KR20110134146A - Composition of external application for whitening and moisturizing skin - Google Patents

Abstract

PURPOSE: An external use skin composition containing horse placenta extract or horse milk is provided to suppress melanogenesis and to ensure skin whitening and moisturizing. CONSTITUTION: An external use skin composition for skin whitening contains 0.001-10.0 wt% of horse placenta extract or horse milk as an active ingredient. The composition is a cosmetic composition. The cosmetic composition is manufactured in the form of an emulsion, lotion, cream, solution, suspension, anhydrous product(oils and glycols), gel, mask, pack, and powder. The cosmetic composition contains acceptable carriers such as alcohol, oil, surfactant, fatty acid, silicon oil, wetting agent, moisturizing agent, stabilizing agent, UV protecting agent, or perfume.

Description

Composition for external application for whitening or moisturizing the skin {Composition of External Application for Whitening and Moisturizing Skin}

The present invention relates to an external composition for skin whitening or skin moisturizing. Specifically, the present invention relates to an external composition for skin whitening or skin moisturizing using enzyme-treated horse placenta extract or horse oil (horse oil).

It is well known that skin whitening is directly related to melanin pigment.

Melanin protects sub-dermal skin organs by blocking UV rays, and protects the damage of proteins and genes in the skin caused by free radicals in the skin. It causes hyperpigmentation such as blemishes and freckles.

Melanin synthesis is achieved by the action of tyrosinase and tyrosinase related proteins present in the melanosomes. Specifically, tyrosine is oxidized by tyrosinase and is converted into dopa (DOPA, 3,4-dihydroxyphenylalanine), followed by dopaoxidase to oxidize the dopa to be dopaquinone. In addition, melanin is produced by the action of tyrosinase-related proteins TRP-1 (5,6-dihydroxy indole-2-carboxylic acid oxidase) and TRP-2 (dopachrome tautomerase).

Therefore, a whitening effect can be expected when the production of melanin is suppressed.

On the other hand, due to the recent environmental pollution caused by the acceleration of the industrial society, various stresses caused by the complicated society, artificial temperature control by air conditioning and heating facilities, aging, ultraviolet rays, etc., the moisture in the stratum corneum of the skin is reduced and the skin becomes rough. The problem of the skin is increasing.

The present invention discloses a cosmetic composition using a protease-treated horse placenta extract or horse oil having skin whitening activity such as melanin production inhibitory activity and skin moisturizing activity.

An object of the present invention to provide a skin external preparation composition for skin whitening using protease-treated horse placenta extract or horse oil.

Another object of the present invention to provide a skin external preparation composition for moisturizing skin using protease-treated horse placenta extract or horse oil.

As the present inventors confirmed in the following Examples and Experimental Examples, horse oil obtained by removing and purifying proteins from horse placenta extract and horse fat meat by the treatment of protease was produced melanin in B16F10 melanoma cells. With inhibitory activity, it was confirmed in the clinical experiment that the skin whitening activity and skin moisturizing activity. The skin whitening activity is obtained by surveying the clinical trial participants in the form of a questionnaire, the skin moisturizing activity is obtained by evaluating the transdermal moisture loss (hereinafter "TEWL") and skin moisture content.

The present invention is provided based on the experimental results as described above.

In one aspect, the present invention relates to a skin external preparation composition for skin whitening comprising a protease-treated horse placenta extract and / or horse oil, and in another aspect, a skin moisturizing skin comprising a horse placenta extract and / or horse oil It relates to an external preparation composition.

In the present specification, the "protease-treated horse placenta extract" refers to those obtained by treating and hydrolyzing protease on horse placenta. Such extract may be in the form of purified form by filtration or in the form of a concentrated liquid or solid form from which the reaction solvent is removed. In order to achieve a sufficient hydrolytic effect, the protease is preferably added at least 0.5 parts by weight, in particular at least 2 parts by weight, based on 100 parts by weight of the horse's placenta. Specifically, the optimal amount of addition will be determined depending on the protease used. The reaction temperature at the time of hydrolysis may be selected to maximize the activity of the protease used. Usually it will be selected within the range of 35 ° C to 55 ° C. If the reaction temperature is too high, there may be a problem of inactivation of the enzyme, if it is too low, the efficiency of hydrolysis may be reduced. Proteases may be any of microorganisms, plants or animals, and there is no particular limitation, but papain, pepsin, trypsin and the like will mainly be used. The following example shows that the horse placenta extract obtained by acid hydrolysis has higher skin whitening and skin moisturizing activity than the horse placenta extract obtained by treatment with protease. It is presumed that this is because various amino acids are decomposed during acid hydrolysis or various bioactive substances such as uronic acid, glycosaminoglycan, sialic acid and epithelial cell promoters are inactivated.

