KR101141168B1 - Whitening composition containing saponification product of sunflower oil as an effective ingredient - Google Patents

Abstract

The present invention relates to a whitening cosmetic composition containing a sunflower oil saponified product as an active ingredient and a composition for preventing and treating skin pigmentation diseases. More specifically, the composition containing the sunflower oil saponified product of the present invention as an active ingredient inhibits the expression of tyrosinase, an enzyme that plays a key role in the production of melanin, at the protein and gene levels, and safely without side effects on the skin. It can be used as a whitening cosmetic composition and a composition for the prevention and treatment of skin pigmentation diseases.

Sunflower oil, saponification, melanin, whitening, pigmentation

Description

Whitening composition containing saponification product of sunflower oil as an effective ingredient}

The present invention relates to a cosmetic composition for skin whitening containing a sunflower oil saponified product as an active ingredient and a composition for preventing and treating skin pigmentation diseases.

The changes in the global environment and the destruction of the ozone layer pose a major threat to the health of modern people, both directly and indirectly. Ultraviolet rays can damage the skin directly and act as a major cause of skin pigmentation. This is combined with whitening, the standard of modern people's beauty, and various solutions have been studied in terms of health as well as beauty. Common skin pigmentation disorders include blemishes, freckles, black spots, birthmarks, and the like, including pigmentation by drugs, hyperpigmentation following inflammation found in healing wounds, and hyperpigmentation as a symptom of dermatitis. . These pigmentation disorders are more common among Asians and Hispanics and are of interest.

Originally, melanin is a black pigment found in animals, plants, microorganisms, etc. It is not essential for growth or development, but it is a substance that enhances the viability and competitiveness of any environment. Especially, melanin in animals has been associated with skin diseases and In addition to dopa melanin produced from tyrosine by tyrosinase, 1,8-dihydroxynaphthalene melanin (DHN melanin; 1,8-Dihydroxynaphtalene melanin), ortho- Diphenol melanin (ο-diphenol melanin), catechol melanin and the like have been reported (Bell AA and Wheeler MH, Annu. Rev. Phytopathol., 24, pp 411-451, 1986). Melanin is an unusually stable pigment found in living organisms, insoluble in almost all solvents, and is difficult to extract or chemically handle. Because of this, there are many unknowns about the chemical aspects of melanin and the progress of research is sluggish.

The process of melanogenesis is clearly shown to occur in organelles called melanosomes of specially differentiated cells called melanocytes.

That is, the skin color is determined according to the content, distribution, and the like of melanin and is related to the number and distribution of melanosomes generated in melanocytes and released to the outside of cells. Hyperpigmentation of the skin can be caused by a number of factors, such as hormonal abnormalities, genetic diseases and UV irradiation after the inflammatory response of the skin, mainly due to melanin pigment synthesis abnormalities and distribution abnormalities.

Treatment methods for skin pigmentation include the use of sunscreens, topical ointment application, chemical dermabrasion, laser treatment and the like.

Among other things, preventing sun exposure is one of the main treatment goals. This is because sun exposure is a major culprit of deterioration or recurrence of pigmentation, regardless of the underlying prior cause. Therefore, the use of broad spectrum sunscreens in outdoor activities is a way to prevent skin pigmentation. It is also recommended to avoid excessive outdoor activities from 10 am to 4 pm.

Ointments used in the treatment of skin pigmentation include bleach hydroquinone, vitamin A ointment, topical steroid ointment, vitamin C, glycolic acid, and azelic acid. Electrophoresis therapy, which infiltrates C, may be performed regularly. Recently, a method of periodically performing dermabrasion (pilling) using TCA or glycolic acid in combination with topical treatment is also widely used. To date, however, there is no perfect way to treat skin pigmentation, and any treatment can easily recur. In addition, hydroquinone is not used in cosmetics because of its high irritation to the skin and stability problems. Therefore, it is necessary to fundamentally understand the melanin production mechanism of the skin and to develop a new treatment that can suppress melanin production.

Accordingly, the present inventors are commonly found around the skin in skin pigmentation diseases (stain, freckles, birthmarks, black spots, pigmentation by drugs, pigmentation after inflammation, etc.), which is a kind of common skin diseases, and shows no toxicity at all. The present invention was completed by confirming that sunflower oil saponification products among substances that can be used for preventing and treating skin pigmentation diseases by inhibiting melanin production process.

An object of the present invention is to provide a cosmetic composition for whitening, containing a sunflower oil saponified product as an active ingredient.

It is also an object of the present invention to provide a composition for the prevention and treatment of skin pigmentation diseases, containing a sunflower oil saponified product as an active ingredient.

