KR102561146B1 - Composition for improving skin condition comprising herb extracts mixture - Google Patents

Abstract

The present invention relates to a composition comprising, as an active ingredient, a composite herbal medicine extract that is safe when applied to the skin and has excellent skin improvement effects. More specifically, the composition according to the present invention is a composition containing peach extract and white confectionery extract as active ingredients, and has excellent effects on skin improvement, including antiglycation effect, skin whitening effect, skin elasticity and wrinkle improvement effect, antioxidant effect and skin trouble improvement effect. In addition, the composition further comprising the white sheep resin extract in the composite extract exhibits an excellent effect in terms of fibroblast proliferation.

Description

Composition for improving skin condition comprising herb extracts mixture

The present invention relates to a composition containing as an active ingredient a composite herbal medicine extract that is safe when applied to the skin and has excellent skin improvement effects.

The skin is an important organ that protects the human body from the external environment and is in charge of various physiological functions such as a barrier function that prevents internal moisture and useful components from leaking out, a body temperature control function, and an excretion function. However, the activity of skin cells is reduced and the skin condition is deteriorated due to the following reasons.

Collagen and elastin present in the dermal layer of the skin have a great effect on the components of the skin by performing functions such as mechanical robustness of the skin, resistance of connective tissue, maintenance of tissue cohesiveness, and support of cell adhesion. It is known that such collagen is reduced by stress, aging, or photoaging caused by ultraviolet irradiation, and, on the other hand, elastin is distorted in its three-dimensional structure by an increased activity of elastin after exposure to ultraviolet rays. As a result, the skin tissue becomes loose and loses its elasticity.

As the cell function of the epidermis is lowered, the metabolism is not smooth, and the exfoliation of the dead skin cells does not occur easily. When UV rays are applied in a state in which the dead skin cells are excessively accumulated, the elasticity is lowered, resulting in the occurrence of wrinkles. Or, if the sebaceous film is not formed on the epidermis, moisture and sebum secretion decreases and the skin becomes dry, resulting in wrinkles.

When the skin is exposed to UV rays, melanin is synthesized to protect the skin. The synthesized melanin is transferred to the keratinocytes of the skin through melanosomes. These keratinocytes turn over every 28 days. Therefore, it is common for the generated melanin to be lost every 28 days by keratinocytes, but if the skin cell cycle is not well regulated due to stress, skin aging, etc., keratinocytes do not drop out in time, resulting in pigmentation such as spots, freckles, and age spots.

In the skin, sebum, sweat components, and cosmetic ingredients are decomposed into highly toxic substances by skin flora, which can cause skin irritation and inflammation. In addition, irritation caused by ultraviolet rays may cause skin trouble by increasing the production of nitric oxide (NO), an inflammatory mediator in the skin. Nitric oxide is mostly produced by iNOS, and iNOS is known to be rapidly induced by stimuli such as LPS and cytokines to produce excess NO.

Reactive oxygen species (ROS), also called toxic oxygen species, are toxic substances generated in cells by physiological processes such as respiration and are constantly produced and destroyed, and exist at about 3-5% in normal conditions. These reactive oxygen species exist as free radicals such as superoxide radical (O2-) and hydroxyl radical (HO+), or in the form of compounds with paired electrons such as hydrogen peroxide (H ₂ O ₂ ) or singlet oxygen (singlet radical). exists as Reactive oxygen species have the advantage of having a biological defense function of sterilizing bacteria in the physiological system, but generally cause harmful effects that cause diseases by causing oxidation in vivo. It has been reported that these reactive oxygen species attack biomolecules, damage cells or tissues, and cause various diseases related to aging or various adult diseases.

Research has been conducted to suppress skin aging caused by the above-mentioned factors, to improve whitening, wrinkles, elasticity or trouble, and to develop substances having antioxidant effects.

Substances having the above effects are widely distributed in the natural world, and have been mainly used as raw materials for foods, cosmetics, and pharmaceuticals using substances derived from plants. However, such materials derived from nature do not have significant effects and must be used in large amounts to obtain significant effects, resulting in toxicity and price increase problems.

