KR20090097550A - Cosmetic composition containing schizandrin and extracts - Google Patents

Abstract

A cosmetic composition containing schizandrin which is isolated from Schizandra chinensis Baillon. is provided to ensure the effects of anti-oxidation, anti-wrinkle, skin whitening, and irritation relief. A cosmetic composition comprises schizandrin and its extract as active ingredient. The schizandrin extract is contained in 10ppm~10000ppm based on the total amount. The schizandrin and its extract is obtained by extracting dried Schizandra chinensis Baillon. with 10-95%(V/V) of ethanol solution at room temperature, filtering suspension, decompress-concentrating under 50°C, and mixing the concentrate in a solvent such as butyleneglycol. The solvent is purified water, methanol, ethanol, glycerin, ethylacetate, butyleneglycol, propyleneglycol, dichloromethane, hexane or its mixture. The cosmetic composition is used in the form of gel, aqueous liquid, cream, essence, oil in water or water in oil emulsion.

Description

Suzandrarin and cosmetic composition containing the extract as an active ingredient {Cosmetic composition containing schizandrin and extracts}

The present invention relates to a cosmetic composition containing Suzandrarin and the extract as a main active ingredient.

Human skin has various physical and chemical changes during the aging process, and the causes are largely divided into internal aging and photo-aging, and research on this has been actively conducted. That is, it may be caused by ultraviolet rays, stress, disease state, environmental factors, wounds, activation of free radicals with age, etc., and when this condition is intensified, it destroys the antioxidant defense network existing in the living body and destroys cells and tissues. Damage to promote adult disease and aging. More specifically, lipids, proteins, polysaccharides, and nucleic acids, which are major constituents of the skin, are oxidized to destroy skin cells and tissues, resulting in skin aging. In particular, the oxidation of protein is caused by the breakdown of collagen, hyaluronic acid, elastin, proteoglycan, and fibronectin, which are the tissues of the skin, causing severe excessive inflammatory reactions and disturbance of skin elasticity. This leads to a situation of reduced immune function.

Therefore, it protects the cell membrane by eliminating the free radicals generated by metabolic process of the body, UV irradiation, and inflammatory reactions.Also, damaged cells should be regenerated by active metabolism to proliferate the skin. You can recover quickly and maintain healthy skin. On the other hand, aging involves not only free radicals but also an enzyme called MMP. In vivo, the synthesis and degradation of extracellular substrates such as collagen is properly regulated, but the synthesis decreases as aging progresses, and the substrate metal, an enzyme that degrades collagen. The expression of proteolytic enzymes (MMP) is promoted, which reduces the elasticity of the skin and forms wrinkles. In addition, these degrading enzymes are also activated by ultraviolet irradiation. 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.

Next, skin aging due to pigmentation may be mentioned, and pigments affecting skin color include melanin, melazoid, carotene, hemoglobin, and the like. The most important thing is melanin, the biggest factor affecting biosynthesis is ultraviolet radiation and hormone secretion in the body. 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 brown and black through a series of oxidation processes, starting with the conversion of tyrosine, an amino acid, from the melanocytes of melanocytes to tyrosinase and 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, and then into the cytoplasm by the phagocytosis of keratinocytes. Accumulate around the nucleus of the site. Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A 6-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-93150), and some extracts are already tyrosinase. It is used as a whitening cosmetic because it has an inhibitory activity, but its stability in cosmetic formulations is poorly 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.

Until now, most of the raw materials used as materials for cosmetics simply inhibited enzyme activity, and there was a problem in that the amount of expression of enzymes could not be controlled. In addition, consumers are increasingly demanding the use of cosmetics with minimal functional side effects such as skin protection, irritation suppression, inflammation suppression, and sun protection while having high quality functional properties according to special composition, and to minimize skin irritation As consumers' response to cosmetics using natural materials instead of using chemical raw materials is increasing, there is a need to develop natural materials as raw materials for cosmetics.

The present invention has been made in order to solve the above problems, an object of the present invention is to prepare the Suzandrarin and extracts extracted from Schisandra chinensis, a deciduous vine of the Magnolia family of several natural products, to confirm the efficacy as a raw material of cosmetics The present invention provides a method of developing a method of using the same and producing a cosmetic having a predetermined functionality by using this raw material.

