KR101427572B1 - A composition for external application of skin for promoting hyaluronic acid production containing ginsenoside Re and ginsenoside compound K - Google Patents

Abstract

The present invention relates to a composition for external application of hyaluronic acid for promoting hyaluronic acid production, which comprises ginsenoside Re and a ginsenoside compound K. More specifically, the present invention relates to a ginsenoside Re and a ginsenoside metabolite ginsenoside compound K (20-O-? -D-glucopyranosyl-20 (S) -protopanaxadiol) to promote the production of hyaluronic acid in human cells.

Ginsenoside, ginseng, saponin, hyaluronic acid, skin dryness relief, skin elasticity enhancement, skin aging prevention

Description

[0001] The present invention relates to a skin external composition for accelerating the production of hyaluronic acid containing ginsenoside Re and a ginsenoside compound K,

The present invention relates to a composition for external application of hyaluronic acid for promoting hyaluronic acid production, which comprises ginsenoside Re and a ginsenoside compound K. More specifically, the present invention relates to a ginsenoside Re and a ginsenoside metabolite ginsenoside compound K (20-O-? -D-glucopyranosyl-20 (S) -protopanaxadiol) to promote the production of hyaluronic acid in human cells.

Hyaluronic acid is a complex polysaccharide composed of amino acid and uronic acid, and is a macromolecular compound having a molecular weight of 200,000 to 400,000 in which N-acetylglucosamine and glucuronic acid are alternately linked in a chain. It is involved in cell division, differentiation and migration, as well as being involved in storage and diffusion of water retention, intercellular space maintenance, cell growth factors and nutrients as main constituents of extracellular matrix. Hyaluronic acid is a component that exists in many vitamins (琉璃 體), joint fluid, cartilage, and skin of animal. It is combined with a large amount of water to become a gel state, and is involved in lubrication of joints and flexibility of skin. It also plays an important role in preventing penetration and penetration of toxins into the skin.

It has been reported that hyaluronic acid is a component existing naturally in all living organisms. It has been reported that more than 50% of hyaluronic acid present in the mammalian body is distributed in the intercellular spaces of the skin, especially the intercellular spaces of the epidermis and the connective tissues of the dermis. Are mainly synthesized by keratinocytes and fibroblasts. Hyaluronic acid makes skin elastic and reduces damage to the lower part of the skin when it is damaged. And important collagen smoothes the movement between cells, and acts as lubricant to smooth movements of bones and joints. Hyaluronic acid also prevents penetration of foreign substances such as bacteria and prevents skin diseases such as infection.

As the age increases, the amount of hyaluronic acid in the skin decreases and the production capacity decreases. Usually, it is the highest at about 18 years old, and it is said to be beautiful and volumey, and to decrease after that. As such, the amount of hyaluronic acid in human skin decreases with aging, which is one of the direct causes of reduced skin elasticity and moisture content due to aging.

Accordingly, the inventors of the present invention have searched for a substance capable of effectively increasing the amount of hyaluronic acid in skin cells, and found that when a mixture of ginsenoside Re and ginsenoside K is injected into human cells, hyaluronic acid And the effect of increasing the synthesis of hyaluronic acid is better than that of each single substance, and completed the present invention.

Accordingly, it is an object of the present invention to provide a composition for external application for skin, which can strengthen the structure of the extracellular matrix component by promoting hyaluronic acid production, suppress skin elasticity deterioration due to aging, and alleviate skin dryness.

The present invention provides a composition for promoting hyaluronic acid production, skin dryness mitigation, skin elasticity enhancement and skin aging prevention composition containing ginsenoside Re and ginsenoside K as an effective ingredient.

The ginsenoside Re and the ginsenoside K of the present invention have the effect of promoting hyaluronic acid production which activates hyaluronic acid production action of skin cells, thereby effectively preventing or preventing skin elasticity reduction, moisture content reduction and progress of aging Can be used. In addition, the effect of accelerating the hyaluronic acid production of the ginsenoside Re and the ginsenoside compound K, which are the active ingredients of the present invention, is better when they are mixed than the respective single substances.

