KR102522196B1 - Skin whitening composition comprising Safflower seed oil hydrolyzate and Saururus chinensis extract - Google Patents

Abstract

The present invention relates to a cosmetic composition containing, as an active ingredient, a mixture obtained by mixing safflower seed oil hydrolyzate and Sambakcho extract in a specific weight part ratio, wherein the cosmetic composition is effective in inhibiting Tyrosinase protein level and MITF mRNA expression. It shows an excellent skin whitening effect as a synergistic effect by the complex action of two mechanisms, such as inhibition of tyrosinase production by inhibition of MITF protein production according to

Description

Skin whitening composition comprising a mixture of safflower seed oil hydrolyzate and Saururus chinensis extract as an active ingredient

The present invention relates to a composition for skin whitening containing a mixture of safflower seed oil hydrolyzate and Sambakcho extract as an active ingredient, and specifically, to a composition having excellent skin whitening effect including safflower seed oil hydrolyzate and Sambakcho extract in a specific weight part ratio.

Human skin color is largely determined by the amount of melanin, hemoglobin, and carotene, among which melanin plays the most important role. Melanin is a pigment present in the epidermal layer of the skin. It is produced by melanocytes present in the epidermal layer of the skin. It blocks ultraviolet rays from the sun to prevent skin damage and catches harmful oxygen and free radicals generated in the skin's body to prevent external damage. It plays a role in protecting the skin from harmful factors. However, excessive exposure to ultraviolet rays or excessive production of melanin due to various environmental factors causes pigmentation and skin troubles such as freckles, age spots, and melasma.

Looking at the melanin synthesis process in detail, an enzyme called tyrosinase present in the endoplasmic reticulum called melanosome in melanocyte produces DOPAauinone using tyrosine as a substrate. , dopaquinone undergoes an autoxidation reaction to become DOPAchirome. Again, dopachrome produces melanin by an enzymatic reaction. The produced melanin is transferred to keratinocytes through melanosomes, where it comes out to the surface of the skin and is lost along with keratin through a keratinization process of about 28 days.

Among them, tyrosinase is an enzyme that determines the production of melanin in the early stage of melanin synthesis and is expressed by a transcription factor called Microphtalmia-associated transcription factor (MITF). MITF plays an important role in the expression of TRP1 and TRP2 enzymes required for melanin formation as well as tyrosinase. The above enzymes can convert L-tyrosine into two types of melanin, eumelanin and pheomelanin, in organelles of melanocytes called 'melanosomes'. to determine skin color or protect skin from UV.

Conventionally, substances having inhibitory activity against tyrosinase, such as hydroquinone, ascorbic acid, kojic acid, and glutathione, are formulated into cosmetics such as ointments and essences for skin whitening. effect or improved skin hyperpigmentation such as melasma and freckles.

However, although hydroquinone shows a certain whitening effect, there is a disadvantage in that the amount of blending should be limited to a very small amount due to severe skin irritation, and ascorbic acid is easily oxidized, and the cosmetic composition containing it may cause discoloration and odor. In addition, thiol-based compounds such as glutathione and cysteine not only have a unique unpleasant odor, but also have problems with percutaneous absorption, and their glycosides and derivatives are also highly polar, making it difficult to use them as cosmetic ingredients. On the other hand, placenta extract and the like have no skin irritation, but have a problem of insufficient whitening effect, and recently, their use has been banned due to the risk of inducing mad cow disease.

The present inventors have devised to solve the problems of the prior art, and when the mixture obtained by mixing naturally obtained safflower seed oil hydrolyzate and Sambakcho extract in a specific weight part ratio is contained as an active ingredient, tyrosinase protein It was confirmed through experiments that an excellent skin whitening effect can be obtained by the synergistic effect of the complex action of two mechanisms, such as suppressing the level and inhibiting the MITF-M protein level to inhibit the production of tyrosinase. invention was completed.

Accordingly, an object of the present invention is to provide a cosmetic composition for skin whitening, containing a mixture of safflower seed oil hydrolyzate and triticale extract as an active ingredient.

In addition, the technical problem to be solved by the present invention is not limited to the above-mentioned technical problem, and other technical problems not mentioned above will become clear to those skilled in the art from the description below. You will be able to understand.

