WO2014175676A1 - Topical composition for skin containing gincenoside rf - Google Patents

Abstract

The present invention relates to a composition containing, as an active ingredient, gincenoside Rf, for providing effects such as improvement of acne and skin troubles, skin clearing, pore tightening, in addition to providing effects such as skin tone improvement, hair growth promotion, improvement of graying hair, anti-dandruff, and antisepsis.

Description

External skin composition containing ginsenoside RF

The present invention can provide acne and skin trouble improvement effect, skin convergence and pore contraction effect by containing ginsenoside Rf, and can provide skin color improvement effect, hair growth improvement, white hair improvement, anti dandruff and antiseptic effect. It relates to a composition.

Human skin is the body's primary protective barrier, which protects the body's organs from changes in temperature and humidity, and from external environmental stimuli such as ultraviolet rays and pollutants. Undergo a change. 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 intensifying environmental pollution, free radicals and free radicals increase, thereby reducing the thickness of the skin, increasing wrinkles, reducing elasticity, Skin color becomes dull, skin problems frequently occur, blemishes, freckles and blotch also increase, the color becomes worse and the skin tone becomes darker.

In order to prevent changes in the skin condition caused by these internal and external factors and to maintain a healthy skin condition, the skin condition is improved by adding bioactive substances obtained from various known animals, plants, and microorganisms to cosmetics. Efforts have been made.

Ginsenoside Rf (Ginsenoside Rf) is a type of saponin contained in ginseng, and its physiological activities such as lipid peroxidation inhibitory and alcohol-induced brain developmental defense, and voltage-dependent calcium channel inhibition (Nah, et al., Proc. Natl. Acad. Sci. USA, 92, 8739-8743, 1995) and anti-pain efficacy (Shin et al., Submited, 1997), but it is known to improve acne and skin troubles of a composition containing ginsenoside Rf as an active ingredient. There have been no reports on the effects, skin convergence and pore contraction effects, skin color enhancement, hair growth, white hair improvement, antidandruff and antiseptic effect.

Therefore, the present inventors have found that ginsenoside Rf can provide acne and skin trouble improvement effect, skin convergence and pore contraction effect, and can improve skin coloration, hair growth, white hair improvement, antidandruff and antiseptic effect. Discovered and completed the present invention.

Therefore, it is an object of the present invention contains ginsenoside Rf can provide acne and skin trouble improvement effect, skin convergence and pore contraction effect, skin color improvement effect, hair growth improvement, white hair improvement, anti dandruff and antiseptic effect It is providing the skin external preparation composition shown.

In order to achieve the above object, the present invention provides a skin external preparation composition for improving acne containing ginsenoside Rf as an active ingredient.

In addition, the present invention provides a skin external preparation composition for improving color and skin tone containing ginsenoside Rf as an active ingredient.

The present invention also provides a topical skin composition for reducing pores containing ginsenoside Rf as an active ingredient.

The present invention also provides a composition for promoting hair growth containing ginsenoside Rf.

The present invention also provides a composition for preventing hair loss containing ginsenoside Rf.

The present invention also provides a composition for anti-dandruff containing ginsenoside Rf.

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

The composition of the present invention can provide anti-inflammatory, acne and skin trouble improvement effect, skin convergence and pore contraction effect by containing ginsenoside Rf, and improves skin complexion, hair growth, white hair improvement, anti dandruff and antiseptic effect. Can provide.

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

Ginsenoside Rf used in the present invention has a structure represented by the following Chemical Formula 1.

Formula 1

Ginsenoside Rf of the present invention can be extracted from plants, can be used synthesized according to methods known in the art, and may be commercially available. Ginsenoside Rf can also be obtained from ginseng extract. The type of ginseng used at this time is not particularly limited, and ginseng, red ginseng, white ginseng, taeguksam, misam and the like can be used. In addition, the ginseng extract is contained not only in the leachate obtained by leaching and transferring from ginseng, but also in the concentrate obtained by partially or fully concentrating the leachate, or in stagnation, whole, regular, liquid extract and ginseng prepared by drying the concentrate again. The chemicals that exert the main effect, of course, include all of the plant itself, and extracts from all parts of ginseng, such as stems, roots, leaves, flowers, fruits, can be used and are not limited to extracts of any particular part. In addition, a method for extracting ginsenoside Rf from ginseng extract may use a known method.

