KR20150030093A

KR20150030093A - Anti-oxidating composition containing Panax ginseng polysaccharides and Green tea polysaccharides

Info

Publication number: KR20150030093A
Application number: KR20130109335A
Authority: KR
Inventors: 이옥찬; 김동현; 황경환; 염명훈; 조준철
Original assignee: (주)아모레퍼시픽
Priority date: 2013-09-11
Filing date: 2013-09-11
Publication date: 2015-03-19
Also published as: KR102052522B1

Abstract

The present invention relates to an antioxidant composition containing a ginseng polysaccharide and a green tea polysaccharide, and more particularly, to a composition for reducing skin wrinkles and improving elasticity by providing an antioxidative effect by mixing a ginseng polysaccharide and a green tea polysaccharide will be.

Description

[0002] Antioxidant compositions containing ginseng polysaccharide and green tea polysaccharide (Panax ginseng polysaccharides and Green tea polysaccharides)

The fact that reactive oxygen species (ROS) is involved in skin aging is well known by many studies. The active acid novel of aging was proposed by Harman in 1956. It is known that the oxidative damage of biocomponents is caused by various active oxygen generated by secondary metabolism during normal metabolism and accumulation of these damages leads to aging and death will be. The damage caused by active oxygen is coped with the defensive ability of the living body, but the defense is not 100% complete, so it is affected by some active oxygen. This harmful effect occurs slowly and, if necessary, chronically through several years or lifetimes, accumulating to diminish the function of cells or tissues of the skin, which is the cause of disease and aging.

Expression of inflammation is a series of reactions to maintain homeostasis in the body, but clinical manifestations may include redness, swelling, fever, pain and dysfunction. Histopathologically, when the inflammation site is observed, infiltration of inflammatory cells mainly consisting of neutrophil and monocyte can be observed at the same time as necrosis of cells, expansion of capillaries, edema and at the same time. However, such a phenomenon does not occur at the same time but occurs in a series of sequences. In addition, many chemical agents (chemical mediators) and their action in the process have been known in recent years. The time course of inflammation is divided into three stages: Hemodynamic changes due to changes in blood flow and blood vessel size (hemodynamic changes), exudation of inflammatory cells, and proliferation of connective tissues. Among these, the part of research that is related to ROS is hemodynamic variation, and it is summarized as follows.

During inflammation, platelet activating factor (PAF) is secreted to increase permeability of blood vessels, increase nitric oxide synthesis, expand blood vessels, and induce adhesion molecules to participate in adhesion and migration of white blood cells. In addition, acute inflammation changes the surface properties of endothelial cells. There is a tendency to do a lot of studies that relate the above series of reactions to ROS.

As such, antioxidation is essential in living organisms. It is not easy to reach the skin when antioxidants are taken as food or supplements. Skin is wrapped around the outermost part of the human body, and it is always in contact with harmful factors such as external environment and ultraviolet rays, but it is a peripheral organ which is not directly connected with life. Therefore, if vitamin or antioxidant is insufficient in body, easy. In other words, from the viewpoint of the human body, it first supplies necessary components to the organs important for survival, such as the five small pieces, and then delivers the remaining ingredients to the skin. Therefore, it is necessary to give antioxidant power to skin.

Therefore, realizing the antioxidant effect on the skin will strongly satisfy the customers' demand for skin antioxidant for cosmetics, and it will become popular in various effects on skin aging, especially antioxidant, skin wrinkle and elasticity improving effect.

Accordingly, the present inventors have searched for a substance having excellent antioxidative activity among natural extracts to solve the above problems. When the ginseng polysaccharide derived from ginseng and the green tea polysaccharide derived from green tea are used simultaneously, excellent antioxidant power is provided by synergistic effect And the present invention has been completed.

Accordingly, it is an object of the present invention to provide a composition capable of providing an excellent antioxidative effect by using a ginseng polysaccharide and a green tea polysaccharide at the same time, and as a result, delaying skin aging.

To achieve the above object, the present invention provides an antioxidant composition for external application for skin or a pharmaceutical composition comprising a ginseng polysaccharide and a green tea polysaccharide as an active ingredient.

The composition of the present invention provides excellent antioxidant efficacy by containing ginseng polysaccharide and green tea polysaccharide, thereby reducing skin wrinkles, improving elasticity, and consequently delaying skin aging.

The composition of the present invention contains a ginseng polysaccharide and a green tea polysaccharide as an effective ingredient, thereby providing an excellent antioxidative effect.

