KR20180052157A - Cosmetic composition for whitening skin comprising extract of Sorbus aucuparia berry or fermented product of Sorbus aucuparia berry - Google Patents

Abstract

The present invention relates to a cosmetic composition for whitening skin comprising a fermented product of Sorbus commixta fruits obtained by fermenting an extract of Sorbus commixta fruits with lactic acid bacteria as an active ingredient. According to the present invention, the cosmetic composition for whitening skin, which comprises the extract of Sorbus commixta fruits or the fermented product of fermented product of the Sorbus commixta fruits inhibits the gene expression of tyrosinase and microphthalmia-associated transcription factor (MITF) more effectively, and inhibits melanin biosynthesis by promoting liver X Receptor (LXR) activity, and thus in conclusion, a skin whitening effect can be shown.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for skin whitening comprising a fermentation product of a fermentation product,

The present invention relates to a skin whitening cosmetic composition containing a hole-sealing material or a hole-sealing material. More specifically, the present invention relates to a cosmetic composition for skin whitening containing a hole-sealing material or a hole-sealing material, To a cosmetic composition for skin whitening.

The factors determining human skin color include melanin, thickness of stratum corneum, peripheral blood vessels and hemoglobin, and carotene. Of these, melanin is the most important factor in determining skin color. The number of melanin-producing cells that produce melanin pigment is constant irrespective of race or skin color. However, the degree of melaninization, the size and number distribution of melanocytes, and the resolution of melanocytes in keratinocytes are different, There is a difference. (Lee, Seungheon, et al., Skin barrier study, Lemoon, 2010)

The amount of biosynthesis increases or decreases in response to various environments surrounding melanocytes during melanin production. Studies of this regulatory structure have centered on the mechanism of ultraviolet-induced skin melanogenesis. The cytokines such as basic fibroblast growth factor (bFGF), alpha-melanocyte stimulating hormone (α-MSH), endothelin-1 (ET-1) and prostaglandin E 2 (PGE 2) are produced and secreted by ultraviolet . Receptors that respond to these factors are present on the cell membrane of melanocytes, and the signal from the receptor affects the microphthalmia-associated transcription factor (MITF). MITF is also affected by LXR (Liver X Receptor), which is influenced by the activation of the extracellular signal-regulated kinase (ERK) pathway as well as the expression itself. Liver X receptor (LXR), a nuclear receptor, is a ligand - activated nuclear receptor that is known to play a central role in maintaining lipid metabolism and cholesterol homeostasis. Recent studies have shown that LXR activation inhibits melanin biosynthesis in melanocytes (Lee CS et al., Liver X Receptor Activation Inhibits Melanogenesis through the Acceleration of ERK-Mediated MITF, Degradation J Invest Dermatol., 2013 ). It has also been reported that the LXR activator inhibits melanin production and exhibits an excellent whitening effect (10-2012-0078675: a skin lightening agent containing liver X receptor activator).

 MITF, which acts as a transcription factor, is involved in not only tyrosinase but also tyrosinase related protein 1 (TRP-1), tyrosinase related protein 2 (TRP-2) induces the transcription of a series of enzymes involved in melanin synthesis such as dopachrome tautomerase (Saha B et al., Activation of the MitF promoter by lipid-stimulated activation of p38-stress signaling to CREB. Pigment Cell Res., 2006 ).

The resulting melanin synthases hydrolyze tyrosine (L-tyrosine) in melanin-producing cells, leading to reactions to L-dopa and L-dopaquinone. Tyrosinase is an enzyme that catalyzes these two reactions, and substances such as hydroquinone and arbutin, which are known as whitening agents, interfere with the reaction, thereby inhibiting melanin production, resulting in a whitening effect. The resulting dopaquinone is converted to black eumelanin by a series of enzymatic actions of DCT (dopachrome tautomerase), TRP-1 (tyrosinase related protein 1) and TRP-2 (tyrosinase related protein 2) Are synthesized.

On the other hand, the hole thread (Sorbus Aucuparia Berry ) is a fruit of the fallen leaves of the fallen leaves, belonging to the Rosaceae. It is harvested in autumn and used in the sun. The sour taste of the hallucinos is a cure for cough and sputum. Previous studies have revealed whitening efficacy of the trunks and bark of the legs (10-2005-0110149: cosmetic composition containing the legume extract), but no whitening effect on the ear shell, which is the fruit of the legs, is described at all.

Accordingly, the present inventors have made intensive researches to overcome the problems of the prior art, and as a result, they have found that a fermentation process using only a positive-working chamber, which is a fruit of a rowan, And it was confirmed that the cosmetic composition for skin whitening using the same can exhibit a skin whitening effect by inhibiting tyrosinase expression and inhibiting melanin biosynthesis, thereby completing the present invention.

