KR20130095949A - Composition of skin external application containing ginsenoside f1 - Google Patents

Abstract

PURPOSE: A composition for skin external application containing ginsenoside F1 is provided to have an excellent antioxidative activity, and to improve not only the overall skin condition but also scalp and hair condition by containing ginsenoside F1. CONSTITUTION: A composition for skin external application contains the ginsenoside F1 presented by the chemical formula 1 as an active ingredient. Per total weight of the composition, 0.001-50 weight% of the ginsenoside is contained. The composition is used for whitening. The composition is used for moisturizing. The composition is used for strengthening a skin barrier function.

Description

Composition for external application containing ginsenoside F1 ginsenoside F1

The present invention relates to an external composition for skin containing ginsenoside F1, and more particularly, by containing ginsenoside F1, while having excellent antioxidant power, skin moisturizing effect, anti-inflammatory effect, acne and atopic skin troubles, etc. In addition to improving the overall skin condition such as improvement, whitening effect, sebum control effect, pore contraction effect, and improvement of complexion through blood circulation, it also provides scalp and hair condition improvement effects such as anti-dandruff effect, hair growth effect and anti-white hair effect. It relates to a composition that can be.

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. 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 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.

The present inventors can provide the ginsenoside F1 whitening, moisturizing effect as well as improve acne and skin troubles, atopic symptoms and improve skin condition such as skin color improvement, sebum control, pore contraction effect The present invention has been completed and found to be able to provide an effect of improving scalp and hair condition such as anti-dandruff, hair growth and anti-hair growth.

Accordingly, it is an object of the present invention to provide an external preparation composition for skin containing ginsenosides F1 which can improve the overall condition of the skin.

In order to achieve the above object, the present invention provides a skin external preparation composition containing ginsenoside F1 as an active ingredient.

The present invention also provides a whitening skin external composition comprising ginsenoside F1 as an active ingredient.

The present invention also provides a moisturizing skin external composition comprising ginsenoside F1 as an active ingredient.

The present invention also provides an external preparation composition for improving acne containing ginsenoside F1 as an active ingredient.

The present invention also provides a topical skin external composition for improving atopy containing ginsenoside F1 as an active ingredient.

The present invention also provides a skin external preparation composition for improving color and skin tone containing ginsenoside F1 as an active ingredient.

The present invention also provides a topical skin composition for reducing pores containing ginsenoside F1 as an active ingredient.

The present invention also provides a skin external composition for controlling sebum containing ginsenoside F1 as an active ingredient.

The present invention also provides a topical anti-dandruff composition containing ginsenoside F1 as an active ingredient.

The present invention also provides a composition for external application for hair growth comprising ginsenoside F1 as an active ingredient.

In another aspect, the present invention provides a composition for preventing skin whitening containing ginsenoside F1 as an active ingredient.

The present invention also provides an external composition for skin using ginsenoside F1 as a natural preservative.

The composition of the present invention has an excellent antioxidant power by containing ginsenoside F1, while improving skin moisturizing effect, anti-inflammatory effect, skin trouble improvement effect such as acne and atopy, whitening effect, sebum control effect, pore contraction effect, blood circulation improvement In addition to improving the overall skin condition, such as improving the complexion through it can provide a scalp and hair condition improvement effect such as anti-dandruff effect, hair growth effect and white hair prevention effect.

Figure 1 shows the hair growth effect after the application of Formulation Example 5 and Comparative Formulation Example 6.

The external preparation composition for skin according to the present invention contains ginsenoside F1 as an active ingredient.

Ginsenoside F1 used in the present invention has a structure represented by the following Chemical Formula 1.

Ginsenoside F1 of the present invention may be extracted from plants, synthesized according to methods known in the art, and may be commercially available. Ginsenoside F1 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, flow extract and mulberry 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 F1 from ginseng extract may use a known method.

Specifically, the ginsenoside F1 may be separated from the ginseng extract by using 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 according to the present invention may contain ginsenoside F1 in an amount of 0.001 to 50% by weight, preferably 0.01 to 30% by weight, more preferably 0.1 to 10% by weight, based on the total weight of the composition. If the content of the ginsenoside F1 is less than 0.001% by weight, the efficacy and effect by the above ingredients are insignificant.

Since ginsenoside F1 is a component having excellent antioxidant power, the composition of the present invention containing ginsenoside F1 provides excellent antioxidant power to the skin.

The composition of the present invention can be used as a composition for whitening, which can provide an excellent whitening effect by inhibiting tyrosinase activity and inhibiting melanin production.