The present invention relates to a pharmaceutical composition for accelerating the production of hyaluronic acid containing ginsenoside Re and a ginsenoside compound K (20-O-? -D-glucopyranosyl-20 (S) -protopanaxadiol) A skin external application composition for alleviation, skin elasticity enhancement and skin aging prevention is provided.

Hereinafter, the present invention will be described in more detail.

Ginsenoside refers to saponin in ginseng. Saponin is a kind of chemical compound called glycoside. Ginsenoside is the glycoside-separated name of ginseng to distinguish it from other plant saponins. It is usually named ginsenoside-Rx. A "," b "," c "," d "," b ", and" b "in accordance with the moving distance (Rf value) of a spot appearing on the TLC, e, f, g, h, and so on. Ginseng saponin has various effects on controlling the body function by affecting the central nervous system, endocrine system, immune system, metabolism system and the like. More than 30 kinds of ginsenosides have been identified from Korean ginseng (including red ginseng) so far, and the pharmacological action of each ginsenoside has been continuously revealed. These ginsenosides may act in similar or opposite ways. For example, G-Rb1, a typical ginsenoside of PPD-saponins, has a suppressive effect on the central nervous system, and G-Rg1, a typical PPT saponin, has a promoting effect.

The ginsenoside Re (Formula 1) used as an active ingredient of the present invention is a main component of Ginsengberry, wherein the ginseng flower of the Ginseng Berry refers to a state in which the green fruit has matured red . Ginsenoside Re plays a role in blood circulation, helps to alleviate women's menopausal symptoms, increases insulin sensitivity, reduces carbohydrate uptake and regulates blood sugar levels. It also protects cells and helps maintain cardiovascular health. In addition, it has various effects such as cholesterol lowering action and liver protecting action.

The ginsenoside Re of the present invention can be obtained according to a conventional method in the art, and the production method thereof is not particularly limited. For example, the dried fresh berry is cut into a pressurized sterilizer and heated at 70 to 150 ° C. for about 2 to 6 hours. Thereafter, water of about 1 to 20 times the weight (kg), C1 to C4 Of lower alcohols or a mixed solvent thereof, preferably methanol, for about 1 hour to 5 hours by a reflux extraction method, is filtered, concentrated under reduced pressure or dried, and then the obtained extract is heated at 40 to 80 ° C To obtain a fresh berry extract. The resulting extract is suspended in water and extracted with a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethyl acetate or a mixed solvent thereof. The remaining aqueous layer is extracted with a polar organic solvent such as butanol, The extract can be obtained by chromatography. If the chromatography is repeatedly performed, the content of ginsenoside can be further increased. The fraction having a higher content can be crystallized in an appropriate solvent system, for example, a solvent system such as water, lower alcohol, lower ketone, chloroform, , Pure ginsenoside Re can be obtained.

The ginsenoside compound K used as an active ingredient of the present invention is 20-O-? -D-glucopyranosyl-20 (S) -protonanaxadiol 20- -protopanaxadiol (Formula 2), and has a structure in which one sugar (glucoside) is attached to protopanaxadiol in the saponin component of ginseng. The ginsenoside K is known to have pharmacological effects such as inhibiting cancer cell proliferation, inhibiting tumor growth, and enhancing the anticancer activity of anticancer drugs. Recently, studies on the metabolites of saponin have been conducted, It is known that the metabolism of intestinal bacteria is the body of its activity rather than saponin itself.

The ginsenoside compound K of the present invention can be obtained according to a conventional method in the art, and the method is not particularly limited. For example, purified saponin prepared from ginseng is dissolved in an aqueous solvent such as water or buffer solution or a mixture of an aqueous solvent and an organic solvent such as water or a buffer solution, Compound K can be obtained by reacting with at least one of the pectinases isolated in the aspergillus.

The present invention provides a composition for external application for skin for promoting hyaluronic acid production, skin dryness alleviation, skin elasticity enhancement and skin aging prevention comprising a complex of ginsenoside Re and ginsenoside K as an active ingredient.