The ginseng (Panax ginseng CA Meyer) polysaccharide used in the present invention is a pectin-like substance having a molecular weight of 34,600, and contains about 60% of galacturonic acid. In addition, arabinose, rhamnose, glucose and And galactose constitute side chains. The ginseng used in the present invention is a plant belonging to the genus Ogawa and Ginseng. In the present invention, ginseng can be used without limitation in its kind and year-round, and both dried and non-dried ginseng can be used. In the present invention, ginseng can be used without limitation in the site, and specifically, roots of ginseng can be used.

The green tea polysaccharide used in the present invention is derived from green tea and is an acidic polysaccharide group, unlike a polysaccharide found in other plants, and is produced by combining sugar components and amino acids produced through photosynthesis. In the present invention, green tea can be used regardless of the type and the number of days of cultivation, and both dried and non-dried green tea can be used. In the present invention, the green tea can be used without limitation in the area, and specifically, the leaves of green tea can be used.

The ginseng polysaccharide and the green tea polysaccharide used in the present invention can be produced by a method known in the art, and the method is not particularly limited.

Specifically, a green tea polysaccharide or a ginseng polysaccharide can be obtained by preparing a ginseng root or green tea leaf extract with water at 38 to 42 캜, concentrating it, removing the impurities with a filter, adding ethanol dropwise thereto and drying with hot air .

The composition of the present invention may contain the ginseng polysaccharide and the green tea polysaccharide in an amount of 0.001 to 10% by weight based on the total weight of the composition, preferably the ginseng polysaccharide and the green tea polysaccharide in an amount of 0.005 to 9.5% 0.09 to 7.5 wt.%, 0.1 to 6.5 wt.%, 0.3 to 6 wt.%, 0.5 to 5.5 wt.%, Or 0.7 By weight to 5% by weight.

In the present invention, the ginseng polysaccharide and the green tea polysaccharide are mixed in a weight ratio of 0.1 to 10: 1. In order to maximize the synergistic effect of the combination of the two components, preferably, the ginseng polysaccharide and the green tea polysaccharide are mixed in a weight ratio of 0.5 to 9.5: 1, 1 to 9: 1, or 1.5 to 8.5: 1 .

The composition of the present invention may be formulated as an external preparation for skin, in particular as a cosmetic composition, and may be formulated containing a cosmetically or dermatologically acceptable medium or base. In addition, the composition of the present invention may be provided in any form suitable for topical application, for example, as a solution, an emulsion obtained by dispersing an oil phase in water phase, an emulsion obtained by dispersing water phase in water phase, a suspension, a solid, a gel, Pastes, foams, or aerosol compositions. Compositions of such formulations may be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may contain, in addition to the above-mentioned substances, other ingredients which can give a synergistic effect to the main effect, to the extent that the main effect is not impaired. The composition according to the present invention may further comprise a humectant, an emollient, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic or inorganic pigment, a perfume, a cold agent or a limiting agent. The compounding amount of the above components can be easily selected by a person skilled in the art within the range not impairing the object and effect of the present invention, and the amount thereof may be from 0.01 to 5% by weight, specifically from 0.01 to 3% by weight based on the total weight of the composition .

In addition, the composition of the present invention can be formulated as a pharmaceutical composition. When the composition according to the present invention is applied to medicines, it may be formulated into oral, parenteral or parenteral dosage forms in solid, semi-solid or liquid form by adding an inorganic or organic carrier compatible with the active ingredient used in the present invention , The pharmaceutical composition according to the present invention may be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, and the like.

Examples of the formulations for oral administration include tablets, pills, granules, capsules, powders, fine granules, powders, emulsions, syrups and pellets. In addition, the formulations for parenteral administration include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like. The composition according to the present invention can be easily formulated into an active ingredient by carrying out the composition according to a conventional method. In this case, a surfactant, excipient, coloring agent, spice, preservative, stabilizer, buffer, suspending agent, have.

The dosage of the active ingredient of the pharmaceutical composition of the present invention will vary depending on the age, sex, body weight, pathological condition and severity of the subject to be treated, route of administration, or judgment of the prescriber. Determination of the appropriate dose based on these factors is well within the level of ordinary skill in the art and its daily dose is, for example, from 0.1 mg / kg / day to 100 mg / kg / day, more specifically from 5 mg / kg / day to 50 mg / / Day, but is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

[Reference Example 1] Preparation of ginseng polysaccharide

1 kg of dried white ginseng was pulverized with a blender, and then 10 L of purified water was added thereto, followed by stirring at 38 to 42 ° C in hot water for 24 hours, and then immersed at 15 ° C for 1 day. Thereafter, the residue and the filtrate were separated by filtration and centrifugation through a filter cloth, and the separated filtrate was concentrated under reduced pressure to 1/10 of the amount of purified water. Four times as much ethanol as the concentrated amount was added dropwise, and the mixture was hot-air dried to obtain 65 g of ginseng polysaccharide.

