WO2017090849A1 - Composition for whitening, anti-inflammation and skin aging inhibition comprising water-soluble pearl powder as active ingredient - Google Patents

Abstract

The present invention relates to a composition for whitening, anti-inflammation and skin aging inhibition comprising a water-soluble pearl powder as an active ingredient. The water-soluble pearl powder or an aqueous solution thereof of the present invention was ionized, and loss of organic matter thereof was minimized. Therefore, the present invention has an excellent effect of inhibiting melanin synthesis, inhibiting inflammatory reactions, and promoting collagen synthesis, and thus can be usefully used for functional cosmetics for whitening, anti-inflammation and skin aging inhibition.

Description

Whitening, anti-inflammatory and anti-aging composition comprising water-soluble pearl powder as an active ingredient

The present invention relates to a composition for inhibiting skin whitening, anti-inflammatory and skin aging comprising pearl powder which is ionized to have a very high water solubility as an active ingredient.

Everyone wants to live a long and healthy life, and spares no time and effort in beauty according to their desire to look pretty and young. As a result, many people are exercising, preventing and treating health through exercise or medicine, and many cosmetic products are produced and sold for the pursuit of beauty. As interest in well-being has increased due to the recent increase in income levels and lifestyle changes, consumers are becoming more and more inclined to prefer high-quality products using natural resources rather than chemical compounds that are currently widely used. In line with the demand, the development of functional cosmetics based on natural resources with intermediate pharmaceutical therapeutic efficacy of cosmetics and pharmaceuticals, such as functional cosmetics and functional foods, has been spotlighted in the cosmetics and health food market.

On the other hand, pearl powder (Pearl Powder) is obtained from pearls cultured in fresh water, and is a raw material obtained by pulverizing pearls into nano-sized fine powder by physical method at cryogenic temperature, is a powder that is hardly soluble in water. Most pearls are composed of calcium carbonate called aragonite, and other minerals such as sodium and magnesium. It also contains small amounts of organic substances such as proteins, vitamins, amino acids, peptides and mucopolysaccharides. Pearl powder has been known for a long time, and has been used for skin care in China, Egypt, and Maya, and is also used for health. Especially in China, freshwater pearls have been harvested from 3000 years ago, and the fine powders have been used for beauty and health. This is because pearl powder promotes skin metabolism, maintains skin tone, reduces pores, reduces skin redness, brightens the skin and makes the skin soft and shiny. In addition, in traditional Chinese medicine, freshwater pearl powder has been used for the treatment of various diseases such as inflammation, epilepsy, high blood pressure, insomnia, and macular degeneration, and also for the purpose of slowing aging and maintaining youth. Currently, pearls are mostly ground into fine powder and applied directly to the skin, used as cosmetic additives, or consumed as food. However, applying pearl powder that does not dissolve in water without pretreatment to the skin as expected does not make the most of the functionality of pearl powder in actual use because calcium or organic ingredients are not well dissolved in water. In addition, when pearl powder is applied to the skin, calcium and some components can be ionized and absorbed by the skin secretions, but most of them are in the form of undissolved powders, and in particular, the intrinsic organic substances of pearls are hardly dissolved. It is difficult to expect to fully utilize the various functionalities of pearl powder. When pearl powder is used on the skin, ionized water-soluble pearl powder or an aqueous solution thereof is required to be dissolved in water while preserving the activity of minerals and organic substances of pearls in order to maximize the skin functionality by the intrinsic ingredients of pearl powder. It is true.