Specifically, the ginsenoside Rf may be separated from the ginseng extract after preparing ginseng extract with 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, and preferably 80% ethanol is used. At this time, the extraction temperature is preferably 10 ~ 80 ℃, it can be extracted for 3 to 24 hours. If the extraction temperature and extraction time is out of the extraction efficiency may decrease or change of components may occur.

The composition of the present invention preferably contains the ginsenoside Rf in an amount of 0.001 to 50% by weight based on the total weight of the composition. This is because when the content of the active ingredient is less than 0.001% by weight, the efficacy and effect by the above ingredients are weak, and when the amount of the active ingredient exceeds 50% by weight, there is a problem of skin safety or formulation.

The composition of the present invention can be used as an external preparation composition for improving acne, which is excellent in antibacterial effect, in particular, antibacterial effect against acne causing bacteria, and also provides an anti-inflammatory effect.

The composition of the present invention can be used as an external skin composition for improving color and skin tone, and when applied to the skin, it smoothly supplies nutrients to the skin by promoting capillaries and promotes blood circulation and inhibits skin aging to improve color and skin tone. The effect is excellent.

The composition of the present invention can be used as a skin external preparation composition for pore reduction, sebum control and skin trouble improvement, which suppresses excessive secretion of sebum when applied to the skin, promotes free radical removal and collagen synthesis to shrink pores. In addition, the effect of suppressing skin problems is excellent by reducing the expression of inflammatory factors.

The composition of the present invention can be used as a composition for promoting hair growth, which promotes hair growth by promoting the transition of the resting hair cycle to the growing hair cycle, and also promotes the production of new hair, as well as promoting healthy hair. It includes growing, and provides the effect of preventing and suppressing the phenomenon of hair falling off from the scalp or the condition of hair growth or thinning.

The composition of the present invention can be used as a composition for preventing white hair, and increases the MITF expression of melanocytes to activate melanocytes and promote melanin synthesis, thereby preventing the induction of white hair and providing an effect of promoting hair loss.

The composition of the present invention can be used as an anti-dandruff skin external composition, which effectively discharges toxins accumulated in the hair and scalp to cleanse the scalp, inhibit the growth and growth of dandruff bacteria, and prevent the scalp inflammatory reaction, and also active It has an excellent antioxidant effect that inhibits the production and action of oxygen, so it can provide the effect of calming and strengthening the scalp and strengthening its natural defense.

The composition of the present invention can be used as a natural preservative composition, and because it is a natural ingredient, it provides an effect that is excellent in preservative effect and harmless 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 emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) and non-obtained by dispersing an oil phase in solution, gels, solids, pasty anhydrous products, aqueous phases. It may be provided in the form of an ionic vesicle dispersant or in the form of a cream, skin, lotion, powder, ointment, spray or cone stick. It may also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions can be prepared according to conventional methods in the art.

In particular, when the topical skin composition of the present invention is used for the prevention of dandruff, hair growth or white hair, it may be formulated as a composition for scalp and hair, and the formulation is not particularly limited, for example, hair tonic, hair nourishing cosmetics, scalp It can be formulated as a treatment, hair treatment, hair shampoo, hair rinse, hair lotion or scalp hair combination treatment and the like.

In addition, the composition according to the present invention is 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 or nonionic. Such as type emulsifiers, fillers, metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics. It may contain adjuvants conventionally used in the cosmetic or dermatology field. Such adjuvants are introduced in amounts generally used in the cosmetic or dermatological arts.

