KR20140126891A - External composition for skin containing Ginsenoside Rf - Google Patents

Abstract

The present invention relates to a composition containing ginsenoside Rf for anti-aging, alleviating skin wrinkles, whitening, moisturizing effect, anti-inflammation, acne and skin troubles, alleviating atopic symptoms, skin astringency, shrinking pores, improving skin complexion, promoting hair growth, mitigating grey hair, anti-dandruff, and anti-corrosion effect.

Description

[0001] The present invention relates to an external composition for skin containing Ginsenoside Rf,

The present invention can provide acne and skin trouble improving effect, skin convergence and pore shrinkage effect by containing ginsenoside Rf, and can provide an effect of improving skin color, hair growth, white moth improvement, anti dandruff and preservative effect ≪ / RTI >

Human skin is the primary protective membrane of the human body. It functions to protect the internal organs from external environmental stimuli such as changes in temperature and humidity, ultraviolet rays, and pollutants. Depending on various internal and external factors, It undergoes change. In other words, internally, secretion of various hormones that regulate metabolism is reduced, the function of immune cells and the activity of cells are lowered, and the biosynthesis of immune proteins and biocompatible proteins necessary for the living body is reduced, and ozone layer destruction As the content of ultraviolet rays reaching the surface of the sunlight increases and environmental pollution is further intensified, free radicals and active noxious oxygen are increased, so that the thickness of the skin decreases, the wrinkles increase, the elasticity decreases Skin color is grayish, skin troubles are frequent, spots, freckles and black spots are increased, bleeding is poor, skin tone is getting dark, and so on.

In order to prevent the change of the skin condition due to the intrinsic and extrinsic factors, and to maintain a healthy skin condition, physiologically active substances obtained from various known animals, plants, microorganisms and the like are added to cosmetics to improve the skin condition There has been effort for.

Ginsenoside Rf is a type of saponin contained in ginseng. It has physiological activities such as inhibition of lipid peroxidation and alcohol-induced brain growth disorder, voltage-dependent calcium channel inhibition (Nah, et al., Proc. Natl. (Shin et al., Submited, 1997). However, it has been reported that the composition containing ginsenoside Rf as an active ingredient has an acne and skin trouble improvement Effect, skin convergence and pore shrinkage effect, skin color improvement effect, hair growth enhancement, white moth improvement, anti-dandruff and preservative effect have not been reported yet.

Accordingly, the present inventors have found that ginsenoside Rf can provide acne and skin trouble improving effect, skin convergence and pore shrinkage effect, skin color improvement effect, hair growth enhancement, white moth improvement, anti dandruff and preservative effect And completed the present invention.

Accordingly, it is an object of the present invention to provide a composition containing ginsenoside Rf, which can provide acne and skin trouble improving effect, skin convergence and pore shrinkage effect, and also has effects of skin color improvement, hair growth, white moth improvement, anti- And a skin external application composition.

In order to achieve the above object, the present invention provides an external composition for skin improvement for acne improvement comprising ginsenoside Rf as an active ingredient.

The present invention also provides a composition for external application for skin color and skin tone improvement comprising ginsenoside Rf as an active ingredient.

The present invention also provides a composition for external application for skin pore reduction comprising ginsenoside Rf as an active ingredient.

In addition, the present invention provides a hair growth promoting composition containing ginsenoside Rf.

The present invention also provides a composition for preventing whey from containing ginsenoside Rf.

In addition, the present invention provides an anti-dandruff composition containing ginsenoside Rf.

The present invention also provides a natural preservative composition containing ginsenoside Rf.

The composition of the present invention can provide anti-inflammation, acne and skin trouble improving effect, skin convergence and pore-shrinking effect by containing ginsenoside Rf, and can improve skin color improvement, hair growth, white moth improvement, anti dandruff and preservative effect .

The composition according to the present invention contains ginsenoside Rf (ginsenoside Rf) as an active ingredient.

The jinenoside Rf used in the present invention has a structure represented by the following formula (1).