In the present specification, a cosmetic composition for skin whitening containing a mixture of safflower seed oil hydrolyzate and triticale extract as an active ingredient is provided.

The mixture may be a mixture of safflower seed oil hydrolyzate and triticale extract in a ratio of 2:0.5 to 1.5 parts by weight.

The safflower oil hydrolyzate is obtained by treating safflower oil with at least one alkali metal hydroxide selected from the group consisting of sodium hydroxide (NaOH), calcium hydroxide (Ca(OH) ₂ ), lithium hydroxide (LiOH) and potassium hydroxide (KOH). It may be decomposed or hydrolyzed by adding an ester hydrolase to safflower seed oil.

The triticale extract is one or more solvents selected from the group consisting of water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol and hydrous glycerin may have been extracted.

The mixture may be included in an amount of 0.001 to 50% by weight based on the total weight of the composition.

The composition may have one or more formulations selected from the group consisting of softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray, and powder.

The cosmetic composition for skin whitening prepared according to the present invention has a complex action of two mechanisms, such as inhibition of tyrosinase protein level and inhibition of tyrosinase production by inhibition of MITF protein production according to inhibition of MITF mRNA expression. As a synergistic effect by , it shows an excellent skin whitening effect.

1 is a result of measuring the linoleic acid content of safflower oil hydrolyzate prepared according to an embodiment of the present invention.
Figure 2 shows the results of the stability test according to the accelerated experiment of the safflower seed oil hydrolyzate and the extract mixture prepared according to an embodiment of the present invention.
Figure 3 shows the results of the cell viability test of the safflower seed oil hydrolysate and the triticale extract (30% ethanol extract, 70% ethanol extract) prepared according to an embodiment of the present invention.
Figure 4 shows the results of measuring the melanin biosynthesis inhibitory effect by UVB irradiation with respect to one embodiment of the present invention.
Figure 5 shows the results of measuring the melanin biosynthesis inhibitory effect by α-MSH stimulation with respect to one embodiment of the present invention.
6 is a measurement of IC ₅₀ concentrations of samples for melanin biosynthesis produced by UV stimulation in Pam212 and B16F0 co-culture conditions for one embodiment of the present invention.
Figure 7 is a measurement of the IC ₅₀ concentration of samples for melanin biosynthesis produced by α-MSH stimulation in the B16F0 cell line with respect to one embodiment of the present invention.
Figure 8 shows the cell viability test results of the safflower seed oil hydrolyzate and the triticale extract mixture according to one embodiment and comparative example of the present invention.
9 is a result of measuring the effect of samples on melanin biosynthesis produced by UV stimulation in Pam212 and B16F0 co-culture conditions of safflower seed oil hydrolyzate and triticale extract mixture according to one embodiment and comparative example of the present invention.
10 is a result of measuring the effect of a sample on melanin biosynthesis produced by α-MSH stimulation in the B16F0 cell line of a mixture of safflower oil hydrolyzate and Sambakcho extract according to one embodiment and comparative example of the present invention.
11 is a result of measuring the protein level inhibitory effect of the melanin synthesis marker MITF-M of a mixture of safflower oil hydrolyzate and Sambakcho extract according to an embodiment of the present invention.
Figure 12 is a result of measuring the mRNA and protein level inhibitory effect of the melanin synthesis marker tyrosinase (Tyrosinase) of the safflower seed oil hydrolyzate and the extract mixture of Sambakcho according to an embodiment of the present invention.
Figure 13 schematically shows the mechanism of action of the mixture of safflower oil hydrolyzate and triticale extract according to an embodiment of the present invention.

Hereinafter, with reference to the accompanying drawings, embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention. However, the present invention may be implemented in many different forms and is not limited to the embodiments disclosed below. In addition, in order to clearly disclose the present invention in the drawings, parts irrelevant to the present invention are omitted, and the same or similar numerals in the drawings indicate the same or similar components.

Specifically, according to one embodiment of the present invention, the cosmetic composition for skin whitening may contain a mixture of safflower seed oil hydrolyzate and triticale extract as an active ingredient.