In addition, in this specification, "horse oil" refers to the fats and oils obtained from the word. These horse oils include milk fat and are from any part of the horse. The horse oil includes a purified form such that phospholipids, proteins, etc. are removed, and its properties may be in a solidified form, or may be in a liquid or powder form.

In addition, the term "active ingredient" as used herein means a component that can exhibit the desired activity alone or in combination with a carrier that is not active itself.

Terms not specifically defined herein follow the Korean dictionary meaning or the meaning generally accepted in the art.

On the other hand, in the following experimental example, when the mixed placenta extract and horse oil were used, the melanin production inhibitory activity was much higher than when used alone, and the participants responded that the whitening activity was high even in actual clinical trials. In addition, the skin moisturizing effect was found to be higher than the moisturizing activity alone.

Therefore, the skin external preparation composition for skin whitening and the external skin composition for moisturizing the skin (hereinafter "composition of the present invention") preferably include both enzyme-treated horse placenta extract and horse oil as active ingredients.

Meanwhile, the composition of the present invention may include any amount (effective amount) of enzyme treated horse placenta extract and / or horse oil as its active ingredient as long as it can exhibit skin whitening activity or skin moisturizing activity. Typical effective amounts will be determined within the range of 0.001% to 20.000% by weight based on the total weight of the composition. Here, the "effective amount" refers to the amount of the active ingredient that can promote skin lightening activity or skin moisturizing activity. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.

The composition of this invention can be grasped | ascertained as a cosmetic composition in a specific aspect.

When the composition of the present invention is conceived as a cosmetic composition, the cosmetic composition can be prepared in various forms, for example, emulsions, lotions, creams (oil-in-water, water-in-oil, multiphase), solutions, suspensions (anhydrous and Aqueous), anhydrous products (oil and glycol based), gels, masks, packs, powders and the like.

The composition of the present invention may include an acceptable carrier in a cosmetic preparation in addition to including the active ingredient.

Herein, "acceptable carrier in cosmetic preparation" means a compound or composition already known and used that can be included in cosmetic preparations or a compound or composition which will be developed in the future and does not have more toxicity than the body can adapt to when contacted with the skin.

The carrier may be included in the composition of the present invention from about 1% to about 99.99% by weight, preferably from about 50% to about 99% by weight of the composition.

However, it is understood that the ratios limit the scope of the present invention in any aspect because the ratios vary depending on the formulation of the cosmetics as described above and their specific application site (face or hand) or their preferred application amount. It should not be.

Meanwhile, examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosity modifiers, emulsions, stabilizers, sunscreen agents, colorants, fragrances, and the like.

Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, sunscreens, colorants, compounds / compositions that can be used as fragrances, etc., which can be used as the carrier are already known in the art. As a result, those skilled in the art can select and use appropriate materials / compositions.

The composition of the present invention may be regarded as a soap composition in another specific embodiment.

When the composition of the present invention is understood as a soap composition, the soap composition of the present invention may be prepared by including an active ingredient thereof in a soap base, and may be prepared by including a skin moisturizer, an emulsifier, a hard water softener, and the like as an additive. .

As the soap base material, vegetable oils such as palm oil, palm oil, soybean oil, perm oil, olive oil, palm kernels, or animal oils such as tallow, pork, sunny and fish oils can be used, and the skin moisturizing agent is glycerin, erythritol and polyethylene glycol. , Propylene glycol, butylene glycol, pentylene glycol, hexyl glycol, isopropyl myristate, silicone derivatives, aloe vera, sorbitol and the like can be used, the emulsifiers include natural oils, wax fatty alcohols, hydrocarbons, natural plant extracts, etc. May be used, and tetrasodium idieti or the like may be used as the water softener.

The soap composition of the present invention may further include an antimicrobial agent, a dispersant, a foam inhibitor, a solvent, a scale inhibitor, a corrosion inhibitor, a perfume, a pigment, a metal ion sequestrant, an antioxidant, a preservative, and the like as an additive.