In order to solve these problems of natural materials, chemically synthesized materials have been developed. These have the advantage of exerting a much superior effect even with a small amount compared to natural substances, but on the other hand, there is a fatal problem that can cause large and small side effects in the human body, so their use is limited.

Accordingly, there is an urgent need to develop a substance derived from a natural product and ensuring stability while exhibiting an excellent skin improvement effect.

The problem to be solved by the present invention is to provide a complex herbal extract derived from natural products and ensuring stability while exhibiting excellent effects on skin improvement, in particular, whitening, wrinkles, elasticity, trouble improvement, and antioxidation.

In order to solve the above problems, the present invention provides a composition for skin whitening, wrinkles, elasticity, trouble, or antioxidant (for example, a cosmetic composition) comprising a peach extract and a white confectionery extract as active ingredients.

In addition, the present invention provides a cosmetic composition for skin whitening, wrinkles, elasticity, trouble, antioxidant, or fibroblast proliferation, comprising peach extract, white confectionery extract, and white sheep resin extract as active ingredients.

[extract]

Peach blossom is peach tree Prunus It is a flower of persica (L.) Batsch. The taste is bitter and the quality is flat. It acts on the stomach meridians, liver meridians, and nerves.

Donga Benincasa, a white confectionery maker It is a seed of cerifera Savi (Gourd family Cucurbitaceae). It has the effect of enriching the lungs, clearing phlegm and cartilage, and urinating well.

Baekyang Suji is a branch of aspen tree Populus davidiana Dode, Populus tomentosa Carr. The bark of the aspen tree is blackish brown, does not crack for a long time, and has no hairs on small branches and winter buds. Leaves are alternate, round or egg-shaped, 2-6 cm long. The edges have shallow sawtooth, the petiole is flat, and the stipules fall early. Distributed in Korea, China and eastern Siberia.

The present inventors found that all of the above extracts have biological activities beneficial to the skin, and among the above extracts, two or more complex extracts, preferably Peach Blossom extract and Baegwain extract; Alternatively, it was found that the composite extracts of Dohwa extract, Baegwain extract, and Baekyang resin extract had synergistic biological activity.

In the present invention, the term "extract" includes an extract obtained by the extraction treatment, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof, and the extract itself and extracts of all formulations that can be formed using the extract.

The extract describes each plant and may include its leaves, stems, barks, roots, flowers or flower buds, fruits, seeds, sap and whole plants in addition to the specified parts.

To prepare the extract, one skilled in the art can use any suitable method known in the art. For example, it may be by a solvent extraction method. A solvent extract can be obtained by crushing the whole plant or any part thereof (eg, in a blender) and then treating the extraction solvent. It may be pulverized after undergoing a drying process (for example, drying at 40 to 70° C. for 15 to 50 hours) before pulverization. In addition, the solvent extract can be prepared in a powder state by additional processes such as distillation under reduced pressure and lyophilization or spray drying.

The type of extraction solvent used is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water; C1 to C4 lower alcohols such as methanol, ethanol, propyl alcohol and butyl alcohol; polyhydric alcohols such as glycerin, butylene glycol, and propylene glycol; and hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether, and dichloromethane; Or a mixture thereof may be used, and preferably, water and lower alcohol may be used alone or in combination of two or more. A solvent extract may be prepared by extracting at least once using the solvent, and a dried extract obtained by distilling the solvent extract under reduced pressure and then freeze-drying or spray-drying may be prepared.

The amount of the extraction solvent may vary depending on the type of extraction solvent used, but may be used, for example, 1 to 20 times, or 5 to 20 times the dry weight of the target plant.

In addition, various extraction processes known in the art, for example, maceration, infusion, percolation, digestion, decoction, hot continuous extraction, aqueous-alcoholic extraction, countercurrent extraction, microwave assisted extraction, ultrasonic extraction, supercritical fluid extraction, phytonic extract (eg, hydro-fluoro-carbon solvent), etc.) may be selected and used, and these may be performed alone or Two or more methods may be used in combination.