The composition of the present invention may be used as a moisturizing skin external preparation composition, which may enhance skin barrier function and induce differentiation of skin keratinocytes. Therefore, it can be usefully used as an external composition for skin to prevent or improve skin dryness, atopic dermatitis, contact dermatitis, or psoriasis caused by incomplete epidermal differentiation.

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, and reduces pores by promoting free radical removal and collagen synthesis. In addition, the effect of suppressing skin problems is excellent by reducing the expression of inflammatory factors. In addition, it is possible to defend against the generation of skin irritation due to its excellent antioxidant power.

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 skin external preparation composition for hair growth, which can promote the growth of hair and prevent hair loss by promoting the transition from the resting hair cycle to the growing hair cycle.

The composition of the present invention can be used as a topical anti-skin composition for white hair, which can significantly increase the expression of MITF in melanocytes, inhibit white hair and promote the induction of black hair.

In addition, ginsenoside F1 used in the external preparation composition for skin of the present invention can provide an effect as a natural preservative.

The external preparation composition for skin of the present invention described above may be formulated as a cosmetic composition, and 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- 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 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 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 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]

Ginsenoside F1 for testing the efficacy of the composition of the present invention was purchased from Ambo laboratory.

Test Example 1 Inhibitory Effect of Reactive Oxygen Species Production

The keratinocytes (F1eratinocyte) isolated from the skin tissue of the person into a 5 × 10 ⁴ are in each well of a 24-well plate was attached for 24 hours. After 16 hours, ginsenoside F1 was treated with 1%. At this time, the control group was not treated with ginsenoside F1 for comparison. After 2 hours, the culture solution was removed, and 100 µl of phosphate buffered saline (PBS) was added to each well. The keratinocytes were irradiated with UV 30mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15 W, SanF1yo DennF1i, Japan). Μl was added. The ginsenoside F1 was retreated and the amount of reactive oxygen species increased by UV stimulation at certain time points was quantified. The amount of ROS was quantified by referring to Tan's method for measuring the fluorescence of DCF-DA (dichlorofluorescin diacetate) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432). Table 1 shows the results of calculating the ratio of the vehicle-treated control group ROS.

Test substance UVB 30 mJ / cm ² Time since survey 0hr 2 hr 3hr Vehicle 100 244 287 UVB + Vehicle 100 325 381 UVB + Ginsenoside F1 100 241 280

In the results of Table 1, ginsenoside F1 according to the present invention effectively inhibits the production of ROS known to cause skin cell damage by ultraviolet rays, and the level of ROS after UV stimulation is almost the same level as when no ultraviolet rays are irradiated. It can be seen that the antioxidant effect is very good enough to inhibit the production of ROS. Therefore, it was confirmed that ginsenoside F1 according to the present invention can prevent pores from widening by inhibiting oxidation and preventing aging, and can improve skin trouble by defending the generation of skin irritation.

Test Example 2 Tyrosinase Inhibitory Effect

Tyrosinase enzyme was extracted from the mushroom (Mushroom) was used as the Sigma (SIGMA). First, the substrate tyrosine was dissolved in distilled water to make a solution of 0.3 mg / ml, and the solution was added to the test tube by 1.0 ml. Then, 1.0 ml of potassium-phosphate buffer solution (0.1 mol concentration, pH 6.8) and 0.7 ml of distilled water were added thereto. Added.

0.2 ml of the sample solution prepared by mixing ginsenoside F1 of the present invention in an appropriate concentration in an ethanol solution was added to the reaction solution and then reacted for 10 minutes in a 37 ° C thermostat. In this case, the control group was prepared by adding only 0.2 ml of solvent instead of each sample solution, and ascorbic acid was used as a positive control group. 0.1 ml of a tyrosinase solution of 2500 units / ml was added to the reaction solution and reacted for 10 minutes in a 37 ° C thermostat. The reaction tube containing the reaction solution was placed in iced water, quenched to stop the reaction, and the absorbance at 475 nm was measured with a photospectrometer. The results are shown in Table 2 below. Each tyrosinase inhibitory effect was calculated by the following equation.

Test substance Tyrosinase inhibition rate (%) Control group (no addition) 0 Ascorbic acid 52 Gin Senocide F1 71

From the results of Table 2, the tyrosinase inhibition rate of the cosmetic composition of Formulation Example 1 containing ginsenoside F1 according to the present invention is much higher than the known tyrosinase inhibitor ascorbic acid, it can be seen that the whitening effect is very excellent.