In the soap composition of the present invention, the soap base or additives may be included in an amount generally used in the art, and the soap base is generally added in an amount of 99.999 wt% to 50 wt% based on the total weight of the soap composition. The additive may be added in an amount of 1% to 20% by weight.

The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier, excipient, and the like in addition to the active ingredient, and may be a topical formulation such as creams, lotions, ointments (semi-solid external preparations), micro-emulsions, gels, pastes, transdermal preparations. (TTS) (such as patches, bandages, etc.) and the like.

As used herein, "pharmaceutically acceptable" means that the subject of application (prescription) is not as toxic as applicable, without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (such as corn starch, potato starch, etc.), cellulose, derivatives thereof (such as sodium carboxymethyl cellulose, ethylcellulose, etc.), malt, gelatin, talc, Solid lubricants (such as stearic acid, magnesium stearate, etc.), calcium sulfate, vegetable oils (such as peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, etc.), alginic acid, emulsifiers (such as TWEENS), wetting agents (Such as sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, water, saline, phosphate buffer solutions, and the like. The carrier may be selected from one or more of suitable pharmaceutical formulations according to the formulation of the pharmaceutical composition of the present invention.

Excipients may be selected and used according to the formulation of the pharmaceutical composition of the present invention, for example, suspending agents and dispersants such as sodium carboxymethyl cellulose, methyl cellulose, hydropropylmethyl cellulose, sodium alginate, polyvinylpyrrolidone, etc. Can be.

The pharmaceutical composition of the present invention may be administered orally or parenterally, preferably topically.

The daily dosage of the pharmaceutical composition of the present invention is usually 0.001 ~ 150 mg / kg body weight range, it can be administered once or divided into several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage may limit the scope of the present invention in any aspect. It should not be understood as.

As described above, according to the present invention, it is possible to provide a composition for skin whitening and a composition for moisturizing skin using an enzyme treated horse placenta extract or horse oil.

Such a skin whitening composition and a skin moisturizing composition may be commercialized as a drug such as cosmetics or soap, ointment, cream.

Hereinafter, the present invention will be described with reference to Examples, Comparative Examples, and Experimental Examples. However, the scope of the present invention is not limited to these examples, comparative examples and experimental examples.

< Example > Preparation of Horse Placenta Extract by Enzyme Treatment and Extraction of Horse Oil

Example 1 Preparation of Horse Placenta Extract by Enzyme Treatment

The frozen horse placenta was thawed to remove the amnion and cord from the placenta, and only placental chorionic tissue was collected and washed several times with water to remove blood. Next, the placental chorionic tissue was finely chopped, and 1 kg of chorionic tissue, 2 L of distilled water and 50 g of protease (papaine, Sigma-Aldich) were added and reacted with stirring at 55 ° C. for 16 hours in a constant temperature water bath. Next, the enzyme reaction was stopped by heating at 95 ° C. for 30 minutes, filtered through a 5 μm filter, the filtrate was concentrated under reduced pressure, water was evaporated, 1% (w / w) activated carbon was added thereto, stirred for 20 minutes, and 5 μm. Filtration again with filter. After filtering with a 0.45㎛ filter lyophilized to prepare a solid horse placenta extract.

Example 2 Extraction of Horse Oil

From the slaughtered horses, the parts (fat meats) with relatively good fat accumulation are selected and washed, and the washed fat meats are mixed with water at a weight ratio of 50:50 and heated at a temperature of 80 to 90 ° C. for about 5 hours. To liquefy fats in fatty meat. The liquefied horse oil was then separated, and the horse oil was treated with 0.1% phosphoric acid and washed with water at 60 to 70 ° C. to remove phospholipids, proteins and other viscous substances contained in the separated horse oil. 2.5% caustic soda was added to remove fatty acids from washed horse oil and washed again with water at 60-70 ° C. The small amount of water present in the horse oil thus obtained is removed, and 5% by weight of acidic clay is added for decolorization and deodorization.Then, warmed to 110-120 ° C and stirred until it is sufficiently stirred, and the stirred horse oil is filtered. press) and filtered to remove acidic clay. Horse oil thus obtained was used in the experiment while refrigerated.

