KR100816267B1 - Cosmetic composition containing cheonsangcho extract as an active ingredient - Google Patents

Abstract

The present invention is a cosmetic composition and the cosmetic composition containing the cheonsangcho extract having an antioxidant effect, skin anti-aging effect, skin wrinkle improvement effect, whitening effect, ultraviolet stimulation effect and moisturizing effect as an active ingredient and the cosmetic composition to human skin It relates to a makeup method characterized in that the coating. According to the present invention, the cheonsangcho extract has not only antioxidant activity but also human fibroblast activity effect, skin fine wrinkle improvement effect, whitening effect, skin damage relieving effect and irritation relieving effect, and various functional cosmetics can be prepared using this substance. have.

Cheonsangcho, aging, wrinkle, whitening, skin irritation, moisture

Description

Cosmetic composition containing cheonsangcho extract as an active ingredient {Cosmetic Composition Comprising the Extract of Rhododendron brachycarpum as Active Ingredient}

The present invention relates to a cosmetic composition containing cheonsangcho as an active ingredient.

Aging can be caused by UV rays, stress, disease conditions, environmental factors, wounds, and aging of free radicals as they age. If this condition is exacerbated, it destroys the antioxidant defenses in vivo and damages cells and tissues. To promote adult disease and aging. In particular, the oxidation of proteins is caused by severe cuts in the connective tissues of collagen, hyaluronic acid, elastin, proteoglycan, fibronectin, and so on, resulting in severe over-inflammatory reaction and skin elasticity. If this worsens, mutations in the DNA can lead to mutations, cancer, and reduced immune function.

Therefore, it protects the cell membrane by eliminating the free radicals generated by metabolic process of the body, irradiation by UV irradiation and inflammatory reaction, and the already damaged cells should be regenerated by active metabolism to proliferate the skin. Can recover quickly and maintain healthy skin. Aging involves not only free radicals, but also an enzyme called matrix metalloprotease (MMP) .In vivo, the synthesis and degradation of extracellular substrates such as collagen are properly regulated, but their synthesis decreases as aging progresses. Expression of matrix metalloproteinase (MMP), which is an enzyme that degrades, is promoted, thereby decreasing the elasticity of the skin and forming wrinkles. Therefore, there is a need for the development of a substance capable of regulating MMP expression that inhibits activation or inhibiting its activity 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.

Among the factors that determine skin color, the most important factor is pigmentation.

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

The biosynthesis of melanin is a type of amino acid tyrosine, which is oxidized by tyrosinase in melanocytes of melanocytes and converted to dihydroxy phenylalanine. (eumelanin) polymer. This biosynthesis process is carried out in a special type of brown intracellular 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 phagocytosis of keratinocytes. Accumulate around the nucleus.

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

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. Although it is used as a whitening cosmetic because of its activity, the stability in cosmetic formulations is low, so it is degraded and colored, or its use is limited due to the occurrence of off-flavor, efficacy at the biological level, unclear effect and safety problems. And the inhibitory effect of tyrosinase has been demonstrated, but the effect is low in experiments similar to the actual biologic 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 the problem of the skin, a lot of cosmetics using natural products have recently been developed to reduce skin irritation caused by various chemicals. Natural materials have less side effects on the skin, and as the consumer's response to cosmetics using natural materials has increased recently, the development value of cosmetics as a raw material for cosmetics is increasing.

The present inventors have studied the possibility of applying to cosmetics in a variety of natural products that have not been known so far, as a result of selecting the Cheoncheoncho of the dicotyledonous Rhododendron and preparing the extract from it, the skin antioxidant effect, collagen synthesis effect, skin wrinkle relief effect As a result of measuring the whitening effect, the skin damage alleviation effect by the ultraviolet ray and the skin stimulation alleviation effect, it was found that the efficacy as a cosmetic product can be expected because it is very excellent.

Accordingly, it is an object of the present invention to provide a cosmetic composition containing an extract of Cheoncheoncho having an antioxidant effect, an anti-aging effect, an anti-wrinkle effect, a whitening effect, a skin irritation-reducing effect and a moisturizing effect as an active ingredient.