[composition]

The composition according to the present invention is a peach extract and baekgwain extract; Or, it contains peach blossom extract, white confectionery extract, and baekyangsuji extract as active ingredients.

In the present invention, the meaning of "included as an active ingredient" means that an extract is added to the extent that can exhibit a skin improvement effect from the composition according to the present invention, and various ingredients are added as subcomponents for skin delivery and stabilization. It means that it can be formulated in various forms.

Each plant extract in the composition according to the present invention may be included in the following ratio. For example, the weight ratio of the peach extract and the white confectionery extract may be in the range of 1: 0.01 to 10, or 1: 0.1 to 10, 1: 1 to 5, or 1: 1 to 3.

In addition, the weight ratio of peach extract, white confectionery extract, and white sheep resin extract is 1: 0.01 to 10: 0.01 to 10, or 1: 0.1 to 10: 0.1 to 10, or 1: 1 to 5: 1 to 5 or 1: 1 to 3: It may be included in the range of 1 to 3.

A composition according to the present invention comprises at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0 .0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025% , 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.00 39%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0 .0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066% , 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.00 80%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0 .0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375% , 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.07 25%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0 .1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000% , 0.5250%, 0.550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.850 0%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2. 2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6. 3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4% 8.5% 8.6% 8.7% 8.8% 8.9% 9.0% 9.1% 9.2% 9.3% 9.4% 9.5% 9.6% 9.7% 9.8% 9.9% 10% 11% 12% 13% 14% 1 5%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% or more of one or more extracts of the present invention.

In addition, in the composition according to the present invention, each extract of the present invention in the final form of the composition is at least about 0.0001% to 90%, 0.001% to 90%, 0.01% to 90%, 0.1% to 90%, 0.1% to 90%, 0.1% to 80%, 0.1% to 70%, 0.1% to 60%, 0.1% to 50%, 0.1% to 40%, 0.1% to 30%, 0.1% to 20%, or 0.1% to 10%.

The % may be calculated based on the total weight of the composition or the total volume of the composition, and the concentration may be adjusted according to the desired effect of the composition or the product into which the composition is incorporated.

Compositions of the present invention may be formulated into all types of vehicles. Examples of suitable vehicles include, but are not limited to, emulsions (e.g., oil in water, water in oil, silicone in water, water in silicone, water in oil in water, oil in water, oil in water in oil, oil in water in silicone, etc.), creams, lotions, solutions (e.g., aqueous and hydro-alcoholic solutions), anhydrous bases (e.g., lipsticks and powders), gels, ointments, pastes, milks, liquids, aerosols, solid forms, or eye jellies.

Compositions of the present invention may also be encapsulated for delivery to a target area such as the skin. Encapsulation techniques may include, but are not limited to, the use of liposomes, vesicles, and/or nanoparticles (including, for example, biodegradable and non-biodegradable colloidal particles comprising polymeric materials into which an ingredient is trapped, encapsulated, and/or absorbed, such as nanospheres and nanocapsules), which can be used as a delivery vehicle to deliver ingredients to the skin, for example.

The composition according to the present invention can be variously commercialized. For example, it can be commercialized as a cosmetic composition, a food composition, and the like.

The cosmetic composition may be prepared in the form of, for example, a general emulsified formulation and a solubilized formulation. For example, lotion such as flexible lotion or nutrient lotion, emulsion such as facial lotion, body lotion, cream such as nourishing cream, moisture cream, eye cream, massage cream, essence, serum, cosmetic ointment, spray, oil gel, gel, pack, sunscreen, makeup base, liquid type, solid type, or spray type foundation, powder, cleansing cream, cleansing lotion, makeup remover such as cleansing oil, cleanser such as cleansing foam, soap, body wash, etc. It is not limited thereto and may be formulated in various forms known in the art.