[Reference Example 2] Production of green tea polysaccharide

1 kg of first processed green tea leaves were pulverized with a blender, 10 l of purified water was added thereto, and the mixture was stirred for 24 hours at 38 to 42 ° C in hot water, and then immersed at 15 ° C for 1 day. Thereafter, the residue and the filtrate were separated by filtration and centrifugation through a filter cloth, and the separated filtrate was concentrated under reduced pressure to 1/10 of the amount of purified water. Four times as much concentrated ethanol as the concentrated amount was added dropwise and the mixture was hot-air dried to obtain 72 g of a green tea polysaccharide.

[Example 1] Preparation of a mixture of ginseng polysaccharide and green tea polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 1: 3.

[Example 2] Preparation of a mixture of ginseng polysaccharide and green tea polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 1: 1.

[Example 3] Preparation of a mixture of ginseng polysaccharide and green tea polysaccharide

The ginseng polysaccharide prepared in Reference Example 1 and the green tea polysaccharide prepared in Reference Example 2 were mixed at a ratio of 3: 1.

[Test Example 1] Inhibitory effect on reactive oxygen species (ROS) production

The keratinocytes isolated from human epidermal tissue (F1 eratinocyte) were added to each well of a 24-well plate at 5 x 10 ⁴ for 24 hours. After 16 hours, the materials of Examples 1 to 3 and Reference Examples 1 and 2 were treated with 1% each. The control group did not do anything for comparison at this time. After 2 hours, the culture solution was removed, and 100 μl of phosphate buffered saline (PBS) was added to each well. This keratinocyte was irradiated with ultraviolet rays of 30 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UVB15W, SanF1yo DennF1i, Japan), then PBS was removed, &Lt; / RTI > Here, the test materials of Examples 1 to 3 and Reference Examples 1 and 2 were treated again and the amount of active oxygen species increased by ultraviolet stimulation was determined at predetermined time intervals. The amount of ROS was determined by referring to Tan's method of measuring the fluorescence of DCF-DA (dichlorofluorescin diacetate) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432) The ratios of the vehicle-treated control to the ROS were calculated. The results are shown in Table 1 below.

Test substance UVB 30 mJ / cm ² Time elapsed after investigation 0hr 2 hr 3hr Vehicle 100 244 287 UVB + vehicle 100 325 381 UVB + Example 1 100 272 311 UVB + Example 2 100 267 304 UVB + Example 3 100 263 297 UVB + Reference Example 1 100 279 319 UVB + Reference Example 2 100 283 326

In the results shown in Table 1, the ginseng polysaccharide and the green tea polysaccharide used in the present invention inhibit the production of ROS which is known to cause skin cell damage by ultraviolet rays, and in particular, the combination of ginseng polysaccharide and green tea polysaccharide, In addition to suppressing the production of ROS very effectively, it can be confirmed that the amount of ROS after ultraviolet stimulation suppresses the production of ROS at almost the same level as that in the case of not irradiating ultraviolet rays, and thus the antioxidant effect is excellent.

Thus, it can be seen that the mixture of the ginseng polysaccharide and the green tea polysaccharide used in the present invention can help prevent and improve skin aging by inhibiting oxidation and preventing the generation of skin irritation.

[Test Example 2] Measurement of inhibitory effect on collagenase expression

The collagenase production inhibitory activities of Examples 1 to 3 and Reference Examples 1 and 2 were measured in comparison with the control groups, tocopherol and EGCG.

(Human Dermal Fibroblasts, neonatal (HDFn)) (lot # 617766, Cascade Biologics Inc.) was added to a 96 well plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal bovine serum. , Portland, Oregon (OR) USA) was added at 5,000 cells / well and cultured until it grew to about 90%. Thereafter, the cells were cultured in a serum-free DMEM medium for 24 hours, treated with the test substances of Examples 1 to 3 and Reference Examples 1 and 2 at a concentration of 10 ^-4 mol for 24 hours in serum-free DMEM medium, Respectively.

The collected cell culture medium was assayed for collagenase production using a commercially available collagenase measuring device (Amersham Pharmacia, USA). First, the cell culture medium, which had been plated on a 96-well plate uniformly coated with primary collagenase antibody, was added and the antigen-antibody reaction was performed in a thermostatic chamber for 3 hours.

