KR20130031988A - Skin external composition containing floral ginsenoside - Google Patents

Abstract

PURPOSE: An external use skin composition containing ginseng flower-derived floral ginsenoside is provided to ensure excellent anti-oxidation, anti-aging, whitening, moisturizing, and anti-inflammation effects. CONSTITUTION: An external use skin composition contains 0.001-50wt% of ginseng flower-derived floral ginsenoside. The floral ginsenoside is selected from the group consisting of floral ginsenoside A, B, C, D, E, F, M, N, O, and P. The composition is used for anti-aging, anti-oxidation, skin whitening, moisturizing, anti-inflammation, and atopic dermatitis treatment.

Description

Skin external composition containing floral ginsenoside

The present invention relates to an external composition for skin containing floral ginsenoside, which is a component of ginseng flower, and more specifically, floral ginsenosides A, B, C, D, which are specific ginsenosides of ginseng flower. It relates to an external composition for skin containing E, F, M, N, O and P, which is excellent in skin antioxidant, anti-aging, whitening, moisturizing and anti-inflammatory effects.

Human skin is changed by a number of internal and external factors as it ages. That is, internally, the secretion of various hormones that regulate metabolism decreases, and the function of immune cells and the activity of cells decreases, thereby reducing the biosynthesis of immune proteins and constituent proteins necessary for living organisms. Due to the increase in the amount of ultraviolet rays that reach the surface of the sun's rays and further increase environmental pollution, free radicals and free radicals, such as free radicals, skin thickness is reduced, wrinkles are increased, elasticity is reduced only In addition, the color of the skin becomes dull, skin problems frequently occur, and various changes such as blemishes, freckles, and black mushrooms also increase.

As aging progresses, symptoms such as changes and decreases in the content and arrangement of collagen, elastin, hyaluronic acid, and glycoproteins, which are constituting skin, appear, and are subjected to oxidative stress caused by free radicals and active harmful oxygen. In addition, by aging or ultraviolet light, cyclooxygenase-2 (Cox-2, cyclooxygenase), an enzyme that produces proinflammatory cytokine, which is known to cause inflammation in most cells that make up the skin Increased biosynthesis, increased biosynthesis of matrix metalloproteinase (MMP), an enzyme that degrades skin tissue by these inflammatory factors, and increased nitric oxide (NO) production by inducible nitric oxide synthase (iNOS) It is known. In other words, the biosynthesis of the substrate material is reduced by the reduction of cellular activity and micro-inflammation due to naturally occurring endogenous aging, and by external factors such as the increase of stress caused by various harmful environments and the increase of free radical species caused by sunlight. As a result, decomposition and degeneration are accelerated, and the skin matrix is destroyed and thinned, resulting in various symptoms of skin aging. Therefore, a lot of research is being conducted on the active ingredient that can prevent and improve the phenomenon of aging.

On the other hand, free radicals generated by various physical, chemical and environmental factors such as enzymes, reducing metabolism, chemicals, pollutants and photochemical reactions in the body are non-selective and irreversible to cell components such as lipids, proteins, sugars and DNA. It has been known to cause various diseases including cell aging or cancer by performing a destructive action. In addition, various peroxides in the body, including lipid peroxides produced as a result of lipid peroxidation by these free radicals, also cause oxidative destruction of cells and cause various functional disorders. Accordingly, antioxidants such as free radical scavengers or peroxide production inhibitors are expected as agents for inhibiting or treating aging and various diseases caused by these oxides.

On the other hand, ginseng is a perennial root crop, which uses the roots of ginseng grown for 4 to 6 years. Recently, however, the use of ginseng has been diversified by researching various parts such as stems, leaves, fruits, flowers, etc. In addition, the demand for ginseng increases due to the trend of well-being, and industrial use and research using parts other than roots are increasing.

Recently, active ingredients of various ginsengs other than ginsenoside, ginseng saponin, have been found, and various physiological activities thereof have been verified. Accordingly, various ginsenosides and active ingredients are being used for the development of health functional foods and cosmetics.

Accordingly, the present inventors conducted a study to find a ginseng ginsenoside having a superior performance than the ginsenosides of the conventional ginseng root, as a result, the ginseng flower-derived floral ginsenoside different from the conventional ginseng ginsenosides is a skin antioxidant, It was found that the anti-aging, whitening and moisturizing effect is excellent and completed the present invention.

Accordingly, it is an object of the present invention to provide an external composition for skin, containing ginseng flower-derived floral ginsenosides having excellent skin efficacy and having excellent skin antioxidant, anti-aging, whitening, moisturizing and anti-inflammatory effects.

In order to achieve the above object, the present invention provides a skin external preparation composition containing a ginseng flower-derived floral ginsenosides.

The ginseng flower-derived floral ginsenoside according to the present invention exhibits excellent skin efficacy against antioxidant, anti-aging, whitening, moisturizing, and anti-inflammatory, compared to ginseng ginsenosides, and the composition containing the same may be used in various fields such as cosmetics and medicines. Do.

The composition of the present invention includes a floral ginsenoside as an active ingredient.

The floral ginsenoside used in the present invention is isolated from the ginseng flower, and is one of the floral ginsenosides A, B, C, D, E, F, M, N, O and P represented by the following Chemical Formulas 1 to 10: More than one species can be used.