The cosmetic composition may further include, in addition to the extract according to the present invention, a conventional component known as a component of a cosmetic composition in the art, optionally selected, as long as the object of the present invention is not impaired. For example, fragrances (synthetic and natural), dyes and color ingredients, absorbents, emulsifiers, stabilizers, lubricants, solvents, humectants (including emollients, humectants, film formers, occlusive agents, and agents that affect the skin's natural moisturizing mechanism), water-repellants, UV absorbers (physical and chemical absorbers such as para Aminobenzoic acid ("PABA") and its corresponding PABA derivatives, titanium dioxide, zinc oxide, etc.), essential oils, vitamins (e.g. A, B, C, D, E and K), trace metals (e.g. zinc, calcium and selenium), anti-irritants (e.g. steroid and non-steroidal anti-inflammatory drugs), anti-microbial agents, antioxidants (e.g. BHT and tocopherol), chelating agents (e.g., disodium EDTA and tetrasodium EDTA), preservatives (e.g., methylparaben and propylparaben), pH adjusting agents (e.g., sodium hydroxide and citric acid), absorbents (e.g., aluminum starch octenylsuccinate, kaolin, corn starch, oat starch, cyclodextrins, talc, and zeolites), skin bleaching and lightening agents (e.g., hydroquinone and niacinamide lactate), humectants (eg glycerin, propylene glycol, butylene glycol, pentylene glycol, sorbitol, urea and mannitol), exfoliants (eg alpha-hydroxy acids and beta-hydroxy acids such as lactic acid, glycolic acid and salicylic acid; and salts thereof), waterproofing agents (eg magnesium/aluminum hydroxy de stearate), skin conditioning agents (e.g., aloe extract, allantoin, bisabolol, ceramide, dimethicone, hyaluronic acid, and dipotassium glycyrrhizate), and thickening agents (e.g., substances capable of increasing the viscosity of the composition, such as carboxylic acid polymers, crosslinked polyacrylate polymers, poly acrylamide polymers, polysaccharides and gums) and silicone-containing compounds such as silicone oils and polyorganosiloxanes.

The food composition may be formulated into, for example, beverages, fortified water, energy drinks, nutritional drinks, solid foods, vitamins, supplements, etc.), but is not limited thereto. Such supplements may include vitamins, minerals, herbs or other botanicals, amino acids, enzymes and metabolites. These adjuvants are suitable for oral consumption and can be administered orally.

It may be provided as a kit comprising a composition according to the present invention. The composition according to the present invention is contained in a container, which may include, but is not limited to, a bottle, a metal tube, a laminate tube, a plastic tube, a dispenser, a pressure vessel, a barrier container, a package, a compartment, a lipstick container, a compact container, a cosmetic pan capable of containing the cosmetic composition, or other types of containers such as, but not limited to, a dispersion or composition or preferred bottle, dispenser, or plastic container that is injected or blow-molded into a package. The kit may include instructions for using the kit or composition, and the instructions may be written on a separate sheet or may be written on the surface of the container or the surface of the container wrapper. Instructions include, but are not limited to, letters, phrases, abbreviations, pictures, or symbols. Instructions may include, for example, instructions on how to use, apply, and maintain the kit or composition. The containers may be distributed according to a pre-determined amount.

Compositions according to the present invention may be provided as topical skin compositions.

In addition, a method of topically applying the composition according to the present invention to the skin can be provided.

In the present invention, the term "topical application" means applying or applying a composition onto the surface of keratinous tissue, and "topical skin composition" includes a composition suitable for topical application or application onto keratinous tissue. Such compositions are typically dermatologically acceptable in that they do not exhibit excessive toxicity, incompatibility, instability, allergic reactions, etc. when applied to the skin. Topical skin care compositions of the present invention may have a selected viscosity to avoid significant dripping or pooling after application to the skin.

[Skin improvement effect]

The composition according to the present invention has biological activity and exhibits excellent effects on skin improvement.

More specifically, the composition according to the present invention has a skin whitening effect.

The skin whitening refers to inhibiting melanin production by inhibiting the melanin formation process.

