KR102155246B1 - Composition for improving skin conditions comprising Dendropanax Morbifera extracts-metal nanoparticles complex - Google Patents

Abstract

The present invention relates to a composition for improving skin condition comprising a hwangchil tree extract-metal salt nanoparticle complex, and specifically, for skin whitening comprising a hwangchil tree extract and a metal salt-containing metal salt nanoparticle complex as an active ingredient Or it relates to a cosmetic composition for preventing skin aging, an external preparation for the skin, a quasi-drug composition, a pharmaceutical composition, and a method for whitening or preventing skin aging.
Hwangchil tree extract-metal salt nanoparticle complex according to the present invention has excellent antioxidant effect, tyrosinase activity inhibitory effect, and melanin production inhibitory effect, and does not show toxicity to cells, so skin whitening, skin aging, skin pigmentation, etc. It can be useful and safely used in the prevention, improvement or treatment of

Description

Composition for improving skin conditions comprising Dendropanax Morbifera extracts-metal nanoparticles complex}

The present invention relates to a composition for improving skin condition comprising a hwangchil tree extract-metal salt nanoparticle complex, and specifically, for skin whitening comprising a hwangchil tree extract and a metal salt-containing metal salt nanoparticle complex as an active ingredient Or it relates to a cosmetic composition for preventing skin aging, an external preparation for the skin, a quasi-drug composition, a pharmaceutical composition, and a method for whitening or preventing skin aging.

Human skin color is genetically determined according to the concentration and distribution of melanin in the skin, but is also affected by environmental or physiological conditions such as solar ultraviolet rays, fatigue, and stress. The chemical action of melanin production has been reported in many papers so far, and tyrosine as a precursor is converted to dihydroxyphenylalanine (DOPA) by tyrosinase, and then It is known that it is finally converted to melanin through a substance called dopaquinone. The melanin synthesized in this way darkens the skin color, and research on a skin whitening method to brighten the skin color by inhibiting melanin synthesis due to health or beauty needs is actively underway, and in relation to it, a skin whitening cosmetic composition containing a watermelon endothelial extract ( Korean Patent Publication No. 10-1325464), a composition for improving skin condition containing plant extracts such as marshmallow, calendula, and St. Mary's (Korean Laid-Open Patent Publication No. 10-2015-0112103) has been developed. .

In addition, commonly known whitening ingredients include substances that inhibit the activity of tyrosinase, such as kojic acid or arbutin, vitamin C (L-Ascorbic acid) or derivatives thereof, and various There are plant extracts. By inhibiting the synthesis of melanin pigments, they can not only realize skin whitening by brightening skin tone, but also improve skin hyperpigmentation such as spots and freckles caused by ultraviolet rays, hormones or genetics. However, when applied to the skin, there is a problem in that the amount of use is limited due to safety issues such as irritation and redness, or the effect is insignificant, so that a practical effect cannot be expected.

The deposition of melanin pigments as well as skin aging are caused by various factors. In particular, radicals of free radicals produced in the body's metabolic process are a representative factor. As radicals accumulate in the skin cells, the skin becomes thinner and less elastic, leading to wrinkles and aging of the skin. To prevent this, antioxidants such as vitamin C, vitamin E, beta-carotene, which are precursors of vitamin A, and retinoic acid, which are derivatives of vitamin A, are actively used. They are in the spotlight as active ingredients in health functional foods and cosmetics. In particular, vitamin C plays an important role in skin whitening by maintaining the synthesis and elasticity of collagen in the skin dermis and preventing melanin pigment from being deposited.

On the other hand, Hwangchil tree is a plant belonging to the umbel family Araliaceae, and essential oil components such as dendropanoxide, saponins, α-Cubebene, β-Elemene, β-cell It is known that various physiologically active ingredients such as linen (β-Selinene), α-Muurolene, Germacrene D, and β-Sitosterol are contained. Accordingly, it is used as a useful medicinal material in traditional medicine, as it exhibits effects such as blood purification, liver function improvement, antioxidant activity, hard tissue (bone and tooth) regeneration, immunity strengthening, nerve stabilization, antibacterial activity, and anticancer activity. However, the whitening effect of Hwangchil wood was not known, and the whitening effect of gold or silver nanoparticles prepared using it was not known.

Under this background, the inventors of the present inventors have conducted a thorough study to develop a substance that promotes skin whitening and prevents skin aging. As a result, the Hwangchil tree extract-metal salt nanoparticle complex prepared using Hwangchil tree leaf extract has an excellent whitening effect as well as By confirming that it exhibits excellent antioxidant activity, the present invention was completed.