The composition of the present invention contains ginseng flower-derived floral ginsenosides in an amount of 0.001 to 50% by weight, based on the form of the external preparation composition. At this time, if the content is less than 0.001% by weight, the following efficacy is insufficient, and if more than 50% by weight, the efficiency of toxicity and efficacy is lowered.

In the present invention, by containing the floral ginsenoside as an active ingredient, it provides a skin external preparation composition for inhibiting skin aging or improving wrinkles having an antioxidant effect, collagen production promotion or MMP-1 inhibitory effect.

In addition, the present invention provides a skin whitening composition for skin whitening having an effect of inhibiting melanin production or improving pigmentation by containing the floral ginsenoside as an active ingredient.

In another aspect, the present invention provides a skin moisturizing or atopic dermatitis composition having an effect of improving skin moisturizing and atopic symptoms by inducing and maintaining normal differentiation of epidermal keratinocytes by containing the floral ginsenoside as an active ingredient. .

In addition, the present invention provides an anti-inflammatory skin external composition for improving skin troubles including acne by controlling the sebum control or anti-inflammatory effect, that is, inhibiting the production of inflammatory factors by containing the floral ginsenoside as an active ingredient.

The external preparation composition for skin according to the present invention may be formulated as a cosmetic or pharmaceutical composition, and may further contain cosmetic or dermatologically acceptable media or bases commonly used in the art. These are all formulations suitable for topical application according to conventional methods in the art, for example emulsions, suspensions, microemulsions, microcapsules, microgranules, obtained by dispersing the oil phase in solutions, gels, solids, pasty anhydrous products, aqueous phases. Alternatively, they may be prepared in the form of ionic (liposomal) and nonionic vesicle dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays, or cone sticks. The external preparation composition for skin according to the present invention may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the composition for external application for skin according to the present invention may be a fatty substance, an organic solvent, a dissolving agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, an ionic type or Nonionic emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics It may further contain an adjuvant commonly used in the cosmetic or dermatological field, such as may be introduced in an amount generally used in the cosmetic or dermatological field.

In addition, the topical skin composition according to the present invention is not particularly limited in the formulation, for example, softening longevity, astringent longevity, nourishing longevity, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing It may be formulated into cosmetic compositions such as foams, cleansing water, packs, powders, body lotions, body creams, body oils or body essences, or pharmaceutical compositions such as suspensions, ointments, lotions, sprays, and the like.

Hereinafter, the present invention will be described more specifically with reference to examples and test examples. These examples are provided only for understanding the contents of the present invention, but the scope of the present invention is not limited to these examples and test examples, and modifications, substitutions, and insertions commonly known in the art may be performed. It may be included in the scope of the present invention.

Example 1 Preparation of Extract Containing Floral Ginsenosides

5 g of water was added to 1 kg of dried ginseng flower (Mayeongmyeon-myeon, Jinan-gun, Jeollabuk-do), and the mixture was extracted under reflux. N-butanol layer was passed through normal phase silica gel column chromatography and reversed phase silica gel column chromatography, and finally, by HPLC, floral ginsenosides A (0.0053%), B (0.057%), C (0.014%) , D (0.0054%), E (0.0014%), F (0.0046%), M (0.0023%), N (0.0016%), O (0.0055%) and P (0.0090%) were prepared.

¹³ C-NMR data of each floral ginsenoside is shown in Tables 1 and 2 below.

¹³ C-NMR data for floral ginsenosides A, B, C, D, E and F One 2 3 4 5 6 C-1 39.6 39.5 39.4 39.4 39.3 39.3 C-2 28.0 28.0 28.1 28.2 26.8 26.8 C-3 78.8 78.9 78.6 78.6 89.0 88.9 C-4 40.4 40.4 40.3 40.4 39.8 39.8 C-5 61.5 61.5 61.8 61.8 56.5 56.5 C-6 80.1 80.1 67.8 67.8 18.5 18.5 C-7 45.3 45.2 47.5 47.5 35.3 35.2 C-8 41.2 41.2 41.2 41.3 40.2 40.2 C-9 50.1 50.3 50.0 49.9 50.5 50.2 C-10 39.8 39.8 39.4 40.4 37.1 37.1 C-11 31.1 31.2 30.9 30.9 32.2 31.1 C-12 70.3 70.5 70.2 70.6 71.1 70.5 C-13 49.3 49.3 49.2 49.2 49.0 49.7 C-14 51.5 51.5 51.4 51.5 51.8 51.6 C-15 30.7 30.6 30.8 30.7 31.4 30.7 C-16 26.8 26.5 26.7 26.4 26.8 26.5 C-17 51.5 52.2 52.1 52.1 54.2 52.3 C-18 17.6 17.6 17.6 17.7 16.0 16.0 C-19 17.6 17.6 17.5 17.5 16.5 16.3 C-20 83.2 83.2 83.3 83.2 73.3 83.2 C-21 22.6 23.2 21.8 23.3 27.9 23.3 C-22 32.9 39.7 32.8 39.9 40.4 39.7 C-23 26.7 126.6 26.7 126.7 127.3 126.6 C-24 89.9 138.0 90.1 138.1 135.8 138.1 C-25 145.8 81.3 146.2 81.3 81.3 81.3 C-26 113.3 25.2 113.3 25.4 25.2 25.2 C-27 17.8 25.4 17.8 25.2 25.3 25.4 C-28 31.8 31.8 32.0 32.0 28.2 28.2 C-29 16.4 16.4 16.5 16.5 16.7 16.8 C-30 17.3 17.1 17.4 17.3 17.1 17.2 C-1 ' 106.0 105.9 98.0 98.2 105.1 106.9 C-2 ' 75.5 75.5 74.9 75.1 83.6 75.8 C-3 ' 79.6 78.1 79.4 78.9 78.1 78.8 C-4 ' 72.0 72.0 72.1 72.2 71.8 72.0 C-5 ' 78.1 79.6 76.4 76.4 78.1 78.3 C-6 ' 63.1 63.1 69.5 68.5 63.0 63.1 C-1 '' 98.1 98.3 105.1 110.2 106.1 98.3 C-2 '' 75.1 75.3 72.2 83.3 77.1 75.3 C-3 '' 79.2 78.0 74.2 78.8 78.4 78.9 C-4 '' 71.8 71.1 68.8 86.2 71.9 71.7 C-5 '' 78.0 78.7 66.0 62.9 78.2 78.2 C-6 '' 63.1 63.2 62.9 63.2

