KR20100016815A - Cosmetic compositions for skin whitening containing sedum sarmentosum extract and arbutin - Google Patents

Abstract

PURPOSE: A cosmetic composition containing Sedum sarmentosum extract and arbutin is provided to suppress melanogenesis, activate anti-oxidation, and ensure skin whitening effect without side effects. CONSTITUTION: A cosmetic composition for skin whitening contains 0.001-30.0 weight% of Sedum sarmentosum extract and 0.1-2.0 weight% of arbutin. The Sedum sarmentosum extract is isolated using water, hydrous or anhydrous low alcohol of 1-3 carbon atoms, acetone, ethylacetate or diethylether. The cosmetic composition is used in the form of solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, powder foundation, emulsion foundation, wax foundation, or spray.

Description

Cosmetic composition for skin whitening containing Sedum sarmentosum extract and Arbutin

The present invention relates to a cosmetic composition for skin whitening using dolmul extract and arbutin together as an active ingredient. Inhibition of oxidative stress produced by the skin by external stress factors such as UV irradiation by antioxidant activity and intracellular radical scavenging effect by sedum extract, and inhibiting melanin production by synergistic action of arbutin and sedum extract By whitening in two stages, synergistic whitening effect is achieved.

In general, the pigmentation phenomenon on the skin is caused by a number of external factors such as ultraviolet rays, medications, pollutants, and irritant skin external preparations and oxidative stress resulting from the internal factors such as hormonal imbalance, inflammatory response factors, and cytokines. Although the factors work in combination, the most important of these is exposure to ultraviolet radiation and oxidative stress.

The skin normally produces melanin to protect the skin from the sun's ultraviolet rays. Melanin is produced from melanocytes, a kind of skin cells, and has a natural screening function that is distributed on the surface of the skin and blocks ultraviolet rays harmful to the human body.

When the skin is exposed to ultraviolet light, very little of it directly affects melanocytes, and most of the ultraviolet light acts on keratinocytes, thereby affecting the production of various signaling substances. In addition, external stressors such as ultraviolet rays promote the generation of oxidative stress factors such as radicals through various pathways. Oxidative stress factors such as signaling materials and radicals increase the activity of an enzyme called tyrosinase in melanocytes in the base layer through complex and complex pathways. Acts on tyrosine, a type of amino acid present in skin tissue, to produce dopa (DOPA), which turns into dopaquinone, forming melanin that causes skin pigmentation Complex reactions are automatic. Meanwhile, the melanin pigment thus produced is widely distributed to the surrounding keratinocytes through dendrite of melanocytes, thereby changing the color of the skin. However, the biological understanding of melanin formation is still unclear, and further studies on dysplasia such as blemishes and blotch, pigmentation caused by ultraviolet rays, and the action of hormones related to pigmentation should be studied.

Hyperpigmentation of the skin often acts as a serious mental burden from the point of view of skin cosmetology and adversely affects normal social activities. Therefore, in addition to the method of blocking ultraviolet rays to prevent pigmentation as described above, by adding a substance that can inhibit the activity of tyrosinase in the dermatological science to the cosmetic, it is possible to improve the pigmentation phenomenon such as spots and freckles Many researches are being made to prevent this.

In particular, Asians, such as Korea, Japan, and China, who consider having white skin as a measure of beauty, are very interested in skin whitening cosmetics, and the desire to expect a substantial skin improvement effect from such whitening cosmetics is very high. strong. Ascorbic acid (JP-A 4-9320), hydroquinone (JP-A-6-192062), kojic acid (JP-A-56-7710), arbutin (JP-A 4-9315) and some Although extracts have tyrosinase inhibitory activity and are used as whitening cosmetics, their use is limited due to poor stability in the prescription system, resulting in discoloration due to deterioration, off-flavor, efficacy at the biological level, unclear effect and safety issues. It's happening. And the inhibitory effect of tyrosinase has been demonstrated, but the effect is low in experiments similar to the actual biologic level. These causes include the induction of melanin production in the skin through various and diverse pathways due to signal transduction agents and oxidative stress factors, produced from melanocytes, and the resulting melanin affecting skin color. The whitening agent that only affects some processes such as tyrosinase is difficult to lead to the actual whitening effect.