After 3 hours, the secondary collagen antibody bound to the chromophore was put into a 96-well plate and reacted for another 15 minutes. After 15 minutes, the color development inducing substance was added and coloration was induced at room temperature for 15 minutes. When the reaction (color development) was stopped by adding 1 M sulfuric acid again, the color of reaction solution was yellow and the degree of yellow was different according to the degree of reaction progress.

The absorbance of a yellow 96 well plate was measured at 405 nm using a spectrophotometer, and the degree of synthesis of collagenase was calculated by the following equation (1). At this time, the reaction absorbance of the cell culture solution of the group not treated with the composition was used as a control.

The collagenase expression level of the test substances in the fibroblasts of the human is measured in terms of collagenase expression level as shown in Table 2 below, and the level of collagenase expression in the untreated group is set at 100 .

matter Expression level of collagenase (%) Untreated group 100 Tocopherol (positive control) 76 EGCG (positive control) 63 Example 1 66 Example 2 67 Example 3 68 Reference Example 1 79 Reference Example 2 77

From Table 2, it can be seen that the mixture of the ginseng polysaccharide and the green tea polysaccharide used in the present invention inhibits collagenase expression in vitro.

In addition, as shown in Table 1, each of Examples 1 to 3 was superior to the collagenase inhibitory effect of Reference Examples 1 and 2, and also exhibited significantly higher collagenase expression than the positive control tocopherol Effect can be confirmed.

[Experimental Example 3] Effect of promoting the production of procollagen

The procoagulant producing ability of Examples 1 to 3 and Reference Examples 1 and 2 was measured in comparison with vitamin C.

Human fibroblasts were plated at 5,000 cells / well in a 96-well plate containing DMEM (Dulbecco's Modified Eagle's Media) medium containing 2.5% fetal bovine serum, cultured until 90% Respectively. Thereafter, the cells were cultured in a serum-free DMEM medium for 24 hours. Then, the test substances of Examples 1 to 3 and Reference Examples 1 and 2 dissolved in a serum-free DMEM medium and vitamin C were added to each well at a concentration of 10 ^-4 mol After the treatment, the cell culture solution was collected. After 24 hours, the amount of procollagen liberated in the medium was measured using a procollagen type-1 C-peptide EIA kit (MK101, Takara, Japan).

The degree of procollagen production in the untreated group was determined as 100, and the degree of procollagen production in the group treated with the test substance was determined. The results are shown in Table 3 below.

matter Procollagen production (%) Untreated group 100 Vitamin C 121 Example 1 120 Example 2 120 Example 3 122 Reference Example 1 110 Reference Example 2 106

As shown in Table 3, it can be confirmed that the mixture of the ginseng polysaccharide and the green tea polysaccharide used in the present invention promotes the production of procollagen in vitro.

In addition, in Examples 1 to 3, the effect of promoting the production of procollagen was remarkably superior to those of Reference Examples 1 and 2, and the effect was confirmed to be equivalent to that of the positive control group, vitamin C.

From this, it can be seen that the mixture of the ginseng polysaccharide and the green tea polysaccharide used in the present invention has an excellent promoting effect on the production of collagen and is excellent in collagen producing ability, thus effectively preventing skin wrinkle formation and providing a wrinkle-reducing effect .

[Test Example 4] Elastase activity inhibition assay

The inhibitory activities of the elastase activity in Examples 1 to 3 and Reference Examples 1 and 2 were measured in comparison with EGCG. The elastase and substrate used were commercially purchased from Sigma Aldrich, USA (Cat. No. E0127).

The elastase activity inhibitory activity was tested by the following test method.

(200 μL) of the above Examples 1 to 3 and Reference Examples 1 and 2 were mixed with 50 μL of a 20 μg / mL elastase type III solution, respectively, in a 96-well plate to prepare 10 mg / L Tris- . EGCG 250 μM was used as a positive control, and negative control group, distilled water, was used. Then, 100 μL of 0.4514 mg / mL N-SUCCINYL-ALA-ALA-ALA-p-NITROANILIDE prepared as the buffer solution was added and reacted at 25 ° C. for 15 minutes. After completion of the reaction, the absorbance at a wavelength of 415 nm was measured. A blank test was carried out in the same manner and corrected.

The calculation method of the inhibitory activity of the elastase activity is shown in the following Equation 2, and the results are shown in Table 4 below.