The ginsenoside Rf of the present invention may be extracted from a plant, and may be synthesized according to a method known in the art, or a commercially available product may be used. Ginsenoside Rf can also be obtained from ginseng extract. The type of ginseng to be used at this time is not particularly limited, and ginseng, red ginseng, white ginseng, taegeuk ginseng and ginseng can be used. In addition, the ginseng extract is not only contained in the leached liquid obtained by leaching and transferring from ginseng but also contained in the concentrate obtained by partially or completely concentrating the leach solution or in the slump, premix, regular season, liquid extract and ginseng prepared by drying the concentrate again It contains all of the plant itself as well as chemicals that exert the main effect. Extracts from all parts of ginseng such as stem, root, leaf, flower, and fruit are available and are not limited to any particular part of the extract. Also, a known method can be used as a method for extracting ginsenoside Rf from ginseng extract.

Specifically, the ginsenoside Rf can be isolated from ginseng by preparing ginseng extract from water or an organic solvent by a method well known in the art. The organic solvent used in the present invention may be selected from the group consisting of ethanol, methanol, butanol, ether, ethyl acetate, chloroform and a mixed solvent of these organic solvents and water, preferably 80% ethanol. At this time, the extraction temperature is preferably 10 to 80 ° C, and the extraction can be performed for 3 to 24 hours. If the extraction temperature and the extraction time are exceeded, the extraction efficiency may be deteriorated or the component may be changed.

The composition of the present invention preferably contains the ginsenoside Rf in an amount of 0.001 to 50 wt% based on the total weight of the composition. If the content of the active ingredient is less than 0.001% by weight, the effect and effect of the ingredient are insufficient, and if it exceeds 50% by weight, there is a problem of skin safety or shape.

The composition of the present invention can be used as a composition for external application for skin for improving acne, and has excellent antimicrobial effect, especially antimicrobial effect against acne causing bacteria, and also provides anti-inflammatory effect.

The composition of the present invention can be used as a composition for external application of skin for improving color and skin tone. It expands capillary blood vessels when applied to skin and promotes blood circulation, thereby smoothly supplying nutrients to the skin and suppressing skin aging, The effect is excellent.

The composition of the present invention can be used as a composition for external application for skin for reducing pores, controlling sebum and improving skin troubles. It can suppress sebum secreted excessively when applied to skin, promote active oxygen elimination and collagen synthesis, , And suppression of skin troubles due to decreased expression of inflammatory factors.

The composition of the present invention can be used as a composition for promoting hair growth, which promotes the transition of the resting hair cycle to the growing hair cycle, thereby promoting hair growth and promoting the generation of new hair, And provides an effect of preventing hair from falling off from the scalp or preventing and suppressing hair tingling or tapering.

The composition of the present invention can be used as a composition for preventing wheat hair. It enhances the expression of MITF in melanocytes to activate melanocytes and promote melanin synthesis, thereby preventing the generation of white moths and promoting the induction of black cohosh.

The composition of the present invention can be used as a composition for external application for skin prevention of dandruff. It can purify the scalp by effectively discharging toxins accumulated in hair and scalp, inhibit proliferation and growth of dandruff to prevent scalp inflammation reaction, It has excellent antioxidant effect which inhibits the production and action of oxygen, so it can calm and strengthen the scalp and provide the effect of strengthening the original defense force.

The composition of the present invention can be used as a natural preservative composition, and since it is a natural component, it has excellent preservative effect and provides a harmless effect to human body.

The composition according to the invention may be formulated containing a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example as a solution, a gel, a solid, a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic form (liposome) In the form of ionic fibrin dispersions, or in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions may be prepared according to conventional methods in the art.

In particular, when the composition for external application for skin of the present invention is used for prevention of dandruff, hair growth or hair loss prevention, it may be formulated as a composition for scalp and hair, and the formulation is not particularly limited, but, for example, hair tonic, A hair treatment, a hair treatment, a hair shampoo, a hair rinse, a hair lotion, or a scalp hair combined treatment.

In addition, the composition according to the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickening agents, gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, Such as fillers, emulsifiers, emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blockers, moisturizers, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics May contain adjuvants commonly used in the field of cosmetics or dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

In addition, the composition of the present invention may contain a skin absorption promoting substance to increase the skin improving effect.

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

[Referential Example 1] Preparation of ginsenoside Rf

The ginsenoside Rf for testing the efficacy of the composition of the present invention was purchased from Ambo Laboratories.

 [Formulation Example 1 and Comparative Formulation Example 1]

Nutrition cream was prepared according to the composition of the following Table 1 (unit: wt%).