Measured at pyridine- d ₅ at 125 MHz and 150 MHz.

¹³ C-NMR data for floral ginsenosides M, N, O and P One 2 3 4 C-1 39.5 39.5 39.2 39.3 C-2 27.8 27.8 26.8 26.7 C-3 78.8 78.8 89.1 89.5 C-4 40.0 40.0 39.7 40.6 C-5 60.9 61.0 56.5 61.8 C-6 79.4 74.8 18.5 67.6 C-7 45.9 46.0 35.1 47.6 C-8 41.3 41.3 40.1 41.2 C-9 50.0 49.7 50.2 49.9 C-10 39.8 39.8 37.0 38.9 C-11 30.9 30.9 30.9 30.8 C-12 70.2 70.3 70.6 70.3 C-13 49.1 49.2 49.7 49.2 C-14 51.5 51.5 51.5 51.4 C-15 30.8 30.8 30.6 30.8 C-16 26.7 26.7 26.4 26.7 C-17 51.8 51.8 52.1 51.7 C-18 17.5 17.5 16.1 17.6 C-19 17.4 17.3 16.3 17.4 C-20 83.4 83.5 83.2 83.5 C-21 22.4 22.4 23.3 22.4 C-22 36.2 36.2 40.0 36.2 C-23 23.1 23.3 126.7 23.3 C-24 126.1 126.0 138.0 123.8 C-25 131.0 131.1 81.3 131.1 C-26 25.8 25.8 25.1 25.8 C-27 17.8 17.8 25.4 17.9 C-28 32.2 32.2 28.2 31.4 C-29 17.6 17.6 16.6 16.8 C-30 17.3 17.4 17.2 17.5 C-1 ' 101.8 101.8 105.1 105.3 C-2 ' 79.0 79.4 83.5 83.5 C-3 ' 78.2 78.5 78.0 78.0 C-4 ' 72.7 72.1 72.0 71.9 C-5 ' 78.2 78.2 78.0 78.1 C-6 ' 63.3 63.3 62.8 63.0 C-1 '' 101.9 101.9 106.0 106.0 C-2 '' 72.3 72.4 77.1 77.0 C-3 '' 72.4 72.4 78.8 78.5 C-4 '' 74.2 74.3 71.7 71.8 C-5 '' 69.5 69.5 78.1 78.1 C-6 '' 18.7 18.7 62.9 63.0 C-1 '' ' 98.1 98.2 98.2 98.2 C-2 '' ' 75.1 75.0 75.2 75.0 C-3 '' ' 78.2 79.1 78.1 79.2 C-4 '' ' 71.8 71.8 71.8 71.9 C-5 '' ' 76.5 76.7 76.4 76.7 C-6 '' ' 68.5 69.2 68.4 69.2 C-1 '' '' 110.6 104.5 110.2 104.5 C-2 '' '' 83.2 72.0 83.2 72.1 C-3 '' '' 78.5 74.1 78.4 74.1 C-4 '' '' 86.2 68.4 86.1 68.4 C-5 '' '' 63.0 65.4 62.8 65.4

Measured at pyridine- d ₅ at 125 MHz and 150 MHz.

Test Example 1 Antioxidant Effect Test (DPPH Test)

In order to determine the antioxidant effect of the floral ginsenosides prepared in Example 1, the organic radical DPPH (1,1-diphenyl-2-picrylhydrazyl; 1,1-diphenyl-2-picryl hydrazyl) It was performed by evaluating the antioxidant capacity by comparing the DPPH antioxidant inhibition effect through the change in absorbance generated by reduction (antioxidant is oxidized). That is, by measuring the degree to which the oxidation of DPPH is suppressed and the absorbance is reduced in comparison with the control group, the concentration of the absorbance of 50% or less compared to the control group is measured. Evaluated as effective antioxidant concentration.

190 μl of 100 μM (in ethanol) DPPH solution and 10 μl of the floral ginsenosides A, B, C, D, E, F, M, N, O, P and the positive control of Example 1 obtained above After the reaction solution was made and reacted at 37 ° C. for 30 minutes, the absorbance was measured at 540 nm. As the positive control group, Trolox, a synthetic antioxidant widely used, was used. The results of DPPH analysis of each substance are shown in Table 3 below, and IC ₅₀ means the sample concentration when the absorbance was reduced by 50% by the added sample.