According to one aspect of the present invention, the present invention provides a cosmetic composition for whitening containing a sunflower oil saponified product as an active ingredient.

Sunflower oil is in the form of triglycerides attached to fatty acids by ester bonding to glycerol, the fatty acid composition of which is more than 90% unsaturated fatty acids such as linoleic acid, oleic acid, arachidonic acid. When the saponification process is heated in an alkaline state, the ester bond is broken and free fatty acid is released. Applying untreated sunflower oil to the skin produces only a small amount of free fatty acids from the action of lipases in triglyceride-type oils.However, externally saponified and applied to the skin can provide large amounts of unsaturated free fatty acids to the skin. have.

Sunflower oil saponification products of the present invention is preferably included from 0.001 to 99.9% based on the total weight of the composition.

According to one embodiment of the present invention, comparing the group treated with sunflower oil or sunflower oil saponified product and the untreated group to investigate whether the sunflower oil or sunflower oil saponified product is toxic to cells, 100 ㎍ / No cell damage could be observed at the concentration of ml. This means that the cytotoxicity of sunflower oil or sunflower oil saponified products is not high and is a safe substance.

The toxicity experiment can be used MTT assay.

In the MTT assay, after treatment with 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) to cells, the amount of the substance was reduced to formazan. It can be done by measuring the relative number of living cells, that is, normal metabolic ability.

The composition of the present invention has the use of skin whitening. The composition of the present invention is to inhibit the production of melanin pigment itself, which is the main mechanism of skin pigmentation. Melanin pigments are produced in melanocytes by various stimuli, and the pigments are transferred and distributed to surrounding keratinocytes, which are manifested in the phenotype of dark brown pigmentation on the skin.

According to one embodiment of the present invention, the present inventors induced melanin production in melanoma cells with IBMX (isobutylmethylxanthine) and added various concentrations of sunflower oil saponification products, resulting in concentration-dependent intracellular melanin content of sunflower oil saponification products. The visual and quantitative reduction of the experimental data was confirmed. In addition, it was confirmed that sunflower oil saponification products reduced the mRNA expression of tyrosinase, an enzyme that plays a key role in the production of melanin, and genes important for melanin production. Genes important for melanin production may be TRP1, TRP2, MITF. That is, the sunflower oil saponified product of the present invention can be confirmed that it is very useful for the prevention and treatment of skin pigmentation disease by inhibiting the production of melanin pigmentation of the skin.

In order to confirm that the composition comprising the sunflower oil saponification product as an active ingredient inhibits the production of melanin pigment, the related enzymes tyrosinase and related genes TRP1, TRP2, and MITF are Western blotting at the protein and gene level, respectively. Western blotting and real time PCR methods can be used.

The components included in the cosmetic composition of the present invention include components conventionally used in cosmetic compositions in addition to sunflower oil saponification products as active ingredients, and include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. And carriers. In addition, the cosmetic composition may further include a skin absorption promoting substance to enhance the effect.

The cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing It may be formulated with hair cosmetics, oils, and the like, but is not limited thereto. More specifically, the cosmetic composition of the present invention skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, Sunscreens, essences, nutritional essences, packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers, emulsions, press powders, loose powders and eye shadows.

When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular, in the case of spray, additionally chloro fluorohydrocarbon, propane Propellant such as butane or dimethyl ether.

 When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

 When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Soluble cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, as a carrier component, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In another aspect, the present invention provides a composition for the prevention and treatment of skin pigmentation diseases, comprising a sunflower oil saponified product as an active ingredient.

The above-mentioned "skin pigmentation disease" includes blemishes, freckles, black spots, birthmarks, pigmentation by drugs, hyperpigmentation appearing as a symptom of post-inflammatory pigmentation or dermatitis found in the wound site healed.

Sunflower oil saponification products of the present invention is preferably included from 0.001 to 99.9% based on the total weight of the composition.

 A pharmaceutically acceptable carrier, excipient, or diluent may be added to the composition of the present invention to form a pharmaceutical unit dosage form.

Pharmaceutically acceptable carriers contained in the composition for the prevention and treatment of skin pigmentation diseases of the present invention are commonly used in the preparation, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, Calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and minerals Oils and the like, but are not limited thereto. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

The composition of the present invention can be administered to various mammals such as oral or parenteral to mammals such as mice, mice, livestock, humans, and the like, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular. It can be administered by injection. It also includes the use of elctrotransfer or microneedle to deliver sunflower oil saponified products into the dermis and epidermis. Preferably, it is applied in a topical application manner by transdermal administration, more preferably by application during parenteral administration.