The present inventors measured the amount of melanin produced by treating the complex extract according to the present invention to melanoma cells, and as a result, it was confirmed that the melanin production inhibitory effect was increased compared to the case of treating each extract, and the skin whitening effect was known. It was found to exert an effect corresponding to arbutin. It was confirmed that the composition according to the present invention has an excellent melanin production inhibitory effect and exhibits an excellent whitening effect.

In addition, the composition according to the present invention has an effect of improving wrinkles and elasticity.

The skin wrinkles refer to fine lines caused by skin deterioration, and may be caused by genes, reduction of collagen and elastin present in the skin dermis, external environment, and the like.

The wrinkle improvement refers to suppressing or inhibiting the formation of wrinkles or alleviating wrinkles already formed.

The skin elasticity is expressed by elastic fibers composed of elastin and collagen fibers present in the dermal layer, and the improvement of elasticity refers to suppressing or inhibiting a decrease in elasticity, maintaining or improving elasticity.

As a result of treating the complex extract according to the present invention, the present inventors confirmed that the collagen synthesis promoting effect and the elastase activity inhibitory effect were increased compared to the case of treating each extract, and the skin collagen synthesis promoting effect was known. It showed an effect corresponding to vitamin C, and it was found that the effect corresponding to quercetin, which is known to inhibit skin elastase activity, was exhibited. It was confirmed that the composition according to the present invention has an excellent collagen synthesis promoting effect and an elastase activity inhibiting effect, and exhibits excellent skin wrinkle and elasticity improvement effects.

In addition, the composition according to the present invention has a trouble improving effect.

The trouble refers to symptoms such as skin irritation and inflammation, and the inflammatory reaction is characterized by pain, fever, redness, swelling, loss of function, etc. caused by external stimuli. Histologically, complex symptoms including dilatation accompanied by increased permeability and blood flow of arterioles, capillaries, and venules, exudation of plasma containing plasma proteins, migration of leukocytes to inflamed areas, etc. are observed, and skin irritation reactions are characterized by erythema, itching, fever, etc.

The improvement of the trouble refers to inhibiting or inhibiting the skin irritation reaction and inflammatory reaction, or alleviating the skin irritation reaction or inflammatory reaction that is already in progress.

The present inventors measured the NO production inhibitory effect by treating the composite extract according to the present invention, and as a result, it was confirmed that the NO production inhibitory effect was increased compared to the case of treating each extract, and the NO production inhibitory effect was known. It was found to exert an effect corresponding to L-NMMA. It was confirmed that the composition according to the present invention exhibits an excellent skin trouble improvement effect through inhibition of NO production.

In addition, the composition according to the present invention has an antioxidant effect.

The antioxidant refers to inhibiting oxidation of cells by free radicals or reactive oxygen species (ROS) that are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, and includes reducing damage to cells caused by removing free radicals or reactive oxygen species.

The present inventors measured the free radical scavenging activity by treating the complex extract according to the present invention, and as a result, it was confirmed that the free radical scavenging activity was increased compared to the case of treating each extract, and the free radical scavenging activity was known. It was found to exert an effect corresponding to vitamin C. Thus, it was confirmed that the composition according to the present invention exhibits an excellent antioxidant effect.

In addition, the composition according to the present invention has an anti-glycation effect.

The glycation refers to a phenomenon in which proteins are destroyed by binding sugars and proteins caused by the presence of excessive sugars. When glycation progresses, it affects collagen and elastin, causing various skin damages in addition to reducing skin elasticity.

As a result of measuring the antiglycation effect by treating the composite extract according to the present invention, the present inventors confirmed that the antiglycation effect was increased compared to the case of treating each extract, and it was found that the antiglycation effect exhibited an effect corresponding to aminoguanidine known. As a result, it was confirmed that the composition according to the present invention exhibits an excellent antiglycation effect.

In addition, the composition according to the present invention has a fibroblast proliferation effect.

Among the compositions according to the present invention, this is the effect shown in the composition containing all of 'dohwa extract, white confectionery extract, and baekyangsuji extract'.

The composition according to the present invention can be used for fibroblast proliferation in vitro or/and in vivo.