DPPH analysis results Test substance IC ₅₀ (ppm) Trolox 46 Example 1 Floral ginsenoside A 20 Floral ginsenoside B 21 Floral ginsenoside C 19 Floral ginsenoside D 20 Floral ginsenoside E 23 Floral ginsenoside F 22 Floral ginsenoside M 21 Floral ginsenoside N 18 Floral ginsenoside O 19 Floral ginsenoside P 21

As can be seen in Table 3, the floral ginsenosides A, B, C, D, E, F, M, N, O and P of Example 1 are compared to trolox, a synthetic antioxidant used as a positive control. The effect was also excellent. Therefore, it was found that the floral ginsenoside according to the present invention has an excellent antioxidant effect.

Test Example 2 Anti-Aging Effect Test I (Measurement of Collagenase Expression Inhibition Effect)

The collagenase production inhibitory activity of the floral ginsenosides obtained in Example 1 was measured in comparison with tocopherol and EGCG. Tocopherol and EGCG are antioxidants and are known to have the function of regenerating the epidermal cells of the skin to prevent aging of the skin.

The test was performed by placing human fibroblasts at 5,000 cells / well in a 96-well microtiter plate containing Dulbecco's Modified Eagle's Media (DMEM) medium containing 2.5% fetal calf serum. Incubate until 90% growth. Then, incubated in serum-free DMEM medium for 24 hours, and then dissolved in the serum-free DMEM medium, the floral ginsenosides of Example 1 A, B, C, D, E, F, M, N, O, P, tocopherol And each of EGCG at 10 ^-4 molarity for 24 hours, and cell culture was collected.

The collected cell culture solution was measured for collagenase production using a commercially available collagenase measuring instrument (Amersham Pharma Co., Ltd., USA). First, the collected cell culture was placed in a 96-well plate uniformly coated with primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours.

After 3 hours, the chromophore-bound secondary collagen antibody was placed in a 96-well plate and reacted for another 15 minutes. After 15 minutes, the coloring stimulant was added, causing color development at room temperature for 15 minutes, and 1M sulfuric acid was added again to stop the reaction (color development). The color of the reaction solution was yellow and the degree of yellow color was different according to the progress of the reaction.

The absorbance of the yellowish 96-well plate was measured at 405 nm using an absorbance meter, and the degree of synthesis of collagenase was calculated by Equation 1 below. At this time, the reaction absorbance of the collected cell culture medium of the group not treated with the composition was used as a control. That is, the expression level of collagenase in the untreated group was set to 100, and the collagenase expression level was calculated in the group treated with the test substance, and the results are shown in Table 4 below.

Test substance Collagenase expression level (%) Untreated group 100 Tocopherol 75 EGCG 60 Example 1 Floral ginsenoside A 71 Floral ginsenoside B 66 Floral ginsenoside C 69 Floral ginsenoside D 65 Floral ginsenoside E 64 Floral ginsenoside F 64 Floral ginsenoside M 67 Floral ginsenoside N 63 Floral ginsenoside O 69 Floral ginsenoside P 68

The lower the expression level of collagenase, the higher the expression inhibiting ability of collagenase and the less the amount of wrinkles generated due to less degradation of collagen in the skin. In Table 4, the floral ginsenosides of the present invention effectively inhibited the expression of collagenase in vitro , and it was confirmed that the expression of collagenase was superior to tocopherol, which is known as an antioxidant. . Therefore, the floral ginsenoside according to the present invention effectively inhibits the expression of collagenase, thereby reducing collagen breakdown in the skin and thus, it was found that the anti-aging effect is excellent.

Test Example 3 Anti-aging Effect Test II (Procollagen Production Promoting Effect Test)

Procollagen production ability of the extract obtained in Example 1 was measured by comparing with vitamin C. Procollagen is a substance that induces collagen production and is necessary for collagen production and anti-aging, and vitamin C is known as an essential ingredient for collagen synthesis.

The test was performed by placing human fibroblasts at 5,000 cells / well in a 96-well microtiter plate containing Dulbecco's Modified Eagle's Media (DMEM) medium containing 2.5% fetal calf serum. Incubate until 90% growth. After culturing in serum-free DMEM medium for 24 hours, and then dissolved in the serum-free DMEM medium, the floral ginsenosides of Example 1 A, B, C, D, E, F, M, N, O, P, vitamin Each C was treated with 10 ^-4 molarity for 24 hours, and then cell culture was collected. After 24 hours, the amount of free collagen in the medium was measured using the procollagen type-1 C-peptide EIA kit (MK101, Takara, Japan).

The degree of procollagen production in the untreated group was set to 100, and the degree of procollagen production in the group treated with the test substance was calculated. The results are shown in Table 5 below.

Test substance Procollagen production level (%) Untreated group 100 Vitamin C 120 Example 1 Floral ginsenoside A 121 Floral ginsenoside B 122 Floral ginsenoside C 119 Floral ginsenoside D 125 Floral ginsenoside E 126 Floral ginsenoside F 124 Floral ginsenoside M 118 Floral ginsenoside N 126 Floral ginsenoside O 120 Floral ginsenoside P 119

The higher the degree of procollagen production, the higher the degree of collagen production, thus preventing the formation of skin wrinkles. In Table 5, the floral ginsenoside in the present invention was confirmed that the degree of promotion of the production of vitamin C and procollagen, which are known as essential ingredients for collagen synthesis in vitro , are comparable or better. Therefore, the floral ginsenosides according to the present invention was found to have an excellent anti-aging effect by effectively promoting the production of procollagen to help the production of collagen in the skin.