Compounding ingredient Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 Ginsenoside Rf 5.00 - Vegetable hydrogenated oil 1.50 1.50 Stearic acid 0.60 0.60 Glycerol stearate 1.00 1.00 Stearyl alcohol 2.00 2.00 Polyglyceryl-10pentastearate and behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 Caprylic / carboxy triglyceride 11.00 11.00 Cyclomedicone 6.00 6.00 Preservative, incense Suitable amount Suitable amount Triethanolamine 0.1 0.1

[Test Example 1]

In order to evaluate the skin blood circulation promoting effect of the cosmetic composition according to the present invention, the degree of blood circulation in the skin was measured using LDPI (Laser Doppler Perfusion Imager). LDPI is a widely used and widely used device for measuring blood circulation in skin. It is a highly sensitive device that can measure not only blood velocity and quantity in the capillaries of the skin, but also flow in the small artery and the parenchyma.

The face was soaked in a constant temperature and humidity chamber, and then adapted for 30 minutes. Initial values were measured using LDPI. First, the initial blood flow in the lower part of the forehead of 30 women with cold hands and feet was measured by LDPI. Table 2 shows the results (skin blood flow changes) of blood flow measured after using Formulation Example 1 and Comparative Formulation Example 1 for one week to the subjects and comparing the initial measured values.

LDPI results before and after using cosmetics - skin blood flow Test substance Percent change of skin blood flow after 1 week use (%) Formulation Example 1 11 Comparative Formulation Example 1 5

From the results of Table 2, it was confirmed that the cosmetic composition according to the present invention significantly increased skin blood flow compared to Comparative Formulation Example 1 which did not contain ginsenoside Rf, and blood circulation was improved by promoting blood circulation . This ultimately suggests that the cosmetic composition containing ginsenoside Rf according to the present invention can contribute to effectively transmit nutrients of the skin and inhibit and retard skin aging.

[Test Example 2] Skin tone improvement effect

In order to examine the skin tone improvement effect of Formulation Example 1 and Comparative Formulation Example 1, 30 subjects were used (one evening / one day application for a total of one week) and then facial stage DM-3 (Moritex, Japan) To evaluate the degree of skin tone improvement. The improvement rate of the skin tone was determined as the brightness and color value of the skin and the change of the brightness and color of the skin. The results are shown in Table 3 below. The larger the change in brightness and color, the better the skin tone.

Test substance Skin tone improvement rate (%) Brightness (mean ± standard deviation) Color (mean ± standard deviation) Formulation Example 1 13 ± 2.52 11 ± 2.33 Comparative Formulation Example 1 5 ± 2.34 5 ± 2.05

In the results of Table 3, Comparative Formulation Example 1 which does not contain ginsenoside Rf according to the present invention shows no significant skin tone improving effect, whereas Formulation Example 1 containing ginsenoside Rf It was confirmed that the skin tone after use was greatly improved.

[Test Example 3] Pore reduction effect

1. Collagen biosynthesis promotes pore reduction effect

The ginsenoside Rf according to the present invention was measured by comparing collagen biosynthesis promoting effect with TGF- ?. First, fibroblasts were seeded at 10 ⁵ per well in 24 wells and cultured until they grew to about 90%. After culturing for 24 hours in serum-free DMEM medium, the ginsenosides Rf and TGF-β of the present invention dissolved in serum-free medium were treated with 10 ng / ml each, and CO ₂ And cultured in an incubator for 24 hours. The supernatant was removed and the procolagen (I) ELISA kit (procollagen type (I)) was used to determine whether procollagen was increased or decreased. The results are shown in Table 4, and the combined performance of the collagen was set to 100 in the non-treatment group.

Test substance Collagen Total Performance (%) Untreated group 100 TGF-beta 183.5 Ginsenoside Rf 187.6

From the results shown in Table 4, it was confirmed that the ginsenoside Rf according to the present invention exhibits a superior collagen synthesis performance higher than that of the positive control group TGF-β. Therefore, it was confirmed that the ginsenoside Rf according to the present invention can increase the amount of collagen produced around the pores, thereby widening the pores.

2. Pore reduction effect

In order to examine the pore reduction effect of Formulation Example 1 and Comparative Formulation Example 1, the following evaluation was made. Twenty male and female subjects with large pore sizes were selected and divided into two groups of 10 persons. The nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were applied to the faces for each group for 4 weeks each day. The evaluation of the effectiveness of the pore reduction was made by a visual evaluation of experts by taking pictures before and after the experiment. The results are shown in Table 5 below (rating scale: 0 - no reduction at all; 5 - very reduced).