Safflower (Carthamus tinctorious L.) is an annual plant belonging to the Asteraceae family and is native to the mountainous regions of Afghanistan or Ethiopia, and is cultivated in China and Tibet. The medicinal parts of safflower include flowers and seeds. It inhibits platelet coagulation, delays bleeding time, and also lowers plasma cholesterol and triglycerides.

Safflower seeds contain phenolic compounds such as serotonin, lignans, and flavonoids, so they have strong antioxidant and antihyperlipidemic effects. In particular, as a phytoestrogen component with estrogen-like activity, it promotes bone formation similar to components known as existing phytoestrogen compounds. By doing so, it is possible to prevent osteoporosis in menopausal women and to form new bone in case of bone defect.

In particular, safflower seed oil contains a large amount of unsaturated fatty acids including linoleic acid and tocopherol, so it is used as a new antioxidant, antithrombotic and antihypertensive material. For this purpose, roasted safflower seeds are milked and used as cooking oil.

On the other hand, in the present invention, safflower oil hydrolyzate is used, and safflower oil hydrolyzate according to an embodiment of the present invention is sodium hydroxide (NaOH), calcium hydroxide (Ca(OH) ₂ ), lithium hydroxide (LiOH) and It may be hydrolyzed by treating at least one alkali metal hydroxide selected from the group consisting of potassium hydroxide (KOH), or hydrolyzed by adding an ester hydrolase to safflower seed oil, and the ester hydrolase is a lipase ( Lipase), for example, lipozymes or lipopans.

As an example, the safflower oil hydrolyzate may be obtained by adding safflower oil to an aqueous sodium hydroxide solution and performing a hydrolysis reaction for about 50 to 70 minutes while stirring in a constant temperature water bath within a temperature range of about 80 to 100 ° C.

Sambaekcho (三白草; Saururus chinensis Baill ) is a perennial herb of the family Sambaek, distributed in Korea, China, and Japan, and is a plant that grows wild in wetlands or beaches. Outpost contains essential oils such as quercetin, quercitrin, isoquercitrin, rutin and water-soluble tannin. It has antibacterial effect, antimutagenic action, reduction of activity of carcinogenic substances, inhibition of growth of cancer cells, lowering of blood pressure, capillary strengthening action, and quercitrin is known to have anti-inflammatory, analgesic and antioxidant effects. In addition, Sambaekcho is effective in treating beriberi, jaundice, jaundice, carbuncle, dropsy, hemorrhoids, acute and chronic urethritis, prostatitis, cystitis, gonorrhea, dysentery, fatigue from heavy labor, aftereffects and muscle pain caused by bruises, and inflammation of the skeletal and bone marrow. It is known to be effective in treating pain caused by

On the other hand, the triticale extract according to an embodiment of the present invention is composed of water, ethanol, methanol, propanol, butanol, acetone, ethyl acetate, hexane, butylene glycol, propylene glycol, hydrous butylene glycol, hydrous propylene glycol and hydrous glycerin It may be extracted with one or more solvents selected from the group, specifically obtained by extraction with 30% aqueous ethanol to 70% aqueous ethanol at room temperature for one week, more specifically with 30% aqueous ethanol at room temperature for one week. It may have been obtained by extraction.

On the other hand, the cosmetic composition for skin whitening according to an embodiment of the present invention contains a mixture of safflower seed oil hydrolyzate and Sambakcho extract as an active ingredient, but the mixture contains safflower seed oil hydrolyzate and Sambaekcho extract in an amount of 2: 0.5 to 1.5 by weight It may be a booby-mixed mixture. On the other hand, when the safflower seed oil hydrolyzate and the triticale extract are mixed within the above weight part ratio range, the mechanism of inhibiting the tyrosinase protein level of the safflower seed oil hydrolyzate and the inhibition of MITF protein production according to the inhibition of MITF mRNA expression in the triticale extract It is possible to optimize the skin whitening effect due to the synergistic action of the mechanism of inhibiting the production of tyrosinase.

In addition, in the cosmetic composition for skin whitening according to an embodiment of the present invention, the mixture may be included in an amount of 0.001 to 50% by weight based on the total weight of the composition.

