KR101072641B1 - Composition for skin whitening containing extract from culture fluid of Actinomycetes - Google Patents

Abstract

In the present invention is Streptomyces (Actinomycetes) strain or Streptomyces (Actinomycetes), an enzyme that relates to a composition for skin whitening comprising the culture extract as an active ingredient, the compositions according to the present invention is involved in melanin biosynthesis tie tyrosinase (tyrosinase) By inhibiting the enzymatic activity of beta-glucosidase, an important enzyme involved in the glycosylation process, the glycosylation of tyrosinase is inhibited to produce melanin. Since the whitening effect is shown by inhibiting, the composition for skin whitening according to the present invention can effectively inhibit melanin production without side effects due to the efficacy of actinomycetes strains or actinomycetes culture extracts.

Actinomycetes Culture Extract, Skin, Whitening, Composition, Melanin, Tyrosinase, Glycosylation, Beta-Glucosidase.

Description

Composition for skin whitening containing extract from culture fluid of Actinomycetes}             

1 is a graph showing the effect of actinomycete culture extract on melanin production of melanocytes, showing how much melanin production is inhibited when treated with 0.01, 0.05, 0.1% actinomycete culture extract.

Figure 2 is a photograph of the melanocytes collected in the test tube after treatment with actinomycete culture extract, showing that the apparent color of the melanosite becomes white when treated with 0.01, 0.05, 0.1% actinomycetes culture extract.

Figure 3 is a protein electrophoresis picture showing the effect of actinomycetes culture extract on the glycosylation of tyrosinase, the upper of the two black lines is a large tyrosinase with a sugar of about 80 kD Shown below is a small tyrosinase of about 60 kD without sugar. a (control), b (0.01% of actinomycetes extract), c (0.05% of actinomycetes extract), d (0.1% of actinomycetes culture extract) were subjected to electrophoresis after hydrolysis of tyrosinase sugars with endoglucosidase. E (control), f (0.01% of actinomycetes extract), g (0.05% of actinomycetes culture extract), h (0.1% of actinomycetes culture extract) are endoglucosidase without hydrolyzing the sugars of tyrosinase. It was electrophoresis.

Figure 4 shows the whitening effect of the actinomycetes culture extract (0.5%) with the naked eye and numerically shown.

5 is a graph showing the whitening effect of the actinomycetes culture solution extract (0.5%) as the L value of the color difference meter.

Figure 6 shows the artificial pigment plate before the sample treatment, quarter quarter 0.5% actinomycetes extract, 2/4 quarter hydroquinone 2%, 3/4 quarter untreated group, 4/4 quarter vehicle Artificial pigment for.

Figure 7 shows the artificial pigment plate after three weeks of sample treatment, quarter quarter 0.5% actinomycetes extract, 2/4 quarter hydroquinone 2%, 3/4 quarter untreated group, 4/4 quarter Is the artificial pigment plate for the vehicle.

The present invention relates to a skin whitening composition, and more particularly to a composition for skin whitening containing actinomycetes strains or actinomycetes culture extract as an active ingredient.

Melanin, which determines human skin color, is made from skin cells called melanocytes and migrates to epidermal cells called keratinocytes. Here, melanin plays an important role such as forming a hat-like structure around the nucleus to protect genes from ultraviolet rays and to remove free radicals to protect intracellular proteins. There are no enzymes that break down melanin in the body, but when keratinocytes break away from the epidermis, they are removed by falling off the skin with melanin. However, if more melanin is produced than necessary, it causes hyperpigmentation such as blemishes, freckles, and spots, resulting in cosmetically bad results. In addition, as the leisure population increases, more people enjoy working outside, and the demand to prevent melanin pigmentation due to ultraviolet rays is increasing. Therefore, it was necessary to develop a whitening agent that prevents excessive melanin production and many efforts have been made.

Until now, the development of whitening agents has been mainly focused on finding a substance that reduces melanin by inhibiting the activity of tyrosinase, a basic and most important enzyme in melanin biosynthesis. The whitening agents developed in this way include kojic acid, arbutin, glutathione, vitamin A, and vitamin C. However, the whitening agent does not have a satisfactory whitening effect.

Therefore, it is an urgent time to develop a whitening agent that is more effective and safer than ever.

