using a conventional solvent, but preferably (a) water, carbon number 1- 4 anhydrous or hydrated lower alcohols (e.g., methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform , hexane, and a solvent extraction method using a solvent selected from the group consisting of mixtures thereof (b) supercritical extraction using reduced pressure and high temperature with carbon dioxide or (c) ultrasonic extraction. In the present invention, preferably, a solvent extraction method using an extraction solvent was used. Yulpi extract in the present invention is a variety of extraction solvents, for example, water, anhydrous or water-containing lower alcohols having 1-4 carbon atoms (eg, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol , glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof can be obtained using at least one extraction solvent selected from the group consisting of, preferably ethanol, 70% (v/v) ethanol or water, and most preferably 70% (v/v) ethanol.

On the other hand, it is obvious to those skilled in the art that the yulpi extract of the present invention can be obtained using not only the above-described extraction solvent, but also an extract exhibiting substantially the same effect using other extraction solvents. In addition, the yulpi extract of the present invention includes not only the extract by the above-described extraction solvent, but also an extract that has undergone a conventional purification and fermentation process. For example, decompression by carbon dioxide, extraction by supercritical extraction by high temperature, extraction by extraction using ultrasound, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity Active fractions obtained through various additional purification and extraction methods, such as separation by means of separation by sex) or extracts by fermentation products using natural conditions or various microorganisms, are also included in the extract of the present invention.

Yulpi extract according to the present invention also includes an extract that has undergone a fermentation process, and the fermented extract of Yulpi ( Castanea crenata ) can be prepared as follows. Yulpi is finely crushed to about 100-500 mesh, then 1-50 g/L of a conventional microbial culture is added, and microorganisms such as yeast strains or lactic acid bacteria are added in an amount of 10,000-100,000 cfu/L. The culture temperature is 30 ~ 37 ℃ cultured under normal microbial culture conditions. At a pH of 5 to 7, culture is performed for about 5 to 10 days under aerobic or usually anaerobic conditions. It can then be obtained through aging and filtration.

In addition, according to an embodiment of the present invention, the cosmetic composition of the present invention may include scoparone and scopoletin, which are isolated compounds separated from the Yulpi extract. Scoparone may be represented by the following Chemical Formula 1.

[Formula 1]

Scopoletin may be represented by Formula 2 below.

[Formula 2]

According to the present invention, the Yulpi extract or a mixture of scoparone and scopoletin is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, and more preferably, the Yulpi extract or a mixture of scoparone and scopoletin. Silver is characterized in that it is contained in an amount of 0.01 to 10% by weight based on the total weight of the cosmetic composition. When the content of the extract is less than 0.001% by weight, wrinkle improvement or whitening effect does not appear, and when it exceeds 30.0% by weight, there is a problem in safety and stability of the formulation, and it is not economical.

According to an embodiment of the present invention, the scofarone and scopoletin are mixed in a weight ratio of 1 to 10:10 to 1, thereby having an excellent photoaging prevention effect.

The ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the active ingredient as an active ingredient, for example, conventional antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances. adjuvants, and carriers. The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , oil, powder foundation, emulsion foundation, wax foundation, pack, massage cream, spray, etc., but is not limited thereto. More specifically, it may be prepared in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylglycol oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, and microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used. When 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, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether. When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether as carrier components Sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester may be used.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, removed, or washed off with water. As a specific example, the soap is liquid soap, powder soap, solid soap, and oil soap, the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, cleansing towel, and cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream , cleansing lotion, cleansing water, and cleansing gel, but not limited thereto. On the other hand, the makeup method of the present invention refers to all makeup methods using the cosmetic composition of the present invention. That is, all methods known in the art using the cosmetic composition belong to the cosmetic method of the present invention. The cosmetic composition of the present invention may be used alone or in combination, 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 can be used according to a conventional method of use, and the number of times of use can be varied according to the skin condition or taste of the user. Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples according to the gist of the present invention.

In the present invention, "extract" is an extract obtained by using Yulpi (Castanea crenata ) as a solvent, and is formed using an extract such as a diluted or concentrated liquid of the extract, a dried product obtained by drying it, a prepared or purified product of the extract, etc. Extracts of all possible formulations are meant.

Hereinafter, the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited by these Examples.

Preparation Example 1 -Yulpi ( Castanea crenata ) extract preparation

Wash 10.0 kg of Yulhu with clean water, dry it, grind it to a constant (100mesh) size, extract under reflux 3 times for 5 hours with 70% (V/V) ethanol aqueous solution, cool, and filter with Whatman #4 filter paper Thus, 1.15 kg of yulpi extract was prepared using a concentrator.