That is, an object of the present invention is to provide a natural extract that is applicable to skin cosmetics and exhibits excellent anti-aging effects such as antioxidant effects, collagen synthesis effects, skin fine wrinkles effects when contained in cosmetics.

In addition, it is an object of the present invention to provide a natural extract exhibiting a whitening effect by ultraviolet irradiation and the like to alleviate skin irritation by an inflammation mediator when applied to cosmetics.

In order to measure the antioxidant effect by using the NBT method and the DPPH method, collagen degrading enzyme activity was measured by measuring the inhibition of MMP expression by MMP inhibition and UV irradiation. The degree of skin irritation was measured by measuring cytotoxicity caused after UV irradiation.

Skin whitening was evaluated by measuring melanin production inhibitory effects such as blemishes, freckles and skin pigmentation.

Suzandrins and extracts of the present invention have excellent anti-aging effects such as antioxidant effect, MMP inhibitory effect, MMP expression control effect by UV irradiation, antioxidant effect, collagenase activity control effect, skin wrinkle improvement effect and skin caused after UV irradiation It is to provide a cosmetic composition exhibiting an effect of relieving irritation.

In addition, it not only exhibits whitening effects such as melanin production inhibitory effect, which causes blemishes, freckles, and skin pigmentation, but also provides acne-bacterial antimicrobial activity and 5 alpha-reductase inhibitory effect.

In the above description of the preferred embodiment of the present invention, those skilled in the art will be able to perform various modifications without departing from the gist of the present invention, and such modifications are within the protection scope of the present invention. The protection scope of the present invention should be construed as described in the claims.

The cosmetic composition according to the present invention for achieving the above object is characterized by containing Shuzanrin extracted from Schisandra chinensis and this extract as the main active ingredient.

Recently, many cosmetic products using natural products have 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 cosmetics as a raw material for cosmetics is increasing. Therefore, 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 Omija natural product, deciduous vine shrub of Magnolia and extracting Suzandrin from them to prepare the extract, As a result of measuring the effect, collagen synthesis effect, skin wrinkle relief effect, whitening effect and skin irritation relief effect was found that it can be expected to be effective as a cosmetic.

Schisandra chinensis is Schizandra chinensis Baillon, a deciduous broad-leaved vine whose leaves are alternate, ovate or obovate, with sharply pointed, jagged, somewhat hairy on the back veins. Flower is reddish white and blooms in June ~ July. Fruits are used for food and hang on the ends of branches. They ripen red in August to September. Fruits are called Schizandrae Fructus, Schizandrae Fructus. Jirisan, Jeonbuk, and Gangwon provinces are 200 ~ 1,600m above sea level, and are wild throughout the country except Chungnam and Chungbuk, and geographically distributed in Japan, Sakhalin, Manchuria, and China. Fruits Schizandrol, Schizandrin A, B, C, angeloylgomisin O, P, epigomisin, pregomoisin, deoxyschizandrin And the like. Suzandrarin contained in Schisandra chinensis has a molecular weight of 432.52 and a structural formula of C ₂₄ H ₃₂ O ₇ . Schisandra chinensis has long been used in herbal medicine as a raw material of medicinal herbs and has the effects of astringent, nourishment, tonic, ginhae medicine, haejudok, thirsty, convergent shovel, ripe ginjin, bosin dyed heart. Human stability with ingestion has already been confirmed.

Suzandrins and extracts used in the invention are contained in the leachate obtained by leaching and transferring from Schizandra chinensis, or the concentrate obtained by partially or fully concentrating the leachate, or the extract extract and extract prepared by drying the concentrate. It contains the chemicals that exert their main effects.

On the other hand, it is preferable that the content of Shuzanrin and the extract according to the present invention is 10 to 10000ppm with respect to the entire cosmetic composition. If the content of the extract is less than 10ppm is almost no skin improvement effect, if more than 10000ppm because the degree of increase in the effect of increasing the content is insignificant and economical.