Tyrosinase, an enzyme that synthesizes melanin, is a glycoprotein and is produced through glycosylation, a glycoprotein synthesis in vivo. If there is a problem in the glycosylation process, and the abnormal part of the tyrosinase sugars, the tyrosinase cannot move to the melanosome, a place of intracellular melanin biosynthesis, and thus the melanin production cannot be achieved.

Many enzymes are involved in the glycosylation process, the most important of which is glucosidase. If the enzyme can inhibit the enzymatic activity, it can inhibit the melanin biosynthesis and thus bring about a whitening effect.

Actinomycetes (放線菌, Actinomycetes) is also known by the generic bangsasanggyun of bacteria belonging to, Streptomyces neck by microorganisms present in the mycelium and sporophyte or the like, soil, plant dongmulche, river, sea water. It is gram-positive, and proliferation is not only division but also conidia or spore formation. Mycobacterium and (Mycobacteriaceae), Tino My bad seseugwa (Actinomycetaceae), and 5 to 8, such as Streptomyces cheat seseugwa (Streptomycetaceae), most of formatting in the ground but also parasitic plants and animals or people.

Previous studies have shown that several microbial species, including actinomycetes, make substances that inhibit the enzyme activity of glucosidase (Fukuda K, Mori H, Okuyama M, Kimura A, Ozaki H, Yoneyama M, Chiba S.). , Biosci.Biotechnol.Biochem . , 66, pp2060-7, 2002; Chang HB, Kim SH, Kwon YI, Choung DH, Choi WK, Kang TW, Lee S, Kim JG, Chun HS, Ahn SK, Hong CI, Han KH., J. Antibiot. (Tokyo) , 55, pp 467-71, 2002).

However, there is no example of developing whitening agents or verifying whitening efficacy using actinomycetes or microorganisms having such characteristics, and patents thereof are not known.

In order to solve the above problems, the present inventors searched for the ability to inhibit the activity of glucosidase enzymes of microorganisms including actinomycetes in order to develop a new whitening material that is more effective and safer than ever before, the actinomycetes culture extract is very The present invention has been completed by finding that it has an excellent inhibitory effect on glucosidase enzyme activity.

Therefore, it is an object of the present invention to provide a novel whitening agent that contains an actinomycetes strain or actinomycetes culture extract and shows an excellent whitening effect but no side effects.

In order to achieve the above object, the present invention provides a composition for skin whitening comprising the Streptomyces (Actinomycetes) strain or Streptomyces (Actinomycetes) culture extract as an active ingredient.

The composition for skin whitening is characterized in that the actinomycetes strain or actinomycetes culture extract that inhibits the enzymatic activity of beta-glucosidase as an active ingredient.

The actinomycetes is Streptomyces genus strain, preferably characterized in that Streptomyces subrutilus ( Streptomyces subrutilus ).

In addition, the actinomycetes strain may be directly isolated from the soil, or purchased and used.

The actinomycetes culture is characterized in that it includes both the culture medium and the culture medium cultured on a liquid medium and the like.

The actinomycetes strain or actinomycetes culture extract may be contained in an amount of 0.001 to 20% by weight based on the total weight of the composition.

In addition, the composition for skin whitening containing the actinomycetes strain or actinomycetes culture extract as an active ingredient is characterized in that the cosmetic composition for skin whitening or a pharmaceutical composition for skin whitening.

Actinomycetes strains or Actinomycetes culture extracts contained as active ingredients in the composition of the present invention are important enzymes involved in the process of glycosylation to synthesize tyrosinase, an enzyme involved in melanin biosynthesis. By inhibiting the enzymatic activity of beta-glucosidase, it inhibits the glycosylation of tyrosinase, thereby preventing melanin production and thus whitening effect.

The present invention provides a cosmetic composition for skin whitening, wherein the actinomycetes strain or actinomycetes culture extract has 0.001 to 20% by weight of the total cosmetic composition. This is because the effect cannot be expected at less than 0.001% by weight, and if it exceeds 20.0% by weight, there is a difficulty in safety or formulation formulation.                     

In addition, the cosmetic composition for whitening according to the present invention, in addition to the above-described strains or extracts, other whitening ingredients, etc., which may give a synergistic effect to the main effect, preferably within the range of not impairing the main effect of the present invention. It may also contain. Examples of such a component include kojic acid, arbutin, iscobic acid derivatives, and the like.