Preparation Example 2 - Preparation of scoparone

After dispersing 10 L of water in 1.0 kg of the yulpi extract obtained in Preparation Example 1, the non-polar component was removed using 1 L of n-hexane (repeated 3 times), and then 10 L of methylene chloride was added to the water layer The fractionated layer was concentrated under reduced pressure to obtain a fraction of 451 g. The active ingredient was separated using a silica gel (60 ~ 120mesh) column, and as a mobile phase, a methanol/methylene chloride mixture (1:20 -> 1:5) was used for separation and purification. A total of 12 fractions were recovered, and 9.1 g was recovered from one fraction having a collagen biosynthesis effect and a melanin synthesis inhibitory effect.

Scoparone: recrystallized from acetone, mp = 144-145℃, UV(methanol) 294 and 345nm, (1H, d, J=9.5, H-3), 3.95 and 3.92 (6H, 2s, 2 * OCH ₃ ). ¹³ C-NMR (CDCl3) 161.54 (C-2), 15.0 (C-5), 150.18 (C-7), 146.50 (C-8a), 143.41 (C-4), 113.65 (C-3), 111.54 (C-4a), 108.10 (C-6 or C-8), 100.10 (C-6 or C-8), 56.40 (2 * -OCH ₃ ), Mass M/Z 207(M+), 206 (100) , 191 (38)

Preparation Example 3 - Preparation of scopoletin

After dispersing 10 L of water in 1.0 kg of Yulpi extract obtained in Preparation Example 1, 1 L of n-hexane was used to remove the non-polar component (repeat 3 times), and then 10 L of ethyl acetate in the water layer The fractionated layer was concentrated under reduced pressure to obtain a fraction of 370 g. The active ingredient was separated using a silica gel (60 ~ 120mesh) column, and as a mobile phase, a methanol-methylene chloride mixture (1:20 -> 1:5) was used for separation and purification. A total of 14 fractions were recovered, and 1.7 g was recovered from one fraction having a collagen biosynthesis effect and a melanin synthesis inhibitory effect.

scopoletin: ¹ H NMR (300, MeOH-d ₆ ) δ: 7.59(1H, d), 6.92(1H, s), 6.85(1H, s), 6.27(1H, d), 3.96(3H, s): ¹³ C NMR (150 MHz, MeOH-d ₆ ): 162.45, 150.72, 150.05, 145.11, 144.09, 112.38, 111.18, 107.95, 103.23, 49.88, 49.46, 48.60: MS (m/z): (M+1) 193.0

Experimental Example 1: Measurement of Elastase Inhibitory Activity

Using N-succinyl-(L-Ala)3-p-nitroanilide (S4760, Sigma) as a substrate, the amount of p-nitroanilide produced from the substrate was measured at 445 nm at 37°C for 20 minutes. That is, prepare each test solution to a certain concentration, take 500 μL of each test tube, and add 500 μL of a solution of porcine pancreas elastase (E1250, Sigma) 2.5 U/mL) dissolved in 2 mM tris-HCl buffer (pH 8.6). N-succinyl-(L-Ala)3-p-nitroanilide (500 μg/mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added and reacted for 20 minutes. The elastase inhibitory activity was calculated according to Equation 1 below as the absorbance reduction rate of the sample solution addition group and the non-additive group.

[Equation 1]

Sample name Concentration (ppm) Elastase inhibition rate (%) Yulpi extract 100 21.11% ± 1.35 10 10.45% ± 2.14 One 5.08% ± 1.82 Scoparon 100 75.21% ± 1.05 10 50.48% ± 1.87 One 28.15% ± 1.29 scopoletin 100 88.73 % ±1.55 10 59.16% ± 1.67 One 30.85 % ±0.97

Experimental Example 2: Tyrosinase activity inhibition experiment

As a substrate, 40 μl of 1.5 mmol/L tyrosine (sigma) dissolved in sodium phosphate buffer (pH 6.8) was used. Each of Preparation Examples 1 to 3 was diluted with 0.05 M sodium phosphate buffer (pH 6.8) to prepare a sample solution at a predetermined concentration. 240 μl of each concentration sample solution was added to 40 μl of tyrosine, and 20 μl of tyrosinase (Sigma, 1500 U/ml) was added to the solution, followed by reaction at 37° C. for 15 minutes, and then at 490 nm. Absorbance was measured. The tyrosinase activity inhibition rate (%) was calculated by the following Equation 2 as the absorbance reduction rate of the experimental group, which was the sample solution addition group, and the control group, which was not added, as shown in Table 2.