In addition, another object of the present invention is to provide a cosmetic method comprising applying a cosmetic composition containing the cheonsangcho extract to human skin.

The present invention provides a cosmetic composition containing an extract of Cheoncheoncho as an active ingredient having an antioxidant effect, an anti-aging effect, an anti-wrinkle effect, a whitening effect, a skin irritation-reducing effect and a moisturizing effect.

Rhododendron brachycarpum included in the cosmetic composition of the present invention is an evergreen evergreen tree growing at high and cold mountain tops, and has a height of 1 to 4 m. The bark is gray with white and leaves are alternated, but at the end of the branch, 5-7 dogs gather together. Flowers bloom in June-July and run in inflorescences at the end of 10-20 branches. Petioles reddish brown with dense hairs. Corollas are funnel-shaped, white or light yellow, with green spots on the inside, with five branches. There are 10 surgeries and 1 pistil. The ovary has dense brown hairs. Fruits are granules, oval-shaped, about 2cm long, ripen brown in September.

Cheonsangcho is a flower made by the freshness of the sky.

In the present specification, the term 'Cheonchocho extract' means that obtained by extracting from various organs or parts (eg, leaves, flowers, roots, stems, branches, shells and seeds, etc.) of Cheoncheoncho, preferably leaves, stems, branches , Bark or root, more preferably leaves, branches or roots, most preferably extracts obtained from leaves.

The cheonchocho extract of the present invention can be prepared according to conventional methods known in the art, i.e., under conditions of conventional temperature and pressure, using conventional solvents.

The cheonsangcho extract of the present invention uses at least one extraction solvent selected from the group consisting of purified water, methanol, ethanol, glycerine, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, chloroform, ethyl ether, butylene glycol and hexane Extraction by using, preferably extracted using ethanol or water, more preferably characterized in that the extraction using 70% ethanol.

On the other hand, it is apparent to those skilled in the art that the cheonsangcho extract of the present invention can be obtained in addition to the above-described extraction solvents, and extracts having substantially the same effects using other extraction solvents.

In addition, the extract of the present invention includes not only the extract by the above-described extraction solvent, but also the extract that has undergone a conventional purification 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 through various purification methods additionally carried out, such as separation by sex)) is also included in the extract of the present invention.

In addition, the cheonsangcho extract of the present invention may be obtained by cold condensation at room temperature, a liquid obtained by heat filtration, and further by concentrating the solvent under reduced pressure or lyophilization.

In addition, the cheonsangcho extract of the present invention is contained 0.001-30.0% by weight, preferably 0.1-10.0% by weight based on the entire cosmetic composition. If the content of the extract is less than 0.001% by weight does not appear to improve the skin, when more than 30.0% by weight of the effect of increasing the effect of the content is insignificant, there is a problem in the safety and stability of the formulation and economical.

According to the present invention, the extract of Cheonsangchocho has antioxidant effect (Experimental Examples 1 and 2), MMP-1 production inhibitory effect (Experimental Example 3), MMP-1 production inhibitory effect by ultraviolet light (Experimental Example 4), collagen synthesis promoting effect ( Experimental Example 5), melanin production inhibitory effect (Experimental Example 6), cytotoxicity mitigating effect by ultraviolet irradiation (Experimental Example 7), inflammatory cytokine expression inhibitory effect (Experimental Example 8), skin elasticity improvement effect ( Experimental Example 9) and excellent moisturizing effect (Experimental Example 10).

Therefore, the cosmetic composition of the present invention comprising the extract of Cheoncheoncho with various functions as an active ingredient has an antioxidant effect, skin anti-aging effect, skin wrinkle improvement effect, whitening effect, skin irritation relief effect and moisturizing effect.

Ingredients included in the cosmetic composition of the present invention as an active ingredient includes components conventionally used in cosmetic compositions in addition to the active ingredient, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, And carriers.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, 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 powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

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.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, 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 provides a cosmetic method comprising applying to the human skin a cosmetic composition containing the cheonsangcho extract of the present invention.