One object of the present invention is to provide a cosmetic composition for skin whitening or skin aging, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

Another object of the present invention is to provide a skin external preparation for skin whitening or skin aging prevention, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

Another object of the present invention is to provide a quasi-drug composition for skin whitening or skin aging, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating skin pigmentation, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

Another object of the present invention is to administer a composition comprising a Hwangchil tree extract and a metal salt-containing Hwangchil tree extract-metal salt nanoparticle complex as an active ingredient to an individual other than humans, skin whitening or skin aging It is to provide a method of prevention.

In order to achieve the above object, one aspect of the present invention provides a cosmetic composition for skin whitening or skin aging, comprising a hwangchil tree extract and a metal salt-containing hwangchil tree extract-metal salt nanoparticle complex as an active ingredient.

In the present invention, it was confirmed that the Hwangchil tree extract-metal salt nanoparticle complex prepared using Hwangchil tree extract has excellent antioxidant effect, tyrosinase activity inhibitory effect, and melanin production inhibitory effect, and further does not show toxicity to cells. .

The term of the present invention, "Hwangchil tree" ( Dendropanax morbifera ) means a plant belonging to the umbel family Araliaceae. Hwangchil tree relaxes the mind and body and suppresses various weights, strengthens the kidneys for men, and treats menstrual irregularities to women, and promotes the growth of immune cells to strengthen the early immune system and biological defense systems. It is used for prevention and treatment. However, it was not known about the whitening effect of the metal salt nanoparticles prepared using the Hwangchil tree extract.

The term "hwangchil tree extract" of the present invention refers to a component or material belonging to the hwangchil tree obtained by a conventional method using a solvent of hwangchil tree, and in the present invention, the hwangchil tree extract may be a hwangchil tree leaf extract.

In addition, in the present invention, the Hwangchil tree extract is an extract obtained by extracting the Hwangchil tree, a dilution or concentrate of the extract, a dried product obtained by drying the extract, a preparation or purified product of the extract, or a mixture thereof. , The extract itself and all formulations that can be formed using the extract are included. The method or extraction solvent for extracting the Hwangchil tree is not particularly limited, and any method or solvent known in the art may be used.

Specifically, the extraction solvent is not particularly limited as long as the Hwangchil tree extract can be prepared, but water, C1 to C4 alcohol or a mixture thereof may be used, specifically ethanol, methanol or water, and , More specifically, distilled water may be used.

As used herein, the term "metal salt" refers to a compound added to prepare metal salt nanoparticles. In the present invention, the metal salt may be a gold salt or a silver salt. Specifically, the gold salt may be tetrachlorogold (III) acid (HAuCl ₄ ), NaAuCl ₃ , or AuCl ₃ , but is not limited thereto, and the silver salt is AgBF ₄ , AgCF ₃ SO ₃ , AgClO ₄ , AgNO ₃ , AgPF ₆ or Ag (CF ₃ COO), but is not limited thereto.

In the present invention, the "metal salt nanoparticles" are made of metal and mean ultrafine particles having a size of 1 to 100 nm. In the present invention, the metal salt nanoparticles may be gold salt nanoparticles or silver salt nanoparticles.

The term "hwangchil tree extract-metal salt nanoparticle complex" of the present invention refers to a form in which the hwangchil tree extract and the metal salt nanoparticles are combined or connected to the metal salt nanoparticles. Since the Hwangchil tree extract-metal salt nanoparticle complex of the present invention contains the physiologically active substance of the Hwangchil tree extract, it may exhibit a better skin whitening or skin aging prevention effect compared to general metal salt nanoparticles prepared by a chemical method. .

For the purposes of the present invention, the Hwangchil tree extract-metal salt nanoparticle complex may be prepared using a Hwangchil tree extract.

In addition, in the present specification, the Hwangchil tree extract-gold salt nanoparticle complex may be referred to as'gold nanoparticles' or D-AuNPs', and the Hwangchil tree extract-silver salt nanoparticle complex is'silver nanoparticles' Alternatively, it may be referred to as'D-AgNPs'.

As used herein, the term "skin whitening" means to brighten skin tone and improve skin hyperpigmentation such as spots and freckles caused by various factors such as ultraviolet rays, hormones, or genetics.

The term "skin aging" as used herein refers to symptoms such as decreased skin elasticity, reduced gloss, wrinkle formation, weakened regeneration, or severe dryness, and "skin aging prevention" inhibits or inhibits symptoms such as skin aging. Or it means relieving the symptoms of skin aging that have already appeared.

In a specific embodiment of the present invention, gold or silver nanoparticles prepared using the Hwangchil tree leaf extract have excellent antioxidant activity (Figure 1), tyrosinase activity inhibitory effect (Figures 2 and 3), and melanin production inhibition It was confirmed that the effect was shown (FIG. 4), and further, it was not toxic to the cells (FIG. 5), and the survival rate of the cells could be increased by scavenging the radicals of the cells reduced by H ₂ O ₂ (FIG. 6). In addition, it was confirmed that the effect of the gold or silver nanoparticles as described above was similar to or better than that of ascorbic acid or arbutin, which are positive controls. This suggests that the cosmetic composition of the present invention comprising the Hwangchil tree extract-metal salt nanoparticle complex can be usefully used for skin whitening or skin aging prevention.