Suitable dosages of the compositions of the present invention may be prescribed in various ways depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of the patient, food, time of administration, route of administration, rate of excretion and response to reaction. have.

The compositions of the present invention may be prepared in unit dosage form by formulating with a pharmaceutically acceptable carrier and / or excipient, according to methods which can be easily carried out by those skilled in the art. It can be prepared by incorporation into a multi-dose container.

The skin external composition containing the sunflower oil saponified product of the present invention may be prepared and used in the form of a skin external preparation such as a cream, gel, patch, spray, ointment, warning, lotion, linen, pasta, or cataplasma. But it is not limited thereto.

The cosmetic composition for whitening and the composition for the prevention and treatment of skin pigmentation disease which use the sunflower oil saponification product of this invention as an active ingredient are characterized by the inhibition of melanin production of skin. More specifically, the cosmetic composition for whitening and the composition for the prevention and treatment of skin pigmentation disease using the sunflower oil saponified product of the present invention is characterized by showing a whitening effect by inhibiting tyrosinase activity.

The whitening cosmetic composition and the composition for the prevention and treatment of skin pigmentation disorders, which are the sunflower oil saponified product of the present invention, inhibit the expression of genes related to the inhibition of tyrosinase activity and the production of melanin pigment and finally the melanin pigment. By suppressing the production, it has an excellent effect in the treatment of pigmentation diseases and can contribute greatly to the skin protective agent and the skin disease treatment industry without side effects.

Hereinafter, the examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, those skilled in the art to which the present invention pertains. Will be self-evident.

Example 1 Preparation of Sunflower Oil Saponified Product

A predetermined amount of 2.5 N KOH dissolved in alcohol in sunflower oil is added and heated at 80 ° C. for 30 minutes to saponify. After the saponification process, the pH is adjusted to 7.4.

Experimental Example 1 Cytotoxicity Experiment

MTT assay was used to investigate whether sunflower oil or sunflower oil saponification products are toxic to cells. After incubation, the cells were treated with 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT), and then the amount of this substance reduced to formazan was measured. It is possible to estimate the relative number of these normal, ie, living cells. After incubation at 37 ° C. for about 30 minutes after MTT treatment, the formazan component was extracted with DMSO and the absorbance was measured at 570 nm with a spectrophotometer.

When treated with sunflower oil or saponified sunflower oil at a concentration of 12.5-100 μg / ml, it showed the same or higher cell viability compared to the untreated group, and no cell damage was observed at the concentration of 100 μg / ml. Could not (see Figure 1).

This means that the cytotoxicity of sunflower oil or sunflower oil saponification products is not high and is a high safety material.

<Experimental Example 2> Melanin production inhibitory effect of saponified sunflower oil (measurement of melanin content in cells)

Melanin production of melanoma cells was induced with IBMX (isobutylmethylxanthine) and treated with various concentrations of sunflower oil or sunflower oil saponification products, and the levels of inhibition of melanin production were measured by comparing the intracellular melanin content. In order to measure the melanin content, melanoma cells after culture were washed with physiological saline, lysed with Lysis Buffer, and treated with 20% TCA. Ethanol and ether were treated again, dried in air, and dissolved in 0.85 M potassium hydroxide and heated for 15 minutes. Absorbance was measured at 440 nm after cooling. For melanin content / cell comparison, the amount of DNA was measured by fluorescence assay using bisbenzimide. Then, the melanin content of DNA was compared between the group treated with the sunflower oil saponified product and the group not treated. After drug treatment, the cells were incubated for 3 days, and the concentration of sunflower oil or sunflower oil saponified product was treated at various concentrations as described in the figure, and IBMX was treated at 0.1 mM concentration. Sunflower oil saponification reduced the intracellular melanin content depending on the concentration (see FIG. 2 and Table 1).

Sunflower oil saponification product treatment concentration (㎍ / ml) Content of melanin (% of control) 0.0 100 12.5 76 25.0 56 50.0 39 100.0 30

Experimental Example 3 Determination of Inhibitory Effects of Sunflower Oil Soaps on Tyrosinase Activity

Tyrosinase activity was measured by DOPA oxidase activity. After obtaining the cell lysate, the reaction solution was treated, and the reaction of DOPA oxidization into DOPA chrome was measured by using a spectrophotometer at 475 nm. At this time, the reaction solution containing a substrate solution (0.1% L-DOPA in 0.1 M sodium phosphate, pH 6.0) and the enzyme solution was used. Each activity result was corrected with the amount of DNA obtained by the DNA assay described in Experimental Example 2. Sunflower oil saponification resulted in a significant reduction in the activity of tyrosinase in a concentration dependent manner (see FIG. 3 and Table 2).