On the other hand, the active ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions in addition to the mixture of safflower seed oil hydrolyzate and triticale extract, for example antioxidants, stabilizers, solubilizers, vitamins , conventional containing agents such as pigments and fragrances, and the like.

In addition, the cosmetic composition containing the mixture of the hydrolyzate of safflower seed oil and the extract of Sambaekcho of the present invention as an active ingredient can be prepared in any conventionally prepared formulation, for example, a solution, suspension, emulsion, paste, gel, It may have one or more formulations selected from the group consisting of cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray.

Example

Hereinafter, the contents of the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of equivalent technical ideas.

manufacturing example

Manufacture of safflower seed oil hydrolyzate (hereinafter referred to as SFH)

After putting 20 g of safflower seed oil in a 500 ml beaker, an aqueous solution obtained by dissolving 20 g of NaOH in 36 g of purified water was added. The hydrolysis reaction proceeded for 60 minutes while stirring this mixed solution in a constant temperature water bath maintained at 90°C. During the hydrolysis reaction, an aqueous solution in which 100 g of NaCl was dissolved in 300 g of purified water was cooled in a refrigerator, and then the solution in which the hydrolysis reaction was completed was slowly added to the brine solution. The resulting solid was filtered under reduced pressure and dried again at room temperature under reduced pressure to prepare 16.4 g of safflower seed oil hydrolyzate.

Manufacture of Sambaekcho extract (hereinafter referred to as SC)

Saururus chinensis was prepared and extracted at room temperature for one week using 30% ethanol aqueous solution and 70% ethanol aqueous solution, respectively. After completion of the extraction, filtration was performed using a filter paper to obtain an extracted solution, and concentration and drying under reduced pressure were performed to obtain the target triticale extract (4.52 g of ethanol 30% aqueous solution (SC30), ethanol 70% aqueous solution). (SC70) 3.95 g) were obtained respectively.

Example 1: Preparation of a mixture of safflower oil hydrolyzate (SFH) and triticale extract (SC)

2.34 g of the prepared Sambaekcho extract (SC30) was completely dissolved in 110 g of 30% 1,3-butylene glycol aqueous solution, and then 4.68 g of the prepared safflower seed oil hydrolyzate was added thereto and dissolved while stirring at 50 ° C. After filtering using a 1.0 μm membrane filter and a 0.45 μm membrane filter sequentially, 100 g of a mixture of safflower seed oil hydrolyzate and Sambacaceae extract in which insoluble matter and microorganisms were completely removed was prepared.

[Experiment 1: Measurement of linoleic acid content]

In order to measure the content of linoleic acid contained in the safflower oil hydrolyzate (SFH) prepared according to the preparation example, analysis was performed under the following conditions, and supelco 37 component FAME Mix (purity 100%) was used as the standard material. The equipment used was a gas chromatograph (GC) Agilent 7890A, and the analysis conditions and results were as follows.

Detector FID Column SPTM-2560 (100m x 0.25mm x 0.20μm) Injection Temp. 225℃ Injection Volume 1.0 μl Detector Temp. 285℃ Oven Temp. 100℃(4min.), 208℃(3℃/min, 5min), 244℃(2℃/min, 15min) Carrier Gas He Column Flow 0.75mL/min. Split Ratio 200:1

sample safflower oil Safflower oil NaOH hydrolyzate Safflower Oil Lipase Hydrolyzate Linoleic acid content (%) 1.3 59.88 57.24

Referring to the results of Table 2 and FIG. 1, in the case of the safflower seed oil hydrolyzate prepared according to the preparation example of the present invention, the linoleic acid content was about 59.88%, and it could be confirmed that it showed a significantly higher content than other oils. there was.

[Experiment 2: Stability test by accelerated test]

A mixture of safflower seed oil hydrolyzate (SFH) and Sambakcho extract (SC30) obtained in Example 1 was placed in an incubator at room temperature (25 ° C) and 45 ° C for 0 month, 1 month, and 2 month intervals, respectively, and the color and pH of the solution changed. As a result of measuring with the naked eye and a pH meter, no color change was observed by visual observation, and it was measured that there was almost no pH change over time (see FIG. 2).