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

Hereinafter, the configuration 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 only for the purpose of illustration to help the understanding of the present invention is not limited to the scope and scope of the present invention by the following examples.

Reference Example 1 Preparation of Ginsenoside Rf

Ginsenoside Rf for testing the efficacy of the composition of the present invention was purchased from Ambo laboratory.

[Formulation Example 1 and Comparative Formulation Example 1]

Nutritional cream was prepared in a conventional manner according to the composition of Table 1 (unit: wt%).

Table 1 Ingredient Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 Ginsenoside Rf 5.00 - Vegetable Cured Oil 1.50 1.50 Stearic acid 0.60 0.60 Glycerol Stearate 1.00 1.00 Stearyl alcohol 2.00 2.00 Polyglyceryl-10 Pentastearate & Behenyl Alcohol & Sodium Stearoyl Lactylate 1.00 1.00 Arachidil Behenyl Alcohol & Arachidil Glucoside 1.00 1.00 Cetylaryl Alcohol & Cetearyl Glucoside 2.00 2.00 PEG-100 Stearate & Glycerolate & Propylene Glycol 1.50 1.50 Caprylic / Carric Triglycerides 11.00 11.00 Cyclomedicon 6.00 6.00 Preservative, incense Quantity Quantity Triethanol amine 0.1 0.1

Test Example 1 Color Improvement Effect

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 a LDPI (Laser Doppler Perfusion Imager; periscan PIM II, Perimed (stochholm, Sweden)). LDPI is a well-known and currently used device for measuring blood circulation in the skin, and is a very sensitive device that can measure not only the speed and amount of blood in capillaries of the skin but also the flow in the small arteries and the venous veins.

After washing the face with soap in a thermo-hygrostat room for 30 minutes, the initial value was measured using LDPI. Initially, initial blood flow in the lower forehead of 30 women with cold hands and feet was measured by LDPI. Then, after the formulation Example 1 and Comparative Formulation Example 1 were used in the test subjects for one week, the result of comparing the blood flow rate measured with the initial measurement value (skin blood flow change) is shown in Table 2 below.

TABLE 2 LDPI Results-Skin Blood Flow Before and After Cosmetic Use Test substance % Change in skin blood flow after 1 week of use Formulation Example 1 11 Comparative Formulation Example 1 5

In the results of Table 2, the cosmetic composition according to the present invention significantly increased the skin blood flow than Comparative Formulation Example 1 containing no ginsenoside Rf, it was confirmed that the blood color is improved through the promotion of this blood circulation. . This ultimately suggests that the cosmetic composition containing ginsenoside Rf according to the present invention can contribute effectively to the skin's nutrient delivery, inhibit skin aging and delay.

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, each of 30 subjects were used (once evening / day application, for a total of 1 week), and then, a Facial Stage DM-3 (Moritex, Japan) device The improvement of skin tone was evaluated using. Skin tone improvement rate was determined by the lightness and color measurement value of the skin and the change in the brightness and color of the skin, the results are shown in Table 3 below. The higher the brightness and color change, the better the skin tone.

TABLE 3 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 containing no ginsenoside Rf according to the present invention did not show significant skin tone improvement effect, while Formulation Example 1 containing ginsenoside Rf as an active ingredient was higher than before use. It was confirmed that the skin tone after use is much improved.

Test Example 3 Pore Reduction Effect

1. Pore reduction effect through promoting collagen biosynthesis

Ginsenoside Rf according to the present invention was measured by comparing collagen biosynthesis promoting effect with TGF-β. First, fibroblasts were seeded by 10 ⁵ per hole in 24 wells and cultured until 90% growth. This was incubated in serum-free DMEM medium for 24 hours, and then treated with 10 g / ml of ginsenoside Rf and TGF-β of the present invention dissolved in serum-free medium, and incubated in a CO ₂ incubator for 24 hours. These supernatants were removed and procollagen increased or decreased using a procollagen type (I) ELISA kit (procollagen type (I); # MK101, TAKARA (Shiga, Japan)). The results are shown in Table 4, and the synthetic ability of collagen was compared with the non-treated group as 100.