A: absorbance at wavelength 415 nm in the absence of the test substance and addition of the enzyme

B: absorbance at a wavelength of 415 nm in the absence of the test substance and no enzyme addition

C: Absorbance at wavelength 415 nm in addition of test substance and enzyme addition

D: Absorbance at wavelength 415 nm in addition of test substance and enzyme-free addition

compound % Inhibition Untreated group 0 EGCG 65 Example 1 71 Example 2 70 Example 3 68 Reference Example 1 62 Reference Example 2 60

In Table 4, the ginseng polysaccharide or green tea polysaccharide used in the present invention inhibited the elastase activity, and in particular, Examples 1 to 3, in which the ginseng polysaccharide and the green tea polysaccharide were mixed, Lt; RTI ID = 0.0 > EGCG < / RTI >

[Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 3]

Nutritive creams were prepared according to the composition of Table 5 below (unit: wt%).

Compounding ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Purified water To 100 To 100 To 100 To 100 To 100 To 100 Example 1 0.1 - - - - - Example 2 - 0.1 - - - - Example 3 - - 0.1 - - - Reference Example 1 - - - 0.1 - - Reference Example 2 - - - - 0.1 - Vegetable hydrogenated oil 1.50 1.50 1.50 1.50 1.50 1.50 Stearic acid 0.60 0.60 0.60 0.60 0.60 0.60 Glycerol stearate 1.00 1.00 1.00 1.00 1.00 1.00 Stearyl alcohol 2.00 2.00 2.00 2.00 2.00 2.00 Polyglyceryl-10pentastearate and behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 1.00 1.00 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 1.00 1.00 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 2.00 2.00 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 1.50 1.50 1.50 1.50 Caprylic / carboxy triglyceride 11.00 11.00 11.00 11.00 11.00 11.00 Cyclomedicone 6.00 6.00 6.00 6.00 6.00 6.00 Preservative, incense Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Triethanolamine 0.1 0.1 0.1 0.1 0.1 0.1

[Test Example 5] Confirmation of improving skin elasticity

In order to confirm the skin elasticity improvement effect in humans, the formulations of Formulation Examples 1 to 3 and Comparative Formulation Examples 1 to 3 were evaluated as follows.

120 healthy women from 30 to 40 were divided into 6 groups each consisting of 20 individuals and one composition was used for each group. This was applied once a day for 12 weeks to the face, and then the skin elasticity was measured using a skin elasticity tester (Cutometer SEM 575, C + K Electronic Co., Germany) was used to measure skin elasticity. The results are shown in Table 6 below. The results in Table 6 indicate the viscoelasticity of the skin elasticity measuring device.

Test substance Skin elasticity effect Formulation Example 1 0.28 Formulation Example 2 0.26 Formulation Example 3 0.25 Comparative Formulation Example 1 0.19 Comparative Formulation Example 2 0.17 Comparative Formulation Example 3 0.10

As shown in Table 6, Formulations 1 to 3 and Comparative Formulation Examples 1 to 2 containing ginseng polysaccharide or green tea polysaccharide showed increased skin elasticity in comparison with the group to which Comparative Formulation Example 3 was applied, It can be confirmed that the skin elasticity of the group to which Formulation Examples 1 to 3 containing the polysaccharide mixture was applied was remarkably increased.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will readily appreciate that many modifications are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of this invention. something to do. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims

A composition for external application for skin for antioxidant comprising ginseng polysaccharide and green tea polysaccharide as an active ingredient.

[2] The composition for external application for skin according to claim 1, wherein the ginseng polysaccharide and the green tea polysaccharide are contained in an amount of 0.001 to 10% by weight based on the total weight of the composition.

The composition for external application for skin according to claim 1, wherein the ginseng polysaccharide and the green tea polysaccharide are mixed in a weight ratio of 0.1 to 10: 1.

The composition for external application for skin according to claim 1, wherein the composition is for reducing skin wrinkles.

The composition for external application for skin according to claim 1, wherein the composition is for improving skin elasticity.

An antioxidant pharmaceutical composition comprising ginseng polysaccharide and green tea polysaccharide as an active ingredient.

[Claim 7] The pharmaceutical composition according to claim 6, wherein the ginseng polysaccharide and the green tea polysaccharide are contained in an amount of 0.001-10 wt% based on the total weight of the composition.

[Claim 7] The pharmaceutical composition according to claim 6, wherein the ginseng polysaccharide and the green tea polysaccharide are mixed in a weight ratio of 0.1 to 10: 1.

The pharmaceutical composition of claim 6, wherein the composition is for reducing skin wrinkles.

The pharmaceutical composition according to claim 6, wherein the composition is for improving skin elasticity.