Therefore, by inhibiting melanin production through blocking some pathways such as tyrosinase and not simply a simple effect, it inhibits various processes including fundamental blocking of melanin production factors such as oxidative stress factors, thereby developing a whitening agent that is more effective than ever before. There is a need for research on the field.

In the present invention, the antioxidant activity and intracellular radical scavenging effect of the oxidative stress generated in the skin by the external stress factors such as ultraviolet irradiation, which is a melanin generating factor, are not merely a skin whitening effect due to melanin production inhibition. By blocking, it aims to effectively block melanin production in melanocytes.

In addition, it is to provide a whitening cosmetic composition having a synergistic whitening effect than the previous whitening agent by complex inhibition of melanin formation. In addition, another object of the present invention is to provide a whitening cosmetic composition that has little irritation to the skin and does not have problems such as stability for cosmetic formulations.

In order to achieve the above object, the skin whitening cosmetic composition according to the present invention is characterized in that it contains a complex consisting of dolmul extract and arbutin as an active ingredient. Skin whitening cosmetic composition of the present invention can effectively inhibit the oxidative stress generated in the skin by external stress factors such as ultraviolet irradiation, such as antioxidant activity and intracellular radical scavenging effect, and effectively inhibit the melanin biosynthesis process to excess of melanin The whitening effect is shown in two stages of inhibiting the production, and the synergistic whitening effect is achieved by combining the whitening effect in the clinic. It is characterized by containing 0.001 to 30.0% by weight and 0.1 to 2.0% by weight arbutin with respect to the whole composition of the present invention.

The present invention also relates to a whitening cosmetic composition having little irritation to the skin and having no problems such as stability on cosmetic formulations through skin irritation-reducing action. The cosmetic composition has a formulation selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. .

The present inventors have developed an antioxidant and intracellular radical scavenging activity, intracellular tyrosinase, to develop a whitening cosmetic having a complex inhibitory effect on melanocyte migration from the melanogenesis process in the skin melanocytes to the keratinocytes The present invention has developed a cosmetic composition of the present invention which has an inhibitory effect of an azease and inhibits melanogenesis, and as yet another object of the present invention, there is little irritation to the skin and as a result of efforts to solve problems such as stability of cosmetic formulations, stone trees The present invention was completed by confirming that the mixed composition of extract arbutin has an excellent effect in solving the above problems.

The cosmetic composition for skin whitening comprising the stone extract of the present invention and arbutin as an active ingredient is obtained by using the dolmul extract with the melanin production inhibitory effect of the arbutin and the dolmul extract synergistically by using together with the dolmul extract and arbutin as an active ingredient Antioxidant activity and intracellular radical scavenging effect are combined to achieve a synergistic whitening effect by combining the whitening effect in the clinic.

In addition, through the skin irritation-reducing effect by the dolmul extract, there is little irritation to the skin and can be usefully used as a cosmetic composition for skin whitening excellent whitening efficacy showing excellent stability of the formulation.

Sedum sarmentosum is a perennial herb with rosacea sedum, with stems extending laterally, with roots at each node, and peduncles standing upright, 15 cm high. The leaves are usually 3 turnsed, without petioles, long oval or lanceolate. The leaves are pointed at both ends and the edges are flat. The flower is yellow and blooms in August ~ September. 5 petals are lanceolate, pointed, longer than calyx. Fruits are goldol (利 咨 果) and there are five carpels (心 皮). Young stems and leaves are soaked in kimchi, and yeonhansoon is namul. It is known to be distributed in mountains throughout Korea, Japan, and China.

The sedum extract of the present invention is conventionally used to extract the active ingredient and prepared by methods well known to those skilled in the art. For example, water, anhydrous or hydrous lower alcohol, acetone, ethyl acetate or diethyl ether having 1 to 3 carbon atoms are added to the medicinal herb powder in an amount of 1 to 15 parts by volume based on the dry weight of the powder, and then the cooling capacitor It is extracted by heating for 5 to 24 hours at a temperature of 50 ~ 100 ℃ in the extractor attached; Water, anhydrous or hydrous lower alcohol, acetone, ethyl acetate or diethyl ether having 1 to 3 carbon atoms was added to the pulverized product in an amount of 1 to 15 parts by volume based on the dry weight of the pulverized product, followed by a temperature of 4 to 25 ° C. Immersion for 3-20 days to obtain an extract; After adding an appropriate amount of water to anhydrous or hydrous ethanol or methanol extract, the resulting precipitate is filtered off, and then added with ethyl acetate, butanol or diethyl ether, mixed well and left to separate the layers, and then the upper layer is separated. The extract is obtained from an extractor with a cooling capacitor.