The active ingredient ginsenoside Re and the ginsenoside compound K of the present invention are contained in an amount of 0.1 to 10% by weight based on the total weight of the composition. If the content is less than 0.1% by weight, the effect is insignificant. If the content is more than 10% by weight, the increase in the effect is insignificant compared with the increase in the content.

The active ingredient of the present invention, ginsenoside Re and ginsenoside K, is contained in a weight ratio of 1: 1 to 20: 1. When the ratio is less than 1: 1 or more than 20: 1, synergistic effects between the active ingredients are not exhibited.

The composition for external application for skin containing a mixture of ginsenoside Re and ginsenoside K according to the present invention as an active ingredient promotes hyaluronic acid production in skin cells, thereby enhancing skin moisturization and skin elasticity, Effect.

The composition for external application for skin of the present invention can be used in the form of an external preparation for skin comprising at least one of a fatty substance, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, Such as fillers, emulsifiers, emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics May contain adjuvants commonly used in the field of cosmetics or dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

The composition for external application for skin is not particularly limited, and the formulation can be appropriately selected according to the purpose. For example, it can be applied to softeners (skin lotion and milk lotion), nutrition lotion, essence, nutrition cream, massage cream, pack, gel, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, A body lotion, a body cream, a body oil, and a body essence, but the present invention is not limited thereto.

The composition for external application for skin of the present invention can be used on the face, especially around the eyes, around the mouth, on the cheeks, or on the forehead, neck, hands, feet, but is not limited thereto.

Hereinafter, the constitution and effects of the present invention will be described in more detail with reference to examples and test examples. However, these examples and test examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples and test examples.

[Test Example 1] Confirmation of cytotoxicity in HaCaT cells by treatment with ginsenoside Re and ginsenoside K

1. Cell line and cell culture

HaCaT cells, a human keratinocyte, were cultured in DMEM medium containing 10% fetal bovine serum (Dulbecco's modified Eagle's Medium, Gibco 1210-0038) and cultured at 37 ° C in 5% CO ₂ Lt; / RTI >

2. Cytotoxicity check

The cultured cell line was trypsinized to make a single cell suspension and 2x10 ⁴ cells per well of a 96-well plate were added and cultured for 24 hours. After that, DMEM medium containing no bovine serum was treated and cultured for 24 hours, and then ginsenoside K and ginsenoside Re were added to media containing no bovine serum at 1, 5, 10, 25, 50 , 75 and 100 μM, respectively. The ginsenoside K was purchased from Ambo Institute (Korea) and purchased from Ginsenoside Re Wako (Japan). After 24 hours, 10% CCK-8 (Cell Counting Kit-8) reagent was added and incubated for 2 hours. Absorbance was measured at 450 nm. The cell viability when the active ingredient was not treated was set at 100%, and the relative cell viability at each concentration was calculated. The results are shown in Figs. Generally, when the cell viability falls below 80%, it can be considered that the cell is cytotoxic.

As can be seen from Fig. 1, the ginsenoside K did not show cytotoxicity in the concentration range of 1 μM or less, and the cytotoxicity was increased at 5 μM or more, and the cell viability was drastically decreased. As can be seen from Fig. 2, ginsenoside Re did not show cytotoxicity in a concentration range of 75 μM or less, and cytotoxicity occurred at 100 μM or more, and cell survival rate was gradually decreased.

Test Example 2 Increase in hyaluronic acid production in HaCaT cells by treatment with ginsenoside Re and ginsenoside K

The cell line cultured in the same manner as in Test Example 1 was trypsinized to prepare a single cell suspension, which was then divided into 1 × 10 ⁵ cells in a 24-well plate and cultured for 24 hours. Thereafter, DMEM medium without bovine serum was treated and cultured for 24 hours. Then, the medium containing no ginsenoside K 500 nM, 1 μM and ginsenoside Re 1 μM , 10 μM, and the ginsenoside K and ginsenoside Re were mixed at a weight ratio of 1: 1, respectively, to 1 μM and 10 μM, respectively. After 24 hours, the cell culture medium was recovered and centrifuged at 10,000 rpm for 3 minutes to remove the precipitated cells. The supernatant was collected and used in an ELISA kit (Echelon Bioscience, Salt Lake, UT) And the amount of hyaluronic acid was measured. The amount of hyaluronic acid in the untreated control group was set at 100%, and the relative amount of hyaluronic acid in each experimental group was calculated. The results are shown in Fig.