Sunflower oil saponification product treatment concentration (㎍ / ml) Tyrosinase Activity (%) 0.0 100 12.5 85 25.0 73 50.0 64 100.0 48

Experimental Example 4 Sunflower Oil In soap products  by Tyrosinase  Reduced protein expression

Protein was extracted from the cells and subjected to Western blotting using SDS-PAGE. Goat anti-murine tyrosinase IgG (Santa-Cruz Biotechnology Inc.) was used, and the protein expression of tyrosinase was developed by the chemi-illuminant system. The amount was measured.

Sunflower oil saponification reduced protein expression of tyrosinase in a concentration dependent manner (see FIG. 4). This means that sunflower oil saponification products do not directly inhibit the activity of tyrosinase but also regulate the amount of expression at the protein level.

Experimental Example 5 Reduction of mRNA Expression of Whitening-related Gene by Sunflower Oil Soap Product

MRNA was extracted from the cells and cDNA was synthesized through reverse transcriptase. Real-time PCR reaction was performed using ABI Prism 7900HT Sequence Detection System (Applied Biosystems).

Sunflower oil saponification reduced mRNA expression of TRP1, TRP2, MITF, which is important for melanogenesis as well as tyrosinase in a concentration dependent manner (see FIG. 5). This means that sunflower oil saponification inhibits melanin production by regulating gene expression.

Figure 1 shows that the sunflower oil or sunflower oil saponified product is not cytotoxic to melanoma cells (B16 melanoma cells) at the concentration used in the experimental example of the present invention. (SU (sunflower oil), sap-SU (sunflower) Oil soap products))

FIG. 2 shows the results of measuring whether melanin pigmentation is induced by treatment with IBMX, and at this time, melanin pigmentation is inhibited by treating sunflower oil or sunflower oil saponification products. The photograph shows the visual comparison of melanin production with the color of cultured cells. The histogram shows the result of comparing the relative melanin production as the ratio of melanin content to the total amount of surviving genes (SU (sunflower oil), sap-SU (sunflower oil soap product))

Figure 3 shows the effect of sunflower oil saponification products on tyrosinase activity, an enzyme that plays a central role in melanogenesis. As a result of measuring the activity in the cell, the concentration of the tyrosinase decreases in a concentration-dependent manner by the sunflower oil saponification product. (sap-SU (sunflower oil soap product))

Figure 4 shows the results of measuring the expression level of tyrosinase. As a result of Western blotting showing the amount of protein, it shows a concentration dependent decrease by sunflower oil saponification products.

Figure 5 shows the realtime RT-PCR results showing the relative amounts of mRNA of Trp1, Trp2, MITF gene related to melanin pigment production in addition to tyrosinase. As with proteins, the higher the concentration of sunflower oil saponification products, the lower the relative value of tyrosinase mRNA, and the other related genes also indicate that the mRNA level is lowered by sunflower oil saponification products. (sap-SU (sunflower oil soap product))

Claims (11)

A whitening cosmetic composition containing a sunflower oil saponified product as an active ingredient. According to claim 1, wherein the cosmetic composition is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack , Soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, milky lotion, press powder, loose powder and eye shadow, cosmetic composition for whitening, characterized in that it takes one. The cosmetic composition for whitening according to claim 1, wherein the skin whitening effect of the composition is due to inhibition of melanin production. The cosmetic composition for whitening according to claim 1, wherein the skin whitening effect of the composition is due to inhibition of tyrosinase activity. Skin selected from blemishes, freckles, black spots, birthmarks, pigmentation by drugs, hyperpigmentation as a symptom of post-inflammatory pigmentation or dermatitis found in cured wounds, containing sunflower oil saponification products as an active ingredient Pharmaceutical compositions for the prevention and treatment of pigmentation diseases. delete 6. The composition for preventing and treating skin pigmentation disorders according to claim 5, wherein the sunflower oil saponification product is contained in an amount of 0.001 to 99.9% based on the total weight of the composition. 6. The composition for preventing and treating skin pigmentation disease according to claim 5, wherein the composition is formulated in a pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient or diluent to the composition. The method of claim 5, wherein the composition is a cream, gel, patch, spray, ointment, warning, lotion, linen, pasta or cataplasma formulations for the prevention and treatment of skin pigmentation diseases Composition. 6. The composition for preventing and treating skin pigmentation disorders according to claim 5, wherein the treatment and prophylactic effect of the skin pigmentation disorders is due to inhibition of melanin production. The composition for the prevention and treatment of skin pigmentation disorders according to claim 5, wherein the therapeutic and prophylactic effect of the skin pigmentation disorders is by inhibiting tyrosinase activity.

Images