< Comparative example > Preparation of Horse Placenta Extracts by Acid Hydrolysis

The frozen horse placenta was thawed to remove the amnion and cord from the placenta, and only placental chorionic tissue was collected and washed several times with water to remove blood. Next, the placental chorionic tissue was finely chopped and hydrolyzed for 5 hours by adding 500 ml of 3N HCl to 1 kg of the chorionic tissue. The hydrolyzed product was filtered through a 5 μm filter, concentrated under reduced pressure, pH was adjusted to 7, and then 1% (w / w) of activated carbon was added thereto, stirred for 20 minutes, and filtered again using a 5 μm filter. After filtering with a 0.45㎛ filter lyophilized to prepare a solid horse placenta extract.

< Experimental Example > Skin whitening activity test and skin moisturizing activity test

Experimental Example 1 Whitening Activity Experiment

Experimental Example 1-1 Whitening Active Cell Experiment

Experimental Example 1-1-1 B16F10 Melanoma Cell Culture

B16F10 melanoma cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 100 units / ml penicillin, and 100 units / ml streptomycin at 37 ° C. and maintained at 5% CO ₂ . Subculture was performed once.

Experimental Example 1-1-2 Cytotoxicity Evaluation

Toxicity to the B16F10 cells of the Example extract was evaluated using the MTT assay. B16F10 cells (2.0 × 10 ⁴ cells / ml) were dispensed into 24 wells and cultured for 18 hours, followed by further treatment for 24 hours by treating samples (40 μg / ml) (extracts of the respective examples and comparative examples). 200 µl of the MTT solution prepared at a concentration of 5 mg / ml PBS was added thereto, followed by further 4 hours of incubation. The culture solution was removed, and 200 µl of DMSO was added to each well, and the absorbance of the formazan produced by MTT reduction was measured at 570 nm.

The results are shown in Table 1 below as a percentage of the control group (untreated group).

Cell survival rate (%) division Cell survival rate (%) Control 100 Horse Placenta Extract of <Example 1> 105.2 Horse oil of <Example 2> 97.4 Horse Placenta Extract of <Comparative Example> 104.7

The results of Table 1 show that the enzyme treated placenta extract of <Example 1>, horse oil of <Example 2> and acid treated horse placenta extract of <Comparative Example> did not show cytotoxicity.

Experimental Example 1-1-3 Melanin Production Inhibitory Activity Experiment

After dispensing B16F10 cells (2.0 × 10 ⁴ cells / ml) into 24 wells, each extract of the above Examples and Comparative Examples (40 μg / ml) or the same mixture of Examples 1 and 2 (40 μg / ml) was added. The cells were treated with α-MSH (50 nM), a melanin-forming stimulant, and cultured for 72 hours. Cells were harvested and 1N NaOH was added to completely lyse the cells, and then absorbance was measured by ELISA at 405 nm.

The results are shown in Table 2 below as a percentage of the control group (α-MSH treated group).

Melanin production inhibition rate (%) division Cell survival rate (%) Control 100 Horse Placenta Extract of <Example 1> 72.3 ± 3.54 * Horse oil of < Example 2> 68.4 ± 2.72 * Mixture of <Example 1> and <Example 2> 46.4 ± 5.25 * Horse Placenta Extract of <Comparative Example> 87.3 ± 2.53 * * p <0.05,

In Table 2, the enzyme treated horse placenta extract of <Example 1>, horse oil of <Example 2> and acid treated horse placenta extract of <Comparative Example> both have melanin production inhibitory activity, The mixture of the horse placenta extract of Example 1 and horse oil of <Example 2> has a high melanogenesis inhibitory activity. In addition, the enzyme-treated horse placenta extract of <Example 1> has higher melanin production inhibitory activity than the acid-treated horse placenta extract of <Comparative Example>.

And when combined with the result of [Table 1], it can be seen that the result of [Table 2] is not the result of cytotoxicity.

Experimental Example 1-2 Whitening Effect Clinical Experiment

For the clinical experiment of the whitening effect was prepared a cosmetic composition with the ingredients and contents as shown in Table 3 below.