[Equation 2]

Sample name Treatment concentration (ppm) Tyrosinase inhibition rate (%) Yulpi extract 100 23.12% ± 1.35 10 15.57% ± 2.04 One - Scoparon 100 67.84% ± 2.13 10 45.90% ± 1.95 One 17.69% ± 0.97 scopoletin 100 87.02% ± 1.38 10 52.95% ± 1.24 One 25.70% ± 1.07

Experimental Example 3: Melanin production inhibitory effect increased by UV

α-MSH used in the UV-stimulated melanin production inhibition experiment was dissolved in 10% DMSO (dimethyl sulfoxide) and used as a 1 mM solution. The mouse-derived B-16 melanoma (ATCC CRL 6323) cell line was inoculated into DMEM medium containing 4.5 g/L of glucose, 10% serum, and 1% antibiotic and passaged in a 75 cm 2 T-flask under 5% CO2 and 37°C conditions. cultured. The subcultured melanoma cell line was cultured for 24 hours in a 75cm2 T-flask, treated with 0.05% trypsin containing 0.02% EDTA to separate the cells, and then inoculated into a 75cm2 T-flask for 6 hours. Further incubation, the number of cells at this time is 1 × 10 ⁷ cells/flask. Here, melanocyte-stimulating hormone was diluted in DMEM medium and added to a final concentration of 100 μg/ml, and fluorescence sunlight in a wavelength range of 240 to 360 nm was irradiated for 1 minute, and Preparation Examples 1, 2 and 3 were mixed with DMEM. After dilution in the medium, the final concentrations of 1, 10, and 100 μg/ml were treated with the melanoma cells, and further cultured at 37° C. for 5 days. After culturing, the medium was obtained and the absorbance was measured at 475 nm to compare the degree of melanin production, and the inhibition of melanin production (%) was calculated using the absorbance value of the cell culture treated with melanocyte-stimulating hormone and the UV wavelength region as a control value. It was calculated according to Equation 3, and the results are shown in Tables 3 and 4.

[Equation 3]

Sample name Treatment concentration (ppm) Melanin production inhibition rate (%) (compared to UV treatment) Yulpi extract 100 25.18% ±1.07 10 13.27% ±1.64 One - Scoparon 100 27.84% ± 2.13 10 25.90% ± 1.95 One - scopoletin 100 30.14% ±0.85 10 19.58% ± 1.17 One -

Sample name Concentration (ppm) Sample name Concentration (ppm) Melanin production inhibition rate (%) (compared to UV treatment) scopoletin 10 Scoparon 10 81.27% ± 1.34 One 55.30 % ±1.85 One Scoparon 10 51.19 % ±2.10 One 37.79 % ±1.15

Experimental Example 4: Recovery of cytotoxicity by UV (cell proliferation)

Keratinocytes were inoculated in a 24-well plate with DMEM (Dulbeco's Modified Eagle's Medium, Gibco) supplemented with 10% FBS at a density of ^{2×10 5} _{cells/well in a humidified 37°C, 5% CO 2} incubator for 1 day. cultured. After exchange with serum-free DMEM, fluorescence sunlight in a wavelength range of 240 to 360 nm was irradiated for 1 minute, and the sample of Preparation Example was treated with a certain concentration (v/v) and cultured for 3 days. In this case, as a control (negative), saline was used instead of the material of Preparation Example. In order to see the cell-generating effect after culture, the degree of cell generation was compared and evaluated using the MTT assay method. The UV-treated group was used as the untreated group, and the cell proliferation effect with the treated group is shown in Tables 5 and 6 below.