The cosmetic method of the present invention refers to all cosmetic methods for applying the cosmetic composition of the present invention to human skin. That is, all the methods known in the art for applying the cosmetic composition to the skin belong to the cosmetic 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 with excellent skin protection effect according to the present invention can be used according to a conventional method of use, the number of times of use can be varied according to the user's skin condition or taste.

When the cosmetic composition of the present invention is a soap, surfactant-containing cleansing or 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 towels 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 including the cheonsangcho extract of the present invention is applied to the skin of a human, the anti-aging, anti-wrinkle effect, whitening effect, moisturizing effect, and skin irritation relief effect may be obtained.

The extract of Cheoncheoncho of the present invention showed an excellent anti-aging effect such as antioxidant effect, MMP inhibitory effect, MMP expression control effect by UV irradiation, fine wrinkle improvement effect and alleviating skin irritation caused by UV irradiation. In addition, the cheonsangcho extract showed a whitening effect such as melanin production inhibitory effect, which is a substance causing blemishes, freckles and skin pigmentation. Therefore, it can be seen that the cosmetic composition containing the extract of Cheoncheoncho has an excellent skin improvement effect such as collagen degrading enzyme activity control effect, skin fine wrinkle improvement effect, anti-inflammatory, whitening and the like with antioxidant effect.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Preparation Example 1 Preparation of Cheonsangcho Extract

The dried and dried leaves of Cheoncheoncho were refluxed three times for 5 hours with an aqueous 70% (V / V) ethanol solution, and then cooled and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C or lower. The extract was prepared by dispersing / dissolving in a 50% butylene glycol solution under reduced pressure and freeze-dried to 1%.

Experimental Example 1 Experiment of Measuring Antioxidant Effect Using NBT Method

In order to confirm the antioxidant effect of the extract of Cheoncheoncho obtained in Preparation Example 1, this experiment was compared with other antioxidants, such as retinol and BHT (Butylated Hydroxytoluene) under laboratory conditions. Was measured.

In order to measure the antioxidant effect, the active oxygen produced by xanthine and xanthine oxidase is measured by the NBT method, and the effect of the test substance removing the active oxygen, that is, the active oxygen scavenging effect, is evaluated. Free radicals are produced by xanthine and xanthine oxidase. This active oxygen reacts with Nitro Blue Tetrazolium (NBT) to measure the active oxygen scavenging rate by measuring the blue color produced by this at a wavelength of 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 Bayer's bottle 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 For the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

⑤ Also, the blank of the sample solution is operated in the same manner using distilled water instead of 6 to measure the absorbance Bo.

The reactive oxygen scavenging ratio was calculated by Equation 1, and the results are shown in Table 1.

Free radical scavenging rate (%) = [1- (St-So) / (Bt-Bo)] X 100

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 addition of sample solution

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

Sample Name Treatment concentration (%) Antioxidative effect(%) Cheonsangcho Extract (Manufacturing Example 1) 0.01 91.3 Retinol 0.01 93.1 BHT 0.01 90.2

As shown in the results of Table 1, all of the samples tested at a concentration of 0.01% was able to confirm the excellent antioxidant effect. It was found that the extract of Cheonsangcho had similar or better antioxidant power than retinol and BHT.

Experimental Example  2: DPPH Antioxidant Effect Measurement Experiment

In this Experimental Example, to measure the antioxidant effect of the extract of Cheonsangcho obtained in Preparation Example 1, the antioxidant activity such as vitamin C was compared in the laboratory conditions and the antioxidant activity was measured using the DPPH method.

The DPPH method measures the antioxidant activity 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 to which the DPPH is reduced by the test substance to reduce the absorbance.

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

As a measuring method

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

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

③ In the blank test, measure the absorbance Bt by using distilled water instead of the sample solution as above.

④ Again, the blank of the sample solution was operated in the same manner using methanol instead of the 0.1 mM DPPH solution to measure the absorbance Bo.