In addition, the present invention provides an external composition for skin, characterized in that the extract is further obtained by concentrating the solvent under reduced pressure or lyophilization.

In addition, the present invention provides a skin whitening cosmetic composition, characterized in that the content of natural dolmulsu is 0.001-30.0% by weight based on the total freeze-dried composition, 0.1 to 2.0% by weight arbutin. When the content of the extract is less than 0.001% by weight, there is almost no skin improvement effect, and when the content of the extract is more than 30.0% by weight, the degree of increase in the effect of increasing the content is insignificant and thus it is not economical.

In addition, in the cosmetic composition of the present invention, when the content of the sedum extract is less than 0.001% by weight, there is a problem in that antioxidative and melanin inhibiting effects are not expected in the cells. There is no increase and there is a problem with formulation stability. In addition, when the content of arbutin in the cosmetic composition of the present invention is less than 0.1% by weight, there is a problem in that tyrosinase inhibitory effect is not expected due to synergistic effect with sedum extract, and when the content exceeds 2.0% by weight, There is no noticeable increase in effectiveness and there is a problem with formulation stability.

In addition, the present invention, the whitening cosmetic composition is sedum extract, characterized in that selected from the lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type and water-in-oil (W / O) type formulation And it provides a skin whitening cosmetic composition containing arbutin as the main active ingredient.

Whitening combinations of dolnamul extract and arbutin as active ingredients may be formulated into compositions such as capsules, liquids or sustained-release agents using pharmaceutically acceptable carriers, and may be formulated as carriers, excipients, binders (e.g., starches, traces). Gercan's rubber, gelatin, molasses, polyvinyl alcohol, polyvinylether, polyvinyl pyrrolidone, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose and carboxymethyl cellulose , Sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, crystalline cellulose, calcium carbonate, sodium bicarbonate and sodium alginate), lubricants (eg magnesium stearate, talc, hydrogenated vegetable oils), colorants and the like. The carriers and excipients include lactose, glucose, sucrose, mannitol, potato starch, corn starch, calcium carbonate, calcium phosphate and cellulose.

In addition to the above, additives such as adjuvants such as stabilizers, dissolution aids, transdermal absorption accelerators, and preservatives may be further added.

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.

Preparation Example 1 Preparation of Stone Sprout Extract

The stems and roots of the dried dolmens were refluxed three times for 5 hours with a 95% (V / V) ethanol solution each time, and the mixture was cooled and then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and lyophilized at 50 ° C. or lower, and the sedum extract was extracted with purified water, ethanol, butylene glycol, and propylene glycol using at least one solvent selected from 0.001 to 30.0% by weight. Was prepared.

Test Example 1 Experiment for Measuring Antioxidant Effect Using NBT Method

In this example, to determine the antioxidant effect of the dolmul extract obtained in Preparation Example 1 was compared with other antioxidants, that is, retinol and BHT under laboratory conditions, and the antioxidant activity was measured using the NBT method.

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, add 0.1ml of sample solution 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 effect was calculated by Equation 1, and the results are shown in Table 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 addition of sample solution

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

As a result, as shown in Table 1, it was confirmed that the excellent antioxidant effect in all the samples tested at 0.1% concentration. Sedum extracts had excellent antioxidant effects similar to retinol and BHT.

Sample Name Treatment Concentration (%) Antioxidative effect(%) Sedum extract 0.1 93 Retinol 0.1 91 BHT 0.1 89

Test Example 2 Intracellular Oxygen Scavenging Effect

In order to determine the inhibitory effect of active oxygen generation generated in the cell by ultraviolet light of the sedum extract obtained in Preparation Example 1, that is, the active oxygen scavenging effect, EGCG was used as a comparative sample, and the following experiment was performed using fluorescent material. .

The cell line used in the experiment was a human keratinocytes HaCaT cell line distributed by Dr. Fusenig of the German Cancer Research Center, which was used per well in a 96-well black plate for fluorescence measurement. Incubate at 2.0 × 10 ⁴ and incubate for 1 day at 37 ° C and 5% CO ₂ using DMEM (Dulbeccos Modification of Eagles Medium, FBS 10%, Gibco, USA) medium supplemented with penicillin / streptomycin The sample was then processed.