Test substance Rating Rating Formulation Example 1 4 Comparative Formulation Example 1 0

In the results of Table 5, Comparative Formulation Example 1 has no effect of reducing pores, but in the case of Formulation Example 1, the effect of reducing pores is visible to the naked eye. The ginsenoside Rf of the present invention reduces the size of pores And the effect was excellent.

Test Example 4 Effect of suppressing sebum secretion

1. Suppression of skin hypersecretion by 5α-reductase inhibition

In order to confirm 5α-reductase inhibitory effect, the ratio of [ ¹⁴ C] testosterone to [ ¹⁴ C] dihydrotestosterone (DHT) in HEK293-5αR2 cells was measured. HEK293 cells were transfected with p3 x FLAG-CMV-5αR2, and cultured in 2.5 × 10 ⁵ cells / well in a 24-well plate (Park et al., 2003, JDS Vol. 31, pp. 191-98). The next day, the medium was changed to a new medium supplemented with an enzyme substrate and an inhibitor. 0.05 [mu] Ci [ ¹⁴ C] testosterone (Amersham Pharmacia biotech, UK) was used as the substrate of the medium.

To confirm the inhibition of 5α-reductase activity, ginsenoside Rf was added and cultured in a 5% CO ₂ incubator at 37 ° C. for 2 hours. At this time, the control group in which ginsenoside Rf was not added was used as a negative control group, and the group in which finasteride was added was used as a positive control group. Then, the culture medium was collected, and the steroid was extracted with 800 쨉 l of ethyl acetate. The upper organic solvent layer was separated, dried, and the remaining residue was again dissolved in 50 쨉 l of ethyl acetate to obtain a silica plastic sheet kiesel gel 60 F254 ) Using ethyl acetate-hexane (1: 1) as the solvent.

After drying the plastic sample in air, the bath system was used to measure the amount of isotope. The dried plastic sheet and x-ray film were put together in a bath cassette, and after 1 week, the testosterone remaining in the film and isotopes of dihydrotestosterone The conversion and inhibition rates were calculated according to the following equations (1) and (2), respectively, and the results are shown in Table 6 below.

Test substance Conversion Rate (%) Inhibition rate (%) Negative control group 48 - Positive control group 27.6 42.5 Ginsenoside Rf 15.4 67.9

In the results shown in Table 6, testosterone is converted into dihydrotestosterone, which binds to the receptor protein in the cytoplasm to enter the nucleus, activates the sebaceous gland, accelerates differentiation, and induces 5α-reductase Was effectively inhibited by ginsenoside Rf, thereby blocking the conversion of testosterone to dihydrotestosterone. It was found to have an inhibitory effect superior to that of finasteride, which is known to inhibit 5α-reductase activity. Thus, it was confirmed that the ginsenoside Rf effectively inhibited the activity of 5? -Reductase, thereby suppressing sebaceous hyperpermeability.

2. Inhibitory effect of sebum secretion

To evaluate the inhibitory effect of sebum secretion of Formulation Example 1 and Comparative Formulation Example 1, the following evaluation was made. 30 men and women who felt that sebum secretion was high were selected and the nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were made every day for 4 weeks on designated areas of the facial skin. The results are shown in Table 7. The results are shown in Table 7 below. The results are shown in Table 7 below. The results are shown in Table 7 below. Respectively.

Test substance Sebum reduction rate (%) After 2 weeks After 4 weeks Formulation Example 1 31 40 Comparative Formulation Example 1 5 5

From the results shown in Table 7, it can be seen that Formulation Example 1 containing the ginsenoside Rf according to the present invention as an active ingredient can effectively inhibit excess sebum secreted from Comparative Formulation Example 1 which does not contain it.

[Formulation Example 2 and Comparative Formulation Examples 2 to 3]

Formulation Example 2 and Comparative Formulation Examples 2 to 3 were prepared according to the ingredients and the contents (% by weight) shown in Table 8 below. More specifically, Formulation Example 2 is a combination of ginsenoside Rf, Comparative Formulation Example 2 does not contain any effective ingredient for acne skin improvement, Comparative Formulation Example 3 is a standard for antibacterial activity, Which contains erythromycin, which is widely used as an acne remedy.