Test Example 4 Whitening Effect Test (Measurement of Melanin Inhibition Effect Using Pigment Cells)

The melanin production inhibitory ability of the floral ginsenosides A, B, C, D, E, F, M, N, O and P obtained in Example 1 was measured in comparison with hydroquinone.

Mel-Ab cells derived from C57BL / 6 mice (Dooley, TP et al, Skin pharmacol, 7, pp 188-200) in DMEM with 10% fetal placental serum, 100nM 12-O-tetradecanoyl Incubated at 37 ° C. and 5% CO ₂ in a medium to which phorbol (tetradecanoylphorbol) -13-acetate and 1 nM cholera toxin were added. The cultured Mel-Ab cells were detached with 0.25% trypsin-EDTA, the cells were incubated at a concentration of 10 ⁵ cells / well in 24-well plates, and then each test material was added for 3 consecutive days. And incubated. As the test substance, hydroquinone and the floral ginsenosides A, B, C, D, E, F, M, N, O and P of Example 1 were used at a concentration of 10 ppm, respectively. At this time, the hydroquinone was used as a positive control group. Then, the culture solution was removed, washed with PBS, and dissolved in 1N sodium hydroxide to measure the absorbance at 400 nm, and the melanin production inhibition rate was calculated according to Equation 2 below and the results are shown in Table 6 below. Dooley's way).

Test substance Melanin production inhibition rate (%) Untreated group 100 Hydroquinone (positive control) 41.1 Example 1 Floral ginsenoside A 40.5 Floral ginsenoside B 39.4 Floral ginsenoside C 41.5 Floral ginsenoside D 40.2 Floral ginsenoside E 39.6 Floral ginsenoside F 42.2 Floral ginsenoside M 40.4 Floral ginsenoside N 41.7 Floral ginsenoside O 40.0 Floral ginsenoside P 41.4

As shown in Table 6, it was confirmed that the floral ginsenosides of the present invention showed melanin production inhibition similar to that of hydroquinone, a known whitening substance. Therefore, the floral ginsenoside according to the present invention was found to have an excellent whitening effect by effectively inhibiting melanin production in the skin.

Test Example 5 Irritation Test

In order to compare the usability of kojic acid, a known whitening substance, and the floral ginsenoside used as an active ingredient in the present invention, the degree of irritation such as stinging, burning, etc. was measured in 15 panels sensitive to irritation such as stinging, burning, etc. Experiment.

The subjects rubbed the ginseng flower extract containing kojic acid (purchased through YM chemical) and floral ginsenoside (Example 1) by applying randomly changing the left and right by 0.5 ml each, and applying 0 to 3.0 by 0.1 point unit. Scored between. The results are shown in Table 7 below.

<Evaluation Criteria>

0 to 0.4: no stimulation

0.5 to 1.0: slightly irritating

1.1 to 2.0: moderate stimulation

2.1 to 3.0: severe irritation

division Kojic acid With floral ginsenosides
Ginseng flower extract
(Example 1) Stinging 0.85 0.18 Burning 0.50 0.24 Average 0.68 0.21

As can be seen in Table 7, in the case of kojic acid, there was some degree of tingling and burning, and there was a sense of irritation that can be felt slightly. On the other hand, the ginseng flower extract containing the floral ginsenoside used in the present invention is almost irritable because it can hardly feel both burning and stinging.

Therefore, it was confirmed that the floral ginsenoside of the present invention has no irritant feeling unlike the kojic acid, thereby providing a better feeling of use.

Test Example 6 Skin Moisturizing Test (Induction Test for Differentiation of Human Keratinocytes)

In order to confirm the skin barrier function and skin moisturizing ability of the floral ginsenosides A, B, C, D, E, F, M, N, O and P, which are the extracts of Example 1, primary cultures were carried out by a test using absorbance. Human keratinocytes were put in a culture flask and attached to the bottom, and the floral ginsenosides A, B, C, D, E, F, M, N, O, and P were added to the culture medium as shown in Table 8 below. In addition, the cells were cultured for 5 days until the cells grew to 70-80% of the floor area. The cells were harvested and washed with PBS (phosphate buffered saline), followed by 10 mM Tris-HCl (Tris-HCl) containing 2% Sodium Dodecyl Sulfate (SDS) and 20 mM Dithiothreitol (DTT). , pH 7.4) 1 ml was added and sonication was performed for 3 minutes and then boiled for 10 minutes. The precipitate was separated by centrifugation at 1200 rpm for 30 minutes and suspended in 1 ml of PBS, and the absorbance at 340 nm was measured.

Separately, a portion of the solution after the sonication was taken to measure the protein content, which was used as a reference for evaluating the degree of cell differentiation. The low calcium (0.03mM) treated group and the high calcium (1.2mM) treated group were negative / positive controls, respectively, and the test results performed by adding test substances to low calcium concentrations are shown in Table 8 below.

