KR102021463B1 - External composition for skin containing Ginsenoside Rg3 - Google Patents

Abstract

The composition of the present invention, by containing ginsenoside Rg3, can provide anti-aging, skin wrinkle improvement, whitening, and moisturizing effect, as well as acne and skin troubles, atopic symptoms, skin convergence and pore contraction, and skin hemoglobin. It relates to a composition that can provide the effect of improvement, hair growth, white hair improvement, antidandruff and antiseptic effect.

Description

External composition for skin containing Ginsenoside Rg3

The present invention can provide acne and skin trouble improvement effect, skin convergence and pore contraction effect by containing ginsenoside Rg3, and can provide skin color improvement effect, hair growth improvement, white hair improvement, antidandruff 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 refers to saponin in ginseng. Saponin is a chemical compound called glycoside. Ginsenoside is a name given to mean glyco-side isolated from ginseng to distinguish it from other plant saponins. It is usually named ginsenoside-Rx and the like. Here, “R” means root (Radix or Root) and “x” means o, a, b, c, d, from bottom to top according to the distance (Rf value) of the spot moving on TLC. They are named in order of e, f, g, and h. Ginseng saponin has a variety of effects on the body's function by affecting the central nervous system, endocrine system, immune system, metabolic system. To date, more than 30 kinds of ginsenosides have been identified and isolated from Korean ginseng (including red ginseng), and the pharmacological action of each ginsenoside has been revealed one after another. These ginsenosides may act similarly or vice versa. For example, G-Rb1, a representative ginsenoside of PPD-based saponins, has an inhibitory effect on the central nervous system, and G-Rg1, a representative of PPT-based saponins, has a promoting effect.

Ginseng saponins are panaxadiol-type Ginsenosides (Ra1, Ra2, Ra3, Rb1, Rb2, Rb3, Rc, Rd, Rg3) and panaxtriol-based ginseng depending on the structure of the saponin aglycon. It can be classified into three types such as panacetic acid (Panaxatriol-type Ginsenosides: Re, Rf, Rgl, Rg2) and oleanoic acid-type Ginsenosides. The content of panaxadiol-based saponins in the ginseng root was the highest at 48-60%, followed by the panaxtriol-based saponins at 30-35%, and trace amounts of oleanoline-based saponins.

The pharmacological action of Rg3, a kind of minor ginsenosides produced during the ginseng process of ginseng, can inhibit cancer cell metastasis, inhibit platelet aggregation and antithrombotic action, relax vasculature, experimental liver injury, and anticancer drug resistance. Although there are patents for oral pharmacological action such as inhibitory effect, the composition of ginsenoside Rg3 as an active ingredient, acne and skin trouble improvement effect, skin convergence and pore contraction effect, skin color improvement effect, hair growth improvement, white hair improvement, No antidandruff and antiseptic effects have been reported.

Therefore, the present inventors can provide ginsenoside Rg3, acne and skin trouble improvement effect, skin converging and pore contraction effect, and can provide skin color improvement effect, hair growth improvement, white hair improvement, anti dandruff and antiseptic effect. The present invention was completed.

Therefore, it is an object of the present invention by containing ginsenoside Rg3 can provide acne and skin trouble improvement effect, skin converging 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 Rg3 as an active ingredient.

In addition, the present invention provides a skin external preparation composition for improving color and skin tone containing ginsenoside Rg3 as an active ingredient.

The present invention also provides a composition for external application for skin pore reduction containing ginsenoside Rg3 as an active ingredient.

The present invention also provides a composition for promoting hair growth containing ginsenoside Rg3.

The present invention also provides a composition for preventing hair loss containing ginsenoside Rg3.

The present invention also provides a composition for anti-dandruff containing ginsenoside Rg3.

The present invention also provides a natural preservative composition containing ginsenoside Rg3.

The composition of the present invention can provide acne and skin trouble improvement effect, skin convergence and pore contraction effect by containing ginsenoside Rg3, and can provide skin color improvement effect, hair growth improvement, white hair improvement, antidandruff and antiseptic effect. Can be.

The composition according to the present invention contains ginsenoside Rg3 (Ginsenoside Rg3) as an active ingredient.