The result of the effect was calculated by Equation 2, the results are shown in Table 2.

Free radical scavenging rate (%) = (1- (St-So) / (Bt-Bo)) X 100

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 (%) Antioxidative effect(%) Cheonsangcho extract (Example 1) 0.01 81.2 vit-C 0.01 83.6

As shown in the results of Table 2, it was confirmed that the Cheoncheoncho extract of the present invention has an excellent free radical scavenging ratio similar to vitamin C at 0.01% concentration.

Experimental Example  3. MMP -1 suppression effect

Fibroblasts (Korean Cell Line Bank, 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, USA) and 37 ° C. After culturing for 24 hours at the serum-free DMEM medium was added to the final concentration of 100 ㎍ / ㎖ Cheonchocho extract of Preparation Example 1 and further incubated for 48 hours in a serum-free DMEM medium containing no cheonchocho extract as a control. 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. At this time, TGF-β (10 ng / ml, Roche, United States) was used as a positive control of MMP-1 production inhibition rate.

Inhibition Rate of MMP-1 Production by Cheonsangcho Extract (%) Test substance % Inhibition of MMP-1 production TGF-β 67.3 Cheonsangcho Extract (Manufacturing Example 1) 66.8

As shown in the results of Table 3, the extract of Cheoncheoncho of the present invention showed a higher MMP-1 production inhibition rate than TGF-β.

Experimental Example  4. Cheonsangcho  By UV induction of extract MMP -1 suppression effect

In general, MMP-1 is known to increase its amount by ultraviolet light. Therefore, the experiment was performed to determine whether the extract of Cheoncheoncho of the present invention inhibits the amount of MMP-1, which is increased by UV light. The experiment was performed in the same manner as in Experiment 3 except that a certain amount of UVA was treated before sample treatment. Cheonsangcho extract used in this experiment was used the extract of Preparation Example 1, but the results of this experiment is not limited thereto. MMP-1 production inhibition rate was calculated according to Equation 3, the results are shown in Table 4 below.

Inhibition rate of MMP-1 production by Cheoncheoncho extract after UV induction (%) Test substance % Inhibition of MMP-1 production TGF-β 75.00 Cheonsangcho Extract (Example 1) 75.7

As shown in the results of Table 4, the inhibition rate of MMP-1 production after UV-induction of the Cheoncheoncho extract of the present invention was better than TGF-β.

Experimental Example  5. Cheonsangcho  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. Serum-free DMEM medium (experimental group) and the serum-free DMEM medium (control group) not containing the extract of Cheoncheoncho were added to the cheonsangcho extract of Preparation Example 1 at a final concentration of 100 μg / ml for 48 hours. 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), and the amount of collagen synthesized therefrom was measured. It was. TGF-β was used as a positive control in this experiment. Collagen biosynthesis increase rate was calculated according to the following equation (4), the results are shown in Table 5.

Enhancement of Collagen Synthesis by Cheonsangcho Extract Test substance Collagen biosynthesis rate (%) TGF-β 78.6 Cheonsangcho Extract (Example 1) 83.8

As shown in the results of Table 5, the collagen synthesis enhancement effect of the extract of Cheoncheoncho of the present invention was significantly higher than TGF-β.

Experimental Example  6: Determination of Melanogenesis Inhibitory Effect Using B16F1 Melanogenic Cells

In order to confirm the whitening effect of the extract of Cheonsangcho obtained in Preparation Example 1, the whitening effect was determined by looking at the degree of inhibition of melanin production on B16F1 melanocytes. B16F1 melanocytes used in this experiment is a cell strain derived from the mouse, and secreted a black pigment called melanin. Samples were treated during the artificial culture of these cells to evaluate the degree of melanin black pigment reduction. B16F1 melanocytes used in the present experiment was used by receiving from the American Type Culture Collection (ATCC).