Table 4 Test substance Collagen Synthesis (%) Untreated group 100 TGF-β 183.5 Ginsenoside Rf 187.6

In the results of Table 4, ginsenoside Rf according to the present invention was confirmed to exhibit a higher level of collagen synthesis than the positive control group TGF-β. Therefore, it was confirmed that ginsenoside Rf according to the present invention can reduce the enlarged pores by increasing the amount of collagen production around the pores.

2. Pore Reduction Effect

In order to determine the pore reduction effect of Formulation Example 1 and Comparative Formulation Example 1 was evaluated as follows. Twenty male and female subjects with large pore sizes were selected and divided into two groups of ten, and the nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were applied to the face each day for 4 weeks. Determination of the effect of pore reduction was made by visual assessment of experts by taking pictures before and after 4 weeks of the experiment. The results are shown in Table 5 below (rating rating: 0-not scaled down at all; 5-scaled down).

Table 5 Test substance Rating Formulation Example 1 4 Comparative Formulation Example 1 0

In the results of Table 5, Comparative Formulation Example 1 does not have a pore reduction effect, but in the case of Formulation Example 1 shows a pore reduction effect that can be visually confirmed, ginsenoside Rf according to the present invention reduces the size of the pores It was found that the effect was excellent.

[Test Example 4] sebum secretion inhibitory effect

1. Inhibition of skin hypersecretion through inhibition of 5α-reductase activity

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

In order to confirm the degree of inhibition of 5α-reductase activity, ginsenoside Rf was added and incubated for 2 hours at 37 ° C. in a 5% CO ₂ incubator. At this time, the ginsenoside Rf was not used as a negative control group, the pinasteride (finasteride) was used as a positive control group. Thereafter, the culture medium was collected, the steroid was extracted with 800 μl ethyl acetate, the organic solvent layer was separated, dried, and the remaining residue was dissolved in 50 μl ethyl acetate. The silica plastic sheet kieselgel 60 F254 ) Was developed using ethyl acetate-hexane (1: 1) as a solvent.

After drying the plastic sample in air, the bath system was used to measure the isotope amount. The isotopic isomer of testosterone and dihydrotestosterone remaining in the film after one week by placing the dried plastic sheet and the x-ray film together in the bath cassette. After the amount was measured, conversion and inhibition rates were calculated according to the following Equations 1 and 2, and the results are shown in Table 6 below.

Equation 1

Equation 2

Table 6 Test substance % Conversion % Inhibition Negative Control 48 - Positive control group 27.6 42.5 Ginsenoside Rf 15.4 67.9

In the results of Table 6, 5α-reductase, which converts testosterone to dihydrotestosterone, binds to a receptor protein in the cytoplasm, enters the nucleus, activates sebaceous gland cells, promotes differentiation, and hypersecretes sebum in sebaceous glands. It was confirmed that ginsenoside Rf effectively inhibits the conversion of testosterone to dihydrotestosterone, and has a superior inhibitory effect than finasteride, which is known to inhibit the activity of 5α-reductase. Therefore, it was confirmed that ginsenoside Rf can suppress sebum hypersecretion by effectively inhibiting the activity of 5α-reductase.

2. Sebum secretion inhibitory effect

In order to determine the sebum secretion inhibitory effect of Formulation Example 1 and Comparative Formulation Example 1 was evaluated as follows. Thirty male and female subjects who felt sebum secretion were selected and the nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were applied to the designated areas of the facial skin every day for 4 weeks. Determination of the effect of sebum reduction was measured using the sebum meter (Sebumeter SM810, C + K Electronic Co., Germany) to measure the average percentage of sebum reduction after 2 weeks and 4 weeks, respectively, and the results are shown in Table 7 Shown in

TABLE 7 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 of Table 7, it can be seen that Formulation Example 1 containing ginsenoside Rf according to the present invention as an active ingredient can effectively inhibit sebum secreted in excess than 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. Specifically, Formulation Example 2 is a mixture of ginsenoside Rf, Comparative Formulation Example 2 does not contain any active ingredients for improving acne skin, Comparative Formulation Example 3 is a standard to be used as a reference for antimicrobial activity It contains erythromycin, which is widely used as an acne treatment.