Ginsenoside Rg3 used in the present invention has a structure represented by the following Chemical Formula 1.

Ginsenoside Rg3 of the present invention can be extracted from plants, may be synthesized according to methods known in the art, and may be commercially available. Ginsenoside Rg3 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 Rg3 from ginseng extract may use a known method.

Specifically, the ginsenoside Rg3 may be isolated from ginseng extract prepared by water or an organic solvent by a method well known in the art in ginseng. 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 Rg3 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 Rg3

Ginsenoside Rg3 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%).

Ingredient Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 Ginsenoside Rg3 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 Arakidil Behenyl Alcohol & Arakidil 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). 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 the 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.

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 13 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 Rg3, it was confirmed that the blood color is improved by promoting the blood circulation. . This ultimately suggests that the cosmetic composition containing ginsenoside Rg3 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 find out the skin tone improvement effect of Formulation Example 1 and Comparative Formulation Example 1, each of 30 subjects were used (once evening / daily 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.

Test substance Skin tone improvement rate (%) Brightness (mean ± standard deviation) Color (mean ± standard deviation) Formulation Example 1 14 ± 3.04 11 ± 2.24 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 Rg3 according to the present invention does not show a significant skin tone improvement effect, while Formulation Example 1 containing ginsenoside Rg3 as an active ingredient 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 Rg3 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 treatment of serum-free DMEM medium and then cultured in non-ginsenoside Rg3 and the TGF-β of the present invention was dissolved in serum-free medium by each of 10ng / ml for 24 hours, CO ₂ The incubator was incubated for 24 hours. These supernatants were removed and procollagen increased or decreased using procollagen type I ELISA kit (procollagen type (I)). The results are shown in Table 4, and the synthetic ability of collagen was compared with the non-treated group as 100.

Test substance Collagen Synthesis (%) Untreated group 100 TGF-β 183.5 Ginsenoside Rg3 197.2

In the results of Table 4, ginsenoside Rg3 according to the present invention was confirmed to exhibit a high level of collagen synthesis ability higher than the positive control TGF-β. Therefore, it was confirmed that ginsenoside Rg3 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).

Test substance Rating Formulation Example 1 3 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 exhibits a pore reduction effect that can be visually confirmed, ginsenoside Rg3 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 Rg3 was added thereto and incubated for 2 hours at 37 ° C. in a 5% CO ₂ incubator. At this time, the ginsenoside Rg3 was used as a negative control group, the pinasteride 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.

Test substance % Conversion % Inhibition Negative Control 48 - Positive control group 27.6 42.5 Ginsenoside Rg3 16.3 66.0

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 Rg3 effectively inhibited the conversion of testosterone to dihydrotestosterone, effectively inhibiting the activity of, and had a superior inhibitory effect than finasteride, which is known to inhibit the activity of 5α-reductase. Thus, it was confirmed that ginsenoside Rg3 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

Test substance Sebum reduction rate (%) After 2 weeks After 4 weeks Formulation Example 1 32 40 Comparative Formulation Example 1 5 5

From the results of Table 7, it can be seen that Formulation Example 1 containing ginsenoside Rg3 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 combination of ginsenoside Rg3, 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.

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 Cured Castor Oil
(hydrogenated castor oil) 0.4 0.4 0.4 Perfume 0.04 0.04 0.04 C Ginsenoside Rg3 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.

Item pH Propionibacterium Acnes Formulation Example 2 5.7 > 48 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 Rg3 has significantly superior antimicrobial activity against the test bacteria.

Test Example 6 Lipogenesis Inhibition Test

3T3-L1 cells, a mouse fibroblast cell line, were loaded with DMEM (Dulbecos modified eagles medium, GIBCO BRL, Life Technologes) medium containing 10% fetal bovine serum (FBS). The well culture plate was 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 Rg3 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 using the differentiated 3T3-L1 adipocytes to evaluate the inhibitory effect of ginsenoside Rg3 on adipocyte accumulation in adipocytes. 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 inhibition of fat accumulation was given by dividing the degree of staining into +++, ++, +, and-, which means that the degree of staining is increased toward +++. The results are shown in Table 10 below.

sample % Inhibition Control +++ Comparison + Ginsenoside Rg3 -

In the results of Table 10, ginsenoside Rg3 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. 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).