The melanin biosynthesis inhibitory effect of B16F1 melanogenesis cells was measured as follows. B16F1 melanocytes were dispensed in 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 sample at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with trypsin-EDTA, the cell number was measured, and the cells were recovered by centrifugation. Quantification of intracellular melanin was carried out 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 and vortexed for 5 minutes to disrupt cells. Melanin was extracted by adding 1N NaOH (10% DMSO) to the cell filtrate obtained by centrifugation (3,000 rpm, 10 minutes), and then the absorbance of melanin was measured at 405 nm with a microplate reader. Production inhibition rate (%) was measured. Melanin inhibition rate (%) of B16F1 melanogenesis cells was calculated by Equation 5, the values are shown in Table 6 below. IC ₅₀ value is the concentration of substance that inhibits melanin production by 50%. In order to confirm the degree of melanin inhibition rate of the extract of Cheonsangcho extract, the same experiment was conducted using hydroquinone and arbutin as positive controls.

% Inhibition = [(A-B) / A] X100

A: Melanin amount in wells without sample

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

Sample Name Treatment concentration (%) Melanin Synthesis Inhibitory Effect (IC50) Cheonsangcho Extract (Manufacturing Example 1) 0.1 0.06% Hydroquinone 0.1 0.03% Arbutin 0.1 0.2%

As shown in the results of Table 6, the melanin synthesis inhibitory effect (IC ₅₀ ) of the cheonsangcho extract of the present invention was found to be 0.06%, showing a similar or superior effect compared to the existing whitening agents hydroquinone, arbutin and the like.

Example  7: Reduction of cytotoxicity by UV irradiation

This example was carried out to evaluate the mitigating effect of cytotoxicity by ultraviolet irradiation of the Cheoncheoncho extract obtained in Preparation Example 1. Fibroblasts (fibroblasts) were placed in a 24-well test plate each ⁵ × ¹ × ¹⁰ 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 10mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B15W, Sankyo Dennki, Japan), and then PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. After treating the Cheoncheoncho extract to be evaluated therein, it was incubated for 24 hours. 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, and the absorbance was measured at 565 nm using a microplate reader. The cell viability (%) was measured by the following Equation 6, and the cytotoxicity relaxation rate by ultraviolet rays was calculated by the following Equation 7.

Cell survival rate (%) = [(St-Bo) / (Bt-Bo)] X 100

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

Reduction rate of cytotoxicity by ultraviolet ray (%) = [1- (St-Bo) / (Bt-Bo)] X 100

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

Cytotoxic alleviation rate (%) Sample Name Treatment concentration Cytotoxic alleviation rate (%) Cheonsangcho Extract 0.013% 45 Cheonsangcho Extract 0.025% 63

As shown in the results of Table 7, the Cheoncheoncho extract of the present invention inhibits cytotoxicity caused by ultraviolet light at 45% at 0.013% concentration, and inhibits 63% at 0.025%, effectively preventing cytotoxicity caused by ultraviolet light. Could know.

Experimental Example  8: Inhibitory Effect of Inflammatory Cytokine Expression by Ultraviolet Irradiation

In this example, in order to evaluate the inhibitory effect of inflammatory cytokine expression expressed by UV irradiation of the extract of Cheoncheoncho obtained in Preparation Example 1, fibroblasts isolated from human epidermal tissue (Fibroblast) were placed on a 24-well test plate in a 5 × 10 ⁴ plate. One by one 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 10mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki Co., Japan), after which the PBS was removed and the cell culture medium (FMEM was not added to DMEM). Medium) 350 μl was added. After treating the Cheoncheoncho extract to be evaluated here, it was incubated for 5 hours. 150 µl of the culture supernatant was taken to quantify IL-1α to determine the inflammatory cytokine expression inhibitory effect of the extract. The amount of IL-1α was quantified using Enzyme-linked Immunosorbent Assay, and the production rate of IL-1α was calculated by Equation 8.

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St-Bo) / (Bt-Bo)] X 100

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 (%) Inhibitory rate of inflammatory cytokine expression (%) Cheonsangcho Extract 0.013% 60 Cheonsangcho Extract 0.025% 73

As shown in the results of Table 8, the extract of Cheoncheoncho of the present invention inhibits the production of inflammatory cytokines, IL-1α, by 60% at a concentration of 0.013%, and inhibits as much as 73% at 0.025%. It can be seen that the concentration effectively protects against inflammation caused by ultraviolet rays.