Test substance Concentration The ability to differentiate in keratinocytes (%) Control group Low Ca ^{2 +} (0.03 mM) 100 Hign Ca ^{2 +} (1.2 mM) 206 Floral ginsenoside A
0.1 ppm 111 0.05 ppm 123 Floral ginsenoside B 0.1 ppm 109 0.05 ppm 125 Floral ginsenoside C 0.1 ppm 107 0.05 ppm 121 Floral ginsenoside D 0.1 ppm 109 0.05 ppm 122 Floral ginsenoside E 0.1 ppm 107 0.05 ppm 123 Floral ginsenoside F 0.1 ppm 110 0.05 ppm 120 Floral ginsenoside M 0.1 ppm 109 0.05 ppm 122 Floral ginsenoside N 0.1 ppm 108 0.05 ppm 119 Floral ginsenoside O 0.1 ppm 109 0.05 ppm 123 Floral ginsenoside P 0.1 ppm 110 0.05 ppm 124

As shown in Table 8, as a result of measuring the amount of CE (Cornified Envelop) produced when keratinocyte differentiation was compared to compare the effect of promoting cell differentiation, Floral ginsenosides according to the present invention was found to promote differentiation in keratinocytes. Therefore, the floral ginsenoside according to the present invention was found to enhance skin barrier function and enhance skin moisturizing ability.

Formulation Example 1 Nutritional Cosmetics

Nutrients were prepared in a conventional manner according to the composition shown in Table 9.

ingredient Content (% by weight) Extract Containing Floral Ginsenosides (Example 1) 5.0 Squalane 5.0 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Caprylic / capric triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount Purified water Balance Sum 100

[Production Examples 1 to 10 and Comparative Production Example 1]

To the composition of Table 10 to prepare a nutrient cosmetics of Preparation Examples 1 to 10 containing a floral ginsenoside and Comparative Preparation Example 1 containing no floral ginsenoside (unit: weight%).

ingredient Preparation Example 1 Preparation Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 Preparation Example 7 Preparation Example 8 Production Example 9 Preparation Example 10 Comparative Preparation Example 1 Floral ginsenoside A 5.0 - - - - - - - - - - Floral ginsenoside B - 5.0 - - - - - - - - - Floral ginsenoside C - - 5.0 - - - - - - - - Floral ginsenoside D - - - 5.0 - - - - - - - Floral ginsenoside E - - - - 5.0 - - - - - - Floral ginsenoside F - - - - - 5.0 - - - - - Floral ginsenoside M - - - - - - 5.0 - - - - Floral ginsenoside N - - - - - - - 5.0 - - - Floral ginsenoside O - - - - - - - - 5.0 - - Floral ginsenoside P - - - - - - - - - 5.0 - Squalane 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Wax 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Sorbitan sesquioleate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Liquid paraffin 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Caprylic / capric triglyceride 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 glycerin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Carboxyvinyl polymer 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Triethanolamine 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance Balance Balance Sum 100 100 100 100 100 100 100 100 100 100 100

[Formulation Example 2] Nourishing cream

Nutritional cream was prepared in a conventional manner according to the composition described in Table 11.

ingredient Content (% by weight) Extract Containing Floral Ginsenosides (Example 1) 5.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 PEG60 hardened castor oil 2.0 Liquid paraffin 10.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount Purified water Balance Sum 100

Reference Example 2 Preparation Examples 11 to 20 and Comparative Production Example 2

The nutritional cream of Preparation Example 11-20 containing floral ginsenoside and Comparative Preparation Example 2 containing no floral ginsenoside were prepared in the composition of Table 12 (unit: wt%).

ingredient Production Example 11 Production Example 12 Production Example 13 Production Example 14 Production Example 15 Production Example 16 Production Example 17 Preparation Example 18 Preparation Example 19 Preparation Example 20 Comparative Production Example 2 Floral ginsenoside A 5.0 - - - - - - - - - - Floral ginsenoside B - 5.0 - - - - - - - - - Floral ginsenoside C - - 5.0 - - - - - - - - Floral ginsenoside D - - - 5.0 - - - - - - - Floral ginsenoside E - - - - 5.0 - - - - - - Floral ginsenoside F - - - - - 5.0 - - - - - Floral ginsenoside M - - - - - - 5.0 - - - - Floral ginsenoside N - - - - - - - 5.0 - - - Floral ginsenoside O - - - - - - - - 5.0 - - Floral ginsenoside P - - - - - - - - - 5.0 - Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Sorbitan sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 PEG60 hardened castor oil 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Liquid paraffin 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 10.0 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Caprylic / capric triglyceride 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Triethanolamine 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance Balance Balance Sum 100 100 100 100 100 100 100 100 100 100 100

[Test Example 7] Atopic dermatitis improvement effect

The effect of improving atopic symptoms was measured as follows. Forty patients who were diagnosed with atopy symptoms (itch, erosion, severe dryness) or had atopy-like symptoms by visual observation were divided into four groups, each of Formulation Examples 1, 2 and Comparative Preparation Examples. 1 and 2 nourishing creams and nourishing creams were provided and applied to the site showing atopic symptoms twice a day for 12 weeks. Efficacy assessments were conducted after 12 weeks of treatment and subjectively judged the improvement. In the questionnaire, the symptoms of atopic dermatitis are divided into "itch", "dry", "keratogenesis", "dandruff", "erythema", "swelling", "skin cracking", and "treat and eczema". In this case, after evaluating the degree of improvement for each symptom, the average of the atopy symptom improvement effect was evaluated for each individual, and the results are shown in Table 13 below.

Test substance Number of respondents Significant improvement Improving No change worse Formulation Example 1 3 7 10 0 Formulation Example 2 7 9 4 0 Comparative Preparation Example 1 2 4 14 0 Comparative Production Example 2 5 9 6 0

In the results of Table 13, the ginseng flower extract containing the floral ginsenoside containing the formulation examples 1 and 2 containing the ginseng flower extract containing the floral ginsenoside of Example 1 according to the present invention It was confirmed that the atopic symptoms were more improved than the group to which Comparative Preparation Examples 1 and 2 were not applied.