In addition, the cosmetic composition according to the present invention may contain, in addition to the above-mentioned extract, other components that can give a synergistic effect to the main effect, preferably within the range of not impairing the main effect of the present invention.

The Suzandrarin and extract may be present in the composition at 10 to 10000 ppm with respect to the whole composition.

In addition, the suzandrins and extracts according to the present invention are extracted with 10-95% (V / V) ethanol aqueous solution at room temperature, cooled and concentrated. The filtrate obtained by filtering the resulting suspension is concentrated under reduced pressure at 50 ° C. or lower, and concentrated under reduced pressure. It is prepared by mixing water in a solvent such as butylene glycol.

The cosmetic composition of the present invention uses a solvent selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures thereof.

The subject of the present invention is also the use of cosmetic compositions used as skin whitening agents, anti-aging agents, skin irritants.

The compositions of the present invention may be in the form of an aqueous, aqueous-alcoholic or oily solution, oil-in-water or oily water or multiple emulsions, aqueous or oily gels, liquid, pasty or solid anhydrous products, oil dispersions in aqueous phase using globules. And more preferably in the form of lipid vesicles in ionic and / or nonionic form.

The composition may be somewhat fluid and may have the appearance of a white or colored cream, ointment, milk lotion, serum, essence, paste or mousse. It may optionally be applied to the skin in the form of an aerosol or may be in the form of a solid such as a stick. It may be used as a skin care product and / or makeup product.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these Examples are only illustrative description of the present invention and the scope of the present invention is not limited to these Examples.

Example  1: from Schisandra Suzandra Lynn  Separation and Extraction Manufacturing

Schisandra chinensis, the fruit of Schisandra chinensis, was ultrasonically extracted twice for 3 hours with 70% (V / V) ethanol aqueous solution, and then cooled and filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure at 50 ° C. or lower to obtain a reduced pressure concentrate. This was purified by preparative high-performance liquid chromatography to isolate Suzurin. In addition, the Suzandrarin extract was prepared by dispersing / dissolving the Suzandrarin in at least one solvent selected from purified water, ethanol, butylene glycol, and propylene glycol so as to contain 10 to 10,000 ppm of Suzanrin.

After preparing the Suzandrin extract, the content of the Suzandrin was confirmed in FIG. 2 using high performance liquid chromatography.

Example 2: NBT Antioxidant Effect Measurement Experiment

In this example, the antioxidant activity of Retinol and BHT was compared by using NBT method in order to measure the antioxidant effect of Suzandrin obtained in Example 1 under laboratory conditions.

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 generated by xanthine and xanthine oxidase, and the active oxygen scavenging rate is measured by measuring the blue color produced by reaction of the reactive oxygen with Nitro Blue Tetrazolium (NBT) at a wavelength of 560 nm. .

As a measuring method,

0.05M Na2CO3 ------------------------------------ 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 CuCl2 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 result of the effect was calculated by Equation 1, the results are shown in Table 1.

Equation 1

% Inhibition = [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 in sample solution

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

As a result of the test, as shown in Table 1, it was confirmed that the antioxidant activity was excellent in the sample tested at 20 ppm concentration of Suzandrarin, and that Suzandrarin extract has a superior antioxidant effect similar to that of retinol and BHT.

TABLE 1

Sample Name Treatment concentration Antioxidative effect(%) Shuzandrin Extract (Example 1) 20 ppm 96 Retinol 0.1% 93 BHT 0.1% 90

Example 3: DPPH Antioxidant Effect Measurement Experiment

In this example, the antioxidant activity of the green tea extract and vitamin E as a comparative sample was measured using DPPH method in order to measure the antioxidant effect of Suzandrin obtained in Example 1.

DPPH method measures the antioxidant activity by reducing power using a free group called 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (DPPH). 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, a 0.1 mM solution of 2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76) 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.

Equation 2

% Inhibition = [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

As shown in Table 2, the Suzandrarin extract had a superior antioxidant effect than the green tea extract and vitamin E at 20ppm concentration.

