KR20060021506A - Cosmetic composition for skin regeneration containing ginsenoside rg3 and its derivatives as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for promoting skin regeneration, and more particularly, to a composition for promoting TGF-β1 synthesis and a cosmetic composition for promoting skin regeneration comprising ginsenoside Rg3 or a derivative thereof as an active ingredient. The composition of the present invention can be effectively used for skin regeneration and wound treatment by promoting TGF-β1 synthesis.

Ginsenoside Rg3, skin regeneration, TGF-β1, wound, cosmetic

Description

Cosmetic composition for promoting skin regeneration comprising ginsenoside Rg 3 or a derivative thereof TECHNICAL FIELD

The present invention relates to a composition for promoting TGF-β1 production and a cosmetic composition for promoting skin regeneration.

Skin regeneration is a tissue response to injury and is a process of tissue recovery, including chemotaxis, differentiation and replication of cells, synthesis of matrix proteins, angiogenesis and wound reconstruction. It is a complex biological process that includes (Steed, DL et al ., Clin. Plast. Surg. , 25: 397 (1998)). One of the representative substances that appear early in the skin regeneration process and regulates the subsequent process is a growth factor, which has a strong effect throughout the skin regeneration process, and regulates the growth, differentiation and metabolism of cells.

Among the growth factors studied so far, platelet-derived growth factor (PDGF) promotes the metabolism of collagen fibers, promotes the migration of fibroblasts and macrophages, and promotes chemotaxis, angiogenesis and differentiation of fibroblasts. (Mustoe, TA, et al ., J. Clin. Invest ., 87: 694 (1991); Lepisto, J. et al ., J. Surg. Res ., 53: 596 (1992)), EGF (epidermal growth factor) acts to increase epithelialization, differentiation of fibroblasts and angiogenesis (Franklin, JD et al ., Plast.Resonst Surg ., 64: 766 (1979); Buckly, A. et al ., Proc. Natl.Acad. Sci. USA , 82: 7340 (1985)).

In addition, basic fibroblast growth factor (bFGF) promotes angiogenesis, epithelialization and collagen deposition, binds to heparin in various forms and has associated effects (Tsuboi, R. et al ., J. Exp. Med . , 172: 245 (1990); Kinsnorth, AN et al ., Br. J. Surg ., 77: 409 (1990)), IGF (insulin-like growth factor) plays a role in promoting cell differentiation and VEGF (vascular endothelial growth factor) is a substance that increases blood vessel permeability and promotes angiogenesis.

In addition, many growth factors and cytokines are known to be involved in skin regeneration, of which TGF-β (transforming growth factor-β) is a substance involved in the growth and differentiation of various cells. TGF-β is present in mammals in three different isoforms, of which TGF-β1 has the strongest activity in the formation of extracellular matrix (ECM) of the dermal layer and is involved in tissue repair and differentiation. TGF-β1 stimulates fibroblasts to produce several ECM components such as Type I and Type III collagen, fibronectin and proteoglycans ( Clin Exp Allergy ., 89 (7): 933-7 (2002). )).

When the skin is damaged, the cell proliferation process is followed by an inflammatory response, which lasts from two days to three weeks after the skin is damaged. At this time, fibroblasts accumulate collagen to fill damaged skin flaws, induce proliferation of fibroblasts, and reconstruct new cells and interstitial components at the wound site.

This multiplication process, also called the granulation process, lasts from two days to three weeks. During this period, fibroblasts accumulate collagen, creating new capillaries and filling them. Small, rounded masses of tissue consisting of capillaries and fibroblasts form at the site of injury, resulting in epithelialization at the edges of the damaged skin. In this series of processes, cell adhesion occurs. Integrin, an adhesion receptor on the cell surface, regulates the growth and differentiation of cells through interaction with extracellular matrix.

One of the extracellular matrices that work with integrins to regulate cell function is fibronectin. Fibronectin is a relatively large protein with a molecular weight of about 220 kDa, which has several motifs, including the RGD motif, which interacts with various integrins to regulate cell function (Hynes RO, Arterioscle Thromb Vasc Biol ., 18: 1363-1370). 1998)).