The manufacturing method of Formulation Example 2 and Comparative Formulation Examples 2 to 3 is as follows. The components of phase A in Table 8 below were completely dissolved, the components of phase B were completely dissolved in a separate melting tank, and the phase B was added to the phase A to be mixed and solubilized. The ingredients of the C phase were added thereto in accordance with the mixing ratios shown in Table 8, followed by mixing and homogenization, followed by filtration to prepare the compositions.

division Formulation Example 2 Comparative Formulation Example 2 Comparative Formulation Example 3 A Deionized Water To 100 To 100 To 100 EDTA-2Na 0.02 0.02 0.02 glycerin 5.0 5.0 5.0 B ethanol 2.0 2.0 2.0 PEG-60 hardened castor oil
(hydrogenated castor oil) 0.4 0.4 0.4 Perfume 0.04 0.04 0.04 C Ginsenoside Rf 5.0 - - Erythromycin - - 5.0

[Test Example 5] Test for antimicrobial activity against acne bacteria

Each of the cosmetic compositions prepared in the formulations of Formulation Example 2 and Comparative Formulations 2 to 3 was tested for antibacterial activity against Propionibacterium acnes (ATCC 6919: medium-BHI broth), which is a cause of acne Respectively.

The test method for the antimicrobial activity against acne bacteria was as follows.

(1) Preparation of test strain

Propionibacterium acnes were cultured in an anaerobic culture inoculated with BHI broth.

(2) Dilution solution preparation

0.15 ml of the above test solution was added to 15 ml of BHI broth (pH 6.8) or LB broth (pH 4.5), and the mixture was used as a diluting solution.

(3) Preparation of sample

The undiluted cosmetic composition prepared from Formulation Example 2 and Comparative Formulation Examples 2 and 3 was directly used as a sample.

(4) Antimicrobial activity test

1) In a 96-well 96-well plate, place the sample in line 1 and add 200 μl of diluted solution.

2) Thoroughly mix the mixture of line 1, and then take 100 μl of the mixture, place it in the second row, mix well, and then repeat the double dilution by adding 100 μl to the third row.

3) After culturing the cells at 32 ° C for 24 hours and 48 hours, determine whether the bacteria are proliferating to the extent of suspension, and determine the minimum concentration without bacterial growth as the MIC (Minimum Inhibitory Concentration) value. If the mixture is opaque and it is difficult to judge the growth of the microorganism, check with a microscope.

The results of the test for antimicrobial activity against acne bacteria are shown in Table 9 below. The MIC was expressed in terms of the concentration of the active ingredient contained in the formulation.

Item pH Propionibacterium acnes Formulation Example 2 5.7 > 41 ppm Comparative Formulation Example 2 5.7 Maximum concentration (no antimicrobial activity) Comparative Formulation Example 3 5.7 > 100 ppm

In the results of Table 9, it can be said that the smaller the ppm concentration in the MIC, the more effective the antimicrobial activity against acne bacteria. In the case of Formulation Example 2, the concentration of ppm And the composition containing senenoside Rf has far superior antibacterial activity to the test bacteria.

[Test Example 6] Lipogenesis inhibition test

3T3-L1 cells, a fibroblast cell line of mice, were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) (Dulbecco's modified eagles medium, GIBCO BRL, Life Technologies) Lt; ⁵ > cells / well in a well culture plate. After 2 days, the cells were replaced with fresh DMEM (containing 10% FBS) medium and cultured for 2 days. The cultured cells were then induced to differentiate into DMEM (containing 10% FBS) containing 1 μg / ml insulin, 0.5 mM IBMX and 0.25 μM dexamethasone, and the ginsenoside Rf and caffeine After 2 days of treatment, the cells were replaced with insulin-containing DMEM and cultured for 5 days. After 5 days, the cells were replaced with normal medium (DMEM, containing 10% FBS) and cultured while observing until the cells morphologically changed into adipocytes.