The present inventors measured the cell proliferation rate by treating the composite extract according to the present invention, and as a result, it was confirmed that the cell proliferation effect was increased compared to the case of treating each extract, and thus the composition according to the present invention showed an excellent fibroblast proliferation effect. Confirmed.

The complex extract according to the present invention is safe for the skin and has excellent effects on skin improvement including antiglycation effect, skin whitening effect, skin elasticity and wrinkle improvement effect, antioxidant effect and skin trouble improvement effect. In addition, it has a fibroblast proliferative effect.

Hereinafter, examples and experimental examples will be described in detail to explain the present invention in detail. However, the examples and experimental examples according to the present invention may be modified in many different forms, and the scope of the present invention should not be construed as being limited to the examples and experimental examples described below. Examples and experimental examples of the present invention are provided to more completely explain the present invention to those skilled in the art.

<Example 1> Preparation of peach extract

After drying and cutting the dohwa well, 100 g of dry weight was put in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a peach extract.

<Example 2> Preparation of white confectionery ginseng extract

After drying the white confectionery well and cutting it, 100 g of dry weight was put in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The chilled extract was filtered through a filter having a pore size of 0.2 μm to prepare a white confectionery extract.

<Example 3> Preparation of Baekyang Resin Extract

After drying well and cutting Baekyang resin, 100 g of dry weight was placed in a flask and extracted by cooling with 1000 g of an extraction solvent (distilled water) for 3 days. The cooled extract was filtered through a filter having a pore size of 0.2 μm to prepare an extract of Baekyang resin.

<Example 4> Preparation of mixed extracts of peach and white confectionery

As in the above-described embodiment, a mixed extract of peach blossom and white confectionery was prepared by injecting the same amount of peach blossom and white confectionery prepared in the form of an extract, respectively.

<Example 5> Preparation of mixed extracts of Dohwa, Baegwajain, and Baekyang resin

As in the above-described example, a mixed extract of Peach blossom, Baekkijain, and Baekyang resin was prepared by injecting the same amount of Peach blossom, Baegwajain, and Baekyang resin prepared in the form of an extract, respectively.

<Experimental Example 1> Anti-glycation effect

In order to confirm anti-glycation efficacy, glycation inhibitory activity was measured using L-arginine and glucose.

First, 1M L-arginine and 1M glucose were dissolved in 1M phosphate buffer (pH 7.4) and prepared by diluting the sample to 50 ppm using 1M phosphate buffer. 1M L-arginine and 1M phosphate buffer were mixed at a ratio of 1:4, and then 80 μl was dispensed into a 96-well plate. Here, 100 μl of 0.01 M aminoguanidine to be used as a positive control and a sample diluted to 50 ppm, respectively, were added thereto. After mixing these samples well, finally, after adding glucose diluted with 1M phosphate buffer so that the final concentration of glucose is 0.1M, the mixture was reacted at 70°C for 4 hours. The degree of glycosylation was measured by measuring the absorbance at 420 nm of the 96-well plate using a spectrophotometer.

The Glycation experimental group of the following formula is an experimental group in which 1M L-arginine and 1M glucose are added to induce glycation. Glycosylation inhibitory activity can be obtained by the following formula.

[Equation 1]

(The 'Glycation experimental group' in Equation 1 means the 'absorbance of the Glycation experimental group'.)

As shown in the results of Table 1, it can be seen that the mixed extract has a more excellent anti-glycation effect compared to aminoguanidin, which is known as an anti-glycation substance.

<Experimental Example 2> Melanin production inhibitory effect

In order to confirm the whitening effect through inhibition of melanin production, according to the method described by Lotan R. et al. (Cancer Res. 40: 3345-3350, 1980), the extract was added to a culture medium of B-16 mouse melanoma cells to measure the total amount of melanin. In the experiment, whitening was evaluated at a non-toxic concentration by first evaluating toxicity to melanoma cells of mice. DMSO was used as a negative control and albutin was used as a positive control.