Conventionally, peptides such as epidermal growth factor, cytokines, interlukin and TGF-β, and madecasol (Asiaticoside) have been used as wound healing agents. . However, peptides are very expensive and unstable, so they are not widely used as wound healing agents. Madecassol has a weak effect and has side effects of contact dermatitis.

The present inventors have diligently tried to solve the above-mentioned needs of the art, and ginsenoside Rg3, which is a saponin component of ginseng, has an excellent effect of promoting TGF-β1 production, and when the composition containing the same as an active ingredient is applied directly to the skin, The present invention was completed by confirming the excellent regeneration effect.

Accordingly, an object of the present invention is to provide a composition for promoting TGF-β1 production comprising ginsenoside Rg3 or a derivative thereof as an active ingredient.

Another object of the present invention is to provide a cosmetic composition for promoting skin regeneration comprising the composition for promoting TGF-β1 production as an active ingredient.

Other objects and advantages of the present invention will become apparent from the following examples and claims.

According to one aspect of the invention, the present invention provides a composition for promoting TGF-β1 production comprising ginsenoside Rg3 or a derivative thereof as an active ingredient.

According to another aspect of the present invention, the present invention provides a cosmetic composition for promoting skin regeneration comprising the composition for promoting TGF-β1 production as an active ingredient.

The composition of the present invention contains ginsenoside Rg3 or a derivative thereof as an active ingredient of ginseng.

Ginseng (ginseng) belongs to the genus Ginseng of Araliace and is a herbaceous plant as a perennial ring-negative sage root. It is produced in many parts of the world, including Korea, China, Japan, and the United States, and is mainly cultivated at 85-140 degrees east, 22-49 degrees north latitude, 70-97 degrees west latitude, and 34-47 degrees north latitude in North America.

As described above, ginseng produced in various parts of the world is different in its composition and shows various physiological and pharmacological effects. However, ginsenosides, the main component of ginseng, are the most contained in Korean ginseng. So far, about 34 ginsenosides have been isolated, and the pharmacological efficacy varies depending on the type of sugar bound to aglycone, the number of sugars bound, or the position of sugar binding. According to the structural characteristics, it is divided into diol, triol, and oleanane, and ginseng contains a large amount of diol-type ginsenosides such as ginsenosides Rb1, Rb2, Rc and Rd, but ginsenoside Rg3 Present in trace amounts.                     

In addition, ginseng contains intrinsic fragrance components, panacen, polyacetylene compounds, polyphenol compounds, flavonoids and vitamins.

Ginseng has long been used for the treatment of various diseases because of its herbal efficacy. Herbal benefits of ginseng include stamina, metabolism, stress relief, diabetes treatment, respiratory disease, digestive tract and anticancer activity.In recent studies, ginseng has anti-complementary activity, anti-ulcer activity, and immune enhancement. It has been reported to have an action, anticancer action and hypoglycemic action.

Among them, the effects related to skin include anti-inflammatory effect ( Korean J. Dermatol., 18 (1): 39-42 (1980); Korean J. Dermatol. 14 (4): 335-339 (1976)) ( Korean J. Dermatol .. 28 (4): 434-440 (1990)), prevention and treatment of acne ( Korean J. Dermatol., 30 (1): 27-33 (1992); Korean J. Dermatol., 28 (4): 434-440 (1990)), antioxidant activity (Proceedings of the 2 ^nd international symposium ginseng, 13-17 (1978)), resilience and hydratable increasing action (Fitoterapia, 57 (1.) : 15- 28 (1986); Fitoterapia ., 57 (4) 217-222 (1986)), collagen degradation inhibitory action ( Mol. Cells ., 9 (5): 476-483 (1999)) and whitening action ( Journal of the society) of cosmetic scientists of korea ., 27 (2): 45-56 (2001)).

Ginsenoside Rg3 or a derivative thereof included in the composition of the present invention is isolated from ginseng, but it is apparent to those skilled in the art that chemically synthesized may have the same effect.

In addition, the derivative of the ginsenoside Rg3 may be a derivative obtained by the addition or substitution reaction of a substituent commonly practiced in the art. For example, derivatives to date have been found to include glucose, rhamnose, xylose or arabinase in the alcoholic hydroxyl groups of protopanaxadiol and protopanaxatriol, which are triterpenoids of the oleanic series. There are compounds in which sugars such as north are ether bonded.