The cosmetic composition of the present invention may contain 0.0001 to 80% by weight, specifically 0.0005 to 70% by weight, and more specifically 0.001 to 60% by weight, based on the total weight of the Hwangchil tree extract-metal salt nanoparticle complex. If the content is less than 0.0001% by weight, it is not preferable because the original intended effect cannot be sufficiently achieved, and if it exceeds 80% by weight, it is uneconomical because no obvious increase in the effect is expected as much as the increasing content. It can cause problems.

In addition, the cosmetic composition of the present invention is used for the skin, softening lotion (nutrition emulsion), astringent lotion, nutritional lotion, nutritional cream, massage cream, essence, pack, patch for skin adhesion, gel for skin adhesion, powder, ointment , Suspension, emulsion, spray, or may be prepared in the form of a cosmetic composition, such as a cosmetic solution, but is not limited thereto. In addition, the cosmetic composition may be made of skin-contacting materials such as cosmetics, soaps, detergents, and fibers.

In addition, the cosmetic composition is a cosmetic composition such as ordinary ingredients, such as oil, water, surfactants, moisturizers, lower alcohols, thickeners, chelating agents, pigments, preservatives, fragrances, etc., which are mixed in general skin cosmetics as needed. It may further include an acceptable carrier, but is not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

Specifically, when the formulation of the present invention is an ointment, paste, cream or gel, as a carrier component, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, Zinc oxide or the like may be used, but is not limited thereto. These may be used alone or in combination of two or more.

In addition, when the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component. In particular, in the case of a spray, additional chlorofluoro It may include a propellant such as lohardrocarbon, propane/butane or dimethyl ether, but is not limited thereto. These may be used alone or in combination of two or more.

In addition, when the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, and the like can be used, and in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or sour. Non-fat fatty acid esters may be used, but are not limited thereto. These may be used alone or in combination of two or more.

In addition, when the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester , Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used, but is not limited thereto. These may be used alone or in combination of two or more.

In addition, when the formulation of the present invention is a soap, as a carrier component, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolyzate, isethionate, a lanolin derivative, an aliphatic alcohol, vegetable oil, glycerol, sugar, etc. May be used, but is not limited thereto. These may be used alone or in combination of two or more.

Another aspect of the present invention provides a skin external preparation for skin whitening or skin aging prevention, comprising a hwangchil tree extract and a metal salt-containing metal salt nanoparticle complex as an active ingredient.

At this time, the terms "Hwangchil tree extract", "metal salt", "Hwangchil tree extract-metal salt nanoparticle complex", "skin whitening", "skin aging" and "prevention" are as described above.

The term "external skin preparation" of the present invention is a concept encompassing all substances generally used for external application of the skin, and non-limiting examples of the external preparation for skin include plasters, lotions, and liniment Liniments, liquid and solutions, aerosols, extracts, ointments, fluid extracts, emulsions, suspensions, capsules ( capsules), creams, soft, hard gelatin capsules, patches or sustained release agents, etc., but are not limited thereto.

The external preparation for skin according to the present invention may be a parenteral administration formulation formulated in a solid, semi-solid or liquid form by adding a commercially available inorganic or organic carrier, excipient, and diluent. The formulation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions, and emulsions, but is not limited thereto.

Carriers, excipients, and diluents that may be included in the external preparation include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

In the composition for external application for skin according to each formulation, components other than the Hwangchil tree extract-metal salt nanoparticle complex of the present invention can be appropriately selected and blended by those skilled in the art without difficulty depending on the formulation or purpose of use of other skin external preparations. Synergistic effects may occur when applied simultaneously with other raw materials.

In a specific embodiment of the present invention, gold or silver nanoparticles prepared using the Hwangchil tree leaf extract have excellent antioxidant activity (Figure 1), tyrosinase activity inhibitory effect (Figures 2 and 3), and melanin production inhibition It was confirmed that the effect was shown (FIG. 4), and further, it was not toxic to the cells (FIG. 5), and the survival rate of the cells could be increased by scavenging the radicals of the cells reduced by H ₂ O ₂ (FIG. 6). In addition, it was confirmed that the effect of the gold or silver nanoparticles as described above was similar to or better than that of ascorbic acid or arbutin, which are positive controls. This suggests that the external preparation for skin of the present invention comprising the Hwangchil tree extract-metal salt nanoparticle complex can be usefully used for skin whitening or skin aging prevention.