Specifically, the sample was added to the medium so that the final concentration was 100 ppm, and arbutin was added to the medium so that the final concentration was 100 ppm, and then the melanoma cells were cultured for 3 days. Thereafter, the cells were treated with trypsin, detached from the culture vessel, centrifuged, and then melanin was extracted. The detached cells were boiled for 10 minutes by adding 1 ml of sodium hydroxide solution (1N concentration) to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to measure the amount of melanin produced.

The amount of melanin was measured by the method of expressing the absorbance per unit cell number (1 × 10 ⁶ cell), the total amount of melanin relative to the control group was calculated as an inhibition rate (%), and the results are shown in Table 2 below. Each experiment was performed three times and represented as an average value.

As can be seen from the results of Table 2, the mixed extract was found to be useful for whitening because it exhibited an excellent effect of reducing the total amount of melanin.

<Experimental Example 3> Anti-inflammatory effect

In order to confirm the anti-inflammatory effect and skin trouble improvement effect, nitric oxide (NO) production inhibition test was performed by the GRIESS method using the RAW264.7 cell line (ATCC number: CRL-2278).

Specifically, RAW264.7 cells, which are mouse macrophages, were subcultured several times, placed in a 24-well plate so that 3×10 ⁵ per well, and then cultured for 24 hours. It was then replaced with a cell medium containing the sample at a final concentration of 10 ppm. At this time, L-NMMA (L-NG-Monomethylarginine), an NO-production inhibitor, was treated as a positive control and cultured for 30 minutes, and 1 μg of LPS (Lipopolysaccharide) was treated as a stimulus and cultured for 24 hours. 100 μl of the supernatant was transferred to a 96-well plate, 100 μl of the GRIESS solution was added, reacted at room temperature for 10 minutes, and the absorbance at 540 nm was measured to determine the NO inhibition effect. The NO production inhibition rate (%) was calculated using Equation 2 below and shown in Table 3 below.

[Equation 2]

NO production inhibition rate (%) = {(absorbance of negative control - absorbance of each extract) / absorbance of negative control} x 100

As can be seen from the results of Table 3, it was found that the mixed extract exhibited excellent activity as a natural substance, although the relative activity was somewhat lower when compared to L-NMMA, a representative anti-inflammatory drug.

<Experimental Example 4> Collagen synthesis promotion effect

The sample was added to the culture medium of human-derived fibroblasts to confirm the effect of promoting type 1 collagen synthesis at the cellular level. The measurement of synthesized collagen was quantified using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immuno Assay KIT). In order to measure the amount of collagen synthesis, the sample was added to the culture medium (DMEM medium) of fibroblasts to a final concentration of 10 ppm, cultured for 48 hours, and then the culture medium was taken and the degree of type 1 collagen synthesis at each concentration with the PICP EIA kit was measured at 450 nm using a spectrophotometer.

For comparison of the effect, the degree of collagen synthesis was measured in the same manner for the samples to which the culture medium of untreated fibroblasts (negative control) and vitamin C (positive control) were added to a final concentration of 52.85 μg/ml. The rate of increase in collagen production was calculated as the ratio of the amount of collagen production relative to the negative control group, and the results are shown in Table 4 below. The experiment was performed 4 times and presented as the average value.

As can be seen from the results of Table 4, collagen synthesis was increased when the mixed extract was treated, and the effect of collagen synthesis was comparable to that of vitamin C, which is generally known to induce collagen synthesis.

<Experimental Example 5> Elastase activity inhibitory effect

The effect of inhibiting the activity of elastase, an enzyme that degrades elastin, was confirmed as follows.

Elastase derived from human leukocytes was used, and a synthetic substrate MeOSuc-Ala-Ala-Pro-Val-pNA was used as a substrate for the elastase. A 100 mM Tris (pH 7.5) solution was used as the buffer solution. Elastase was finally used at 0.2 mU using a buffer solution. In addition, the synthetic substrate of Elastase was diluted with a buffer solution to a final concentration of 0.5 mM after making a 100 mM solution using DMSO. At this time, as a positive control group, Quercetin, known as an elastase inhibitor, was set at a concentration of 10 ppm. The elastase inhibitory candidate was added to a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and reacted at room temperature for 20 minutes. The absorbance was measured at 405 nm at 1-minute intervals using a spectrophotometer, and the slope of the absorbance versus time was determined to determine the activity of the enzyme. The elastase inhibition rate was calculated as follows.