At present, there are several methods for producing an ionized calcium solution from shells including pearls. Since 95% or more of shellfish powder such as pearl powder is present as calcium carbonate, the solubility in water is very low, and it is not possible to prepare an directly ionized aqueous solution with water alone. Therefore, a method of preparing a water-soluble shell powder is a method of dissolving calcium, which is a main component, by ionizing calcium and minerals from the shell, and are specialized processes for preparing shell-derived calcium agent. Conventional methods include the plastic working method used in the conventional limestone or shell processing, a method using a strong acid or a strong alkali, and a method using a weak acid such as a high concentration of acetic acid. However, all of the above methods focus only on dissolving calcium as easily as possible from the shell powder. In this way, it is difficult to prepare an ionized pearl powder solution in which water-soluble organic substances contained in pearl powder are well preserved. In addition, in the above methods, the obtained calcium ion solution is dried at a high temperature of 50 to 200 ° C. for a long time to prepare a powder, and this drying process causes degeneration of the water-soluble organic substance contained in the ionized calcium solution. As a result, the ionized water-soluble pearl powder prepared by the above method does not have any special functionalities other than calcium.

Therefore, the inventors of the present invention have developed a method of inducing ionization of calcium while minimizing the loss of water-soluble organic substances contained in pearl powder by inducing ionization with low concentration of acetic acid and lyophilizing the ionized solution. The present invention was completed by confirming the skin whitening effect, anti-inflammatory effect and skin aging inhibitory effect of the aqueous solution of water-soluble pearl powder.

An object of the present invention is to provide a composition for inhibiting whitening, anti-inflammatory and skin aging comprising an ionized water-soluble pearl powder or an aqueous solution thereof as an active ingredient.

Another object of the present invention to provide a method for producing a water-soluble pearl powder and a water-soluble pearl powder raw material prepared thereby.

Accordingly, the present invention provides a composition comprising an ionized water-soluble pearl powder or an aqueous solution thereof as an active ingredient. The composition containing the water-soluble pearl powder (WSPP) of the present invention as an active ingredient includes a cosmetic composition, a pharmaceutical composition, a topical skin composition or a food composition.

Hereinafter, the present invention will be described in detail.

The water-soluble pearl powder of the present invention, unlike conventional methods focused on recovering only calcium from pearl powder, induces ionization of pearl powder with a weak acid of 0.01 to 10%, preferably 0.2 to 5% at room temperature, thereby inducing calcium carbonate and organic matter. The material was dissolved and the solution lyophilized to minimize loss of functional organics. In the present invention, the weak acid refers to an acid having a pH of 4 to 6.5, and the type of the weak acid is not limited, but acetic acid is preferred.

The water-soluble pearl powder of the present invention or an aqueous solution thereof was ionized, and organic material loss was minimized. Functional organic materials minimized loss in the pearl powder inhibits the activity of tyrosinase, inhibits melanin production, has a whitening effect, and inhibits the production of NO, inhibits inflammation, and has excellent skin soothing (anti-inflammatory) effect. Since it promotes production, it has excellent skin aging inhibitory effect. Therefore, the water-soluble pearl powder according to the present invention can be usefully used in cosmetics, medicines, external skin preparations, quasi-drugs, health foods.

First, the composition of the present invention can be used as a cosmetic composition for whitening, skin soothing (anti-inflammatory) and skin aging inhibiting. In this case, the formulation of the cosmetic may be, for example, but not limited to, lotion, essence, lotion, gel, nourishing cream, patch, mask sheet, massage cream, lipstick, foundation or powder. In addition, when used as a cosmetic, in addition to the water-soluble pearl powder according to the present invention, additives commonly used in the formulation of cosmetics, for example, purified water, humectants, oils, surfactants, higher alcohols, thickeners, chelating agents, pigments, fatty acids, It may further include one or more additives selected from the group consisting of antioxidants, preservatives, waxes, pH adjusting agents and fragrances.

As the moisturizing agent, glycerin, propylene glycol, butylene glycol, hexylene glycol, diglycerin, sorbitol, betaine, glycerin-26 and the like may be used.

The oils include mineral oil, cyclomethicone, squalane, octyldodecyl myristate, olive oil, macadamia nut oil, glyceryl octanoate, castor oil, ethylhexyl isononanoate, dimethicone, cyclopentasiloxane, Sunflower seed oil and the like can be used.