Test sample was added and cultured for 24 hours, and then washed with HCSS (HEPES-buffered control salt solution) to remove the remaining medium, and DCFH-DA (2 ', 7'-dichlorodihydro-fluorescein diacetate, Molecular Probes prepared with 20 μM in HCSS) , USA) was added thereto, incubated for 20 minutes at 37 ° C. and 5% CO ₂ , followed by washing with HCSS. Thereafter, 100 μl of the treated HCSS was added to each concentration, and the fluorescence of DCF (dichlorofluorescein) oxidized with active oxygen was measured with a fluorescence plater (Ex; 485 nm, Em; 530 nm). After UVB (20mJ / cm ² ) was irradiated and the sample was measured by a fluorescent plate reader (Ex; 485 nm, Em; 530nm).

As can be seen in Table 2, the sedum extract showed an active oxygen scavenging effect, and it was found that there was an active oxygen scavenging effect similar to that of EGCG, which is widely used as a comparative sample.

Sample Name Free radical scavenging effect (%) 0.01% by weight sedum extract 63 EGCG 61

Test Example 3 Inhibition of Tyrosinase Activity

Tyrosinase was isolated and purified from mushrooms and purchased from Sigma. Substrate L-tyrosine (Sigma) was dissolved in 0.05M sodium phosphate buffer (pH 6.8) to make a 0.1mg / ㎖ solution was used. Sedum extract was used after dissolving in an appropriate concentration in 0.05M sodium phosphate buffer (pH 6.8). 0.5 ml of L-tyrosine solution was placed in a test tube, and 0.5 ml of dolmul extract sample solution was added thereto, and left for 10 minutes in a 37 ° C thermostat. Thereafter, 0.5 ml of 200 unit / ml tyrosinase was added to each sample solution, and the mixture was reacted at 37 ° C for 10 minutes. The test tube containing the reaction solution was placed on ice and quenched to stop the reaction, and then the absorbance at 475 nm was measured with a spectrophotometer. As a control, 0.5 ml of buffer solution was used instead of the sample. The tyrosinase activity inhibition rate of each sample was calculated according to Equation 2.

Enzyme Inhibition Rate (%) = 100-[(Comparative Absorption / Control Absorbance)] X 100

As a result of testing the inhibitory effect of tyrosinase enzyme activity, the use of 0.01% dolnamul extract alone and 0.1% arbutin alone showed relatively weak enzyme activity inhibition.Albutin 0.1% and 0.01% dolnamul extract were used simultaneously. In the case of tyrosinase enzyme activity inhibitory effect was significantly increased than when used respectively, it was confirmed that it has a very good tyrosinase inhibitory effect (Table 3).

Sample Name Tyrosinase inhibition rate (%) Control - Sedum Extract 0.05% by weight 61% 0.01% by weight sedum extract 34% Arbutin 0.1 wt% 28% 0.1% by weight arbutin + 0.01% by weight sedum extract 65%

Test Example 4: Measurement of melanin production inhibitory effect

In order to confirm the whitening effect of the dolmul extract and Arbutin obtained in Preparation Example 1, the melanin synthesis inhibitory effect on B16F10 melanocytes was measured. The melanin synthesis inhibitory effect of B16F10 melanocytes was measured as follows. B16F10 melanocytes were dispensed in 6-well plates at a concentration of 2 x 10 ⁶ per well, cells were attached, and the samples were 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. Quantification of intracellular melanin was performed by slightly modifying 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 (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. Percent inhibition of production was measured. Melanin inhibition rate (%) of B16F10 melanocytes was calculated by the 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

The results of testing the melanogenesis inhibitory effect of B16F10 melanocytes showed a slight inhibitory effect when 0.01% of dolmul extract alone was used. However, when 0.2% arbutin and 0.01% of dolmul extract were used simultaneously, The production inhibitory effect was significantly increased, it was confirmed that it has a very good melanin production inhibitory effect (Table 4).