TABLE 2

Sample Name Treatment concentration Antioxidative effect (%) Shuzandrin Extract (Example 1) 20 ppm 87 Green Tea Extract 0.01% 35 Vitamin E 0.01% 31

Example 4: in in vitro MMP -One inhibition assay

Substrate metalloproteinase (MMP-1) inhibitory activity was determined in a biochemical model based on the use of collagen and gelatin conjugated to purified collagenase and its substrate, fluorescein (EnzChek ™ gela) The collagenase kit (Molcula Probes), collagenase purified from Clostridium histories, was supplied into the EnzChek ™ gelatinase / collagenase kit. The reaction buffer consisting of DQ-collagen conjugated with Sane and 0.05M Tris-HCl, 0.15M NaCl, 5mM CaCl2 and 0.2mM sodium azide (pH 7.6) was found in the EnzChek (TM) gelatinase / collagenase kit (Morcula). Probes) were dissolved in the reaction buffer, which was tested at 4; 2; 0.4; 0.2; 0.1% (w / v) Dilutions of the test extract were 25 ug / ml of DQ-collagen and 0.1U / ml Incubated with collagenase at room temperature for 15 minutes, 45 minutes, and 120 minutes.

In each experimental condition, the control corresponding to the collagenase and DQ-collagen mixture was also incubated in the same manner.

Also in each experimental condition a sample called Blank, hereinafter 'enzyme free blank', was incubated in the presence of DQ-collagen and in the absence of collagenase. Each experiment was performed three times.

After 15, 45 and 120 minutes, the signal corresponding to the degradation of DQ-collagen was measured with a fluorometer (excitation: 485 nm, emission 505 nm).

The fluorescence value of each sample was measured based on the 'enzyme free blank' fluorescence value. Results are expressed as percent variance for fluorescence units per sample and control.

In conclusion, the Susanzan extract tested at 0.04 to 4% (v / v) under selected experimental conditions retained anticollagenase / gelatinase activity in a dose dependent manner.

As shown in Table 3, Suzandrarin inhibited 86% of collagen degrading activity of Clostridium collagenase when treated with 0.04%, which was superior to the inhibitory effect of green tea extract.

TABLE 3

Experimental Sample % Inhibition (treatment concentration) Suzandrin Extract 58 (10ppm) 86 (20ppm) Retinol 0 (0.02%) 6 (0.04%) Green Tea Extract 51 (0.02%) 63 (0.04%)

Example  5: after UV irradiation Suzandra Lynn  by MMP -1 expression inhibition evaluation

In this example, ELISA was performed to measure the concentration of MMP-1 after UV irradiation and sample addition of the Suzurinrin obtained in Example 1.

Human dermal fibroblasts are irradiated with UVA at an energy of 5 J / cm 2 using a UV chamber. UV irradiation dose and culture time were established through preliminary experiments to maximize the expression of MMP in fibroblasts. Negative controls were wrapped in tinfoil and maintained at the same time in the environment of UVA. UVA emission was measured using a UV radiometer. The cells during the UVA irradiation are intact in the previously dispensed medium and irradiated with UVA and then exchanged with the medium containing the sample to recover the medium after incubation for 24 hours and coated on a 96-well plate. The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) is treated and reacted at 37 ° C. for 60 minutes. After reacting the secondary antibody anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) for about 60 minutes, the alkaline phosphatase substrate solution (1mg / mlρ-nitrophenyl phosphate in diethanolamine buffer solution) was reacted at room temperature for 30 minutes and then the microplate reader was reacted. Absorbance is measured at 405 nm. As a control, the one without addition of the sample is used.

As shown in Table 4, for MMP-1, which is induced at the time of ultraviolet irradiation, Suzandranine showed more than 20% inhibition rate compared to the control group, which was superior to the inhibition rate of retinol used as a control group. The result is.

TABLE 4

Test group Treatment concentration MMP-1 expression inhibition rate (%) Control - - Suzandrin Extract 20 ppm 29 Retinol 0.1% 21

Example  6: B16F1 Melanosite  Experiment to measure melanin production inhibitory effect

In this example, the whitening effect was determined by looking at the degree of inhibition of melanin production against B16F1 melanocytes in order to confirm the whitening effect of Suzandrrin obtained in Example 1.