KR 10-2005-0110149 A

Accordingly, it is a main object of the present invention to provide a cosmetic composition for skin whitening comprising a hole-sealer extract or a hole-sealer fermented product having a skin whitening effect by inhibiting tyrosinase expression and inhibiting melanin biosynthesis.

According to one aspect of the present invention, there is provided a skin whitening cosmetic composition comprising as an active ingredient an extract of a hole-sealing material or a fermentation product of a hole-sealing material.

Hole chamber ( Sorbus Aucuparia Berry ) is a fruit of the broad-leaved broadleaf tree belonging to the rose family. It is distinguished from the stem and bark extracts of the legs , which were conventionally used as cosmetics. The inventors of the present invention have confirmed that melanin production inhibitory ability of the positive hole, which is a fruit, is higher than that of the trunk and bark of the legs. In addition, in the case of fermented yeast fermented product, inhibition of biosynthesis of tyrosinase and melanin, And the extract of the hole-sealing material, thus showing a superior skin-whitening effect. Thus, the present invention has been completed.

In the present invention, the above-mentioned hole-chamber extract can be extracted with any solvent conventionally used for extracting plants, herbal medicines and fruits, preferably water, lower alcohol having 1 to 4 carbon atoms, glycerin, ethyl acetate, butyl Propylene glycol, dichloromethane, or a nucleic acid.

As the extraction method using the above-mentioned extraction solvent, conventional methods of extracting plants such as ethanol extraction, reflux cooling extraction, ultrasonic extraction and supercritical extraction can be used. In the embodiment of the present invention, But is not limited to this.

In the present invention, the fermentation product of the positive-working chamber is characterized in that the fermentation of the positive-working chamber extract is carried out with lactic acid bacteria.

In the present specification, the term "fermentation" means a process in which a microorganism decomposes an organic substance using an enzyme contained therein. The fermentation may be carried out by selecting or applying the conventional methods known in the art of the present invention. For example, the fermentation may be carried out at a temperature of 30 ° C to 45 ° C, specifically 35 ° C to 38 ° C for 30 hours to 140 hours, 130 hours. Through such fermentation, the skin whitening effect of the fermentation product of the hole furnace can be further enhanced.

In the present invention, the fermentation product of the positive-working chamber refers to a fermentation product obtained by fermenting the positive-working-chamber extract with a microorganism. The microorganism may be a lactic acid bacteria, preferably Lactobacillus sp., Leuconostoc sp. Or Bifidobacteria sp., But is not limited thereto. It is not.

In the present invention, the above-mentioned hole-chamber extract or fermented product of the hole-sealing material contains 0.01 to 90% by weight based on the total weight of the cosmetic composition. If it is outside the above range, formulation is not easy and it is not preferable.

In the present invention, the composition is characterized by exhibiting a skin whitening effect by inhibiting tyrosinase expression and inhibiting melanin biosynthesis.

According to the experimental example of the present invention, the positive-working chamber extract has a higher inhibitory rate of melanin production than the raspberry stem and bark extract, decreases the amount of melanin in a melanin-producing cell in a concentration-dependent manner, and inhibits tyrosinase expression in a concentration- Can be confirmed. These results indicate that the fermented product of the present invention can inhibit the synthesis of tyrosinase and melanin, thereby exhibiting a skin whitening effect (see Experimental Example 3-5).

In addition, according to the experimental example of the present invention, the fermented product of the positive-working chamber decreased the amount of melanin in the melanin-producing cells in a concentration-dependent manner more than that of the fermented untreated positive-working chamber extract and inhibited tyrosinase expression in a concentration- . These results indicate that the fermented product of the present invention can inhibit the synthesis of tyrosinase and melanin more effectively than the fermented non-fermented airtight chamber extract (Example 3-5 Reference).

Furthermore, the fermented product of the present invention inhibits skin melanin synthesis by inhibiting the protein expression of tyrosinase and DCT (dopachrome tautomerase). Specifically, the fermented product of the positive-working chamber and the positive-working chamber is involved in the production of melanin Tyrosinase, and DCT, respectively, and it was confirmed that the fermentation of the hole-sealed cells decreased the expression of tyrosinase and DCT more effectively than that of the hole-chamber extract. These results indicate that the fermented yeast of the present invention can inhibit tyrosinase through inhibition of the genes, resulting in skin whitening (see Experimental Example 6).