The surfactant includes sorbitan sesquinolate, polysorbate 60, glyceryl stearate, lipophilic glyceryl stearate, sorbitan oleate, sorbitan stearate, die-cetyl phosphate, sorbitan stearate / shoe Cross-Cocoate, Glyceryl Stearate / Polyethylene Glycol-100 Stearate, Ceteareth-6 Oliveate, Lakydyl Alcohol / Behenyl Alcohol / Arachidyl Glucoside, Polypropylene Glycol-26-butes-26 / Polyethylene glycol-40 hydrogenated castor oil and the like can be used. Cetyl alcohol, stearyl alcohol, octyldodecanol, isostearyl alcohol and the like may be used as the higher alcohol. Xanthan gum, carbomer, magnesium aluminum silicate, cellulose gum, dextrin palmitate may be used as the thickener. As the chelating agent, disodium idieti, tetrasodium idieti and the like may be used. As the dye, Blue No. 1, Red No. 40, Yellow No. 4, Yellow No. 5, etc. may be used. As the fatty acid, stearyl acid, myristic acid, palmitic acid, isostearic acid, lauric acid, or the like may be used. Tocopheryl acetate, butylated hydroxytoluene, etc. may be used as the antioxidant. As the preservative, methyl paraben, butyl paraben, propyl paraben, phenoxyethanol, imidazolidinyl urea, chlorphenesin and the like may be used. Carnauba wax, candenilla wax, beeswax, braze and the like may be used as the wax. Triethanolamine, citric acid and the like may be used as the pH adjusting agent. As the fragrance, natural fragrance, combination fragrance, or the like may be used.

Cosmetic composition comprising a water-soluble pearl powder or an aqueous solution thereof according to the present invention may be included in an amount of 0.001 to 90% by weight, preferably 0.001 to 30% by weight relative to the total weight of the cosmetic composition. However, the composition as described above is not necessarily limited thereto, and may vary depending on the state and progress of the application target.

In addition, the composition of the present invention can be used as a pharmaceutical composition for inhibiting skin aging and anti-inflammatory. The pharmaceutical composition may include, but is not limited to, oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations such as powders, gels, ointments, creams, or sterilizations. It can be formulated and used in the form of injectable solutions. The composition of the present invention may further comprise suitable antibiotics, carriers, excipients or diluents commonly used in the manufacture of a medicament.

Specifically, the carrier, excipient or diluent may be lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose. , Polyvinyl pyridone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, lofil hydroxybenzoate, talc, magnesium stearate and mineral oil can be used. In the case of formulation, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants may be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin, or the like. You can mix. In addition to simple excipients, lubricants such as magnesium styrate and talc may also be used. Oral liquid preparations include suspensions, solvents, emulsions, syrups, and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, and the like. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.

The dosage of the water-soluble pearl powder according to the present invention may vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration, and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, it is preferable to administer the water-soluble pearl powder of the present invention at 0.0001 to 100 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect. The compositions of the present invention can be administered by various routes. All modes of administration can be expected, for example by oral, intravenous, intramuscular or subcutaneous injection.

In addition, the composition of the present invention can be used as a skin external preparation for inhibiting skin aging. The external preparation for skin refers to a preparation applied on the skin and may be a form of a cosmetic or a medicament. In this case, the formulation of the external preparation for skin may be a cream, gel, ointment, liquid, powder or spray, but is not limited thereto. In the case of the external preparation for skin according to the present invention, the water-soluble pearl powder according to the present invention may be included in an amount of 0.001 to 90% by weight, preferably 0.001 to 30% by weight, for a preferable effect.

In addition, the composition of the present invention can be used as a functional food for inhibiting skin aging or anti-inflammatory.

The health functional food formulation may be, for example, various foods, beverages, gums, teas, vitamin complexes, health functional foods, and the like.

The water-soluble pearl powder according to the present invention may be added in an amount of 0.01 to 90% by weight, preferably 0.001 to 30% by weight, based on the total weight of the food when used in the manufacture of the health functional food, and when used in the manufacture of the beverage The amount of the water-soluble pearl powder of the present invention may be added in a ratio of 0.02 to 10.00 g, or 0.3 to 1.0 g based on 100 ml of the total beverage.