B16F1 melanocytes used in this example 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 melanin black pigment reduction. The B16F1 melanocytes used in this example were used after being sold from ATCC (American Type Culture Collection, Accession No .: 6323).

The melanin biosynthesis inhibitory effect of B16F1 melanocytes was measured as follows. B16F1 melanocytes were dispensed in 6-well plates at a concentration of 2 x 10 ⁶ per well, and cells were attached and then incubated for 72 hours by treating the samples 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.

Determination of intracellular melanin was performed by 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 measure the absorbance of melanin at 405 nm with a microplate reader, and then quantify the melanin. Melanin production inhibition rate (%) was measured. Melanin inhibition rate (%) of B16F1 melanocytes was calculated by the equation (3), IC ₅₀ value is the concentration of the substance inhibits 50% melanin production.

Equation 3

% Inhibition = [(A-B) / A] x 100

A: Melanin amount in wells without sample

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

As a result of testing the melanin production inhibitory effect of B16F1 melanocytes, as shown in Table 5, the IC ₅₀ value of Suzandranine is 0.06%, which is similar to or superior to that of the existing whitening agents, hydroquinone, arbutin, oil-soluble licorice extract, and lettuce extract. It can be seen that indicates.

TABLE 5

Sample Name Treatment concentration Melanin production inhibitory effect (IC ₅₀ ) Shuzandrin Extract (Example 1) 20 ppm 0.06% Hydroquinone 0.1% 0.04% Arbutin 0.1% 0.18% Lettuce extract 0.1% 0.04% Soluble Licorice Extract 0.1% 0.03%

Example  7: Reduction of cytotoxicity by UV irradiation

This example was carried out to evaluate the moderating effect of cytotoxicity caused by ultraviolet irradiation of Suzudrrin obtained in Example 1. Fibroblasts (fibroblasts) were placed in a 24-well test plate each 1x10 ⁵ and attached for 24 hours. Each well was washed once with PBS and 500 ul 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, Japan), and then PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. It was incubated for 24 hours after treatment with Suzandrarin to be evaluated here. After 24 hours, the medium was removed, and 500 µl of the cell culture medium and 60 µl of the 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 (0.04 N) was added. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, followed by measurement of 565 nm absorbance in a microplate reader. Cell survival rate (%) was measured by the equation (4) and cytotoxicity relaxation by ultraviolet light was calculated by the equation (5).

Equation 4

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

Bo: 565 nm absorbance of wells that only reacted with 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

Equation 5

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

Experimental results As shown in Table 6 Suzudrin extract can be seen that the cytotoxicity of the ultraviolet light by 10% to reduce the cytotoxicity of 53% at 10ppm concentration can be seen to effectively protect the cytotoxicity by ultraviolet light even at low concentrations.

TABLE 6

Sample Name Treatment Concentration (ppm) Cytotoxic alleviation rate (%) Shuzandrin Extract (Example 1) 10 53 20 78

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

This Example is to evaluate the degree of inhibition of inflammatory cytokine expression expressed by ultraviolet irradiation in order to confirm the anti-inflammatory effect of Suzandrarin obtained in Example 1.

Fibroblasts isolated from human epidermal tissue (Fibroblast) were placed in a 24-well test plate each 5X10 ⁴ 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 treatment with Suzandrarin 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 Suzandrarin extract. The amount of IL-1α was quantified using Enzyme-linked Immunosorbent Assay (ELISA), and the production rate of IL-1α was calculated by Equation 6.

Equation 6

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

Bo: amount of IL-1α produced in wells not irradiated with UV

Bt: amount of IL-1α produced in wells that were not irradiated with UV light

St: Production of IL-1α in Wells Treated by Ultraviolet Irradiation

As shown in Table 7, Suzandrarin inhibits 52% production of inflammatory cytokine-induced inflammatory cytokine IL-1α at 10ppm concentration, thereby effectively preventing the inflammation caused by UV rays at low concentration.