In addition, it is evident in consideration of the state of the art that any derivative which exhibits a skin regeneration effect (through promoting TGF-β1 production) among the derivatives of ginsenoside Rg3 is included in the scope of the present invention.

Referring to the following specific preparation of the present invention, the preparation method of ginsenoside Rg3 or a derivative thereof is as follows: Ginseng is washed with purified water, dried and crushed into small pieces, and then the dry weight of Add 10 times lower alcohol. After extracting for a certain period of time at a certain temperature, filtered and dried with a rotary vacuum evaporator. Ginsenoside Rg3 or a derivative thereof is separated from the dried ginseng extract powder using prep LC.

According to a preferred embodiment of the present invention, the content of ginsenoside Rg3 or a derivative thereof is 0.0001-10.0% by weight, preferably 0.001-3% by weight, based on the total weight of the composition. If the content is less than the expected effect is difficult to appear, if the content is exceeded the effect is no longer increased.

In the composition of the present invention, the term 'skin regeneration' refers to the process of repairing skin tissue against skin damage, and includes chemotaxis, differentiation and replication of cells, synthesis of matrix proteins, angiogenesis and reconstruction of wounds, etc. Is a complex biological process.

The components included in the composition of the present invention include components conventionally used in the composition in addition to the ginsenoside Rg3 or a derivative thereof as an active ingredient, and include, for example, conventional agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings. Phosphorus aids and carriers.

The compositions of the present invention may be prepared in any formulations commonly prepared in the art, including, for example, solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, It may be formulated as an oil, powder foundation, emulsion foundation, wax foundation, spray, and the like, but is not limited thereto. More specifically, when the composition of the present invention is used as a cosmetic composition, it is prepared in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder Can be.

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

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

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

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

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

The cosmetic composition of the present invention has a skin regeneration use. In particular, the composition of the present invention promotes the expression of TGF-β1 in skin cells, thereby exhibiting excellent skin regeneration effect, and also excellent wound healing effect. This fact is clearly described in the following examples.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Preparation Example 1 Preparation of Ginsenoside Rg3

Six years old ginseng was washed with purified water, dried and broken into small pieces, and then 70% lower alcohol was added 1-10 times the dry weight of ginseng. After extracting at 15-35 ° C. for 5 days, it was filtered through a 300 mesh filter cloth, and again filtered through a filter paper of Whatman No. 5, dried on a rotary vacuum evaporator, to obtain a dry weight of 30.4 g of ginseng extract. Ginsenoside Rg3 was isolated from the dried ginseng extract powder using prep LC (Waters Co., Ltd. USA, Delta prep 4000). In the conditions of the flap elsi, acetonitrile: water (50:50) was used as a mobile phase and the UV detector (210 nm) was passed through a 30 cm x 10 cm column filled with octadecyl silica at a flow rate of 5 ml / min. Separated using. Next, qualitative analysis of the separated ginsenoside Rg3 was performed using HPLC (Waters Co., Ltd. USA, Waters 2690 Separations Module, Waters 2487 Dual λ Absorbance Detector), and as a result, the purity of ginsenoside Rg3 was 90%. It was above.

Experimental Example 1: Promoting TGF-β1 Production

TGF-β1 production promoting effect was measured using ginsenoside Rg3 prepared in Preparation Example 1.

Normal fibroblasts of human (normal human fibroblast) 10% right in 48-well microtiter plates containing DMEM medium supplemented with fetal serum-containing 1 × 10 ⁵ cells / well concentration and inoculated with, 5% CO ₂ concentration of the CO ₂ The incubator was incubated at 37 ° C. for 24 hours. The normal fibroblasts of the humans were separated from the skin tissue obtained after circumcision of the newborn in the hospital by a conventional cell separation method. Ginsenoside Rg3 was used after dissolving in an appropriate concentration in PBS buffer. A medium in which ginsenoside Rg3 prepared in Preparation Example 1 was added to a final concentration of 0-30 μg / ml was added to DMEM medium containing no serum, and the fibroblast culture medium was replaced with this medium and cultured. After incubation, cell cultures were harvested 24 hours, 48 hours and 72 hours.