When using the composition according to the present invention as a health functional food, it may contain other ingredients in addition to the compound as an essential ingredient, it may contain a variety of flavors or natural carbohydrates that can be added as an additional ingredient. Examples of the natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The natural carbohydrate is not limited thereto, but may be included in a ratio of about 1 to 20 g, preferably about 5 to 12 g, per 100 ml of the composition of the present invention. Compositions of the present invention include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and neutralizers (such as cheese, chocolate), pectic acid and salts thereof, alginic acid and salts thereof, Organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the composition of the present invention may contain a pulp for the production of natural fruit juices and vegetable drinks. These components can be used independently or in combination, and the proportion of such additives is not particularly limited.

The present invention provides a method for producing a water-soluble pearl powder and a water-soluble pearl powder produced thereby.

First, the method for producing a water-soluble pearl powder of the present invention, 1) ionizing the pearl powder with a weak acid of low concentration; And 2) lyophilizing the ionized pearl powder.

The pearl powder of step 1) is preferably nano-pearl powder pulverized into particles of 90 to 110nm size because the powder is large, the ionization reaction is too weak to low concentration acetic acid, it is difficult to ionize. The low concentration of weak acid is preferably 0.01 to 10%, preferably 0.2 to 5% of low concentration acetic acid. Using the acetic acid concentration of 0.2% to 5%, the calcium carbonate in the pearl powder used is dissolved in water with calcium ions, and the water-soluble organic substances bound to the pearl powder in the process are added to the ionized pearl powder solution. You can make it melt. In addition, since acetic acid used in the ionization process is a weak acid and its concentration is low, it is possible to minimize the destruction of the water-soluble organic components contained in the pearl powder, it is also possible to obtain functionality by these organic components. In addition, after step 1), the ionized pearl powder solution in which calcium carbonate is dissolved may be filtered through a filter paper, and then the ionized pearl powder solution may be recovered.

The freeze-drying process of step 2) is a process for maximally preserving the functionality of the organic components in the ionized pearl powder solution. When using the previous high temperature drying method, it is difficult to preserve the organic components in the solution. However, when the freeze-drying method of step 2) is used, this technical limitation is overcome. After the step 2), the water-soluble pearl powder has a very high solubility in water of 250 to 350mg / ml, when the water is dissolved to obtain a water-soluble pearl powder solution (PPS),

The water-soluble pearl powder or the pearl powder aqueous solution prepared by the production method of the present invention is excellent in the preservation of the organic components, it is preferable that the content of the protein and amino acid in the components of 0.01 to 0.1%. The pearl powder aqueous solution is preferably neutral pH 6.0 to 7.0, there is no acid odor, it may be a colorless transparent liquid.

The water-soluble pearl powder of the present invention or an aqueous solution thereof was ionized, and organic material loss was minimized. Therefore, it is excellent in the effect of inhibiting melanin synthesis, suppressing inflammatory reactions and promoting collagen synthesis, and can be usefully used in functional cosmetics for whitening, anti-inflammatory and skin aging inhibition.

1 is a view showing a method for producing a water-soluble pearl powder (WSPP) or an aqueous solution (PPS) thereof of the present invention.

2 is a diagram showing the tyrosinase inhibitory ability of the aqueous solution of water-soluble pearl powder (PPS) of the present invention (1: Arbutin 1mM addition; 2: 0.001mg / ml of high-temperature dry water-soluble pearl powder aqueous solution; 3: 0.01mg of high-temperature water-soluble pearl powder aqueous solution) / ml; 4: aqueous solution of high temperature dried water-soluble pearl powder 0.1mg / ml; 5: aqueous solution of freeze-dried water pearl powder (PPS) 0.001mg / ml; 6: aqueous solution of freeze-dried water-soluble pearl powder (PPS) 0.01mg / ml; 7: 0.1 mg / ml lyophilized water-soluble aqueous pearl powder (PPS); 8: distilled water; the experiment was repeated three times, the significance of the control and P <0.01.