[Equation 3]

The elastase inhibition rate was calculated using Equation 3 and shown in Table 5 below.

As can be seen from the results of Table 5, when the mixed extract was treated, the effect was relatively weaker than that of the positive control group, but it was found to exhibit an excellent elastase activity inhibitory effect. Therefore, it was found that the mixed extract can be used for skin regeneration and wrinkle improvement.

<Experimental Example 6> Antioxidant effect

The free radical scavenging ability of the mixed extract was measured by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method (Blois, Nature 181, 1190, 1958). DPPH is a relatively stable free radical and exhibits maximum absorption at 517 nm when present in a radical state, and loses absorbance when the radical is scavenged. DPPH was used from Sigma, and was dissolved in methyl alcohol at a concentration of 0.15 mM.

First, 100 μl of the mixed extract or vitamin C as a positive control was added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, left at room temperature for 30 minutes, and absorbance at 517 nm was measured using a microplate reader (BioTek EL-340).

Table 6 shows the results of expressing the concentration of the extract as IC ₅₀ when the absorbance of the treated sample becomes half of that of the control group. This experiment was repeated 3 times.

As shown in Table 6, the mixed extract had very strong free radical scavenging activity compared to vitamin C, and thus it was confirmed that the antioxidant effect was excellent.

<Experimental Example 7> Cell proliferation effect

The cell proliferation effect was tested in the following way. Human fibroblast cells were inoculated and cultured in a 96 well plate (96 well plate, Corning), then Examples 1 to 8 were applied at a concentration of 1% for 2 days, and cell proliferation was evaluated by performing the MTT test method.

More specifically, human fibroblasts were counted using a hemocytometer at the same rate of 5×10 ³ cells/well in a 24-well plate and then dispensed. After culturing for 48 hours in DMEM containing 10% FBS and culturing 40 to 50% of the surface area of the culture vessel, the sample was replaced with FBS-free DMEM and further cultured for 24 hours. After culturing, 50 ul of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT, Sigma M5655, USA) solution (2.5 mg/ml) was added and incubated for an additional 3 hours. Thereafter, the cell culture medium was discarded entirely, and 200 ul of dimethyl sulfoxide (DMSO, Sigma D2650, USA) was treated with 200 ul per well and stirred, and then 100 ul each was taken into 96 wells and the absorbance was measured at 570 nm by Enzyme-Linked Immunosorbent Assay (ELISA). Serum-free medium was used as a control group, and serum (serum) 1% was added as a control group. The experiment was repeated three times each, and the results are shown in Table 7.

As shown in the results of Table 7, it can be seen that the mixed extract according to the present invention has an excellent cell activity effect through cell proliferation. In addition, it was found that Example 5, in which the Baekyang resin extract was mixed with the mixed extract of Example 4, exerted an additional synergistic effect in terms of cell proliferation.

Claims (6)

A cosmetic composition for skin wrinkles, elasticity, trouble, or anti-oxidation comprising peach extract and white confectionery extract as active ingredients. The cosmetic composition according to claim 1, wherein the weight ratio of the peach blossom extract and the white confectionery extract is 1: 0.1 to 10. A cosmetic composition for fibroblast proliferation, characterized in that it comprises peach extract, white confectionery extract, and white sheep resin extract as active ingredients. [Claim 4] The cosmetic composition according to claim 3, wherein the weight ratio of the peach blossom extract, the white confectionery extract, and the baekyang resin extract is 1: 0.01 to 10: 0.01 to 10. The cosmetic composition according to claim 1 or 3, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C4 lower alcohol, or mixtures thereof. The cosmetic composition according to claim 1, wherein the cosmetic composition is antiglycation, promotes collagen synthesis, inhibits elastase activity, or scavenges free radicals.