The preparation method of Formulation Example 2 and Comparative Formulation Examples 2-3 is as follows. The components of phase A in Table 8 were completely dissolved, and the components of phase B were completely dissolved in a separate dissolution tank, and then mixed and solubilized by adding phase B to phase A. The ingredients of phase C were added thereto according to the blending ratios described in Table 8 to homogenize the mixing and then filtered to prepare the compositions.

Table 8 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 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 Antibacterial Activity Test against Acne Bacteria

Test the antimicrobial activity against the propionibacterium Acnes (ATCC 6919: medium-BHI broth) of the acne-causing strain with each of the cosmetic composition prepared in the composition of Formulation Example 2 and Comparative Formulation Examples 2-3 It was.

The antibacterial test method for acne bacteria was as follows.

(1) Test bacteria preparation

Propionibacterium acnes was used as a culture broth inoculated in BHI broth and anaerobic culture.

(2) dilution solution preparation

0.15 ml of the test bacteria was added to 15 ml of BHI broth (pH 6.8) or LB broth (pH 4.5) and mixed well as a dilution solution.

(3) sample preparation

The cosmetic composition stock solutions prepared from Formulation Example 2 and Comparative Formulation Examples 2-3 were used as samples.

(4) antimicrobial activity test

1) Insert the sample to the starting concentration in the first well of 96-well cell culture tube (96 well plate) and add 200 μl of the total amount into the dilution solution.

2) Mix the mixed solution in the first row well, take 100μl into the second row, mix well, and then add 100μl to the third row and perform double dilution.

3) After incubation for 24 hours and 48 hours at 32 ℃, determine the growth of bacteria to the extent of suspension and determine the minimum concentration without growth of bacteria as MIC (Minimum Inhibitory Concentration) value. If the mixed solution is opaque and it is difficult to determine the growth of bacteria, check through a microscope.

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

Table 9 Item pH Propionibacterium Acnes Formulation Example 2 5.7 > 41 ppm Comparative Formulation Example 2 5.7 Highest concentration (no antimicrobial activity) Comparative Formulation Example 3 5.7 > 100 ppm

In the results of Table 9, the smaller the ppm concentration in the MIC can be said to be an effective material for the antimicrobial activity against acne bacteria, in the case of Formulation Example 2 ppm concentration compared to Comparative Formulation Example 3 using a known acne treatment erythromycin It can be seen that the composition containing the ginsenoside Rf has significantly superior antimicrobial activity against test bacteria.

Test Example 6 Lipogenesis Inhibition Test

A mouse fibroblast cell line, 3T3-L1 cells, was loaded with DMEM (Dulbeco's modified eagle's medium, GIBCO BRL, Life Technologes) medium containing 10% fetal bovine serum (FBS). The well culture plates were attached at 1 × 10 ⁵ cells / well. After 2 days, it was again exchanged with fresh DMEM (containing 10% FBS) medium and incubated 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 ginsenoside Rf and caffeine. After 2 days of treatment 50μM was exchanged for DMEM containing insulin and incubated for 5 days. After 5 days, the cells were exchanged with normal medium (DMEM, containing 10% FBS) and cultured while observing until the cells changed to fat cells.

Sudan III staining (S4136, sigma-aldrich) was performed on the differentiated 3T3-L1 adipocytes to evaluate the inhibitory effect of ginsenoside Rf on adipocyte accumulation. Adipose cells were fixed at room temperature with 4% paraformaldehyde (pH 7.2) in phosphate buffer, washed with PBS (phosphate buffered saline), stained with Sudan III, and photographed for visual comparison. As a control group, only the medium without the test substance or the comparative substance was used, and 50 μM of caffeine was treated with another control group. The degree of fat accumulation inhibition was given by dividing the degree of staining into +++, ++, +, and-, and this means that the degree of staining is increased toward +++. The results are shown in Table 10 below.