In the present invention, the composition may be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, , A soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser. For example, softening longevity, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body essence , And may also be a cosmetic composition having transdermal delivery formulations such as lotions, ointments, gels, creams, patches or sprays. The cosmetic composition of the present invention may contain other adjuvants in addition to the carrier, for example, preservatives, antioxidants, stabilizers, solubilizers, vitamins, pigments and perfumes.

As described above, the fermented yeast cells obtained by fermenting the extracts of the positive-pressure chamber or the positive-working chamber with lactic acid bacteria inhibit the expression of tyrosinase or DCT and inhibit the production of melanin in the melanin-producing cells.

Fig. 1 shows the results of human melanocyte-induced inhibition of melanin secretion by the extracts of Rumexii and Saccharomyces cerevisiae.
FIG. 2 shows a result of observing that the fermented product of the positive-working chamber exhibits a higher inhibitory effect on the melanin formation than the positive-working chamber extract.
FIG. 3 shows the result of observing that the fermented product of the positive-working chamber has a higher rate of suppression of tyrosinase expression than the positive-working chamber extract.
FIG. 4 shows the results of observing that the fermented product of the positive-working chamber has a higher inhibitory rate of DCT expression than the positive-working chamber extract.

Hereinafter, the present invention will be described in more detail with reference to Examples. These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

Comparative Example  One: Hole chamber  Preparation of extract

It was purchased from a specialized company called Oriental Herb which is specialized in delivering medicines to Oriental Medical Clinic and used as a sample of the present invention.

 In order to prepare the hole chamber extract, 100 g of the positive hole was washed with flowing water, dried naturally, partially crushed by a crusher, and subjected to compression extraction using a juicer. The extracts were sterilized using a 0.2 μm syringe filter for sterilization.

COMPARATIVE EXAMPLE 2: Preparation of Rectal Stem and Bark Extract

Rice stalks and bark were purchased from a specialized company called Oriental Herb, which professionally delivers medicines to the oriental clinic, and used as a sample of the present invention.

 In order to prepare Rake Extract, 100 g of Rake stems and bark were washed with running water, air dried, partially crushed using a crusher, and compressed by a juicer. Extracted Rectal Extracts were sterilized using a 0.2 um syringe filter for sterilization.

Example 1: Preparation of a fermentation product of a positive hole (F-SAF)

It was purchased from a specialized company called Oriental Herb which is specialized in delivering medicines to Oriental Medical Clinic and used as a sample of the present invention.

The fermented product of the positive-working chamber was inoculated with 10 ⁷ -10 ⁹ cells / mL of Lactobacillus sp. CXHB-4 in a liquid medium containing Comparative Example 1 (the hole-sealing material extract) and cultured for 5 days (120 hr) , And the cells were cultured while maintaining the conditions of pH 6.5 to 7.5, temperature 35 to 38 ° C, aeration amount 1 VVM. The fermentation broth was centrifuged at 10,000 xg for 30 minutes to obtain a final fermentation product in which the polymer precipitate and microbial cells were removed.

Experimental Example 1: Measurement of melanin production inhibitory effect of human melanocytes by the hole-chamber extract

The inhibitory effect of melanocyte on human melanocytes was examined by using the positive-pressure-room extract (Comparative Example 1), Rake Stem and Bark Extract (Comparative Example 2), and H89 (10 uM), which is known to inhibit melanin production, To inhibit melanin production.

The human epidermal melanocyte cells (obtained from Cascade Biologics) were cultured in Medium 254 medium containing Human Melanocyte Growth supplement purchased from the same company. Human epidermal melanocytes were inoculated in a 6-well plate at 1 × 10 ⁵ cells per well and cultured at 37 ° C in 5% CO ₂ until cells were attached to the bottom of the wells at about 80%. After the culture, the medium was removed, the sample was diluted to a proper concentration, treated with the medium, and incubated at 37 ° C in 5% CO _{2 for} 5 days while changing the medium every other day. The concentration range of the extract was determined to be 10 μg / ml, 50 μg / ml without cytotoxicity. After the treatment, the medium was removed, washed with PBS (phosphatized buffer saline), and treated with trypsin to recover the cells. The recovered cells were counted using a hematocytometer, centrifuged at 5,000 ~ 10,000 rpm for 10 minutes, and the supernatant was removed to obtain cells. The cells were dried at 60 ° C., and 100 μl of a 1 M sodium hydroxide solution containing 10% DMSO was added thereto to obtain intracellular melanin in a 60 ° C. thermostatic chamber. Absorbance was measured at 490 nm with a microplate reader to determine the amount of melanin per cell constant. The results are shown in Fig.