[Experiment 3: Cell viability measurement experiment]

cell culture

Pam212 cells are mouse keratinocytes, B16F0 cells are mouse melanoma cell lines, and adhere to the culture dish to proliferate. After diluting 6 × 10 ⁵ Pam212 cells and 3 × 10 ⁵ B16F0 cells in a 100 mm culture dish with 10 ml of DMEM containing 10% FBS, they were subcultured every 48 hours and used for experiments.

Cell viability measurement

The MTT method was used to measure the viability of cells by a colorimetric method using absorbance. Pam212 cells were dispensed at 5 × 10 ³ cells/well in a 96-well culture plate, B16F0 cells were dispensed at 2.5 × 10 ³ cells/well in a 96-well culture plate, and after 24 hours, the medium of the 96-well culture plate was removed. Fresh DMEM 10% FBS medium and serially diluted samples were treated respectively. After 72 hours, the medium of the 96-well culture plate was removed and the MTT reagent was reacted with the medium at a concentration of 500 ug/ml for 1 hour. After removing the MTT solution, 200 ul of DMSO was added to dissolve formazan crystals, and absorbance was measured at a wavelength of 590 nm.

Specifically, in order to examine the cytotoxicity of each sample on the mouse melanoma B16F0 cell line and the mouse keratinocyte cell line Pam212, tetrazolium, a yellow water-soluble substrate, was converted to blue-purple water-insoluble formazan by mitochondrial dehydrogenase action in living cells. The maximum concentration that does not exhibit cytotoxicity was confirmed using the MTT method using the function of reducing to . In order to confirm the presence or absence of toxicity to B16F0 and Pam212 cells, cell viability was measured using the MTT method after culturing the cells for 72 hours for each sample. After treatment with each concentration of SC30, SC70 and SFH according to Preparation Example, MTT cell viability was measured in B16F0 and Pam212 cells. As a result, when the survival rate of the control group was set to 100%, the non-toxic concentration was confirmed as the maximum concentration that could be treated. As a result, as shown in FIG. 3, it was confirmed that 100 ug/ml for SC30, 10 ug/ml for SC70, and 100 ug/ml for SFH could be set as the maximum concentration.

[Experiment 4: Measurement of the amount of melanin]

Melanin assay by UVB irradiation (Co-cullture system)

After growing B16F0 3.5 × 10 ⁴ cells/well in a 6-well plate and Pam212 1 × 10 ⁴ cells/well cells in a 6-well plate insert for 24 hours, the medium was removed and enough PBS was added to lightly cover the cells. It was irradiated with an intensity of 2 mJ/cm ² that does not affect the cell growth rate using a UV-crosslinker. After removing the PBS, the medium containing the sample was added, cultured for 72 hours, and melanin was measured by measuring the absorbance at 405 nm.

Specifically, in order to measure the effect of samples on melanin biosynthesis produced by UV stimulation in Pam212 and B16F0 co-culture conditions, melanin biosynthesis was induced by stimulating UV in keratinocytes and melanocytes co-culture conditions similar to skin conditions. After that, the effect of SC30 and SC70 extracts on melanin biosynthesis was confirmed. After stimulation with UV 2mJ/cm ² under B16F0 and Pam212 co-culture conditions, each sample was treated for 72 hours at a concentration that was not toxic to cells, and the amount of melanin synthesized in cells at an absorbance of 405 nm and released into the medium was measured. As a result of the experiment, SC30 and SC70 inhibited melanin biosynthesis (see FIG. 4).

α-MSH stimulation-induced melanin assay

After dispensing B16F0 2.5 × 10 ³ cells/well in a 96-well plate, the cells are grown for 24 hours. After the addition of a medium containing 100nM α-MSH and the sample, the mixture was cultured for 72 hours, and the amount of melanin was calculated by measuring the absorbance at 405 nm.

Specifically, in order to measure the effect of the sample on melanin biosynthesis produced by α-MSH stimulation in the B16F0 cell line, B16F0 cells were treated with the sample at each concentration for 72 hours, and the sample was synthesized in the cells at an absorbance of 405 nm and transferred to the medium. The amount of melanin released was measured. As a result of the experiment, it was confirmed that the samples prepared according to Preparation Example (SC30, SC70, SFH) inhibit melanin biosynthesis by α-MSH stimulation (see FIG. 5).