Actinomycetes culture extract according to the present invention, it is suitable to apply the eluate, eluent, eluent or dry powder of the eluent as the whitening cosmetic formulation, but can also be used as a general internal or external formulation.

The composition of the present invention can be used in a variety of cosmetics and the like for the skin whitening effect. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, skins, and the like, cleansing agents, face washes, soaps, and essences.

The cosmetic for whitening to which the composition containing the actinomycetes strain or actinomycetes culture extract of the present invention is added may take the form of a solution, an emulsion, a viscous mixture, or the like.

That is, the whitening cosmetic of the present invention is not particularly limited in the formulation, for example, latex, cream, lotion, essence, pack, gel, powder, foundation, lotion, ointment, ger patch, beauty liquid, cleansing foam, cleansing Formulations such as creams, cleansing water, soaps, sprays and the like.

In the cosmetic composition of each formulation, other components in addition to the above strains or extracts can be appropriately selected by those skilled in the art without difficulty according to the formulation or use purpose of the other cosmetics.

Such formulations may contain skin absorption promoting substances to increase the whitening effect.

In addition, the cosmetic of the present invention may include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients.

Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, purified water and the like.

Moreover, the compounding component which may be added other than this is not limited to this, Moreover, Although all said components can be mix | blended within the range which does not impair the objective and effect of this invention, Preferably it is 0.01-5 weight% with respect to a total weight, More preferably, 0.01 to 3 weight% is mix | blended.

The present invention also provides a pharmaceutical composition for skin whitening, wherein the actinomycetes strain or actinomycetes culture extract has 0.001 to 20% by weight of the total pharmaceutical composition.

Pharmaceutical compositions comprising actinomycetes strains or actinomycetes culture extract of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

The pharmaceutical composition comprising the strain or extract according to the present invention may be in the form of powder, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, ointments, creams, etc. It may be used in any form suitable for pharmaceutical preparations, including suppositories and sterile injectable solutions.

 The composition according to the invention can be administered to mammals such as rats, mice, livestock, humans by various routes, such as parenteral, oral, and all modes of administration can be expected, for example oral, rectal or It can be administered by intravenous, muscle, subcutaneous, intrauterine dural or intracerebroventricular injection. At this time, as a parenteral route, transdermal administration is preferable, and topical application is the most preferable.

The dosage of the preparation varies depending on the subject's age, sex, weight, symptoms, and method of administration, but in the case of external preparations, 1.0 to 3.0 ml per day may be applied 1 to 5 times a day to continue for more than one month. . In addition, the oral dosage form may vary depending on the age, sex, and weight of the patient, but may be administered once to several times in an amount of 0.1 to 100 mg / kg. The dosage may also be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

On the other hand, actinomycetes strains or actinomycetes culture extract of the present invention has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of prevention.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the present invention is not limited thereto.                     

<Example: Preparation of an external preparation composition for skin whitening containing actinomycetes culture extract as an active ingredient>

Example 1 Production of Actinomycetes Culture Extracts Having Efficacy of Inhibiting Beta-glucosidase Activity

1-1. Actinomycetes Culture

Streptomyces subrutilus KCCM 40337, ATCC 27467 (hereinafter referred to as Strain 1), Streptomyces lavendulae KCCM 32799, ATCC 8664; 2) and Streptomyces lavendulae SEN-158 (ATCC 31434; hereinafter strain 3) were incubated in OM agar medium for 10 days, and the spores were used for spawning medium (200 ml). / 1 ℓ Earlenmeyer flask; Yeast extract 4 g, Malt extract 10 g, Glucose 4 g / ℓ) and incubated for 3 days at 30 ℃, 120 rpm and then culture medium (Yeast extract 6 g, Malt extract 6 g, Soypeptone 10 g, 40 g / l of Glucose) were used to incubate in a 30 L fermentation bath. At this time, the culture conditions in the fermentation tank was 20 L medium, temperature 30 ℃, stirring speed 350 rpm, aeration amount 1.0 vvm, initial pH 7.0. Incubation for 4 days under the conditions described above gave each actinomycete culture (318 U / ml) having beta-glucosidase inhibitory activity.

1-2. Production of Actinomycetes Culture Extracts

Each actinomycete culture (Culture broth) (1) obtained in Example 1-1 was extracted and purified in the following manner.                     