As shown in FIG. 1, it was confirmed that the amount of melanin in the human melanocytes was reduced in a concentration-dependent manner by the treatment with the extract of the hole chamber of Comparative Example 1. On the other hand, Comparative Example 2, which is a trunk extract and bark extract, showed an inhibitory effect on the production of melanin, but the inhibition rate was lower than that of Comparative Example 1. Therefore, it was confirmed that the extract of the hole-sealing material has a whitening effect on human cells.

EXPERIMENTAL EXAMPLE 2 Promotion of Promoter Activity of Liver X Receptor Response Element (LXRE)

Experiments were conducted to investigate the mechanism of suppression of the hole - chamber extract which reduces the amount of melanin. It has been reported that activation of the liver X receptor (LXR) inhibits melanogenesis, and it is possible to determine whether the LXRE promoter activated by the LXR protein can activate the LXRE promoter luciferase reporter Respectively. Transfection was induced by inserting LXRE reporter DNA into murine melanoma (B-16 F10) cells using superfect, and the extract was treated 24 hours after transformation. After 16 hours, the cells were collected and luminescence was measured at 450 nm using a Luminometer (Berthold, Germany). (Planta medica 2005; 71: 338-343). As a positive control, TO901317 known as activator of LXR was used. The results are shown in Table 1 below.

sample Treatment concentration Relative luciferase activity Untreated group 1007.5 + 14.5 Positive control - T0901317 20uM 3932.5 ± 90.5 Comparative Example 1 - 10 mu g / ml 1181.2 + - 24.9 Comparative Example 1 - 50 mu g / ml 1331.5 ± 53.1 Comparative Example 2 - Rectal stem and bark extract 10 mu g / ml 1061.9 ± 8.0 Comparative Example 2 - Rectal stem and bark extract 50 mu g / ml 1076.2 + - 43.5

As shown in Table 1, it was observed that the efficacy was different depending on the site of the bean curd. In the case of the stem and bark extract of the bean curd, Comparative Example 2 had no activation effect of the LXR protein, In the case of the extract, Comparative Example 1, LXR protein was activated in a concentration-dependent manner. Therefore, through this experiment, it was proved that the positive hole has an excellent whitening effect.

Experimental Example 3: Measurement of inhibitory effect on melanin formation of pigment cells by fermentation of a hole-sealed chamber

The inhibitory effect of melanin production by Comparative Example 1 and Example 1 was measured using mouse melanoma cells (B16 melanoma cells) and compared with the effect of inhibiting melanin formation by arbutin (100 ppm), which is known to inhibit melanin production Respectively.

In this experiment, murine melanoma (B16 F10) cells were suspended in DMEM containing 10% FBS, and inoculated on a 6-well plate at 1 × 10 ⁵ cells per well, and cultured until adhered to the well bottom. For the induction of melanin production, 0.1 μM α-MSH was added, and then samples were added at different concentrations and cultured for 3 days. The concentration range of the material treatment was set at 0.1% and 1%. As a positive control for melanin production, arbutin was treated at 100 ppm. The cultured cells were washed with phosphate buffered saline (PBS) and recovered with trypsin. The recovered cells were counted with a hematocytometer and collected at 1 × 10 ⁶ cells / mL for each treatment group, and centrifuged at 1000 rpm for 10 minutes to obtain cells. The recovered cells were dried at 60 ° C for 1 hour, and 100 μl of a 1 M NaOH solution containing 10% DMSO was added to obtain melanin in the cells. The melanin inhibition rate was evaluated by measuring the absorbance at 490 nm using a microplate reader. The results are shown in Fig.

As shown in FIG. 2, it was observed that the amount of melanin was decreased in a concentration-dependent manner by the treatment with the substance. In particular, in Example 1, whitening efficacy was superior to that of arbutin. Therefore, it was proved through this experiment that the fermentation product of the hole-sealing material has the most excellent whitening effect.

Experimental Example 4: Measurement of intracellular tyrosinase gene expression by a fermentation product

The inhibitory effect of tyrosinase, an enzyme important for melanin production, in Comparative Example 1 and Example 1 was confirmed by using mouse pigment cells (B16 melanoma cells).