Ingredients and content (% by weight) of lotion ingredient Preparation Example 1 Production Example 2 Production Example 3 Preparation Example 4 Horse Placenta Extract of <Example 1> 5.0 - - - Horse oil of <Example 2> - 5.0 - - Equivalent Mixture of <Example 1> and <Example 2> - - 5.0 - Horse Placenta Extract of <Comparative Example> - - - 5.0 Complex fruit acid 2.0 glycerin 3.0 1.3-butylene glycol 3.0 Fiji 1500 0.5 Allantoin 0.1 DL-Panthenol 0.3 EDTA-2Na 0.02 Benzopinone-9 0.04 Sodium hyalonate 3.0 ethanol 7.0 Octyldodec-16 0.2 Polysorbate 20 0.2 Preservative, fragrance, coloring a very small amount Purified water Balance Sum 100

Here, the cosmetic composition of the comparative example was used instead of the extract of the examples and comparative examples, the cosmetic composition containing the amount of purified water.

The whitening effect test divided 100 women in their 20's and 30's into 5 groups (20 in each group), and allowed each group to use for 8 weeks according to the usual usage method. Excellent (5 points), excellent (4 points), normal (3 points), bad (2 points), very bad (1 point)), and the results are shown in Table 4 below as average scores. It was.

Whitening effect evaluation result division result Production Example 1 3.84 Preparation Example 2 3.95 Preparation Example 3 4.68 Preparation Example 4 3.52  Comparative example 3.12

The results of [Table 4] are similar to the results of [Table 2], but the case of the cosmetic composition containing the placental extract of the enzyme-treated horse placenta extract of <Example 1> and <Example 2> (Preparation Example 3) showed the highest whitening activity.

Experimental Example 2 Evaluation of Moisturizing Effect

100 women in their 20s to 30s are divided into 5 groups (20 in each group), and each group is allowed to use for 8 weeks according to the usual method of use. The composition was allowed to apply for 8 weeks according to the usual method of use.

After 8 weeks, TEWL (higher in dry skin) (Tewameter TM300, C + K electronic GmbH. Germany) and moisture content (lower in dry skin) (Corneometer CM825, C + K electronic GmbH. Germany).

The results are shown in [Table 5] and [Table 6] below with mean ± standard deviation.

TEWL division 0 parking Week 8 Preparation Example 1 17.27 ± 5.42 16.12 ± 4.52 Production Example 2 19.03 ± 4.72 17.31 ± 5.93 * Production Example 3 18.42 ± 4.23 14.16 ± 3.63 * Preparation Example 4 17.68 ± 5.29 16.79 ± 5.83 Comparative example 18.52 ± 6.32 18.26 ± 4.15 * p <0.05

Moisture content division 0 parking Week 8 Preparation Example 1 37.2 ± 6.26 38.3 ± 7.15 Production Example 2 37.3 ± 8.93 38.5 ± 5.42 Production Example 3 37.6 ± 4.28 42.5 ± 7.32 * Preparation Example 4 38.2 ± 7.36 38.9 ± 10.2 Comparative example 37.4 ± 6.54 37.5 ± 5.29 * p <0.05

Looking at the results of the <Table 5> and <Table 6>, the cosmetic lotion composition containing the enzyme-treated horse placenta extract of <Example 1>, horse oil of <Example 2> and acid-treated horse placenta extract of <Comparative Example> (Preparation Examples 1 to 4, respectively) Although there was no statistically significant difference, the TEWL decreased and the water content tended to increase. And in the case of the cosmetic composition of the comparative example does not contain the extract did not have a special effect on improving the skin moisturizing.

And the lotion composition (Preparation Example 3) containing both the horse placenta extract of <Example 1> and horse oil of <Example 2> showed statistical significance in the skin moisturizing effect.

Claims (8)

A skin external preparation composition for skin whitening comprising at least one of an extract and horse oil obtained by hydrolyzing a horse placenta with a protease.
The method of claim 1,
The protease is a papaya skin external composition, characterized in that the papaya.
The method according to claim 1 or 2,
The composition for skin whitening for skin whitening, characterized in that the composition is a cosmetic composition.
The method according to claim 1 or 2,
The composition for skin whitening for skin whitening, wherein the composition is a soap composition.
A skin external preparation composition for moisturizing skin comprising at least one of extract and horse oil obtained by hydrolyzing horse placenta with protease.
The method of claim 5,
The proteolytic enzyme composition for external skin moisturizing skin, characterized in that papa.
The method according to claim 5 or 6,
The composition for external skin moisturizing skin, characterized in that the composition is a cosmetic composition.
The method according to claim 5 or 6,
The composition for external skin moisturizing skin, characterized in that the composition is a soap composition.