First, 20 l of each culture medium was filtered using diatomaceous earth as a filter aid to remove the cells, and then the pH was adjusted to 3.0 using hydrochloric acid. Then, 2% (w / v) activated carbon powder was added and stirred for 30 minutes. After filtration to remove activated carbon, the filtrate (activated carbon treated liquid (2)) was diion PK-208 column (φ6cm × 50cm, H + form), which is a cation exchange resin. Was adsorbed at 100 ml / min to adsorb the active portion and washed with about 10 L of ionized water until the eluate had a pH of 6 to 7, followed by 20 ml / min using 0.5 N NH ₄ OH. The eluted solution was distilled under reduced pressure at 40 ° C. or lower, and the pH was neutralized to 7.0. The active fraction (cationic exchange resin treatment solution 3) was collected at an activated carbon column at a rate of 2 ml / min. Adsorption and washing with 3 to 5 L of ionized water It was eluted with ethanol, methanol eluent (activated carbon column treatment solution 4) was obtained for each actinomycete culture extract of the present invention by drying and vacuum distillation. At this time, the overall purification yield is shown in Table 1 below.

Activity (units / ml) Liquid amount (ml) Total active
(× 10 ⁵ units) yield(%) Strain 1 Actinomycetes Culture (1) 318 20,000 63.6 100 Activated Carbon Treatment Liquid (2) 308 20,000 61.6 96.9 Cation exchange resin treatment liquid (3) 1,394 2,000 27.9 43.9 Activated Carbon Column Treatment Liquid (4) 2,692 362 9.7 15.3 Strain 2 Actinomycetes Culture (1) 450 20,000 90.0 100 Activated Carbon Treatment Liquid (2) 423 20,000 84.6 94 Cation exchange resin treatment liquid (3) 1,448 2,000 28.9 32.1 Activated Carbon Column Treatment Liquid (4) 2,784 401 11.2 12.4 Strain 3 Actinomycetes Culture (1) 412 20,000 82.4 100 Activated Carbon Treatment Liquid (2) 399 20,000 79.8 96.8 Cation exchange resin treatment liquid (3) 1,401 2,000 28.0 33.9 Activated Carbon Column Treatment Solution (4) 2,388 396 9.5 11.5

Example 2. Preparation of whitening composition containing actinomycetes culture extract

Using the Streptomyces subrutilus KCCM 40337, ATCC 27467 culture extract of the actinomycete culture extract obtained in Example 1-2, the composition for whitening was prepared in the composition of Table 2 below.

As a method for producing a composition for whitening, first, raw materials 1 to 6 in Table 2 were mixed, dissolved at 70 ° C., and dissolved in an aqueous phase. The mixture was added to the mixture, stirred with a homomixer (Tokushu Kika, Japan), and then emulsified. The raw material 15 was added thereto to increase, and then the bubbles were removed, followed by cooling to room temperature.

Comparative Example 1.

The following raw materials 1-6 of Table 2 were mixed, it melt | dissolved at 70 degreeC, it was made into the water phase, and raw materials 8-14 were melt | dissolved at 70 degreeC, and it was set as the oil phase. Then, the oil phase was added to the aqueous phase, stirred first with a homomixer (Tokushu Kika, Japan) and emulsified, and the raw material 15 was added to increase. After removing bubbles, the mixture was cooled to room temperature.

ingredient Example 2 Comparative Example 1  1. Purified water Up to 100 Up to 100  2. Glycerin 8.0 8.0  3. Butylene Glycol 4.0 4.0  4. Hyaluronic Acid Extract 5.0 5.0  5. Beta Glucan 7.0 7.0  6. Camo 0.1 0.1  7. Actinomycetes Culture Extract 0.5 -  8. Caprylic Capric Triglycerides 8.0 8.0  9. Squalane 5.0 5.0  10. Cetearyl Glucoside 1.5 1.5  11.Sorbitan Stearate 0.4 0.4  12. Cetearyl Alcohol 1.0 1.0  13. Preservative Quantity Quantity  14. Incense Quantity Quantity  15. Triethanolamine 0.1 0.1

The present inventors performed the following experiment to prove the effect of the composition for skin whitening according to the present invention.

Experimental Example 1. Effect of actinomycetes culture extract on melanin production of human melanoma cells

1-1. Effect of each actinomycete strain

In order to determine the effect of each actinomycete extract of Example 1-2 on the melanin production of human melanoma cells, the following experiment was performed.