As can be seen from FIG. 3, the ginsenoside K and ginsenoside Re increased the amount of hyaluronic acid, respectively. However, when the above-mentioned active ingredients were mixed and used, the hyaluronic acid Of the total.

Therefore, the mixture of ginsenoside K and ginsenoside Re of the present invention can exhibit excellent skin dryness, skin elasticity and skin aging prevention effect by significantly increasing the amount of hyaluronic acid.

 [Example 1 and Comparative Example 1]

Comparative Example 1 containing only ginsenoside Re and compound K in a weight ratio of 1: 1 and ginsenoside K alone were produced according to the composition shown in Table 1 below.

Ingredients Example 1
(weight%) Comparative Example 1
(weight%) Purified water Balance Balance glycerin 3.0 3.0 Butylene glycol 3.0 3.0 Liquid paraffin 7.0 7.0 Beta Glucan 7.0 7.0 Carbomer 0.1 0.1 A mixture of ginsenoside Re and compound K (1: 1) 3.0 - The ginsenoside compound K - 3.0 Caprylic / capric triglyceride 3.0 3.0 Squalane 5.0 5.0 Cetearyl glucoside 1.5 1.5 Sorbitan stearate 0.4 0.4 Polysorbate 60 1.2 1.2 antiseptic Suitable amount Suitable amount incense Suitable amount Suitable amount Pigment Suitable amount Suitable amount Triethanolamine 0.1 0.1

[Test Example 3] Skin moisturizing effect

The cosmetic composition of Example 1 and Comparative Example 1 was applied twice daily for four weeks to 50 adult men and women who were thought to be dry or dry skin. The transepidermal water loss (TEWL) was measured at 2 weeks and 4 weeks after application, and moisture was measured at constant temperature and humidity (24 ℃, relative humidity 40%). The results are shown in Table 2 below.

<TEWL measurement method>

Principle: The amount of water evaporating from the skin is calculated according to the area and time, and the amount of water evaporating is read by the temperature and humidity electronic sensor to measure the moisture of the skin.

1. Environmental conditions: The temperature and humidity are kept constant and only the minimum number of persons other than the measurer and the subject are left. (It blocks the flow of air as much as possible.)

2. Place a TEWL probe on the subject's measurement site.

3. Place the probe lightly on the measurement site (not to leave marks on the skin) and make a 90 degree angle with the ground.

4. Measure the value at which the TEWL value of the skin becomes constant as time goes by. (Usually takes 1 to 3 minutes)

5. Be careful not to let the temperature of the measurer pass by holding the handle of the probe as far away as possible.

6. Calculate the amount of water (g / m ² / h) that evaporates per hour per area.

division Comparative Example 1
(Compound K) Example 1
(Ginsenoside Re + compound K) 0 weeks 16 g / m ² / h 17 g / m ² / h 4 weeks 14 g / m ² / h 13 g / m ² / h

As shown in Table 2, Comparative Example 1 containing the ginsenoside K reduced the amount of water evaporating from the skin by 2 g / m ² / h. However, Example 1, in which the active ingredients were mixed and applied, The evaporation amount was reduced by 4 g / m ² / h, and the amount of moisture evaporated from the skin could be reduced by about 2 times as compared with Comparative Example 1. Therefore, it was found that the mixture of ginsenoside K and ginsenoside Re of the present invention is excellent in skin moisturizing effect by suppressing water evaporation.