Another aspect of the present invention provides a quasi-drug composition for skin whitening or skin aging prevention, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

At this time, the terms "Hwangchil tree extract", "metal salt", "Hwangchil tree extract-metal salt nanoparticle complex", "skin whitening", "skin aging" and "prevention" are as described above.

The term "quasi-drug" of the present invention refers to items that are less effective than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals, for example, according to the Pharmaceutical Affairs Act. Quasi-drugs exclude items used for pharmaceutical purposes, and include products used to treat or prevent diseases of humans/animals, and products that have mild or no direct action on the human body.

The quasi-drug composition of the present invention may be prepared as one or more selected from the group consisting of a body cleanser, foam, soap, mask, ointment, cream, lotion, essence, and spray, but is not limited thereto.

When the Hwangchil tree extract-metal salt nanoparticle complex according to the present invention is used as a quasi-drug additive, the composition may be added as it is or may be used with other quasi-drugs or quasi-drug components, and may be appropriately used according to a conventional method. The mixing amount of the active ingredient may be appropriately determined according to the purpose of use.

In a specific embodiment of the present invention, gold or silver nanoparticles prepared using the Hwangchil tree leaf extract have excellent antioxidant activity (Figure 1), tyrosinase activity inhibitory effect (Figures 2 and 3), and melanin production inhibition It was confirmed that the effect was shown (FIG. 4), and further, it was not toxic to the cells (FIG. 5), and the survival rate of the cells could be increased by scavenging the radicals of the cells reduced by H ₂ O ₂ (FIG. 6). In addition, it was confirmed that the effect of the gold or silver nanoparticles as described above was similar to or better than that of ascorbic acid or arbutin, which are positive controls. This suggests that the quasi-drug product of the present invention containing the Hwangchil tree extract-metal salt nanoparticle complex can be usefully used for skin whitening or skin aging prevention.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating skin pigmentation, comprising a hwangchil tree extract and a metal salt-containing metal salt-metal salt nanoparticle complex as an active ingredient.

In this case, the description of the terms "Hwangchil tree extract", "metal salt" and "Hwangchil tree extract-metal salt nanoparticle complex" is as described above.

As used herein, the term "skin pigmentation" refers to a hyperpigmented macule caused by an increase in melanin on the skin, nails, or the mucous membrane surrounding the oral cavity or nasal cavity, and is also referred to as hyperpigmentation.

The term "prevention" of the present invention means any action of inhibiting or delaying skin pigmentation by administration of the pharmaceutical composition of the present invention.

The term "treatment" of the present invention means any action in which skin pigmentation is improved or beneficially altered by administration of the pharmaceutical composition of the present invention.

In a specific embodiment of the present invention, gold or silver nanoparticles prepared using the Hwangchil tree leaf extract have excellent antioxidant activity (Figure 1), tyrosinase activity inhibitory effect (Figures 2 and 3), and melanin production inhibition It was confirmed that the effect was shown (FIG. 4), and further, it was not toxic to the cells (FIG. 5), and the survival rate of the cells could be increased by scavenging the radicals of the cells reduced by H ₂ O ₂ (FIG. 6). In addition, it was confirmed that the effect of the gold or silver nanoparticles as described above was similar to or better than that of ascorbic acid or arbutin, which are positive controls. This suggests that the pharmaceutical composition of the present invention containing the Hwangchil tree extract-metal salt nanoparticle complex can be usefully used for preventing or improving skin pigmentation.

The pharmaceutical composition of the present invention is in the form of tablets, pills, powders, granules, capsules, suspensions, liquids, emulsions, syrups, aerosols, sterile injectable solutions, etc. according to conventional methods for preventing and treating skin pigmentation. It can be formulated as.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations are prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. Can be. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc may be used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc., and various excipients, such as humectants, sweeteners, fragrances, and preservatives, in addition to water and liquid paraffin, which are commonly used simple diluents. Can be used.

Formulations for parenteral administration may include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized formulations, suppositories, and the like. As the non-aqueous solvent and the suspension solvent, vegetable oils such as propylene glycol, polyethylene glycol, olive oil, and injectable esters such as ethyl oleate may be used.

In addition, the pharmaceutical composition of the present invention may further include a carrier, excipient or diluent. Carriers, excipients or diluents include lactose, textrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, hydroxy Mineral oils such as propyl methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, flophilhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and silicon dioxide can be used. have.

The specific dosage of the pharmaceutical composition according to the present invention depends on factors such as formulation method, patient's condition and weight, patient's sex, age, degree of disease, drug type, route and duration of administration, excretion rate, response sensitivity, etc. It can be selected in various ways by those skilled in the art, and the dosage and frequency do not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, and humans through various routes. All modes of administration can be expected and can be administered, for example by oral, intravenous, intramuscular or subcutaneous injection.