Table 10 sample % Inhibition Control +++ Comparison + Ginsenoside Rf -

From the results of Table 10, ginsenoside Rf used in the present invention can be seen that not only the amount of fat accumulated in adipocytes, but also has a superior lipid synthesis inhibitory effect than caffeine, a known lipid synthesis inhibitor. . Therefore, sebum is reduced by inhibiting lipid synthesis, thereby suppressing the occurrence of acne.

[Test Example 7] Test for acne improvement, sebum secretion and irritation

Thirty people with acne were divided into three groups of ten people each and subjects corresponding to each group were allowed to use the cosmetic composition prepared in Formulation Example 2 and Comparative Formulation Examples 2-3 for one month. The acne improvement scale ranged from 1 to 5 points, with 1 being ‘no’, 3 being ‘normal’ and 5 being ‘very yes’. Experimental results are shown in the average score of 10 people in Table 11 below.

Acne disappearance was based on the number of days the extinction was read, acne recurrence was based on the results after one month. Sebum secretion is reduced from 1 to 5 points, with 1 being ‘no’, 3 being ‘normal’ and 5 being ‘very yes’. Experimental results are shown in the average score of 10 people in Table 11. The presence or absence of skin irritation was determined as (number of irritants) / (total number of test subjects).

Table 11 Inflammatory Acne Improvement Cotton acne extinction Acne recurrence Reduce sebum secretion Irritation 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, Formulation Example 2 did not recur acne compared to Comparative Formulation Example 2, it can be seen that there is an excellent effect on the overall acne improvement. On the other hand, Comparative Formulation Example 3 containing an antimicrobial activity standard shows an acne-improving effect, but due to its strong skin irritation in use, it may not be suitable for long-term use, but the composition according to the present invention has no irritation for long-term use. Even appeared to be appropriate.

Formulation Example 3 and Comparative Formulation Example 4

To prepare a shampoo in the composition of Table 12. Specifically, the surfactant and ethylene glycol distearate are added to purified water, heated to 80 ° C., uniformly dissolved, and then slowly cooled to 40 ° C. under stirring, and the active ingredient and preservative according to the present invention are added to the mixture. A viscosity modifier, a perfume, and a hair conditioner were added and mixed, followed by cooling to room temperature under stirring.

Table 12 Ingredient (% by weight) Formulation Example 3 Comparative Formulation Example 4 Ammonium Lauryl Sulfate 10 10 Polyoxyethylene lauryl ammonium sulfate 5 5 Cocoamidopropylbetaine 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 Methylparaben 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 Example 8 Dandruff Reduction Effect Test

Twenty-four males aged 19 to 35 years with relatively high dandruff were selected and used in two groups of 12 shampoos for each of Formulation Example 3 and Comparative Formulation Example 4 for one month as follows. .

Before the start of the test, triturate with a normal shampoo, and collect the dandruff accumulated for 2 days after the shampoo, and the weight of the collected dandruff and trituration once every 2 days with the shampoos of Formulation Example 3 and Comparative Formulation Example 4, and 2 days after the end of the test. The weight of accumulated dandruff was evaluated. At this time, the accumulated dandruff was collected directly from the scalp with a vacuum suction device, to obtain a rate of dandruff reduction based on Equation 3 below and the results are shown in Table 13 below.

Equation 3

Table 13 Formulation Example 3 Comparative Formulation Example 4 Dandruff reduction rate (%) 66.5 0.7 55.6 -0.5 59.6 -1.1 72.6 1.5 66.3 2.2 72.3 1.3 68.9 6.3 63.8 3.6 71.2 3.3 59.8 4.6 68.3 3.7 52.6 4.2 medium 64.79 2.5 SD 6.59 2.2

In the results of Table 13, it can be seen that the formulation of Example 3 containing ginsenoside Rf shows an excellent anti-dandruff effect.