 Figure 3 is a diagram showing the ability to inhibit the MITF expression of the aqueous solution of aqueous pearl powder (PPS) of the present invention (1: distilled water treatment, 2: distilled water and IBMX treatment; 3: 0.10 mg / ml water-soluble pearl powder aqueous solution (PPS) and IBMX; 4: 0.1 mg / ml water-soluble aqueous pearl powder (PPS) and IBMX treatment).

Figure 4 is a diagram showing the effect on the production of NO in the macrophages of water-soluble aqueous pearl powder (PPS) of the present invention (top: the amount of NO in LPS-treated macrophages; bottom: cell survival rate in LPS-treated macrophages; 1: Distilled water treatment; 2: distilled water and LPS treatment; 3: dexamethasone and LPS treatment; 4: aqueous aqueous pearl powder solution 4 mg / ml and LPS treatment; 5: aqueous aqueous pearl powder solution 2 mg / ml and LPS treatment; 6: aqueous aqueous pearl powder solution 1 mg / ml and LPS treatment; all experiments were repeated three times, and the significance of the inhibitory effect of LPS and aqueous pearl powder solution on the cells treated with LPS and distilled water was P <0.05).

Figure 5 is a diagram showing the effect of increasing the TGF-β expression of the aqueous solution of water-soluble pearl powder (PPS) of the present invention (1: distilled water and LPS treatment, 2: 4 mg / ml and aqueous solution of water-soluble pearl powder; all experiments 3 times) Was repeated, the significance of the transcription amount of the TGF-β gene of LPS and aqueous solution of water-soluble pearl powder to the control group was P <0.05.

Figure 6 is a diagram showing the collagen gene expression promoting effect of the aqueous solution of water-soluble pearl powder (PPS) of the present invention (1: distilled water treatment, 2: 4 mg / ml water-soluble pearl powder aqueous solution; all experiments were repeated three times, Significance level of collagen gene expression in aqueous solution of water-soluble pearl powder is P <0.05).

Hereinafter, examples will be described in detail to help understand the present invention. However, the following examples are merely to illustrate the content of the present invention is not limited to the scope of the present invention. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

Example 1 Preparation of Water-Soluble Pearl Powder

In order to make a water-soluble pearl powder material with complex skin function, it is processed by physical grinding method at low temperature, and the nano freshwater pearl powder (Chinese food grade nano pearl powder), which is pulverized into particles of approximately 100 nm size, has a low concentration of 0.2-5%. The reaction was conducted at room temperature to induce ionization of the pearl powder, thereby preparing a pearl powder solution in which calcium carbonate was dissolved. Then, the ionized water-soluble pearl powder solution was filtered through a filter paper to recover the ionized pearl powder solution, and then lyophilized to prepare a water-soluble pearl powder (WSPP) of the present invention. Thereafter, the lyophilized water-soluble pearl powder was dissolved in water to prepare a water-soluble pearl powder aqueous solution (PPS), and the manufacturing process of the water-soluble pearl powder or its aqueous solution is shown in FIG.

The water-soluble pearl powder (WSPP) prepared by the method of the present invention has a very high solubility in water (300 mg / ml), and when dissolved by adding water, a water-soluble pearl powder aqueous solution (PPS) was obtained. Most of the components were ionized calcium carbonate or Bradford method, and as a result of confirming that the organic matter is preserved at 0.05% (500mg / kg) content of protein and amino acids, the functionality of these organic components can be expected. Therefore, using water-soluble pearl powder ionized with 1% acetic acid of the present invention or an aqueous solution thereof, the functionalities such as the whitening effect, anti-inflammatory effect and skin aging inhibitory effect of Examples 2 to 4 were confirmed.