Another aspect of the present invention is a skin whitening or skin aging comprising administering a composition comprising a hwangchil tree extract and a metal salt-containing metal salt nanoparticle complex as an active ingredient to an individual other than a human. Provides a method of prevention.

At this time, the descriptions of the terms "Hwangchil tree extract", "metal salt", "Hwangchil tree extract-metal salt nanoparticle complex", "skin whitening", "skin aging" and "prevention" are as described above.

The term "administration" of the present invention means introducing the composition of the present invention to a patient by any suitable method.

As used herein, the term "individual" refers to all animals such as rats, mice, livestock, including humans, which have or may develop skin pigmentation or skin aging. Specifically, it may be a mammal including a human.

The administration route of the composition may be administered through any general route as long as it can reach the target tissue. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, but are not limited thereto. Due to the nature of the composition of the present invention that is effective in improving skin whitening or skin aging, the route of administration of the composition may be applied to the skin and administered.

The composition of the present invention may be administered daily or intermittently, and the number of administrations per day may be administered once or divided into 2 to 3 times. In addition, the composition of the present invention may be used alone or in combination with other drug treatments for skin whitening or skin aging. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and can be easily determined by a person skilled in the art.

In a specific embodiment of the present invention, gold or silver nanoparticles prepared using the Hwangchil tree leaf extract have excellent antioxidant activity (Figure 1), tyrosinase activity inhibitory effect (Figures 2 and 3), and melanin production inhibition It was confirmed that the effect was shown (FIG. 4), and further, it was not toxic to the cells (FIG. 5), and the survival rate of the cells could be increased by scavenging the radicals of the cells reduced by H ₂ O ₂ (FIG. 6). In addition, it was confirmed that the effect of the gold or silver nanoparticles as described above was similar to or better than that of ascorbic acid or arbutin, which are positive controls. This suggests that the method for whitening or preventing skin aging of the present invention using a composition containing the Hwangchil tree extract-metal salt nanoparticle complex can achieve a desired effect to an excellent degree.

Hwangchil tree extract-metal salt nanoparticle complex according to the present invention has excellent antioxidant effect, tyrosinase activity inhibitory effect, and melanin production inhibitory effect, and does not show toxicity to cells, so skin whitening, skin aging, skin pigmentation, etc. It can be useful and safely used in the prevention, improvement or treatment of

1 is a graph showing the DPPH radical scavenging activity of gold nanoparticles (D-AuNPs) or silver nanoparticles (D-AgNPs) prepared using a hwangchil tree leaf extract according to an embodiment of the present invention. Vit. C (vitamin C) refers to ascorbic acid, which is a positive control.
Figure 2 is a graph showing the inhibition rate of mushroom tyrosinase activity in vitro of the D-AuNPs or D-AgNPs. DE stands for Hwangchil tree leaf extract, and Arbutin stands for arbutin, a positive control.
3 is a graph showing the tyrosinase activity of cells reduced by the D-AuNPs. α-MSH (α-Melanocyte-stimulating hormone) refers to melanin formation stimulating hormone, DE refers to Hwangchil tree leaf extract, and Arbutin refers to arbutin, a positive control. ### means a negative control for comparing tyrosinase activity, and *, ** and *** mean that statistical significance is p <0.05, p <0.01, and p <0.001 with respect to the negative control. .
4 is a graph showing the amount of melanin in cells reduced by the D-AuNPs. α-MSH stands for melanin formation stimulating hormone, DE stands for Hwangchil tree leaf extract, and Arbutin stands for arbutin, a positive control. ## means a negative control for comparing tyrosinase activity, and *, ** and *** mean that statistical significance is p <0.05, p <0.01, and p <0.001 with respect to the negative control.
Figure 5 is a graph showing the cytotoxicity of the D-AuNPs, showing the survival rate of HDF (Human dermal fibroblast) or B16 (murine melanoma B16BL6) cells when D-AuNPs are treated. DE refers to Hwangchil tree leaf extract, and RA (Retinoic acid) refers to retinoic acid, a positive control. A-C relates to HDF, and D-F relates to B16 cells; A and D refer to the results of treatment with D-AuNPs for 24 hours, B and E for 48 hours treatment with D-AuNPs, and C and F refer to the results of treatment with D-AuNPs for 72 hours.
6 is a graph showing the cell radical scavenging activity of the D-AuNPs, showing the activity of increasing cell viability against cells damaged by the radical. Pre-treatment refers to pretreatment of H ₂ O ₂ , Co-treatment refers to co-treatment of H ₂ O ₂ , and Post-treatment refers to post-treatment conditions of H ₂ O ₂ . AA refers to ascorbic acid as a positive control, and DE refers to Hwangchil tree leaf extract. ### means a negative control for comparing tyrosinase activity, and *, ** and *** mean that statistical significance is p <0.05, p <0.01 and p <0.001 with respect to the negative control. .