This experiment murine melanoma (B-16 F1) were inoculated the cells of 10% of FBS (fetal bovine serum) is the 6-well plate in the DMEM medium 5 × 10 ⁵ pieces per well 5% CO _2, 37 ℃ Were cultured under the condition that the cells were attached to the bottom of the well by about 80% or more. For the induction of melanin production, 0.1 μM α-MSH was treated, followed by addition of 1% sample, followed by incubation for 24 hours. As a positive control, 100 ppm of arbutin was treated. The cultured cells were washed with phosphate buffered saline and recovered by adding 1 ml of Trizol. The mixture was subjected to voltexing by adding 0.2 ml of chloroform (1/5 ratio of Trizol), and the supernatant was obtained by centrifugation at 4 ° C and 14,000 rpm for 15 minutes. 0.5 ml of isopropanol was added to the supernatant, followed by reaction for 10 minutes, followed by centrifugation at 14000 rpm for 15 minutes to obtain a precipitate. The supernatant is removed and washed twice with 200 μl of 100% EtOH. The precipitate was then completely dried at room temperature and dissolved in 20 to 30 μl of DEPC water. RNA was dissolved in DEPC and cDNA was synthesized using the same amount of RNA per group. The synthesized cDNA was used as a target and cyanogen dyes SYBR GREEN master mix was used to perform real - time PCR (PCR). Finally, gene expression of tyrosinase was evaluated. The results are shown in Fig.

As shown in Fig. 3, it was observed that the expression of tyrosinase was decreased by the treatment of the material. In the case of Example 1, the effect of suppressing the expression was better than that of Comparative Example 1. Therefore, through this experiment, it was proved that the fermentation product of the hole-spinning machine had better whitening effect than the extract of the hole-spinning machine.

EXPERIMENTAL EXAMPLE 5: Expression of DCT gene in cells by fermentation of a positive-working chamber

The effect of inhibiting the expression of DCT (dopachrome tautomerase), which is a factor controlling the synthesis of melanin in the melanin of Comparative Example 1 and Example 1, was confirmed by using mouse pigment cells (B16 melanoma cells).

This experiment murine melanoma (B-16 F1) were inoculated the cells of 10% of FBS (fetal bovine serum) is the 6-well plate in the DMEM medium 5 × 10 ⁵ pieces per well 5% CO _2, 37 ℃ Were cultured under the condition that the cells were attached to the bottom of the well by about 80% or more. For the induction of melanin production, 0.1 μM α-MSH was treated, followed by addition of 1% sample, followed by incubation for 24 hours. As a positive control, 100 ppm of arbutin was treated. The cultured cells were washed with phosphate buffered saline and recovered by adding 1 ml of Trizol. 0.2 ml Chloroform (1/5 ratio of Trizol) was added, and the supernatant was obtained by voltexing and centrifugation at 14000 rpm for 15 minutes at 4 ° C. 0.5 ml of isopropanol was added to the supernatant, followed by reaction for 10 minutes, followed by centrifugation at 14000 rpm for 15 minutes to obtain a precipitate. The supernatant is removed and washed twice with 200 μl of 100% EtOH. The precipitate was then completely dried at room temperature and dissolved in 20 to 30 μl of DEPC water. RNA was dissolved in DEPC and cDNA was synthesized using the same amount of RNA per group. Real-time PCR (PCR) was performed using SYBR GREEN master mix, which is a cyanine dye, with the synthesized cDNA as a target, and the degree of gene expression of MITF was finally evaluated. The results are shown in Fig.

As shown in FIG. 4, it was observed that the expression of DCT transcription factor was reduced by the treatment of the material. Particularly, in Example 1, an excellent inhibitory effect was shown as compared with Comparative Example 1. Therefore, through this experiment, it was proved that the fermentation product of the hole-spinning machine had better whitening effect than the extract of the hole-spinning machine.

Claims (7)

A cosmetic composition for whitening skin, which contains an extract of a hole-sealing material or a fermentation product of a hole-sealing material as an active ingredient.
2. The cosmetic composition for skin whitening according to claim 1, wherein the hole-extracting material is extracted with water, a lower alcohol having 1 to 4 carbon atoms, glycerin, ethyl acetate, butylene glycol, propylene glycol, dichloromethane or a nucleic acid.
[Claim 7] The cosmetic composition for skin whitening according to claim 1, wherein the fermentation product is fermented with a lactic acid bacterium.
The cosmetic composition for skin whitening according to claim 3, wherein the lactic acid bacterium is Lactobacillus sp., Leuconostoc sp. Or Bifidobacteria sp.
[Claim 2] The cosmetic composition for skin whitening according to claim 1, wherein the extract of the hole-sealing material or the fermentation product of the hole-sealing material comprises 0.01 to 90% by weight based on the total weight of the cosmetic composition.
[Claim 2] The cosmetic composition for skin whitening according to claim 1, wherein the extract of the positive-pressure chamber or the fermentation product of the positive-working chamber exhibits a skin whitening effect by inhibiting tyrosinase expression and inhibiting melanin biosynthesis.
The composition according to claim 1, wherein the composition is at least one selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritional cream, a moisturizing cream, a hand cream, Wherein the at least one cosmetic composition is at least one selected from the group consisting of a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

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