In addition, in order to measure the effect of improving atopic symptoms on floral ginsenosides A, B, C, D, E, F, M, N, O, and P, respectively, they were diagnosed with atopic symptoms (itch, erosion, severe dryness). Or, 200 people in their 10's and 50's with atopic-like symptoms by visual observation were divided into 10 groups, and provided with nutrients of Preparation Examples 1-10, respectively, twice a day at the site showing atopic symptoms. Application was made for a week. Evaluation of efficacy was carried out in the same manner as described above, the results are shown in Table 14 below.

Test substance Number of respondents Test substance Significant improvement Improving No change worse Production Example 1 3 7 10 0 Production Example 2 4 7 9 0 Production Example 3 3 6 11 0 Production Example 4 3 7 10 0 Production Example 5 5 8 7 0 Production Example 6 3 5 12 0 Production Example 7 4 6 10 0 Production Example 8 4 5 11 0 Production Example 9 3 6 11 0 Production Example 10 5 8 7 0 Comparative Preparation Example 1 2 4 14 0

Looking at the results of Table 14, as in the case of applying the formulation examples 1 and 2 containing the ginseng flower extract containing the floral ginsenoside of Example 1, the floral ginsenosides A, B, C, D, E, Even using Preparation Examples 1 to 10 containing F, M, N, O, and P, respectively, it can be confirmed that atopic symptoms were improved overall.

Test Example 8 Increasing Epidermal Lipid (Total Ceramide) Production in Human Skin

In order to evaluate the effect of the ginseng flower extract containing floral ginsenoside of Example 1 on the increase of lipid synthesis in real human skin, Carbopol ETD 2020 0.5%, TEA 0.45%, test substance 5%, and deionized 100 was used with water. Twelve adults and 20 μl of ginseng flower extract containing the control group and the floral ginsenoside of Example 1 were placed twice daily on the forearm with the diameter of 50 ml polypropylene cornical tube on the forearm. During application, each subject was divided into two groups, and the amount of lipid produced on day 3 and day 11 was extracted and analyzed in the following manner. Wash the forearm with tap water, tape stripping once with Scotch 810 Magic tape, and cut 50 ml polypropylene conical tubes as a reservoir using 1 ml of cyclohexane Add cyclohexane / ethanol (4: 1), mix for about 1 minute, transfer it to a new tube, and again treat 1 ml of cyclohexane / ethanol (1: 1) in the same manner as above. Collected. Thereafter, the mixture was dried using nitrogen gas at 50 ° C., and then dissolved in 500 μl of chloroform / methanol (2: 1), and each test material was prepared using CAMAG's Automated TLC sampler-4. (Thin Layer chromatography) was applied to the plate, and developed using a developing solvent having a composition ratio as shown in Table 15 using AMD (Automated Multiple Development).

chloroform Methanol water Acetic acid Hexane Diethyl ether Petroleum ether Solvent movement distance
(cm) Primary 40 10 One - - - - 6.0 Secondary 190 9 - - - - - 6.5 Third - - - 2 12 3 - 7.5 4th - - - - - - 100 Top

After the TLC plate was developed, the size of each band was measured at a wavelength of 570 nm using a TLC scanner-II (CAMAG: densitometer). The ceramide production amount of each treatment group which converted the ceramide production amount of the untreated group into 100% is shown in Table 16 below.

Test substance 3 days after processing (%) 11 days after processing (%) Control group Untreated group 100 100 Control group Vehicle 98 94 Control group Glycerin (5%) 102 108 Floral ginsenoside A 104 111 Floral ginsenoside B 107 114 Floral ginsenoside C 105 112 Floral ginsenoside D 106 113 Floral ginsenoside E 104 113 Floral ginsenoside F 107 115 Floral ginsenoside M 108 115 Floral ginsenoside N 106 114 Floral ginsenoside O 105 114 Floral ginsenoside P 107 116

In the results of Table 16, three days after the treatment with floral ginsenosides, ceramide production was increased compared to the negative control (non-treated vehicle) and ceramide production was increased compared to the positive control (glycerine). After 11 days, it was confirmed that the lipid producing ability was much better than the negative control group (carrier) and the positive control group (glycerine). This demonstrates the ceramide increase by the floral ginsenosides according to the present invention.

As the production of ceramide increases, the formation of the stratum corneum increases, making the stratum corneum firm and flexible, thereby retaining its function as a skin barrier. Therefore, it can be seen that the floral ginsenoside according to the present invention is effective in restoring skin barrier function.

[Test Example 9] Acne and skin trouble improvement effect

Participants in the experiment were 20 women with severe skin problems including acne. After washing the face with soap in a constant temperature and humidity room for 30 minutes, the skin trouble was measured by comparing the number of acne and the number with inflammation. .

The test subjects were allowed to use the nutritional longevity and nutrition cream of Formulation Examples 1 and 2 and Comparative Preparation Examples 1 and 2 for 4 weeks, respectively, and then compared the degree of acne and skin problems, and the results are shown in Table 17 below. It was.

Evaluation item Formulation Example 1
(5 people) Formulation Example 2
(5 people) Comparative Production Example 1
(5 people) Comparative Production Example 2
(5 people) Skin trouble improvement rate 60% 80% 25% 55%

As shown in the results of Table 17, the formulation of Examples 1 and 2 containing the ginseng flower extract containing the floral ginsenoside of Example 1 according to the present invention for 4 weeks as a result of reducing the number of existing acne and accompanied by inflammation Trouble improved.