In order to evaluate the inhibitory effect of ginsenoside Rf on the fat accumulation in adipocytes, the above-described 3T3-L1 adipocytes were subjected to S-III staining (S4136, sigma-aldrich). The adipocytes were fixed in 4% paraformaldehyde (pH 7.2) in phosphate buffer at room temperature, washed with PBS (phosphate buffered saline), stained with means III and then photographed and compared visually. The control group was fed with the test substance or the test substance without the reference substance, and 50 μM of caffeine was treated with another comparative group. The degree of inhibition of fat accumulation was graded by dividing the degree of staining by +++, +, +, -, which means that the degree of staining is greater toward +++. The results are shown in Table 10 below.

sample % Inhibition Control group +++ Comparative group + Ginsenoside Rf -

In the results of Table 10, it can be seen that the ginsenoside Rf used in the present invention not only has a small amount of fat accumulated in adipocytes but also has a lipid synthesis inhibitory effect better than caffeine, which is a known lipid synthesis inhibitor . Therefore, lipid synthesis is inhibited, and sebum is reduced, so that occurrence of acne can be suppressed.

[Test Example 7] Test for improvement of acne and reduction of sebum secretion and stimulation

30 persons who had acne were divided into 3 groups of 10 persons, and subjects corresponding to each group were allowed to use the cosmetic composition prepared in Formulation Example 2 and Comparative Formulation Examples 2 to 3 for one month. Acne improvement scale is from 1 point to 5 points, 1 point is 'No', 3 points are 'Normal' and 5 points are 'Very agree'. The experimental results are shown in Table 11 below with an average score of 10 persons.

The acne extermination period was based on the number of days of disappearance readings, and the results were based on the results after 1 month with or without recurrence of acne. The reduction of sebaceous secretion was from 1 point to 5 points, 1 point was 'No', 3 points were 'Normal' and 5 points were 'Very agree'. The experimental results are shown in Table 11 below with an average score of 10 persons. The presence or absence of skin irritation was evaluated as (number of stimuli) / (total number of test subjects).

Inflammatory acne improvement Acne-bloating period Recurrence of acne Decrease sebaceous glands Presence or absence of stimulation Formulation Example 2 4.1 3 days radish 4.3 0/10 Comparative Formulation Example 2 2.1 13th U 2.0 0/10 Comparative Formulation Example 3 4.2 2 days radish 4.1 9/10

In the results of Table 11, it can be seen that Formulation Example 2 has no recurrence of acne as compared with Comparative Formulation Example 2, and has excellent effects on improvement of acne as a whole. On the other hand, Comparative Formulation Example 3 containing the antibacterial activity standard substance shows the effect of improving acne but it is not suitable for long-term use due to strong skin irritation in use. However, since the composition according to the present invention has no irritation, .

[Formulation Example 3 and Comparative Formulation Example 4]

A shampoo was prepared with the composition shown in Table 12 below. Specifically, the surfactant and ethylene glycol distearate were added to purified water, heated to 80 DEG C to dissolve uniformly, and then gradually cooled to 40 DEG C with stirring. To the mixture were added the active ingredient according to the present invention and a preservative, A viscosity adjusting agent, a perfume, and a hair conditioning agent were mixed and stirred, followed by cooling to room temperature.

Component (% by weight) Formulation Example 3 Comparative Formulation Example 4 Ammonium lauryl sulfate 10 10 Polyoxyethylene lauryl ammonium sulfate 5 5 Cocoamidopropyl betaine 2 2 Ethylene glycol distearate 1.5 1.5 Cocoyl monoethanolamide 0.8 0.8 Ginsenoside Rf 5.0 - Polyquaternium-10 0.2 0.2 Blue No. 1 0.0002 0.0002 Yellow No. 4 0.0001 0.0001 Methyl paraben 0.1 0.1 Spices 0.8 0.8 Citric acid 0.1 0.1 Dimethicone 1.0 1.0 water to 100 to 100

[Test 8] Dandruff reduction test

Twenty-four men aged 19 to 35 years with a relatively large number of dandruff were selected and the dandruff reduction rate was measured after using the shampoo of each of Formulation Example 3 and Comparative Example 4 for 12 months in two groups in the following manner .

Before the start of the test, the hair was smeared with a normal shampoo, and the dandruff accumulated for 2 days after hair removal was sampled, and the dandruff weight was sampled once every two days with the shampoo of each of Formulation Example 3 and Comparative Formulation Example 4, The weight of accumulated dandruff was compared and evaluated. The accumulated dandruff was collected directly from the scalp using a vacuum suction device, and the dandruff reduction rate was calculated according to the following equation (3). The results are shown in Table 13 below.