According to R & D system, the manufacturer's policy by using ELISA kit (Quantikine ^TM high sensitivity ELISA kit) of (R & D system, USA), followed by measuring the amount of TGF-β1 produced in a cell culture fluid collected over time. All samples used to measure the amount of TGF-β1 provided in the kit were used. First, the collected cell culture solution was added to a 96-well microplate to which TGF-β1 antibody was uniformly applied to allow antigen-antibody reaction to occur at 37 ° C for 1 hour. Cell cultures in the wells were removed and washed three times with PBS buffer. In each well, a secondary antibody conjugated with chromophore horseradish peroxidase was added to allow antigen-antibody reactions to occur for 1 hour. Next, tetramethylbenzidine was developed in each well, and reacted at room temperature for 15 minutes. Then, 1 N sulfuric acid solution was added to stop the reaction. Absorbance was measured at a wavelength of 450 nm with a spectrophotometer. A standard curve was prepared using the standard solution, and the measured absorbance was substituted into the standard curve to calculate the amount of TGF-β1 generated in the cell culture solution to which each sample was added. The TGF-β1 production accelerating rate was calculated by substituting the calculated amount of TGF-β1 production (Table 1) into Equation 1 below.

Ginsenoside Rg3 (μg / ml) TGF-β1 (pg / ml) 24 hours 48 hours 72 hours 0 33.7 47.7 65.5 10 54.6 92.0 130.8 20 80.1 114.8 157.5 30 105.4 125.5 158.8

In the above equation,

A: amount of TGF-β1 produced in the cell culture medium to which the sample was added,

B: TGF-β1 production amount of cell culture medium without sample

As shown in Table 1, the amount of TGF-β1 production of fibroblasts was increased in proportion to the treatment concentration and treatment time of ginsenoside Rg3, and the greatest effect was obtained when the ginsenoside Rg3 was treated at 30 ㎍ / ml. However, at higher concentrations, the effect no longer increased.

Based on the above experimental example, using a ginsenoside Rg3 having a TGF-β1 production promoting effect to propose a composition of various formulations that can provide a skin regeneration promoting effect. Although this is described in detail in the following formulation example, the composition of the present invention is not limited only to the following examples.

Formulation Example 1: Softener (Skin Lotion)

In the following ingredients and contents as shown in Table 2, the flexible longevity containing ginsenoside Rg3 prepared in Preparation Example 1 was prepared according to a conventional method.                     

ingredient Content (% by weight) Ginsenoside Rg3 1.0 glycerin 5.0 1,3-butylene glycol 3.0 PEG 1500 1.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium hyaluronate 5.0 ethanol 10.0 Octicdodeceth-16 0.2 Polysorbate 20 0.2 incense 0.02 Uniside-U 13 0.01 Green # 13 0.001 Distilled water Remaining amount Sum 100

Formulation Example 2: Converging Cosmetic Water

Converging cosmetic water containing ginsenoside Rg3 prepared in Preparation Example 1 was prepared according to a conventional method with the ingredients and contents shown in Table 3 below.

ingredient Content (% by weight) Ginsenoside Rg3 1.0 glycerin 2.0 1,3-butylene glycol 2.0 Allantoin 0.2 DL-panthenol 0.2 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium hyaluronate 3.0 ethanol 15.0 Polysorbate 20 0.3 Witch Hagel Extract 2.0 Citric acid a very small amount incense 0.02 Uniside-U 13 0.01 Green # 13 0.001 Distilled water Remaining amount Sum 100

Formulation Example 3: Cream

A cream containing ginsenoside Rg3 prepared in Preparation Example 1 was prepared according to a conventional method with the ingredients and contents shown in Table 4 below.

ingredient Content (% by weight) Ginsenoside Rg3 2.0 Lipophilic monostearate 2.0 Cetearyl Alcohol 2.2 Stearic acid 1.5 Beeswax 1.0 Polysorbate 60 1.5 Sorbitan stearate 0.6 Cured Vegetable Oil 1.0 Squalane 3.0 Mineral oil 5.0 Trioctanoine 5.0 Dimethicone 1.0 Sodium magnesium silicate 0.1 glycerin 5.0 Betaine 3.0 Triethanolamine 1.0 Sodium hyaluronate 4.0 incense 0.01 Uniside-U 13 0.02 Green # 3 0.003 Distilled water Remaining amount Sum 100