Example 2 Confirming the Whitening Effect of Water-Soluble Pearl Powder

1. Confirmation of tyrosinase inhibitory ability

In order to confirm the skin whitening effect of the aqueous solution (PPS) made of water-soluble pearl powder, it was confirmed that the inhibitory ability of tyrosinase important for the synthesis of melanin pigment. More specifically, the tyrosinase inhibitory activity of the aqueous pearl powder in the tyrosinase activity reaction was compared with the inhibitory ability of arbutin, a typical whitening functional substance. Aqueous solutions of water-soluble pearl powder were added to the tyrosinase enzyme reaction solution to 0.1, 0.01, and 0.001 mg / ml to investigate the degree of inhibition of tyrosinase enzyme activity. Activity for tyrosinase was mixed with 0.9 ml, 0.1 ml of 0.1 M phosphate buffer (pH6.8) and 1 ml of 1.5 mM L-tyrosine in each concentration sample, and 0.1 ml of tyrosinase (1,500 U / ml) was added at 37 ° C. After reacting for 10 minutes, the tyrosinase enzyme activity was measured by measuring the absorbance at 475 nm. The tyrosinase inhibitory effect (inhibition rate) of the pearl powder solution was calculated by comparing the enzyme activity value thus measured with the tyrosinase enzyme activity of the reaction obtained by adding distilled water (vehicle) instead of the aqueous aqueous pearl powder solution. The tyrosinase inhibitory activity of the pearl powder solution was compared with the tyrosinase enzyme inhibitory activity of 1 mM of arbutin (Sigma Chemical Co. St. Louis, USA), a representative whitening substance. The tyrosinase inhibitory effect of the water-soluble pearl powder of the present invention is shown in FIG. 2.

As shown in Figure 2, the aqueous water-soluble pearl powder solution was confirmed to inhibit the tyrosinase enzyme action in a concentration proportion of about 20%. In addition, when treated with an aqueous solution of water-soluble pearl powder at 0.1mg / ml, it was confirmed that the reaction solution treated with arbutin and tyrosinase inhibitory ability is almost similar, it was confirmed that the pearl powder solution can interfere with the skin melanin pigment synthesis. Therefore, it was confirmed that a whitening effect exists in the water-soluble pearl powder of the present invention. In addition, the effectiveness of the freeze-drying method was compared with the activity of the freeze-dried aqueous solution of water-soluble pearl powder (PPS) obtained by the method of the aqueous solution of water-soluble pearl powder dried at 80 ° C. without lyophilizing the water-soluble pearl powder solution obtained by ionization. . As a result, although some tyrosinase inhibitory ability was exhibited even in the aqueous solution of high temperature dried water-soluble pearl powder (2, 3, 4 of FIG. 2), the effect was the effect of the aqueous solution of lyophilized water-soluble pearl powder (PPS) (FIG. 2, 5). , 7) was found to fall significantly. These results show that the freeze-drying method used in the present method is a method of minimizing the destruction of organic components in the water-soluble pearl powder.

2. Water Soluble Pearl Powder (PPS) MITF ( microphthalmia -inhibition of associated transcription factor expression

In order to reconfirm the skin whitening effect of the pearl powder solution, the effect of inhibiting MITF expression was measured using B16F0 cells, which are melanoma cell lines obtained from mice. To determine whether aqueous solution of aqueous pearl powder (PPS) inhibits MITF expression in B16F0 cells, after treatment of the cells with IBMX (3-isobutyl-1-methylxanthine), the aqueous solution of aqueous pearl powder interferes with the promotion of MITF transcription of IBMX. (Chaing et al., Hydroalcoholic extract of Rhodiola rosea L. and its hydrolysate inhibit melanogenesis in B16F0 cells by regulating the CREB / MITF / tyrosinase pathway.Food Chem Toxicol. 2014 65: 129-139). First, B16F0 cells were treated with a water-soluble aqueous pearl powder sample and 100 μM of IBMX, followed by incubation for 24 hours. In order to examine the extent of MITF gene expression in the cells thus treated, RNA was extracted from the cells treated according to the above conditions, and RT-PCR was performed. 5-CAGGCTAGAGCGCATGGACTT-3 and 5-CTCCGTTTCTTCTGCGCTCAT-3 primers were used to amplify the MITF gene, and 5-CCTCGCCTTTGCCGATCC-3 and 5-GACTGACTACCTCATGAAGATCC-3 primers were used to amplify the β-actin gene. The RT-PCR results are shown in FIG. 3.