[Formulation Example 1 and Comparative Formulation Example 1]

Nutritional cream was prepared in a conventional manner according to the composition of Table 3 (unit: wt%).

Compounding ingredient Formulation Example 1 Comparative Formulation Example 1 Purified water To 100 To 100 Gin Senocide F1 0.1 - 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-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 Cyclomedicone 6.00 6.00 Preservative, incense Suitable amount Suitable amount Triethanolamine 0.1 0.1

[Test Example 3] melanin production inhibitory effect using B16 / F10 melanoma cells

B16 / F10 melanoma cells (Korean cell line) at a constant concentration using a sample containing 0.001% by weight of Formulation Example 1 and Comparative Formulation Example 1 and a sample containing kojic acid instead of ginsenoside F1 in Formulation Example 1, respectively. After culturing for 3 days, the culture medium was removed, washed with PBS, the cells were dissolved with 1N NaOH, and the absorbance was measured at 405 nm. The cells without test substance were used as a control group, and the degree of inhibition of melanin production of each test substance was measured by comparing with the melanin content of the control group. Melanin inhibition rate was calculated according to Equation 2 and the results are shown in Table 4.

Test substance Melanogenesis inhibition rate (%) Control group (no addition) 0 Kojic acid 53 Gin Senocide F1 70

From the results of Table 4, it can be seen that the melanin production inhibition rate of the cosmetic composition of Formulation Example 1 containing ginsenoside F1 according to the present invention is much higher than kojic acid, a known melanin production inhibitor, and the whitening effect is very excellent.

Test Example 4 Measurement of Effect of Increased Skin Moisturizing Power

In order to measure the effect of ginsenoside F1 on the increase in skin moisturizing power, Formulation Example 1 and Comparative Formulation Example 1 were used, and evaluated as follows.

Twenty male and female adults in their 40s to 50s, classified as dry skin, were divided into two groups of 10 people for two groups, Formulation 1 and Comparative Formulation Example 1, respectively. Skin moisture meter at constant temperature and constant humidity (24 ° C, 40% relative humidity) before start of application, after 1 week, 2 weeks, 4 weeks after application, and after 2 weeks (total 6 weeks) after application was stopped. Skin moisture content was measured by (Corneometer CM825, C + F1 Electronic Co., Germany). The results are shown in Table 5 below. The results in Table 5 show the percentage increase of the measured value after a certain period of treatment based on the skin moisture meter value measured immediately before the start of the test.

Test group Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Formulation Example 1 33 37 35 33 Comparative Formulation Example 1 30 32 32 15

In the results of Table 5, when the Comparative Formulation Example 1 was applied, it showed a water increase rate of about 30% up to 4 weeks after the application was made, but after stopping the application, the moisture content of the skin decreased, whereas Ginsenoside F1 In the case of application of Formulation Example 1 containing, it can be confirmed that even after the application was stopped, the skin moisture increase rate was more than 30%. Through this, the composition of the present invention containing ginsenoside F1 can be seen that the skin moisturizing effect and excellent.

Test Example 5 Measurement of Keratinocyte Differentiation Promoting Effect

In order to examine the differentiation promoting effect of keratinocytes of ginsenoside F1, the amount of CE (Cornified Envelop) generated during the differentiation of keratinocytes was measured using absorbance.

First, the keratinocytes of the primary cultured human isolated from the epidermis of the newborn were put in a culture flask and attached to the bottom, and then treated with ginsenoside F1 at 5 ppm concentration in the culture medium. Incubate for 5 days until growth. At this time, the low calcium (0.03mM) treated group and the high calcium (1.2mM) treated group were used as negative and positive controls, respectively. Then, the cultured cells were harvested and washed with PBS (Phophate buffered saline), followed by 10 mM Tris-HCl buffer (Tris-HCl, containing 2% SDS (sodium dodecyl sulfate) and 20 mM Dithiothreitol (DTT). pH 7.4) 1 ml was added to sonication, boiling and centrifugation, and the precipitate was suspended in 1 ml of PBS again to measure absorbance at 340 nm. Separately, a portion of the solution after the sonication was taken to measure the protein content and used as a reference when evaluating the degree of cell differentiation. The results are shown in Table 6 below.

Test substance The ability to differentiate in keratinocytes (%) Low calcium (0.03 mM) solution
(Negative control) 100 High calcium (1.2 mM) solution
(Positive control group) 210 Gin Senocide F1 320

As shown in Table 6, when the ginsenoside F1 was treated, it was confirmed that the effect of promoting differentiation of keratinocytes was excellent.