First, HM3KO cells (Y. Funasaka, Department of dermatology, Kobe university school of medicine, 5-1 Kusunoki-cho 7-chrome, Chuo-ku, Kobe 650, Japan), human melanoma cells, were 10% fetal calf serum. Into MEM (Minimum Essential Medium) medium was incubated under 37 ℃, 5% CO ₂ conditions. The cells thus cultured were placed in a 75 cm ² flask so that the number of cells was 3 × 10 ⁵ per flask, and the cells were allowed to adhere to the base wall overnight, and then the Streptomyces subroutine obtained in Example 1-2 from the next day. Streptomyces subrutilus KCCM 40337, ATCC 27467 (hereinafter referred to as strain 1), Streptomyces lavendulae KCCM 32799, ATCC 8664 (hereinafter referred to as strain 2), Streptomyces lavendulae SEN- Each actinomycetes extract of 158 (ATCC 31434; hereinafter strain 3) and kojic acid (Sigma) were respectively ground to a new medium containing 0.05%. As a control, a medium containing a vehicle (ethanol: propylene glycol: water = 5: 3: 2) used to dissolve the actinomycetes extract was used. The new medium containing the sample was changed every 1 to 2 days and cultured until the cells were filled in the flask. When the cells were grown, the cells were collected and compared with the cell color of the control group and the actinomycetes extract treatment group, and the amount of melanin produced by the HM3KO cells was measured. The amount of melanin was determined by measuring the absorbance at 490 nm after completely dissolving the cells with 1N sodium hydroxide (NaOH), and when the amount of melanin in the control group was 100%, the amount of melanin in each actinomycetes extract treated group was calculated as% of the control group. It is shown in Table 3 below.

As a result, as can be seen in Table 3, the highest melanin production inhibition rate of human melanoma cells of Streptomyces subrutilus KCCM 40337, ATCC 27467. On the other hand, each actinomycetes culture extract and kojic acid showed no cytotoxicity at all experimental concentrations.

Strain 1 Strain 2 Strain 3 Kojic acid Control group % Melanin production relative to the control 32.9 45.5 50.1 85.7 100

1-2. Effect of Streptomyces Subrutirus Culture Extract

According to the results of Experimental Example 1-1, to determine the effect of the concentration of Streptomyces subrutilus KCCM 40337, ATCC 27467 culture extract in the actinomycetes culture extract, Streptomyces of Example 1-2 Seth subrutirus culture extract was treated in the concentration of 0.1, 0.05 and 0.01% as in Experimental Example 1-1 to determine the effect on melanin production of human melanoma cells.

As a result, as shown in FIG. 1, 0.05% of kojic acid decreased melanin production by 17.5%, whereas 0.01, 0.05, and 0.1% of streptomyces subrutirus culture extracts inhibited melanin production by 63.4, 64.4, and 62.1%, respectively. It showed an inhibitory effect. In addition, looking at Figure 2 it can be seen that the apparent color of the melanocytes treated with the Streptomyces subrutirus culture extract was white compared to the control.

Experimental Example 2 Effect of Actinomycetes Culture Extract on Glycosylation of Tyrosinase in Human Melanoma Cells

To investigate the effect of actinomycetes culture extract on the glycosylation of tyrosinase in human melanoma cells, the following experiment was performed.

In the same manner as in Experiment 1-1, HM3KO cells were grown in the medium treated with the Streptomyces subrutilus KCCM 40337, ATCC 27467 culture extract of Example 1-2, and then the cells were collected and cell lysed. The solution (2% CHAPS in 50 mM Hepes and 200 mM NaCl, pH 7.5, protease inhibitors) was added and sonicated, followed by centrifugation for 10 minutes at 4 ° C and 12000 rpm. Cells and melanin were separated and removed, and only the supernatant was taken. Then, endoglycosidase H (125 units) was added to this supernatant (protein amount: 20 g) for 1 hour at 37 ° C. After enzymatic reaction, the proteins in the supernatant were separated by size by electrophoresis.