Analyzing the above experimental results, it was confirmed that melanin production was increased by stimulation by UV in the co-culture system or stimulation by α-MSH in B16F0, but SC30, SC70 and SFH according to the manufacturing example reduced it.

IC of samples for melanin biosynthesis produced by UV stimulation under Pam212 and B16F0 co-culture conditions ₅₀ concentration measurement

By selecting SC30, SC70, and SFH having excellent melanin production inhibitory effect by UV stimulation, the sample concentration (IC ₅₀ ) when the melanin synthesis inhibition rate of the sample was 50% was confirmed. After stimulation with UV 2 mJ/cm ² under B16F0 and Pam212 co-culture conditions, each sample was treated with various concentrations that were not toxic to cells for 72 hours, and melanin synthesized by cells at an absorbance of 405 nm and released into the medium The amount of was measured. As a result, the concentration of 50% inhibition of melanin (IC ₅₀ ) by UV stimulation was 5.9 ug/ml for SC30, 0.7 ug/ml for SC70, and 22 ug/ml for SFH (see FIG. 6).

IC of samples for melanin biosynthesis produced by α-MSH stimulation in B16F0 cell line ₅₀ density

SC30 and SC70, which have excellent melanin inhibitory effect by α-MSH stimulation, were selected to confirm the sample concentration (IC ₅₀ ) when the melanin synthesis inhibition rate of the sample was 50%. The B16F0 cell line was stimulated with α-MSH, samples of various concentrations were treated for 72 hours, and the amount of melanin synthesized in the cells and released into the medium was measured at an absorbance of 405 nm. As a result, IC ₅₀ , which is the concentration of 50% inhibition of melanin by α-MSH stimulation, was confirmed to be 22 ug/ml for SC30 and 2 ug/ml for SC70 (see FIG. 7 ).

Comparative Examples 1-1, 1-2 and 1-3

A mixture was prepared in the same manner as in Example 1, but SFH and SC70 were mixed at a weight part ratio of 2:0.5 (Comparative Example 1-1) and 2: mixed at a weight part ratio of 0.1 (Comparative Example 1-2 ), SFH and SC30 were mixed at a weight part ratio of 2: 0.5 (Comparative Example 1-3).

[Experiment 5: Cell viability measurement experiment]

Cell viability measurement

The MTT method was used to measure the viability of cells by a colorimetric method using absorbance. Pam212 cells were dispensed at 5 × 10 ³ cells/well in a 96-well culture plate, B16F0 cells were dispensed at 2.5 × 10 ³ cells/well in a 96-well culture plate, and after 24 hours, the medium of the 96-well culture plate was removed. Fresh DMEM 10% FBS medium and serially diluted samples were treated respectively. After 72 hours, the medium of the 96-well culture plate was removed, and the MTT reagent was reacted with the medium at a concentration of 500 ug/ml for 1 hour. After removing the MTT solution, 200 ul of DMSO was added to dissolve formazan crystals, and absorbance was measured at a wavelength of 590 nm.

Specifically, in order to examine the cytotoxicity of each sample on the mouse melanoma B16F0 cell line and the mouse keratinocyte cell line Pam212, tetrazolium, a yellow water-soluble substrate, was converted to blue-purple water-insoluble formazan by mitochondrial dehydrogenase action in living cells. The highest concentration that does not exhibit cytotoxicity was confirmed using the MTT method using the function of reducing to . In order to confirm the presence or absence of toxicity to B16F0 and Pam212 cells, cell viability was measured using the MTT method after culturing the cells for 72 hours for each sample. Example 1, the mixture of Comparative Examples 1-1 to 1-3 was cultured for 72 hours and cell viability was measured using the MTT method. As a result, when the survival rate of the control group was set at 100%, treatment without toxicity The maximum possible concentration was confirmed (see FIG. 8).