Test Example 6 Measurement of Skin Barrier Function Restoration Effect

In order to measure the effect of ginsenoside F1 on the recovery of damaged skin barrier function due to skin damage, the following experiment was performed. Ten adult men and women's upper arms were damaged by a tape stripping method for 7 days while applying two groups of Formulation Example 2 and Comparative Formulation Example 2, each prepared by the composition of Table 7 below. The recovery of transdermal moisture loss (TWEL) was measured once a day using a Vapometer (Delfin, Finland). Comparative Formulation Example 2 is a vehicle as the negative control group. The experimental results are shown in Table 8 below. The results in Table 8 were compared on the basis of the 100% difference before and after the barrier damage.

Compounding ingredient Formulation Example 2 Comparative Formulation Example 2 Purified water 69 70 Propylene glycol 30 30 Gin Senocide F1 One -

Test group TWEL change (%) Before processing 1 day 2 days 3 days 4 days 5 days 6 days Formulation Example 2 100 124.5 123.1 121.5 119.8 114.6 109.2 Comparative Formulation Example 2 100 121.4 112.7 98.3 70.5 62.3 43.5

As can be seen in Table 8, when treated with Comparative Formulation Example 2 that does not contain ginsenoside F1, the amount of transdermal moisture loss gradually increases over time, while the ginsenoside F1 is rapidly treated. At this rate, the loss of transdermal moisture returns to normal and the barrier damage is recovered.

Test Example 7 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 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.

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 for 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 9 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 15 Comparative Formulation Example 1 5

In the results of Table 9, the cosmetic composition according to the present invention significantly increased the skin blood flow than Comparative Formulation Example 1 containing no ginsenoside F1, 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 F1 according to the present invention can contribute effectively to the skin's nutrient delivery, inhibit skin aging and delay.

[Test Example 8] 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 brightness and color measurement value of the skin and the skin brightness and color change value, the results are shown in Table 10 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 16 ± 3.24 14 ± 2.34 Comparative Formulation Example 1 5 ± 2.34 5 ± 2.05

In the results of Table 10, Comparative Formulation Example 1 containing no ginsenoside F1 according to the present invention did not show significant skin tone improvement, whereas Formulation Example 1 containing ginsenoside F1 as an active ingredient was used more than before use. It was confirmed that a lot of later skin tone is improved.

Test Example 9 Pore Reduction Effect

1. Pore reduction effect through promoting collagen biosynthesis

Ginsenoside F1 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 ng / ml of ginsenoside F1 and TGF-β of the present invention dissolved in serum-free medium and CO ₂ And cultured in an incubator 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 11, and the synthetic ability of collagen was compared with the non-treated group as 100.

Test substance Collagen Synthesis (%) Untreated group 100 TGF-beta 183.5 Gin Senocide F1 198.2

In the results of Table 11, ginsenoside F1 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 F1 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 12 below (rating rating: 0-not scaled down at all; 5-scaled down).

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

From the results of Table 12, 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 F1 according to the present invention reduces the size of the pores It was found that the effect was excellent.

Test Example 10 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 HEF1293-5αR2 cells was measured. HEF1293 cells were transfected with p3 × FLAG-CMV-5αR2 and cultured in a well plate at 2.5 × 10 ⁵ cells per well (ParF1 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, UF1) was used as the substrate of the medium.

To confirm the degree of inhibition of 5α-reductase activity, ginsenoside F1 was added and incubated for 2 hours at 37 ° C. in a 5% CO ₂ incubator. At this time, the ginsenoside F1 was added as a negative control group, the 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 Fiesieselgel 60 F154 was used. ) 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 isotope 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, the conversion and inhibition rates were calculated according to Equations 3 and 4, respectively, and the results are shown in Table 13 below.