TABLE 7

Sample Name Treatment concentration (ppm) Inhibitory rate of inflammatory cytokine expression (%) Shuzandrin Extract (Example 1) 10 52 20 80

Example  9 and Comparative example  1: improve skin elasticity

In this example, the cosmetic preparation containing Suzandrrin extract obtained in Example 1 was prepared, and the skin elasticity improvement effect was evaluated in humans by performing a comparative experiment with Comparative Example 1.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 8. First, the phase b) recorded in Table 8 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Example 9, Comparative Example 1). The cream prepared in Example 9 was applied to the right side of the face and the cream prepared in Comparative Example 1 was applied twice a day for three consecutive months to 15 test subjects (women aged 20-35 years).

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. The experimental results are described in Table 9 below as the ΔR8 value of the cutometer SEM 575. The R8 value represents the nature of the viscoelasticity of the skin.

As shown in Table 9, it can be seen that the skin elasticity improving effect of the experimenter who applied the cream containing Suzandrarin extract.

TABLE 8

Raw material Example 11 Comparative Example 1    end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2  I Suzandrin Extract 20 ppm - Propylene glycol 5 5 Quantity Quantity Quantity

Note) Unit: Weight%

TABLE 9

Experimental product Skin elasticity effect (ΔR8) Example 9  0.27 Comparative Example 1  0.14

 n = 15, p <0.05

Example  10 and Comparative example  2

In this example, the cosmetics containing Suzandrrin extract obtained in Example 1 were prepared, and the skin whitening effect of humans was evaluated by comparative experiments with Comparative Example 2.

The cosmetics used in the comparative experiments are in the form of a cream, the composition of which is shown in Table 10. First, the phase b) recorded in Table 10 is heated and stored at 70 ° C. To this, a) phase is added, and after pre-emulsification, it is emulsified uniformly with a homomixer, and then cooled slowly to prepare a cream (Example 10, Comparative Example 2). For 15 experimenters (women aged 20-35 years), the cream prepared in Example 10 was applied to the right side of the face, and the cream prepared in Comparative Example 2 was applied to the left side of the face twice a day for 2 consecutive months. . After completion of the test, the change of the brightness of the color (ΔL) was measured by using an image analyzer and the color change of the skin on both left and right sides of the face using a chromameter (Minolta CR300). Subjective visual observation was performed and the effect was measured according to the following classification. The results are shown in Table 11 below. At this time, the degree of whitening efficacy was classified into the following seven ratings.

Evaluation criteria for whitening efficacy

-3: very worse -2: worse -1: slightly worse 0: no change

1: slightly improved 2: improved 3: highly improved

As shown in Table 11, it can be seen that the whitening effect is excellent in the facial skin of the tester coated with the cream containing Suzandrarin extract.

TABLE 10

Raw material Example 12 Comparative Example 2    end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2  I Suzandrin Extract 20 ppm - Propylene glycol 5 5 Quantity Quantity Quantity

Note) Unit: Weight%

TABLE 11

Change in brightness of skin color (ΔL) Objective evaluation of the skilled person Subjective Evaluation of the Subject Example 10 Comparative Example 2 Example 10 Comparative Example 2 Example 10 Comparative Example 2 medium 5.24 2.23 2.9 1.8 2.9 1.5

1: Schizandrin molecular structure

Figure 2: Quantification of Schizandrin by HPLC

Claims (8)

Suzandrin (Schizandrin) and a cosmetic composition comprising the extract as the main active ingredient. The method of claim 1, The Suzandrin extract is a cosmetic composition, characterized in that it contains 10ppm to 10000ppm of the total composition. The method according to claim 1 or 2, The extraction solvent is a cosmetic composition, characterized in that selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane, hexane and mixtures thereof. The method according to claim 1, wherein Cosmetic composition, characterized in that used for skin whitening. The method according to claim 1, wherein Cosmetic composition, characterized in that used for improving skin wrinkles. The method according to claim 1, wherein Cosmetic composition, characterized in that it is used for skin irritation relief. The method according to claim 1, wherein Cosmetic composition, characterized in that it is used for antioxidant. According to claim 1 to 3, characterized in that it is selected from the formulation of the lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) or water-in-oil (W / O) type, ointment. Cosmetic composition.

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