As shown in FIG. 3, the aqueous aqueous pearl powder solution (PPS) was found to inhibit the transcription of the MITF gene increased by IBMX concentration in proportion.

In addition, it was confirmed that the transcription of the MITF gene was lower in the cells treated with the aqueous solution of 0.1 mg / ml aqueous pearl powder (first line of FIG. 3). The above results show that the water-soluble pearl powder prepared according to the present method inhibits the transcription factor activity required for tyrosinase expression, thereby more effectively inhibiting melanin synthesis.

Example 3 Confirmation of skin soothing (anti-inflammatory) effect of water-soluble pearl powder

1. NO NO inhibition and cytotoxicity

Eczema, rash, itching, acne and hot flashes are one of the main causes of skin irritation. Therefore, since anti-inflammatory substances can show skin calming effect, the inhibitory effect of water-soluble pearl powder aqueous solution (PPS) was confirmed using an in vitro test for inflammatory reactions. More specifically, in order to determine whether the water-soluble pearl powder of the present invention can exhibit skin sedation by inhibiting the skin inflammatory response, the water-soluble pearl powder for the production of nitric oxide (NO) in the mouse RAW264.7 macrophage line in culture. The effect of the aqueous solution was investigated. After treating the RAW264.7 macrophage line in culture with an aqueous aqueous pearl powder solution, it was confirmed that NO production induced by LPS (lipopolysaccharide, endotoxin) was reduced in these cells. First, 1 ⁶ / ml of RAW 264.7 cells were inoculated in a culture dish, and then cultured for one day, and treated with 1, 2, 4 mg / ml of an aqueous solution of aqueous pearl powder, and then 1 µg of cells were treated with LPS after 1 hour. Treated to / ml. Then, incubated for 12 hours at 37 ℃, CO ₂ incubator. The NO produced in the medium was converted to nitrite using Griess reagent, and then the culture medium was reacted with an equal amount of 0.1% N- (1-Naphtyl) ethylene diamine dihydrochloride and 5% phosphoric acid / 1% Sulfanilamide solution. After that, absorbance was measured at 550 nm to compare the production of nitrite. In addition, since the production of NO is affected by the number of living cells, the survival rate of RAW264.7 cells used in the experiment was also examined by the MTT assay whether the aqueous solution of aqueous pearl powder. After washing RAW264.7 cells treated with LPS, and replaced with 150μl of NO measuring medium, 15μl of MTT was added and cultured to confirm the formation of purple precipitate. After about 5 hours, 150 µl of the cell lysis solution was added to the culture, and the cells were reacted overnight at 37 ° C to lyse the cells. Then, absorbance was measured at 595 nm. The results are shown in FIG. 4.

As shown in Figure 4, it was confirmed that the pearl powder aqueous solution reduced NO production in RAW264.7 cells in proportion to the concentration. The effect was similar to the inhibitory effect of dexamethasone (dexamethasone) of 100mM well known as an inhibitory function. In addition, in the case of cell viability experiments, it was confirmed that the aqueous aqueous pearl powder solution did not appear to affect the survival of RAW264.7 cells at any concentration used. Therefore, the aqueous solution of water-soluble pearl powder inhibits the NO production necessary for the progression of the inflammatory reaction without affecting the survival of the macrophage line, and is believed to inhibit the inflammatory response.

2. Identification of cytokine gene expression

In order to confirm how the NO production inhibitory effect of the water-soluble pearl powder (PPS) material of Example 3-1 was expressed, the cells were treated with LPS-treated RAW264.7 cell culture solution at 4 mg / ml, and then Transcriptional activity of various cytokine genes expressed was confirmed using quantitative real-time PCR. More specifically, β-actin using 5-TGACGTCACTGGAGTTGTACGG-3 and 5-GGTTCATGTCATGGATGGTGC-3 primers and 5-CCTCGCCTTTGCCGATCC-3 and 5-GACTGACTACCTCATGAAGATCC-3 primers were used to amplify the TGF-β gene by the real-time gene amplification reaction. Corrected by comparison with the expression level of the gene. The results are shown in FIG. 5.