[Experiment 6: Experiment to measure the effect on melanin biosynthesis]

Effect of samples on melanin biosynthesis produced by UV stimulation in Pam212 and B16F0 co-culture conditions of Example and Comparative Example mixtures

Based on the mixtures of Example 1 and Comparative Examples 1-1 to 1-3, after stimulation with UV 2 mJ/cm ² under B16F0 and Pam212 co-culture conditions, the effect of inhibiting melanin synthesis by the mixture was confirmed. As a result, IC ₅₀ , the concentration of 50% inhibition of melanin by UV stimulation, was 5ug/ml of SFH and 1.3 ug/ml of SC70 in the case of a mixture (Comparative Example 1-1) in which the ratio of SFH and SC70 was 2: 0.5, and the ratio was 2: 0.1 In the case of the phosphorus mixture (Comparative Example 1-2), SFH was 11.7 ug/ml and SC70 was 0.6 ug/ml. In the case of a mixture with a ratio of SFH and SC30 of 2: 1 (Example 1), IC ₅₀ shows SFH 6.5 ug / ml and SC70 0.6 ug / ml, and in the case of a mixture with a ratio of 2: 0.5 (Comparative Example 1-3), the IC ₅₀ showed SFH 12.3 ug/ml and SC70 3.1 ug/ml (see FIG. 9).

Effect of samples on melanin biosynthesis produced by α-MSH stimulation in the B16F0 cell line of Example and Comparative Example mixtures

Based on the mixtures of Example 1 and Comparative Examples 1-1 to 1-3, the effect on melanin biosynthesis generated in α-MSH in the B16F0 cell line was confirmed. B16F0 cells were treated with the samples at each concentration for 72 hours, and the amount of melanin synthesized in the cells and released into the medium was measured at an absorbance of 405 nm. As a result of the experiment, IC ₅₀ , which is the concentration of 50% inhibition of melanin by α-MSH stimulation, was 5.9 ug/ml of SFH and 1.5 ug/ml of SC70 for the mixture of Comparative Example 1-1, and SFH for the mixture of Comparative Example 1-2 19.3 ug/ml, SC70 1 ug/ml. In the case of the mixture of Example 1, the IC ₅₀ was SFH 18.5 ug/ml and SC70 9.3 ug/ml, and in the case of the mixture of Comparative Examples 1-3, the IC ₅₀ was SFH 19.6 ug/ml and SC70 4.9 ug/ml ( see Figure 10).

Referring to the above experimental results, it can be seen that the melanin IC ₅₀ value is lower when used as a mixture of SFH and SC70 and SFH and SC30 than when SFH, SC70, and SC30 are used alone, respectively, Accordingly, it can be confirmed that the SC70/SFH or SC30/SFH mixture exhibits a more excellent whitening effect than the single agent.

In particular, in the case of a mixture containing 30 ug/ml of SFH and 15 ug/ml of SC30 (mixed at a weight part ratio of 2:1), such as the mixture according to Example 1, within a range that does not exhibit cytotoxicity, UV Alternatively, it was confirmed that melanin production by α-MSH stimulation could be inhibited by more than 90% (see Table 3 below).

[Experiment 7: Western blot]

Identification of the protein level of the melanin synthesis marker MITF-M

In order to confirm the protein level of the melanin synthesis marker MITF-M of the mixture according to Example 1, quantitative analysis was performed through Western blotting. In the case of SFH alone treatment, the increased MITF-M protein amount was not suppressed by α-MSH stimulation, but it was confirmed that the MITF-M protein amount was suppressed when SC30 alone treatment or the mixture treatment of Example 1 was performed (Fig. 11).

Determination of mRNA and protein levels of Tyrosinase, a melanin synthesis marker

The effect of the mixture of Example 1 on the mRNA and protein levels of Tyrosinase, which is the most important protein for melanin biosynthesis, was quantitatively analyzed through RT-PCR and Western blotting. As a result, treatment with SFH alone suppressed the amount of Tyrosinase protein, which was increased by αMSH stimulation, but did not suppress the mRNA level. However, both SC30 alone and the mixture of Example 1 inhibited both Tyrosinase mRNA and protein levels. This result indicates that SFH alone is involved in Tyrosinase protein inhibition, and that SC30 inhibits melanin synthesis by inhibiting MITF-M protein production according to inhibition of MITF mRNA expression and thus inhibition of Tyrosinase production. In particular, in the case of the mixture of Example 1, two mechanisms, such as a mechanism of directly inhibiting the level of tyrosinase protein and a mechanism of suppressing tyrosinase production by inhibiting MITF protein production according to inhibition of MITF mRNA expression, It shows an excellent skin whitening effect as a synergistic effect by a complex action (see FIG. 12).