Hereinafter, the present invention will be described in more detail through the following examples. These examples are for illustrative purposes only and are not to be construed as limiting the scope of the present invention by these examples.

Manufacturing example 1. Method for producing gold or silver nanoparticles using Hwangchil tree extract

Manufacturing example 1-1. Method for producing Hwangchil tree extract

Dendropanax Morbifera ) In order to prepare gold or silver nanoparticles by an eco-friendly method using an extract, Hwangchil tree extract was first prepared.

Specifically, the leaves of the Hwangchil tree were washed with deionized water, cut into small pieces, and then dried. 5 g of the dried leaves were placed in a 1 liter beaker containing distilled water and heated for 30 minutes. After cooling at room temperature, it was centrifuged at 5,000 rpm for 10 minutes, and filtered through filter paper to obtain a hwangchil tree leaf extract.

Manufacturing example 1-2. Gold nanoparticles (D- AuNPs ) Manufacturing method

In order to prepare gold nanoparticles (D-AuNPs) by an eco-friendly method using Hwangchil tree extract, the following method was performed.

Specifically, 5 ml of the Hwangchil tree leaf extract prepared in Preparation Example 1-1 was mixed with 45 ml of deionized water, and the final concentration was 1 mM, so that tetrachlorogold(III) acid (HAuCl _4· 3H ₂ O ) An aqueous solution was added to the solution. After incubating the solution at 80° C. for 1 hour, it was confirmed whether gold nanoparticles were generated by observing the color change of the reaction mixture. Thereafter, the gold nanoparticles were collected by centrifugation at 16,000 rpm for 20 minutes, washed several times with deionized water, and dried and used.

Manufacturing example 1-3. Silver nanoparticles (D- AgNPs ) Manufacturing method

In order to prepare silver nanoparticles (D-AgNPs) in an eco-friendly way, the following method was performed.

Specifically, 5 ml of the hwangchil tree leaf extract prepared in Preparation Example 1-1 was mixed with 45 ml of deionized water, and the final concentration was 1 mM, and an aqueous silver nitrate (AgNO ₃ ) solution was added to the solution. After incubating the solution at 80° C. for 1 hour, it was confirmed whether silver nanoparticles were generated by observing the color change of the reaction mixture. Thereafter, the silver nanoparticles were recovered by centrifugation at 16,000 rpm for 20 minutes, washed several times with deionized water, and dried and used.

Example 1. Gold or silver nanoparticles (D- AuNPs Or check the antioxidant activity of D-AgNPs)

In order to confirm the effect of improving the skin condition of the gold or silver nanoparticles (D-AuNPs or D-AgNPs) prepared through Preparation Example 1, the antioxidant activity was first confirmed.

Specifically, DPPH radical scavenging activity was confirmed. 180 µl of 0.1 mM methanol solution of DPPH was added to 20 µl of D-AuNPs or D-AgNP solutions having different concentrations of 10 to 500 µg/ml, and absorbance at 517 nm using a UV-Vis spectrophotometer after 30 minutes The value was measured, and DPPH radical scavenging activity was calculated through Equation 1 below. Ascorbic acid was used as a positive control.

[Equation 1]

[(Absorbance of DPPH-absorbance of sample-absorbance of control)/absorbance of DPPH]×100

At this time, the absorbance of the sample is the absorbance of the DPPH solution containing nanoparticles, and the absorbance of the control is the absorbance of the D-AuNPs or D-AgNPs solution.

As a result, as can be seen in Figure 1, it was confirmed that D-AuNP or D-AgNP exhibited DPPH radical scavenging activity in a dose-dependent manner, and it was confirmed that it did not show much difference from the DPPH scavenging activity of ascorbic acid used as an antioxidant. .

Example 2. D- AuNPs Or D- AgNPs Confirmation of tyrosinase activity inhibitory effect in vitro

In order to confirm the effect of improving the skin condition of D-AuNPs or D-AgNPs prepared in Preparation Example 1, the effect of inhibiting tyrosinase activity in vitro was confirmed.

Specifically, it was confirmed that the inhibitory effect on the activity of mushroom tyrosinase (mushroom tyrosinase). After dissolving 0.05 ml of 2 mM L-DOPA in a phosphate buffer solution (50 mM, pH 6.8), 0.05 ml of D-AuNP, D-AgNP or Hwangchil tree leaf extract (DE) sample was put into a 96-well microplate and mushroom tiro. 0.05 ml of synase (200 U/ml) were mixed. The mixture was incubated at 37° C. for 30 minutes, and the optical density was measured at 492 nm using an ELISA reader (Bio-Tek Instrument, Vermont, USA), and the tyrosinase activity inhibition rate was calculated through Equation 2 below. Arbutin was used as a positive control.