Formulation Example 3 Comparative Formulation Example 4 Dandruff reduction rate (%) 66.5
55.6
59.6
72.6
66.3
72.3
68.9
63.8
71.2
59.8
68.3
52.6 0.7
-0.5
-1.1
1.5
2.2
1.3
6.3
3.6
3.3
4.6
3.7
4.2 medium 64.79 2.5 SD 6.59 2.2

From the results of Table 13, it can be seen that Formulation Example 3 containing ginsenoside Rf exhibits excellent anti-dandruff effect.

[Test Example 9] Test for preventing the itchiness of the scalp

Twenty-four male and female twenty-four to twenty-five to fifty-year-olds who felt severe scalp itching were selected and divided into two groups of twelve persons. The shampoo of each of Formulation Example 3 and Comparative Formulation Example 4 was used once every three days for two weeks. Then, scalp itching The prevention effect was evaluated through the following evaluation criteria, and the results are shown in Table 14 below.

[Evaluation standard]

Very good - 5 points

Excellent - 4 points

Usually - 3 points

Poor - 2 points

Very bad - 1 point

division Formulation Example 3 Comparative Formulation Example 4 The itching effect of scalp 4.5 2.3

From the results shown in Table 14, it can be seen that Formulation Example 3 containing ginsenoside Rf exhibits a more excellent effect in preventing scalp itching.

Test Example 10 Evaluation of Potassium Ion Channel Activity Increase Effect

Minoxidil, a remedy for depilation, is known as a potential mitochondrial potassium ion channel opener (K _ATP channel opener) and is a representative drug used for the treatment of androgenetic alopecia. In order to evaluate the mechanism of minoxidil, cell proliferation was inhibited by treatment with tolbutamide (SIGMA AlDRICH, T0891) blocking K _ATP channel in fibroblasts constituting the dermis of the scalp, The restored test method was used.

To evaluate the function of this composition as a K _ATP channel opener, the fibroblast cell line NIH3T3 (mouse embryonic fibroblast cell line) cell line was used in the present invention. This cell line is a cell line obtained by natural immortalization of fibroblasts isolated from NIH Swiss mouse embryo by the 3T3 protocol. The cells were cultured in DMEM (Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS for 24 hours in an incubator maintained at 37 ° C in 5% CO ₂ . NIH3T3 was added to a 96-well plate, cultured in a 37 ° C incubator for 24 hours, treated with 2.5 mM tolbutamide, and 10 minutes after the positive control, minoxidil 10 μM and ginsenoside Rf were added at 2.5 ppm, 5 ppm and 10 ppm , And cell proliferative activity was measured using a WST-1 kit (Roche) 48 hours after the drug treatment. The results are shown in Table 15 below.

division Cell proliferation (%) Untreated Control (Control) 100 Minoxidil 132 Ginsenoside Rf (2.5 ppm) 115 Ginsenoside Rf (5 ppm) 121 Ginsenoside Rf (10 ppm) 132

In the results of Table 15, it can be seen that when the ginsenoside Rf is treated, proliferation of fibroblasts is restored and the cell proliferative capacity is increased depending on the concentration of the treated ginsenoside Rf. When ginsenoside Rf is treated at 10 ppm , It can be confirmed that the proliferation of the cells is restored to the level when minoxidil is treated.

[Test Example 11] Test for promoting the production of melanin of ginsenoside Rf

Melanocytes (melan-a) were added to a 24-well microtiter plate at 50,000 cells / well in RPMI medium supplemented with 5% fetal bovine serum, 100 IU penicillin G and 0.2 μM TPA Respectively. The next day, the ginsenoside Rf as the test substance was treated at a final concentration of 10 ppm or 50 ppm, and 0.1% DMSO was used as a negative control, 100 μM IBMX was used as a positive control, and the cells were incubated at 37 ° C. for 3 days. After incubation, the wells were washed with PBS, and 100 μl of 1N NaOH was added thereto, and melanin in the cells was dissolved. The absorbance of dissolved melanin was measured at 405 nm using a microplate reader. The results of comparing melanin production promoting effect of ginsenoside Rf with the control group are shown in Table 16 below.

sample Amount of melanin synthesis (%) DMSO (0.1%) 100 IBMX (100 [mu] M) 120 Ginsenoside Rf (10 ppm) 112 Ginsenoside Rf (50 ppm) 123

From the results shown in Table 16, it can be seen that ginsenoside Rf promotes melanin synthesis of melanocyte to increase melanin production and exhibit excellent melanin production promoting effect.