In summary, as confirmed through the above experiments, when UV stimulates keratinocytes in the epidermis, keratinocytes secrete α-MSH, ET-1, and SCF, which are paracrine factors that can stimulate the melanin synthesis signal transduction system. . These proteins bind to each receptor in melanocytes to increase MITF mRNA expression, and the increased MITF-M binds to the promoter of Tyrosinase production to increase Tyrosinase protein production, thereby promoting skin melanin synthesis. On the other hand, when SFH alone is applied, Tyrosinase protein level is directly suppressed, and when SC30 is applied alone, both MITF-M protein and Tyrosinase mRNA are suppressed. In the end, the mixture of Example 1 suppresses the Tyrosinase protein level due to the combined effect of the two agents, suppresses both the mRNA of Tyrosinase and the amount of MITF-M protein, an important transcription factor for Tyrosinase production, and inhibits melanin synthesis. It can be confirmed that it exhibits an excellent whitening effect (see FIG. 13).

[Experiment 8: Clinical trial on skin brightness improvement effect]

For 12 healthy adult women (average age 51.25), the cream formulation containing the mixture of Example 1 was applied to the face, and the skin brightness of the hyperpigmentation area after 2 weeks and 4 weeks of application, respectively ( The averages of individual increase rates (%) for L*) and L* values were measured, respectively, and are shown in Tables 4 and 5 below.

2 weeks after application 4 weeks after application Individual authentication rate of L* value (%) 0.099 0.295

Looking at the results of Table 4, the L* value showed an increase rate of 0.099% and 0.285%, respectively, after 2 weeks and 4 weeks after application compared to before application of the product. On the other hand, as a result of analyzing the proportion of study subjects whose L* value increased, it was confirmed that 75.000% and 100.000% of study subjects showed an increase in L* value after 2 weeks and 4 weeks after application, respectively (Table below). see 5).

2 weeks after application 4 weeks after application Ratio of study subjects with increased L* value (%) 75.000 100.000

As described above, in the case of a mixture (SS-Complex) containing safflower seed oil hydrolysate (SFH) and Sambakcho extract (SC30) in a specific weight part ratio according to the present invention, the mechanism of inhibiting Tyrosinase protein level and the production of MITF protein It was confirmed that the mechanism of inhibiting Tyrosinase mRNA can be simultaneously performed, effectively suppressing melanin production through two pathways, thereby exhibiting a strong synergistic effect related to skin whitening.

In the foregoing, although specific embodiments of the present invention have been described and shown, the present invention is not limited to the described embodiments, and it is common knowledge in the art that various modifications and variations can be made without departing from the spirit and scope of the present invention. It is self-evident to those who have Therefore, such modifications or variations should not be individually understood from the technical spirit or viewpoint of the present invention, and modified embodiments should fall within the scope of the claims of the present invention.

Claims (6)

Contains a mixture of safflower seed oil hydrolyzate and triticale extract as active ingredients,
The mixture is a mixture of safflower oil hydrolyzate and triticale extract in a ratio of 2: 0.5 to 1.5 parts by weight,
The safflower oil hydrolyzate is prepared by adding sodium hydroxide (NaOH) aqueous solution or lipase hydrolase to safflower seed oil obtained by milking roasted safflower seeds, stirring at a temperature of 80 to 100 ° C., and performing a hydrolysis reaction for 50 to 70 minutes is obtained,
The triticale extract is extracted for 7 days with 30% ethanol aqueous solution to 70% ethanol aqueous solution at room temperature, a cosmetic composition for skin whitening. delete delete delete According to claim 1,
The mixture is contained in 0.001 to 50% by weight based on the total weight of the composition, a cosmetic composition for skin whitening. According to claim 1,
The composition is one or more formulations selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, and sprays. Having a cosmetic composition for skin whitening.

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