In addition, 50 women with severe skin problems, including acne, to measure the effects of acne and skin problems on floral ginsenosides A, B, C, D, E, F, M, N, O and P, respectively. After washing the face with soap in a constant temperature and humidity room, it was adapted for 30 minutes, and the degree of skin trouble was measured by comparing the number of acne and the number with inflammation.

The test subjects were allowed to use the nutrient cosmetics of Preparation Examples 1 to 10 for 4 weeks, and then the degree of acne and skin problems was compared, and the results are shown in Table 18 below.

Evaluation item Skin trouble improvement rate Production Example 1 65% Production Example 2 60% Production Example 3 55% Production Example 4 65% Production Example 5 60% Production Example 6 60% Production Example 7 55% Production Example 8 50% Production Example 9 55% Production Example 10 65% Comparative Preparation Example 1 25%

Looking at the results of Table 18, as in the case of applying the formulation examples 1 and 2 containing the ginseng flower extract containing the floral ginsenoside of Example 1, the floral ginsenosides A, B, C, D, E, Even using the preparation examples 1 to 10 containing F, M, N, O and P, respectively, it can be confirmed that there is an improvement in acne and skin troubles.

Test Example 10 Anti-inflammatory Effect: Inhibitory Effect of Biosynthesis on Tumor Necrosis Factor (TNF-α) Induced by Lipopolysaccharide

Human keratinocytes were cultured in a 12 well plate incubator at a concentration of 10 ⁵ , treated with LPS, and then replaced with a medium containing 1, 10 and 100 ppm of ginseng flower extract containing the floral ginsenoside of Example 1 . After culturing for 6 to 24 hours, the cells were harvested, and the amount of TNF-α produced was quantified using ELISA (Pharmingen 555212) method. Table 19 shows a comparison with the measured value of the control group. At this time, the control group was an untreated group cultured without treatment with an extract containing floral ginsenosides.

Test substance TNF-α biosynthesis (%) Example 1 Floral Ginsenoside A 100 ppm 10 10 ppm 32 1 ppm 62 Floral Ginsenoside B 100 ppm 13 10 ppm 40 1 ppm 68 Floral Ginsenoside C 100 ppm 12 10 ppm 37 1 ppm 66 Floral Ginsenoside D 100 ppm 11 10 ppm 33 1 ppm 61 Floral Ginsenoside E 100 ppm 13 10 ppm 34 1 ppm 65 Floral Ginsenoside F 100 ppm 12 10 ppm 36 1 ppm 61 Floral Ginsenoside M 100 ppm 10 10 ppm 33 1 ppm 64 Floral ginsenoside N 100 ppm 11 10 ppm 35 1 ppm 66 Floral ginsenoside O 100 ppm 12 10 ppm 31 1 ppm 60 Floral Ginsenoside P 100 ppm 13 10 ppm 33 1 ppm 61 Control group 100

In the results of Table 19, the floral ginsenosides A, B, C, D, E, F, M, N, O and P according to the present invention concentration-dependently reduced the biosynthesis of TNF-α by the proinflammatory factor LPS By suppressing the inflammatory response that can occur due to this it could contribute to the improvement of skin problems.

Examples of the formulation of the composition are described below, but are not intended to limit the present invention but merely to be described in detail. The extract containing the floral ginsenoside used at this time is the ginseng flower extract which passed the chromatography in Example 1, and the floral ginsenosides A, B, C, D, E, F, M, N, O and P It is all inclusive.

[Formulation Example 3] Massage cream

To prepare a massage cream in a conventional manner according to the composition described in Table 20 below.

ingredient Content (% by weight) Extract Containing Floral Ginsenosides (Example 1) 5.0 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 PEG60 hardened castor oil 2.0 Liquid paraffin 40.0 Squalane 5.0 Caprylic / capric triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount Purified water Balance Sum 100

[Formulation Example 4] Pack

To prepare a pack in a conventional manner according to the composition described in Table 21 below.

ingredient Content (% by weight) Extract Containing Floral Ginsenosides (Example 1) 5.0 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 PEG 12 nonyl phenyl ether 0.3 Polysorbate 60 0.3 Preservative, coloring, fragrance Suitable amount Purified water Balance Sum 100

Those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims (10)

A composition for external application of skin containing a floral ginsenoside derived from ginseng flower. The composition for external application for skin according to claim 1, wherein the floral ginsenoside is at least one selected from the group consisting of floral ginsenosides A, B, C, D, E, F, M, N, O and P. The composition for external application for skin according to claim 1, wherein the ginseng flower-derived floral ginsenoside is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. The external preparation composition for skin according to any one of claims 1 to 3, wherein the composition is for antioxidant. The external preparation composition for skin according to any one of claims 1 to 3, wherein the composition is for anti-aging. The external preparation composition for skin according to any one of claims 1 to 3, wherein the composition is for skin whitening. The external preparation composition for skin according to any one of claims 1 to 3, wherein the composition is for moisturizing the skin. The composition for external application for skin according to any one of claims 1 to 3, wherein the composition is for anti-inflammatory. The composition for external application for skin according to any one of claims 1 to 3, wherein the composition is for improving atopic symptoms. The composition for external application for skin according to any one of claims 1 to 3, wherein the composition is for improving skin troubles.