Experimental Example  9. Cheonsangcho  Containing extract Cosmetic  Skin elasticity improvement effect

In this Experimental Example, the cosmetics containing the extract of Cheonsangcho obtained in Preparation Example 1 (Example 1) were observed to improve skin elasticity in humans. The comparison was conducted and evaluated. The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 9 below. For 20 experimenters (20-35 year old female), the cream of Example 1 was applied to the right side of the face and the cream of Comparative Example 1 was applied to the left side of the face twice a day for 2 consecutive months.

The skin elasticity improvement effect after the completion of the experiment was measured using a skin elasticity measuring instrument (cutometer SEM 575, C + K Electronic Co., Germany) before and after using the product for 2 months. Experimental results are described in Table 10 as ΔR8 value of Cutometer SEM 575, where R8 value represents the nature of the viscoelasticity of the skin.

ingredient Example 1 Comparative Example 1 Cheonsangcho extract lipophilic monostearate glycerine cetearyl alcohol stearic acid beeswax polysorbate 60 sorbitan stearate cured vegetable oil squalane mineral oil trioctanoin dimethicone sodium magnesium silicate glycerin betaine triethanolamine sodium hyaluronate preservative, fragrance, pigment distilled 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 -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 Sum 100 100

The content units in the table are by weight.

Experimental product Skin elasticity effect (△ R8) Example 1 0.25 Comparative Example 1 0.11

As shown in the results of Table 10, it can be seen that the skin elasticity improving effect of the experimenter applying the cream containing the cheonsangcho extract is about twice as compared to the comparison group applied the cream containing the cheonsangcho extract.

Experimental Example 10. Moisturizing effect of the cosmetic containing Cheonsangcho extract

In order to evaluate the moisturizing power of the cosmetics prepared in Experimental Example 9 (Example 1 and Comparative Example 1), the moisture retention ability of the skin to which each cosmetic was applied was measured as follows.

In the constant temperature and humidity room of 22 ° C. and 45% RH, each of the cosmetics of Examples and Comparative Examples was divided into half of 24 test subjects while applying the product of Example to one side and the product of Comparative Example to the other (2 times / day). ) The water content after 4 weeks was measured.

As a measuring device, a moisture content measuring instrument (Corneometer CM825, Courage + Khazaka, Germany) was used to measure the moisturizing power by measuring the electric capacity of the skin according to the moisture content of the skin. The results are shown in Table 11 below.

Water content Use week Example 1 Comparative Example 1 0 Week 23.31 ± 9.34 23.44 ± 11.36 4 Week 37.71 ± 8.09 26.28 ± 5.58 4 Week-0Week 14.40 ± 1.25 2.84 ± 5.78

As shown in the results of Table 11, the cosmetic composition (Example 1) containing the cheonsangcho extract of the present invention was superior to the skin moisturizing effect compared to the cosmetic composition (Comparative Example 1) containing no cheonsangcho extract.

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

Claims (5)

A cosmetic composition comprising an Rhododendron brachycarpum extract as an active ingredient having an antioxidant effect, an anti-aging effect, an anti-wrinkle effect, a whitening effect, a skin irritation-reducing effect, and a moisturizing effect. The cosmetic composition according to claim 1, wherein the cheonsangcho extract is contained in an amount of 0.001-30.0 wt% based on the total composition. The extract of claim 1, wherein the cheonsangcho extract is selected from the group consisting of (a) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, chloroform, butyl acetate and 1,3-butylene glycol Cosmetic composition, characterized in that extracted by using a solvent extraction method, (b) supercritical extraction method or (c) ultrasonic extraction method using a solvent. According to claim 1, wherein the cosmetic composition is selected from cosmetic composition or ointment of the lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type or water-in-oil (W / O) type formulation. Cosmetic composition characterized in that. delete