[Equation 2]

Tyrosinase activity inhibition rate (%) = [(A-B)/(C-D)]×100

Here, A is the absorbance of the reaction mixture containing the test sample (D-AuNP, D-AgNP or DE) and mushroom tyrosinase, B is the absorbance of the reaction mixture with the test sample but no mushroom tyrosinase, C is the mushroom The absorbance of the reaction mixture with tyrosinase but no test sample, and D is the absorbance of a well containing only L-DOPA.

On the other hand, all experiments according to the present invention were independently performed three times, and all data were expressed as mean ± standard deviation (S.D.). The average value of the group treated with the sample (D-AuNP, D-AgNP or positive control) was compared with the average value of the group without treatment, and their statistical significance was confirmed using Student's t-test. I did.

Through the above results, it was found that D-AuNP or D-AgNP exhibited an in vitro tyrosinase inhibitory activity similar to arbutin, and thus would exhibit an excellent skin whitening effect.

Example 3. Confirmation of skin whitening effect of D- AuNPs

Example 3-1. Confirmation of the inhibitory effect of cell tyrosinase activity

In order to confirm the effect of improving the skin condition of D-AuNPs prepared in Preparation Example 1, the inhibitory effect on the activity of cells against tyrosinase was confirmed.

Specifically, mouse melanoma-forming cells, B16 (murine melanoma B16BL6) cells, were cultured in a 100 mm plate at a density of 4 × 10 ⁵ cells/well, and treated with 100 nM α-MSH (α-Melanocyte-stimulating hormone). Incubated for 24 hours. Thereafter, the cells were treated with D-AuNPs at a concentration of 1-100 μg/ml and cultured for 24 hours. After that, the medium was removed and lysed in a phosphate buffer (pH 6.9) containing 1% Triton X-100. The lysate was freeze-thaw at -80°C for 15 minutes and then kept at room temperature for 10 minutes. Thereafter, after centrifugation at 12,000 g for 15 minutes, a substrate solution (15 mM L-DOPA contained in 50 mM phosphate buffer solution (pH 7.1)) was added to 90 μl of the lysate supernatant and incubated at 37° C. for 1 hour. , Absorbance was measured at 475 nm using an ELISA reader.

As a result, as can be seen in Figure 3, it was confirmed that D-AuNPs showed superior cellular tyrosinase inhibitory activity than arbutin or hwangchil tree leaf extract. Specifically, 100 μg/ml In the case of treatment with a concentration of arbutin, the tyrosinase activity of B16 cells is about 80%, 25 μg/ml When the concentration of Hwangchil tree leaf extract is treated, the tyrosinase activity of B16 cells is about 70%, compared to 25 ㎍/㎖ Concentration When treated with D-AuNPs, it was confirmed that the tyrosinase activity of B16 cells was about 40%.

Through the above results, it was found that D-AuNP exhibited remarkably superior cellular tyrosinase inhibitory activity than arbutin or hwangchil tree leaf extract even at a low concentration, and thus it was found that it would exhibit excellent skin whitening effect.

Example 3-2. Confirmation of cell melanogenesis inhibitory effect

In order to confirm the effect of improving the skin condition of D-AuNPs prepared in Preparation Example 1, the effect of inhibiting the production of melanin in cells was confirmed.

Specifically, mouse melanin-forming cells, B16 cells, were cultured in a 100 mm plate at a density of 4 × 10 ⁵ cells/well, and cultured for 24 hours while being treated with 100 nM α-MSH. Thereafter, the cells were treated with D-AuNPs at a concentration of 1 to 100 μg/ml and cultured for 24 hours. After removing the medium and obtaining the cells, the cells were dissolved in 500 μl of 1 N NaOH containing 10% DMSO and dissolved at 80° C. Stored at for 1 hour. Thereafter, the amount of melanin produced was confirmed by measuring the absorbance at 475 nm using an ELISA reader. At this time, the amount of melanin was confirmed by comparing it with a standard curve for synthetic melanin (0 to 100 µg/ml), and the amount of melanin generated was expressed as a percentage of the negative control without sample treatment.

As a result, as can be seen in Figure 4, it was confirmed that D-AuNPs exhibited superior melanin production inhibitory activity than arbutin or hwangchil tree leaf extract. Specifically, 100 μg/ml In the case of treatment with a concentration of arbutin, the amount of melanin in B16 cells is about 80%, compared to 25 ㎍/㎖ Concentration When processing D-AuNPs, it was confirmed that the amount of melanin in the generated B16 cells was about 70%.

Through the above results, it was found that D-AuNP exhibited excellent skin whitening effect, such as exhibiting superior melanogenesis inhibitory activity than arbutin even at low concentrations.