Inflammatory Acne Improvement Cotton acne extinction Acne recurrence Reduce sebum secretion Irritation Formulation Example 2 4.3 3 days radish 4.5 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.

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 Rg3 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.

Formulation Example 3 Comparative Formulation Example 4 Dandruff reduction rate (%) 82.7
86.1
85.6
59.6
77.6
89.1
87.1
69.8
73.5
79.3
83.2
87.6 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 80.1 2.5 SD 8.70 2.2

In the results of Table 13, it can be seen that in the case of Formulation Example 3 containing ginsenoside Rg3 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

division Formulation Example 3 Comparative Formulation Example 4 Itching effect of scalp 4.3 2.3

In the results of Table 14, it can be seen that the formulation of Example 3 containing ginsenoside Rg3 shows a better effect in preventing scalp itch.

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 10 minutes later, 2.5 ppm, 5 ppm, and 10 ppm of 10 μM of minoxidil and ginsenoside Rg3 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.

division Cell proliferation (%) Untreated Control 100 Minoxidil 132 Ginsenoside Rg3 (2.5 ppm) 114 Ginsenoside Rg3 (5 ppm) 123 Ginsenoside Rg3 (10 ppm) 129

In the results of Table 15, it can be seen that when the ginsenoside Rg3 is treated, the proliferation of fibroblasts is restored, and the cell proliferation ability is increased depending on the concentration of the treated ginsenoside Rg3, and the ginsenoside Rg3 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 Rg3

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 cells were treated with ginsenoside Rg3 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. The result of comparing the melanin production promoting effect of ginsenoside Rg3 with the control is shown in Table 16 below.

sample Melanin Synthesis (%) DMSO (0.1%) 100 IBMX (100 μM) 120 Ginsenoside Rg3 (10 ppm) 110 Ginsenoside Rg3 (50 ppm) 120

In the results of Table 16, it can be seen that ginsenoside Rg3 promotes melanin synthesis of melanocytes to increase melanin production, showing an excellent melanin production promoting effect.

[Test Example 12] Effect of Ginsenoside Rg3 on the Melanosite-induced Expression of MITF and Tyrosinase

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. Rg3 was treated with 10 ppm, and then incubated at 37 ° C. for 24 hours, 48 hours, and 72 hours to obtain protein. 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.

MITF Tyrosinase IBMX Ginsenoside Rg3 IBMX Ginsenoside Rg3 24hr 121 94 160 150 48hr 98 110 149 153 72hr 224 119 298 182

In the results of Table 17, it can be seen that ginsenoside Rg3 increases the expression of MITF and tyrosinase protein in melanocytes.

Test Example 13 Evaluation of Antibacterial Activity of Ginsenoside Rg3

Antimicrobial experiments were conducted to evaluate the antimicrobial activity of ginsenoside Rg3. The specific experimental method is as follows.

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa strains used in the experiments 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 1/10) of the strain 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 Rg3 was added to the first row of 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. The mixture was well mixed in the first row, and then 100 μl of the mixed solution was added to the second row and mixed well.

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.

Sample Minimum Inhibitory Concentration (MIC),% Sedonas
Aruginosa Staphylococcus aureus Escherichia Coli Candida Albicans Aspergillus Niger Ginsenoside Rg3 0.18 0.05 0.142 > 3 > 1

In the results of Table 18, it can be seen that ginsenoside Rg3 exhibits antimicrobial activity against various strains, through which it can be predicted that ginsenoside Rg3 can act as a natural preservative or antimicrobial agent in the composition.

Claims (13)

Cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is for improving acne. A cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is antibacterial. Cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is for improving color and skin tone. A cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is for reducing pores. Cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is for improving skin troubles. Cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is for preventing white hair. A cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is anti-dandruff cosmetic composition. A cosmetic composition comprising ginsenoside Rg3 as an active ingredient, wherein the composition is a natural preservative. The cosmetic composition according to any one of claims 1 to 8, wherein the ginsenoside Rg3 is represented by the following Chemical Formula 1.
[Formula 1]

. The cosmetic composition according to any one of claims 1 to 8, wherein the ginsenoside Rg3 is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition. delete delete delete