Test Example 9 Scalp Itching Prevention Test

Twenty-four men and women aged 25 to 45 who feel severely itchy scalp were selected and divided into two groups of 12 people each to use the shampoo of Formulation Example 3 and Comparative Formulation Example 4 once every three days for two weeks. 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

Normal-3 points

Poor-2 points

Very bad-1 point

Table 14 division Formulation Example 3 Comparative Formulation Example 4 Itching effect of scalp 4.5 2.3

In the results of Table 14, it can be seen that the case of the formulation example 3 containing ginsenoside Rf shows a better effect in preventing scalp itching.

Test Example 10 Evaluation of Potassium Ion Channel Activity Increase Effect

Hair loss treatment of minoxidil is known as mitochondrial potential potassium ion channel openers (K _ATP channel opener), a representative drug used in the treatment of androgenetic alopecia. To evaluate the mechanism of minoxidil, the treatment of toltamide (SIGMA AlDRICH, T0891), which blocks the K _ATP channel in fibroblasts constituting the dermis of the scalp, inhibits cell proliferation, and then opens potassium ion channels to allow cell proliferation. A recovering test method was used.

In order to evaluate the function of the composition as a K _ATP channel opener, in the present invention, a mouse embryonic fibroblast cell line (NIH3T3) cell line was used. The cell line is a cell line in which the fibroblast cell line isolated from NIH Swiss mouse embryo was naturally immortalized by 3T3 protocol. The cell line was incubated in DMEM (Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS for 24 hours in an incubator maintained at 5% CO ₂ , 37 ℃. NIH3T3 was placed in a 96-well plate and incubated in a 37 ° C. incubator for 24 hours, treated with 2.5 mM tolbutamide, and after 10 minutes, 10 μM of minoxidil and ginsenoside Rf of 2.5 ppm, 5 ppm and 10 ppm, respectively, were added. The cell proliferation was measured using the WST-1 kit (Roche) after 48 hours of drug treatment. The results are shown in Table 15 below.

Table 15 division Cell proliferation (%) Untreated 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, the proliferation of fibroblasts is restored, and the cell proliferation ability is increased depending on the concentration of the treated ginsenoside Rf, and the concentration of ginsenoside Rf is 10 ppm. In the case of treatment with, it can be confirmed that the proliferation of cells is restored to the level when minoxidil is treated.

Test Example 11 Melanin Promoting Effect Test of Ginsenoside Rf

Add melan-a to 50,000 cells / well in a 24-well microtiter plate in medium containing 5% fetal bovine serum, RPMI medium, 100 IU penicillin G and 0.2 μM TPA in RPMI medium. Busy. The next day, the divided cells were treated with ginsenoside Rf at a final concentration of 10 ppm or 50 ppm as a test substance, 0.1% DMSO as a negative control, and 100 μM IBMX as a positive control, followed by incubation at 37 ° C. for 3 days. After incubation, the wells were washed with PBS, 100 μl of 1N NaOH was added, and the melanin in the cells was lysed. The absorbance of the dissolved melanin was measured at 405 nm using a microplate reader (Synergy2, BioTek (VT, USA)). The result of comparing the melanin production promoting effect of ginsenoside Rf with the control is shown in Table 16 below.

Table 16 sample Melanin Synthesis (%) DMSO (0.1%) 100 IBMX (100 μM) 120 Ginsenoside Rf (10 ppm) 112 Ginsenoside Rf (50 ppm) 123

In the results of Table 16, it can be seen that ginsenoside Rf promotes melanin synthesis of melanocytes, thereby increasing melanin production, showing an excellent melanin production promoting effect.