Sample name Concentration (ppm) Cell proliferation effect (%) (compared to UV treatment) Yulpi extract 100 101.82% ± 3.51 10 - One - Scoparon 100 113.10 % ±1.27 10 105.18% ± 2.01 One - scopoletin 100 105.44 % ±2.13 10 102.27 % ±1.09 One -

Sample name Concentration (ppm) Sample name Concentration (ppm) Cell proliferation effect (%) (compared to UV treatment) scopoletin 10 Scoparon 10 157.07 % ±1.95 One 130.91 % ±2.41 One Scoparon 10 129.53% ± 2.07 One 119.41 % ± 1.61

Experimental Example 5 : Collagen biosynthesis

Inoculate fibroblasts in a 24 well plate with DMEM (Dulbeco's Modified Eagle's Medium, Gibco) with 10% FBS and Penicillin at a density of ^{2×10 5} _{cells/well, humidified at 37°C, 5% CO 2} It was cultured for 1 day in an incubator. After exchange with serum-free DMEM, fluorescence sunlight in a wavelength range of 240 to 360 nm was irradiated for 1 minute, and the sample of Preparation Example was treated at a certain concentration (v/v), and 1×10 ⁵ pieces/ml in 24 wells. 500 μl of each plate was aliquoted and cultured for 24 hours. Quantification of collagen type I in fibroblasts was carried out by immunosorbent assay (ELISA) using 'Procollagen Type I c-peptide EIAkit (TaKaRa, Japan)'. The cultured cell culture supernatant was carefully collected. First, 100 ul each of the culture supernatant cultured for 48 hours was added to a strip coated with an antibody binding to 'Procollagen Type I c-peptide' and reacted at 37°C for 2 hours. After the reaction, it was washed three times using a washing buffer to remove unreacted materials, and then 100 ul of blocking buffer was added thereto, followed by reaction at 37° C. for 2 hours. After the reaction, it is washed three times using a washing buffer to remove unreacted substances, and a solution in which POD is conjugated to another antibody binding to 'Procollagen Type I c-peptide' (Antiobody-) POD conjugate solution) 100 ul each was added and reacted at 37° C. for 2 hours. After the reaction, wash 3 times using a washing buffer to remove unreacted substances, put an equal amount of substrate solution, react for 30 minutes, and then add a stop solution (1N H2SO4) and conduct the experiment. ended. The absorbance of the strip at which the experiment was completed was measured at 450 nm, and compared with the collagen standard, the amount of newly synthesized collagen was measured as the amount of PICP. The amount of PICP ^{was converted into ng/2x10 4} cells to show the synthesis effect of collagen Table 8 shows.

Sample name Concentration (ppm) Collagen synthesis amount (ng/2x10 ⁴ )
(Compared to UV treatment) Yulpi extract 100 24.9 10 20.1 One 13.5 Scoparon 100 33.7 10 20.9 One 10.5 scopoletin 100 19.4 10 10.1 One -

Sample name Concentration (ppm) Sample name Concentration (ppm) Collagen synthesis amount (ng/2x10 ⁴ )
(Compared to UV treatment) scopoletin 10 Scoparon 10 47.2 One 40.9 One Scoparon 10 33.7 One 25.2

Experimental Example 6 : MMP-1 production inhibitory effect

Inoculate fibroblasts in a 24-well plate with DMEM (Dulbeco's Modified Eagle's Medium, Gibco) supplemented with 10% FBS and Penicillin at a density of ^{2×10 5} _{cells/well in a humidified 37°C, 5% CO 2} incubator. incubated for one day. After exchange with serum-free DMEM, fluorescence sunlight in a wavelength range of 240 to 360 nm was irradiated for 1 minute, and the sample of the preparation example was treated at a certain concentration (v/v), and 1 × 10 ⁵ pieces/ml in 24 wells. 500 μl of each plate was aliquoted and cultured for 24 hours. Collect the supernatant of each well and measure the amount (ng/ml) of newly synthesized MM P-1 using an MMP-1 assay kit (Amersham, USA), and according to Equation 4 below, MMP- compared to the UV treatment group 1 The production inhibition rate (%) was calculated, and the results are shown in Tables 9 and 10.

[Equation 4]

Sample name Concentration (ppm) MMP-1 production inhibition rate (%)
(Compared to UV treatment) Yulpi extract 100 31.21% 10 18.18% One - Scoparon 100 33.25% 10 26.11% One - scopoletin 100 29.01% 10 12.79% One -

Sample name Concentration (ppm) Sample name Concentration (ppm) MMP-1 production inhibition rate (%)
(Compared to UV treatment) scopoletin 10 Scoparon 10 67.92% One 55.04% One Scoparon 10 39.52% One 30.75%

Prescription Example 1 - Cream

A prescription example of a cream among cosmetics containing the yulpi extract and its separated compounds made through the above preparation examples is shown in Table 11 below, and is not limited to this formulation example. More specifically, it can be prepared in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a shampoo, a treatment, a scalp ampoule, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder. have.