Sample Name Melanin Synthesis Inhibitory Effect (%) Control - Sedum Extract 0.1% by weight 35.2 Arbutin 0.2 wt% 56.2 Arbutin 0.2% by weight + 0.1% Sedum Extract 61.5

Test Example 5: Whitening effect test on human skin

Emulsion Example 1 containing all the dolmul extract and Arbutin obtained in Preparation Example 1 and latex Comparative Examples 1 and 2 containing dolmul extract and arbutin alone, and Comparative Example 3 containing no dolmul extract and arbutin It was prepared as shown in Table 5 and subjected to the whitening effect test on human skin.

The whitening effect test on human skin was carried out by attaching an opaque tape with six holes of 1.5cm in diameter to the inside of the upper arm of 20 healthy women, and then applying 1.5 ~ 2 times the minimum erythema of each subject. (UVB) was irradiated to induce skin blackening. Example 1 and Comparative Examples 1 to 3 prepared above were applied twice daily, and after 2 months, the contrast of the skin was measured using a chromameter (Minolta CR300). The determination of the effect was determined by obtaining a "L" value representing the contrast of the skin (unburned Korean skin color generally shows a value of 50-70). When the test material is applied, the L value gradually increases, and the comparison between the test materials is expressed as the change in brightness (ΔL) value of the color (final L value-the value before application of the test material). The greater the ΔL value, the greater the whitening effect. The ΔL values according to the experimental results are summarized in Table 6 below.

Ingredient (% by weight) Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 Stearyl alcohol 8 8 8 8 Stearic acid 2 2 2 2 Stearic Acid Cholesterol 2 2 2 2 Squalane 4 4 4 4 2-octyldodecyl alcohol 6 6 6 6 Polyoxyethylene Alcohol Ester 3 3 3 3 Glyceryl Monostearic Acid Ester 2 2 2 2 Purified water Remaining amount Remaining amount Remaining amount Remaining amount Sedum extract 0.2 0.2 - - Arbutin 0.5 - 0.5 -

division Example 1 Comparative Example 1 Comparative Example 2 Comparative Example 3 ΔL value 4.63 2.89 3.12 1.06

From the results of Table 6, it was confirmed that the ΔL value of Example 1 containing both the dolmul extract and arbutin showed a higher value than the other comparative examples.

Test Example 6 Inhibitory Effect of Prostaglandin Biosynthesis by UV Irradiation

This experimental example evaluated the prostaglandin biosynthesis inhibitory effect in order to confirm the skin inflammation inhibitory effect of the dolmul extract obtained in Example 1. Keratinocytes isolated from human epidermal tissues were placed in 24-well test plates 5 × 10 ⁴ and attached for 24 hours. After replacement with medium containing no FBS and incubation for 18 hours, prostaglandin biosynthetic enzyme (prostaglandin E2 synthetase, present in keratinocytes) was treated with aspirin to achieve a final concentration of 50 uM on the keratinocytes. Or cyclooxygenase (hereinafter referred to as COX) activity was removed. Two hours after aspirin treatment, each well containing keratinocytes was washed twice with PBS and 100 μl of PBS was added to each well. The keratinocytes were irradiated with UV 30mJ / cm ² using a UVB lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then PBS was removed and each cell was prepared with keratinocyte growth media (Clonetics). , Biowhittacker, MD, USA) 250 μl was added. After treatment with the sedum extract to be evaluated here was incubated for 16 hours. An appropriate amount of the culture supernatant was taken to quantify biosynthesized PGE2 (prostaglandin E2) for 16 hours to determine the prostaglandin inhibitory effect of sedum extract. The amount of PGE2 was quantified using enzyme immunoassay, and experimental results showed that sedum extract effectively inhibited the production of prostaglandins by UV light. In particular, the produced PGE2 is known to be mainly produced by the COX-2 enzyme, and thus the experiment shows that the sedum extract inhibits the induction or activity of the COX-2 enzyme (Table 7).

Sample Name PGE2 Inhibition Rate (%) (-) UV B control (+) UV B - (+) UV B + Sedum Extract 0.05% by weight 46% UV B + Sedum Extract 0.01% by weight 21%

Claims (3)

Cosmetic composition for skin whitening containing sedum extract and arbutin as an active ingredient. The cosmetic composition for skin whitening according to claim 1, which contains 0.001 to 30.0 wt% of dolmul extract and 0.1 to 2.0 wt% of arbutin with respect to the whole composition. The cosmetic composition according to claim 1 or 2, wherein the cosmetic composition comprises a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation and Cosmetic composition for skin whitening, characterized in that it has a formulation selected from the group consisting of a spray.