Example 4. Confirmation of skin safety of D- AuNPs

Example 4-1. Cytotoxicity check

In order to confirm the skin safety of D-AuNPs prepared through Preparation Example 1, toxicity to cells was confirmed.

Specifically, cytotoxicity was confirmed by measuring cell viability through the MTT assay. Human dermal fibroblast (HDF) and mouse melanin-forming cells, B16 cells, were inoculated in a 96-well plate at a density of 1×10 ⁵ cells/well and cultured for 24 hours, and had a cell density of 90%. At this time, 1 to 100 μg/ml of D-AuNP or RA (Retinoic acid, positive control) was treated and cultured for 24, 48 and 72 hours. Thereafter, 10 μl of MTT assay solution (5 mg/ml in PBS) was added to each well, incubated at 37° C. for 3 hours, and finally, 100 μl DMSO was added to dissolve the formed formazan crystals, and its absorbance Was measured at 570 nm with an ELISA reader.

As a result, as can be seen in Figure 5, for both human and mouse cells, D-AuNP does not show toxicity to cells even when cultured for 72 hours at a high concentration of 100 μg/ml, but rather increases cell viability. I did.

Through the above results, since D-AuNP does not show cytotoxicity, it was found that it can be useful and safely used as an active ingredient of a cosmetic composition.

Example 4-2. Confirmation of radical scavenging activity of damaged cells

In order to confirm the skin safety of D-AuNPs prepared in Preparation Example 1, the radical scavenging activity against cells damaged by H ₂ O ₂ , that is, the regeneration ability of cells was confirmed.

Specifically, human skin fibroblasts were inoculated into a 96-well plate at a density of 1 × 10 ⁵ cells/well and cultured at 37° C. for 24 hours. Cells were treated with Hwangchil tree leaf extract or 5, 10, and 25 ㎍/㎖ of D-AuNP, where H ₂ O ₂ was each pre-treatment, co-treatment, and post-treatment. treatment). As a pretreatment condition, cells were treated with a sample (Hwangchil tree leaf extract or D-AuNPs), and then H ₂ O ₂ was treated for 1 hour, and as a co-treatment condition, H ₂ O ₂ and the sample were simultaneously treated for 1 hour. As a post-treatment condition, H ₂ O ₂ was treated for 1 hour, rinsed, and then the sample was treated. Thereafter, the MTT assay was performed by the method according to Example 4-1. Ascorbic acid (100 μg/ml) was used as a positive control.

As a result, as can be seen in Figure 6, it was confirmed that D-AuNPs can increase the survival rate of cells by scavenging the radicals of the cells reduced by the H ₂ O ₂ treatment. Specifically, under all conditions of pre-treatment, co-treatment, and post-treatment, in the case of the negative control group treated with only H ₂ O ₂ , the viability of cells is about 80%, whereas in the case of D-AuNPs, the viability of cells is It was confirmed that it was about 90%, which was similar to the cells treated with ascorbic acid as a positive control.

Through the above results, it was found that D-AuNP can recover and regenerate radicals in cells damaged by H ₂ O ₂ , and thus, it was found that it would exhibit excellent improvement effects on skin aging caused by radicals.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. In this regard, the examples and experimental examples described above are illustrative in all respects and should be understood as non-limiting. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description and equivalent concepts are included in the scope of the present invention.

Claims (9)

Hwangchil tree extract-a cosmetic composition for skin whitening or skin aging, comprising a complex of gold nanoparticles as an active ingredient. The composition of claim 1, wherein the hwangchil tree extract is a hwangchil tree leaf extract. The method of claim 1, wherein the cosmetic composition is a softening lotion (nutrition emulsion), astringent lotion, nutrient lotion, nutrition cream, massage cream, essence, pack, skin adhesion patch, skin adhesion gel, powder, ointment, suspension, emulsion , The composition that is a formulation selected from a spray or cosmetic solution. The method of claim 1, wherein the cosmetic composition further comprises one or more carriers selected from the group consisting of oil, water, surfactant, moisturizer, lower alcohol, thickener, chelating agent, colorant, preservative and fragrance, Composition. Hwangchil tree extract-containing gold nanoparticle complex as an active ingredient, skin external preparation for the prevention or treatment of hyperpigmentation. Hwangchil tree extract-quasi-drug composition for the prevention or treatment of skin hyperpigmentation, comprising a gold nanoparticle complex as an active ingredient. Hwangchil tree extract-a pharmaceutical composition for the prevention or treatment of skin hyperpigmentation, comprising a complex of gold nanoparticles as an active ingredient. Hwangchil tree extract-comprising the step of administering a composition comprising the gold nanoparticle complex as an active ingredient to an individual other than human, dermatological pigmentation prevention or treatment method. The composition for external application for skin according to claim 5, wherein the skin hyperpigmentation syndrome is a spot or freckles.

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