[Test Example 4] Effect of improving skin wrinkles

Experiments were carried out as follows to examine the wrinkle-reducing effect of Example 1 and Comparative Example 1. 160 women in their 20s and 40s were divided into eight groups each consisting of 20 individuals. The nutritional creams of Example 1 and Comparative Example 1 were applied to the eye area designated for 8 weeks, twice a day, morning and evening, respectively, And the state of the wrinkles at the designated region was measured with a visiometer (SV60, Courage + Khazaka electronic GmbH, Germany) as an image analyzer. The results are shown in Table 3, which is the value obtained by subtracting the wrinkle depth before coating from the wrinkle depth after 4 weeks of application and the wrinkle depth before application at the wrinkle depth of 8 weeks of application. That is, the negative value indicates that the wrinkle improving effect is higher.

division Comparative Example 1
(Compound K) Example 1
(Ginsenoside Re + compound K) 0 weeks to 4 weeks -0.027 -0.046 0 weeks to 8 weeks -0.031 -0.053

As can be seen from Table 3, the ginsenoside K was able to reduce the wrinkles of the skin to some extent. However, in the case of Example 1 where the active ingredients were mixed together, when the ginsenoside K was used alone The effect of improving skin wrinkles was higher. Therefore, it was found that the mixture of the ginsenoside K and ginsenoside Re of the present invention is excellent in skin wrinkle improving effect and skin aging suppressing effect.

 [Formulation Example 1] Nutritional lotion

Nutrition lotion was prepared according to a conventional method according to the composition shown in Table 4 below.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Carbomer 0.1 Complex of ginsenoside Re and compound K 0.05 Caprylic / capric triglyceride 8.0 Squalane 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Triethanolamine 0.1

[Formulation Example 2] Nourishing cream

Nutritive creams were prepared according to the compositions listed in Table 5 below in a conventional manner.

ingredient Content (% by weight) Purified water Balance glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Complex of ginsenoside Re and compound K 3.0 Caprylic / capric triglyceride 3.0 Squalane 5.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Triethanolamine 0.1

[Formulation Example 3] Massage cream

Massage creams were prepared in a conventional manner according to the compositions shown in Table 6 below.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Complex of ginsenoside Re and compound K 1.0 Caprylic / capric triglyceride 3.0 Wax 4.0 Cetearyl glucoside 1.5 Sesquioleic acid sorbitan 0.9 Vaseline 3.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount paraffin 1.5

[Formulation Example 4] Pack

The packs were prepared in a conventional manner according to the composition shown in Table 7 below.

ingredient Content (% by weight) Purified water Balance glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Complex of ginsenoside Re and compound K 0.5 Nonyl phenyl ether 0.4 Polysorbate 60 1.2 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount ethanol 6.0

[Formulation Example 5] ointment in external preparation for skin

Ointment was prepared in a conventional manner according to the composition shown in Table 8 below.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Complex of ginsenoside Re and compound K 1.0 Caprylic / capric triglyceride 3.0 Squalane 1.0 Cetearyl glucoside 1.5 Sorbitan stearate 0.4 Cetearyl alcohol 1.0 antiseptic Suitable amount incense Suitable amount Pigment Suitable amount Wax 4.0

FIG. 1 is a graph showing cytotoxicity after treating ginsenoside K with HaCaT skin cells. FIG.

FIG. 2 is a graph showing cytotoxicity after treating ginsenoside Re with HaCaT skin cells. FIG.

FIG. 3 is a graph showing the amount of hyaluronic acid produced after treatment of HaCaT skin cells with the ginsenoside K, ginsenoside Re, and the mixture thereof by ELISA.

Claims (6)

delete A composition for external application for skin for relieving dryness comprising ginsenoside Re and a ginsenoside K (20-O-? -D-glucopyranosyl-20 (S) -protopanaxadiol) as an active ingredient, Wherein the ginsenoside Re and the ginsenoside compound K are mixed at a concentration ratio of 1: 1. delete delete The composition for external application for skin according to claim 2, wherein the ginsenoside Re and the ginsenoside K are contained in an amount of 0.1 to 10% by weight based on the total weight of the composition. delete

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