[Test Example 12] Promotion of MITF and tyrosinase expression in melanocytes of ginsenoside Rf

The cells were divided into 6-well microtiter plates at a density of 500,000 cells / well using a 501mel cell line. DMSO 0.1% as a negative control, IBMX 100 μM as a positive control group, and ginsenoside Rf was treated at 10 ppm, and cultured at 37 DEG C for 24 hours, 48 hours, and 72 hours to obtain a protein. The proteins thus obtained were subjected to western blotting using MITF and tyrosinase antibodies. Protein extraction and Western blotting were routinely performed by standard methods used by those skilled in the art. After Western blotting, the results were compared with the negative control group of 100 and are shown in Table 17 below.

MITF Tyrosinase IBMX Ginsenoside Rf IBMX Ginsenoside Rf 24hr 121 97 160 153 48hr 98 111 149 155 72hr 224 119 298 186

From the results in Table 17, it can be confirmed that ginsenoside Rf elevates MITF and tyrosinase protein expression in melanocytes.

[Test Example 13] Evaluation of antibacterial activity of ginsenoside Rf

An antimicrobial test was conducted to evaluate the antimicrobial activity of ginsenoside Rf. Specific experimental methods are as follows.

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa strains used in the experiment were obtained from Tryptic Soy Broth, Candida albicans and Aspergillus niger strains were cultured in Sabouraud Dextrose broth. The culture was added to each medium at a ratio of 1/100 (Candida albicans 1/10 diluted diluted solution was used as a test solution. Aspergillus niger, a spore suspension prepared to have a concentration of 2 × 10 ⁸ cfu / ml was used as the test strain.

0.15 ml of the above-mentioned test solution was added to 15 ml of each medium, and well-mixed solution was used as a diluting solution.

In 96 well plate (96 well plate), 16 μl of 10 ppm of ginsenoside Rf was added to each sample, and 184 μl of the diluted solution was added. In the remaining wells, 100 μl of the dilution solution was added. 100 μl of the mixed solution of No. 1 line was mixed well and 100 μl of the mixture was added to the No. 2 line. After mixing well, 100 μl of the mixture was added to the No. 3 line and diluted 2-fold.

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were incubated in a 32 ° C thermostat with Candida albicans, Aspergillus niger (Aspergillus niger), Staphylococcus aureus niger) were incubated in a 25 ° C incubator.

After 48 hours, the microbial proliferation was determined by suspension and microscope to determine the minimum inhibitory concentration (MIC) value, and the results are shown in Table 18 below.

Sample Minimum inhibitory concentration (MIC), unit% Sedona Monas
Aruzinosa Staphylococcus aureus Escherichia coli Candida Albikans Aspergillus niger Ginsenoside Rf 0.2 0.04 0.128 > 3 > 1

From the results shown in Table 18, it can be confirmed that the ginsenoside Rf exhibits antibacterial activity against various strains, and it can be predicted that ginsenoside Rf can act as a natural preservative or an antimicrobial agent in the composition.

Claims (13)

A composition for external application for skin comprising ginsenoside Rf as an active ingredient. 2. The composition for external application for skin according to claim 1, wherein the ginsenoside Rf is represented by the following formula (1).
[Chemical Formula 1]

The composition for external application for skin according to claim 1 or 2, wherein the ginsenoside Rf is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. The composition for external application for skin according to claim 1 or 2, wherein the composition is for improving acne. The composition for external application for skin according to claim 1 or 2, wherein the composition is antibacterial. The composition for external application for skin according to claim 1 or 2, wherein the composition is for improving color and skin tone. The composition for external application for skin according to claim 1 or 2, wherein the composition is for shrinking pores. The composition for external application for skin according to claim 1 or 2, wherein the composition is for sebum control. The composition for external application for skin according to claim 1 or 2, wherein the composition is for improving skin troubles. The composition for external application for skin according to claim 1 or 2, wherein the composition is for promoting hair growth. The composition for external application for skin according to any one of claims 1 to 3, wherein the composition is anti-whiplash. The composition for external application for skin according to claim 1 or 2, wherein the composition is anti-dandruff. The composition for external application for skin according to claim 1 or 2, wherein the composition is a natural preservative.