As shown in Figure 5, it was confirmed that the expression of TGF-β in the cells treated with water-soluble pearl powder significantly increased more than 2.5 times compared to the TGF-β expression of the control cells (P <0.01). TGF-β is a cytokine that inhibits the expression of iNOS (induced NO synthase) by IL-1 in cells (Boche, D., Cunningham, C., Docagne, F., Scott, H. and Perry, VH. (2006) TGF-beta1 regulates the inflammatory response during chronic neurodegeneration. Neurobiology Disease, 22, (3), 638-650.), NO synthesis inhibitory effect of water-soluble pearl powder is associated with increased TGF-β expression. Seems to be. As a result, it is assumed that the water-soluble pearl powder contains a substance that acts on macrophages important for the inflammatory response, increases the expression of TGF-β, and inhibits NO synthesis. Therefore, it was confirmed that the water-soluble pearl powder aqueous solution (PPS) of the present invention has a skin soothing effect.

Example 4 Confirmation of Skin Aging Inhibitory Effect of Water-Soluble Pearl Powder

In order to confirm whether the water-soluble pearl powder in the skin can increase the collagen synthesis to inhibit skin aging, in vitro assay using fibroblast culture was confirmed whether the aqueous solution of water-soluble pearl powder (PPS) increased the expression of collagen gene. After treating water-soluble pearl powder to 4 mg / ml in NIH3T3 cell culture medium, which is a fibroblast of mouse, the expression of collagen gene expressed in the cells was confirmed using a quantitative real-time PCR reaction. More specifically, in order to amplify the collagen gene, primers of mice COL1A1 (type I collagen gene) 5-AAACCCGAGGTATGCTTGATCTGTA-3 and 5-GTCCCTCGACTCCTACATCTTCTGA-3` were used, and 5-CCTCGCCTTTGCCGATCC-3, 5-GACTGACTACCTCATGAAGATCC-3 The primers were corrected by comparison with the expression level of β-actin gene. The results are shown in FIG. 6.

As shown in Figure 6, it was confirmed that the expression of collagen gene in the cells treated with water-soluble pearl powder increased significantly by 2.4 times compared to the cells treated with water as a control. In Example 3, it was confirmed that the water-soluble pearl powder material promotes the transcription of TGF-β, and TGF-β also acts as an important collagen synthesis promoter. In summary, the water-soluble pearl powder or its aqueous solution promotes TGF-β expression and collagen gene expression in fibroblasts of the skin to increase collagen synthesis, thereby inhibiting skin aging.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (9)

1. Cosmetic composition for inhibiting whitening, anti-inflammatory and skin aging comprising a water-soluble pearl powder or an aqueous solution thereof.
2. The cosmetic composition according to claim 1, wherein the water-soluble pearl powder is ionized with acetic acid of 0.2% to 5%.
3. The cosmetic composition of claim 1, wherein the water-soluble pearl powder is lyophilized.
4. 1) ionizing the pearl powder with a weak acid of 0.2% to 5%; And

   2) lyophilizing the ionized pearl powder; Method for producing a water-soluble pearl powder and an aqueous solution thereof comprising a.
5. According to claim 4, wherein the size of the pearl powder of step 1) is characterized in that 90 to 110nm, the manufacturing method.
6. The method of claim 4, wherein the weak acid of step 1) is acetic acid.
7. According to claim 4, After the step 1) and before the step 2), characterized in that it further comprises the step of filtering the ionized pearl powder.
8. The method of claim 4, wherein the water-soluble pearl powder has a whitening effect, an anti-inflammatory effect, and an anti-aging effect.
9. Water-soluble pearl powder or water-soluble pearl powder aqueous solution prepared by the method of claim 4.

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