Formulation Example 4: Essence

An essence containing ginsenoside Rg3 prepared in Preparation Example 1 was prepared according to a conventional method with the ingredients and contents shown in Table 5 below.

ingredient Content (% by weight) Ginsenoside Rg3 2.0 glycerin 10.0 Betaine 5.0 PEG 1500 2.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethyl cellulose 0.1 Sodium hyaluronate 8.0 Carboxy Vinyl Polymer 0.2 Triethanolamine 0.18 Octyldodecanol 0.3 Octyldodeceth-16 0.4 ethanol 6.0 incense 0.02 Uniside-U 13 0.01 Green # 3 0.001 Distilled water Remaining amount Sum 100

Experimental Example 2: Wound Healing Effect

Sprague-Dawley rat males (approximately 200 g in weight) (central laboratory animals, Korea) were anesthetized with pentobarbital sodium to shave the hairs on the back and disinfect the skin with iodine tincture. Using a leather punch with an inner diameter of 1 cm, two symmetrical round excisions were made on the back skin. From the day of wound development, hydrocortisone was subcutaneously administered daily at a dose of 50 mg / kg once daily to delay wound healing. The cream of Formulation Example 3, the cream (placebo) which replaced ginsenoside Rg3 of Formulation Example 3 with purified water, and the cream (comparative example) which replaced the ginsenoside Rg3 of Formulation Example 3 with the same amount of Asianticoside, the day of wounding 200 mg was applied once a day for 1 wound at every day. After 8 days after measuring the area of the wound, it was compared with the untreated control, and substituted the measured wound area in the following equation 2 to confirm the wound healing promotion rate.

Test substance Kill Mouse Wound area (mm2) Wound healing promotion rate (%) Control 10 58.9 ± 3.8 - Placebo 10 57.0 ± 2.3 1.7 Comparative example 10 49.3 ± 3.2 16.3 Formulation Example 3 10 38.4 ± 2.8 34.8

As shown in Table 6 above, the formulation containing ginsenoside Rg3 significantly reduced the wound area compared to the untreated control (P <0.01), and compared with Asiaticoside, which is widely used for wound healing purposes. Compared to the example formulation, the rate of wound healing promotion was very high (P <0.05). In addition, the placebo healing effect was not observed in the placebo formulation containing no ginsenoside Rg3, and it was found that components other than ginsenoside Rg3 included in the formulation did not affect the wound healing. Therefore, it can be seen that the ginsenoside Rg3 of the present invention is also excellent in the wound healing effect.

In conclusion, the ginsenoside Rg3 of the present invention increases the expression of TGF-β1. TGF-β1 promotes skin regeneration by promoting the production of several ECM components, including Type I and III collagen and proteoglycans, in fibroblasts, and promotes fibronectin production in fibroblasts to increase cell adhesion at the wound site. Increase Therefore, ginsenoside Rg3 of the present invention increases the expression of TGF-β1, it is believed to exhibit excellent skin regeneration promoting and wound healing effect.

As described in detail above, the present invention provides a composition for promoting TGF-β1 production and a cosmetic composition for promoting skin regeneration, comprising ginsenoside Rg3 or a derivative thereof as an active ingredient. Ginsenoside Rg3 of the present invention increases the expression of TGF-β1 in fibroblasts, and when applied directly to the skin a composition containing it as an active ingredient, it is excellent in promoting skin regeneration and wound healing, cosmetics for skin regeneration and It can be usefully used as a wound healing agent.

Having described the specific part of the present invention in detail, it is apparent to those skilled in the art that the specific technology is merely a preferred embodiment, and the scope of the present invention is not limited thereto. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (4)

A composition for promoting TGF-β1 production, comprising ginsenoside Rg3 or a derivative thereof as an active ingredient. A cosmetic composition for promoting skin regeneration comprising the composition of claim 1 as an active ingredient. The composition according to claim 1 or 2, wherein the content of ginsenoside Rg3 or a derivative thereof is 0.0001-10% by weight based on the total weight of the composition. The composition of claim 1 or 2, wherein the composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and A composition characterized by having a formulation selected from the group consisting of sprays.