Experimental Example 12 Effect of Ginsenoside Rf on the Melanosite in Promoting MITF and Tyrosinase Expression

Dispense 500,000 cells / well in a 6-well microtiter plate using a 501mel cell line, in each hole 0.1% DMSO as a negative control, 100 μM as a positive control, and ginsenoside as a test group. Rf was treated with 10 ppm, and the protein was obtained after incubation at 37 ° C. for 24 hours, 48 hours, and 72 hours. The protein thus obtained was subjected to western blot using MITF and tyrosinase antibody. Protein extraction and Western blot were usually performed by standard methods used by those skilled in the art. After western blot the results are shown in Table 17 below by comparing the negative control to 100.

Table 17 MITF Tyrosinase IBMX Ginsenoside Rf IBMX Ginsenoside Rf 24hr 121 97 160 153 48hr 98 111 149 155 72hr 224 119 298 186

In the results of Table 17, it can be seen that ginsenoside Rf increases the expression of MITF and tyrosinase protein in melanocytes.

Test Example 13 Evaluation of Antimicrobial Activity of Ginsenoside Rf

Antimicrobial experiments were conducted to evaluate the antimicrobial activity of ginsenoside Rf. The specific experimental method is as follows.

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa strains used in the experiment were tested on Tryptic Soy Broth. Candida albicans and Aspergillus niger strains were cultured in Sabouraud Dextrose Broth. Incubate 1/100 (Candida albicans) in each medium The strain was diluted 1/10) was used as the test bacteria. Aspergillus niger used a spore suspension prepared to be 2 × 10 ⁸ cfu / ml as the test cell solution.

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

16 µl of 10 ppm of ginsenoside Rf was added to a 96-well plate, and 184 µl of the diluted solution was added thereto. The remaining wells were added with 100 μl of dilution solution. After mixing well the mixture of the first row, 100 μl was taken in the second row, mixed well, and then diluted 100 times by taking 100 μl again in the third row.

Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger, in a 32 ° C. thermostat. niger) was incubated in a 25 ℃ thermostat.

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

Table 18 Sample Minimum Inhibitory Concentration (MIC),% Sedona Monas Aruginosa Staphylococcus aureus Escherichia Coli Candida Albicans Aspergillus Niger Ginsenoside Rf 0.2 0.04 0.128 > 3 > 1

In the results of Table 18, it can be seen that ginsenoside Rf exhibits antimicrobial activity against various strains, through which it can be predicted that ginsenoside Rf can act as a natural preservative or antimicrobial agent in the composition.

Claims (23)

Skin external composition comprising ginsenoside Rf as an active ingredient. The external preparation for skin composition according to claim 1, wherein the ginsenoside Rf is represented by the following Chemical Formula 1. [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 of skin according to claim 1 or 2, wherein the composition is for antibacterial use. The external preparation composition for skin according to claim 1 or 2, wherein the composition is for improving color and skin tone. The external preparation composition 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 controlling sebum. The external preparation composition 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 claim 1 or 2, wherein the composition is for preventing white hair. The composition for external application for skin according to claim 1 or 2, wherein the composition is for antidandruff. The composition for external application for skin according to claim 1 or 2, wherein the composition is for a natural preservative. Use of the external preparation for skin for acne improvement comprising ginsenoside Rf as an active ingredient. Use as an antibacterial use of an external composition for skin containing ginsenoside Rf as an active ingredient. Use as an active ingredient for improving skin color and skin tone of a skin external preparation composition comprising ginsenoside Rf as an active ingredient. A use for externally reducing pores of a skin external preparation composition comprising ginsenoside Rf as an active ingredient. Use as a sebum control of the external composition for skin containing ginsenoside Rf as an active ingredient. Use as a skin trouble improvement of the external composition for skin containing ginsenoside Rf as an active ingredient. Use of the external preparation composition for skin growth containing ginsenoside Rf as an active ingredient for promoting hair growth. Use of the external preparation for skin on the skin for containing white ginsenoside Rf as an active ingredient. Use as anti-dandruff of external composition for skin, comprising ginsenoside Rf as an active ingredient. Use of the external preparation for skin as a natural preservative for ginsenoside Rf.