Tyrosinase among the proteins isolated by electrophoresis as described above was confirmed by the immune response using the antibody. That is, tyrosinase, in which sugar residues are normally formed, appears as a glycoprotein having a size of about 80 kD due to enzymatic degradation of sugar residues by endoglycosidase H, and an enzyme of glucosidase involved in the glycosylation process. Tyrosinase, in which activity is inhibited and a normal sugar residue is not formed, is hydrolyzed by endoglycosidase H, resulting in a protein of about 60 kD.

Referring to Figure 3, the tyrosinase of the control group (a) is not degraded by the endoglycosidase and appears to be large in size (about 80 kD), whereas the streptomyces subrutirus culture extract 0.01 (b) , Tyrosinase of melanocytes treated with 0.05 (c) and 0.1 (d)% was found to be completely degraded by endoglycosidase and appeared to be small (about 60 kD) in size.

These results show that the actinomycetes culture extract can inhibit the enzymatic activity of tyrosinase by causing problems with glycoprotein formation of tyrosinase.

On the other hand, when the endoglucosidase was not treated, the control group (e), streptomyces subrutirus culture extract 0.01 (f), 0.05 (g), 0.1 (h)% of all the size of the sugar attached (about 80 kD We show tyrosinase of).

Experimental Example 3. Whitening effect of actinomycetes culture extract using brown guinea pig

The following experiment was performed to confirm the whitening effect of the actinomycetes culture extract of the present invention in animal experiments.

First, the Streptomyces subrutilus KCCM 40337, ATCC 27467 culture extract of the actinomycetes culture extract of Example 1-2 of the present invention was used as a vehicle (vehicle, ethanol: propylene glycol): water = 5: 3: 2) in a concentration of 0.5% to prepare a sample.

Next, the hair on the back of the brown guinea pig, weighing around 500 g, was removed and anesthetized with pentobarbital (30 mg / kg) and burned with 500 mJ UV light (UVA lamp (TL 20W / 09 UV Phillips). 3), 7 UVB lamps (TL 20W / 12 UV Phillips)). The burned area was a circle (artificial pigment plaque) having a diameter of about 1 cm, and the same site was irradiated with ultraviolet rays three times a week. After the last UV irradiation, from the next day, morning and evening, 0.5 μl of Streptomyces subrutirus culture extract and 2% of hydroquinone (Sigma) prepared for each artificial pigment plate were applied for 3 weeks. . The whitening effect during the application of the sample was based on the non-treated group (NT) based on the artificial pigment plate (control group) treated with vehicle only, the artificial pigment plate treated with actinomycetes culture extract, and the hydrochromon treated artificial pigment plate once a week. Visually, the relative color difference was quantified. The digitization is 0.5 for the whitening effect that only the expert can know, 1 for the whitening effect that anyone can know, 3 for the whitening effect as white as the original skin color, 2 for the whitening effect between 1 and 3, and 2 whitening than the original skin color Is given by 4 points. The whitening effect of the six animals in three weeks was averaged and labeled, and the results are shown in FIG. 4.

In addition, the untreated group, the artificial pigment plate treated with the vehicle only (control group), the artificial pigment plate treated with the streptomyces subrutirus culture extract of Example 1-2, and the artificial pigment plate treated with hydroquinone The color was determined using a color meter (chromometer), and the average value (L value) of the numerical value (L value) obtained by quantifying the contrast of the artificial color plate was shown in FIG. 5. On the other hand, the L value is gradually increased when the test substance is effective, and the comparison between the test substances is the value of ΔL, that is, the value of the final L minus the initial L value (ΔL) (final L value-the test substance). Value before application), the larger the ΔL value, the greater the whitening effect.

As a result, as a result of visual evaluation, 0.5% of the streptomyces subrutirus culture extract extract was 0.46, and 2% of the hydroquinone was 0.65 compared to the control group, which showed a whitening effect (see FIG. 4). In the experimental results using the color difference meter, the streptomyces subrutirus culture extract showed a large L value compared to the control group for 3 weeks and showed a similar whitening effect as compared to 2% of hydroquinone. (See Figure 5).

The artificial pigment plate before the sample treatment and the artificial pigment plate after the sample treatment for 3 weeks are shown in FIGS. 6 and 7, respectively.