ingredient content
(weight%) Formulation Example 1 Formulation Example 2 Formulation Example 3 Preparation Example 1 (1000 ppm) Preparation Example 2 (1000 ppm) Preparation Examples 2 and 3
(Scophalon 1ppm and
scopoletin 10ppm) glycerin glycerin glycerin 3.0 betaine betaine betaine 10.0 Fiji 1500 Fiji 1500 Fiji 1500 5.0 allantoin allantoin allantoin 2.0 DL-Panthenol DL-Panthenol DL-Panthenol 0.1 E.D.T.A-2Na E.D.T.A-2Na E.D.T.A-2Na 0.3 Benzophenone - 9
hydroxyethyl Benzophenone - 9
hydroxyethyl Benzophenone - 9
hydroxyethyl 0.02
cellulose cellulose cellulose 0.04 Sodium Hyaluronate Sodium Hyaluronate Sodium Hyaluronate 0.1 Carboxyvinyl Polymer Carboxyvinyl Polymer Carboxyvinyl Polymer 8.0 triethanolamine triethanolamine triethanolamine 0.2 Octyldodecanol Octyldodecanol Octyldodecanol 0.18 Octyldodeces -16 Octyldodeces -16 Octyldodeces -16 6.0 preservatives, fragrances, colors preservatives, fragrances, colors preservatives, fragrances, colors a very small amount
Distilled water Distilled water Distilled water remaining amount Sum 100

Example 1 : Anti-wrinkle effect of cosmetics containing Yulpi extract or an isolated compound thereof

In Formulation Examples 1 to 3, Preparation Examples 1 to 3, which are active ingredients, were replaced with purified water and evaluated using Comparative Formulation Example 1. 100 women aged 30-40 years were randomly divided into 5 groups, and after washing the cream of Formulation Examples 1 to 4 and Comparative Example twice every morning and evening, an appropriate amount of the cream was applied continuously for 2 months centered on both cheeks. The skin elasticity improvement effect of each subject was evaluated using a Cutometer (C+K, Germany) to evaluate the elasticity improvement effect. The experimental results are shown in Table 12 below.

division before use 2 weeks of use 4 weeks after use Formulation Example 1 24.10 30.24 31.23 Formulation Example 2 26.21 31.13 33.57 Formulation Example 3 20.93 29.62 33.04 Comparative Formulation Example 1 25.61 27.33 29.46

Example 2 : Whitening improvement effect of cosmetics containing Yulpi extract or its isolated compound (Scoparon)

For 10 women aged 25 to 45 with dark skin color, Preparation Examples 1 to 3, which are the active ingredients of Formulation Examples 1 to 3, were replaced with purified water and used as Comparative Formulation Example 1 for evaluation. 12 places on the subject's arms are marked by 1 cm each, and UV B is irradiated 3 times at intervals of 1 day with a light amount of 0.7 MED, and after inducing pigmentation for 7 days, each cosmetic of Formulation Examples 1 to 4 is applied. Then, the remaining one was used as a control in which the active ingredient was replaced with purified water. Cosmetics were applied twice a day for 2 months. The skin color was measured after 2 months using the Minolta CR 300, a measuring device, and the change in skin color was calculated and the results are shown in Table 13.

division ΔL value Formulation Example 1 1.98 Formulation Example 2 2.42 Formulation Example 3 2.95 Comparative Formulation Example 1 1.27

Claims (6)

Yulpi ( Castanea crenata ) extract or its isolated compound scoparone (scoparone) and scopoletin (scopoletin) a cosmetic composition for preventing skin photoaging containing a mixture as an active ingredient. The cosmetic composition for preventing photoaging of skin according to claim 1, wherein the mixture of scoparone and scopoletin is mixed in a weight ratio of 1-10:10-1. The cosmetic composition for preventing photoaging of the skin according to claim 1, wherein the skin photoaging prevention inhibits wrinkle formation or melanin biosynthesis due to UV-induced skin damage. The cosmetic composition for preventing photoaging of skin according to claim 1, wherein the cosmetic composition contains yulpi extract or a mixture of scofarone and scopoletin, which are isolated compounds thereof, in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. . According to claim 1, wherein the yulpi extract is water, anhydrous or hydrated lower alcohol having 1-4 carbon atoms, acetone, ethyl acetate and butyl acetate, characterized in that the extraction is extracted with an extraction solvent selected from the group consisting of, skin photoaging prevention cosmetic composition. According to claim 1, wherein the cosmetic composition is composed of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray. A cosmetic composition for preventing photoaging of skin, characterized in that it has a formulation selected from the group.