Experimental Example 4. Whitening effect of the composition prepared according to Example 2 and Comparative Example 1

After attaching an opaque tape with six holes of 1.5 cm in diameter to the upper forearm of a subject, 12 to 1.5 times the UVB of each subject's minimum erythema (Minimal Erythema Dose). Irradiate the skin (3 UVA lamps (TL 20W / 09 UV Phillips) and 7 UVB lamps (TL 20W / 12 UV Phillips)) to induce blackening of the skin, apply the test substances, and after 2 months The contrast of the skin was measured. Each test substance was applied to the external preparation for skin according to Comparative Example 1 and Example 2 twice daily (morning and evening) at 10 μl per artificial pigment plate.

At this time, 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). If the test substance is effective, the L value is gradually increased, and the comparison between the test substances is the ΔL value, that is, the value of the final L minus the initial L value (ΔL) (final L value-application of the test substance). Previous value). The larger the ΔL value, the greater the whitening effect. The results are shown in Table 4 below.

As shown in Table 4, the ΔL value of the composition containing the actinomycetes culture extract is 2.47, it can be seen that the high value when compared with the ΔL value 1.61 of the comparative example.

Example 2 Comparative Example 1 ΔL 2.47 1.61

Hereinafter, a formulation example of the cosmetic composition will be described, but the present invention is only intended to be described in detail and not intended to limit it.                     

Soap manufacturers

Actinomycetes culture extract of Example 1-1 ......... 1-30 (%)

Maintenance ...

Sodium hydroxide ...............

Sodium Chloride .........................................

Spices Quantity

The whole amount was 100 with purified water, and soap was prepared by the above blending ratio.

Lotion manufacturers

Actinomycetes culture extract of Example 1-1 ............. 3.00 (%)

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Water Soluble Collagen (1% Aqueous Solution) ........................ 1.00

Sodium Citrate ... 0.10

Citric Acid ........................................... 0.05

Licorice Extract ......... 0.20

1,3-butylene glycol ... 3.00

The whole amount was 100 with purified water, and a lotion was prepared by the above blending ratio (%).

Cream manufacturers

Actinomycetes culture extract of Example 1-1 ....................... 1.00 (%)

Polyethylene Glycol Monostearate ............... 2.00

Self-emulsifying glycerin monostearate ......................... 5.00

Cetyl Alcohol ... 4.00

Squalene ......................................... 6.00

Glyceryl tri2-ethylhexanoate ......... 6.00

Sphingose Sugar Lipids ......................................... 1.00

1.3-Butylene Glycol ... 7.00

The total amount was 100 with purified water, and a cream was prepared at the blending ratio (%).

Pack manufacturer

Actinomycetes culture extract of Example 1-1 ....................... 5.00 (%)

Polyvinyl Alcohol ... 13.00

L-ascorbic acid-2-phosphate magnesium salt ......... 1.00

Lauroylhydroxyproline ............... 1.00

Water Soluble Collagen (1% Aqueous Solution) ............... 2.00

1,3-butylene glycol ... 3.00

Ethanol ......................................... 5.00

The total amount was 100 with purified water, and a pack was prepared at the blending ratio (%).                     

Essence

Actinomycetes culture extract of Example 1-1 ........ 2.00 (%)

Hydroxyethylene Cellulose (2% Aqueous Solution) ... 12.00

Xanthan gum (2% aqueous solution) ....................... 2.00

1,3-butylene glycol ......... 6.00

Dark glycerin ... 4.00

Sodium Hyaluronate (1% Aqueous Solution) ......................... 5.00

The whole amount was 100 with purified water, and a cosmetic liquid was prepared at the blending ratio (%).

Preparation of Capsules

Actinomycetes culture extract of Example 1-1 ........ 100 mg

Lactose ......................................... 50 mg

Starch ......................................... 50 mg

Talc ......................................... 2 mg

Magnesium Stearate ...

The capsules were prepared by mixing the above components and filling the gelatin capsules according to a conventional method for preparing capsules.

 As can be seen from the above, the cosmetic composition for skin whitening according to the present invention contains the actinomycetes strain or actinomycetes culture extract has the effect of effectively inhibiting melanin production without side effects.

Claims (6)

A composition for skin whitening containing Streptomyces subrutilus strain as an active ingredient. A composition for skin whitening containing a culture extract of Streptomyces subrutilus strain as an active ingredient. delete delete The composition for skin whitening according to claim 1 or 2, wherein the strain or culture extract is contained in an amount of 0.001 to 20% by weight based on the total weight. The composition for skin whitening according to claim 1 or 2, wherein the composition is a cosmetic composition.

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