Test substance Conversion Rate (%) Inhibition rate (%) Negative control group 48.0 - Positive control group 27.6 42.5 Gin Senocide F1 13.5 62.3

In the results of Table 13, 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 F1 effectively inhibited the activity of blocking the conversion of testosterone to dihydrotestosterone, and was shown to have a superior inhibitory effect than finasteride known to inhibit the activity of 5α-reductase. Thus, it was confirmed that ginsenoside F1 can inhibit 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. 30 men and women who felt that sebum secretion was high were selected and nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were made daily for four weeks in designated areas. Determination of the effect of sebum reduction was measured using the sebum meter (Sebumeter SM810, C + F1 Electronic Co., Germany) and measured the average percentage of sebum reduction after 2 weeks and 4 weeks, respectively, and the results are shown in Table 14 Shown in

Test substance Sebum reduction rate (%) After 2 weeks After 4 weeks Formulation Example 1 47 51 Comparative Formulation Example 1 5 5

From the results of Table 14, it was found that Formulation Example 1 containing ginsenoside F1 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 3 and Comparative Formulation Examples 3 to 4]

Formulation Example 3 and Comparative Formulation Examples 3 to 4 were prepared according to the ingredients and the contents (% by weight) shown in Table 15 below. Specifically, Formulation Example 3 is a combination of ginsenoside F1, Comparative Formulation Example 3 does not contain any active ingredients for acne skin improvement, Comparative Formulation Example 4 is a standard to be used as a reference for the antimicrobial activity As an anti-acne treatment, it contains erythromycin.

The preparation method of Formulation Example 3 and Comparative Formulation Examples 3-4 is as follows. The components of phase A in Table 15 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 15, homogenized, mixed and filtered to prepare the present compositions.

division Formulation Example 3 Comparative Formulation Example 3 Comparative Formulation Example 4 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 Gin Senocide F1 5.0 - - Erythromycin - - 5.0

Test Example 11 Antibacterial Activity Test against Acne Bacteria

Antibacterial activity was tested against propionibacterium acnes (ATCC 6919: Medium-BHI broth), which is an acne-causing strain, with each cosmetic composition prepared in the formulation of Formulation Example 3 and Comparative Formulation Examples 3-4. .

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 3 and Comparative Formulation Examples 3 to 4 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 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 antimicrobial activity test results for acne bacteria are shown in Table 16 below. MIC is expressed in terms of the concentration of the active ingredient contained in the formulation.

Item pH Propionibacterium Acnes Formulation Example 3 5.7 > 30 ppm Comparative Formulation Example 3 5.7 Highest concentration (no antimicrobial activity) Comparative Formulation Example 4 5.7 > 100 ppm

The smaller the ppm concentration in MIC, the more effective the antimicrobial activity against acne bacteria. In the case of Formulation Example 3, the concentration of ppm was significantly lower than that of Comparative Formulation Example 4 using erythromycin, a known acne treatment agent. It can be seen that the composition containing the side F1 has a much better antimicrobial activity against the test bacteria.

Test Example 12 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 F1 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 F1 on adipocyte accumulation in adipocytes. Adipose cells were fixed at room temperature with 4% paraformaldehyde (pH 7.2) in phosphate buffer, washed with PBS (phsphate 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 17.

sample % Inhibition Control group +++ Comparative group + Gin Senocide F1 -

As shown in Table 17, the ginsenoside F1 used in the present invention not only has a small amount of fat accumulated in adipocytes, but also has a superior lipid synthesis inhibitory effect than caffeine, a known lipid synthesis inhibitor. have. Therefore, lipid synthesis is inhibited, and sebum is reduced, so that occurrence of acne can be suppressed.

[Test Example 13] Test for acne improvement, sebum secretion and irritation

Thirty people with acne were divided into three groups of 10 people, and subjects corresponding to each group were allowed to use the cosmetic composition prepared in Formulation Example 3 and Comparative Formulation Examples 3 to 4 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 expressed in the average score of 10 people in Table 18 below.

Acne disappearance was based on the number of days the extinction was read, acne recurrence was based on the results after one month. 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'. Experimental results are shown in the average score of 10 people in Table 18 below. 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 3 4.7 2 days radish 4.5 0/10 Comparative Formulation Example 3 2.1 13th U 2.0 0/10 Comparative Formulation Example 4 4.2 2 days radish 4.1 9/10

As shown in Table 18, Formulation Example 3 did not recur acne compared to Comparative Formulation Example 3, it can be seen that there is an excellent effect on the overall acne improvement. On the other hand, Comparative Formulation Example 4 containing an antimicrobial activity standard, although showing an acne-improving effect, there is a risk of skin irritation for long-term use due to the strong irritation in use, the composition according to the present invention is not irritating appear.

Test Example 14 Inflammation Improvement Effect

1. Inhibitory Effect of Prostaglandin Production

The anti-inflammatory effect was evaluated as the inhibitory effect of the production of prostaglandins. Ginsenoside F1 was used to measure the effect on macrophages. First, aspirin was added to macrophages taken from the abdominal cavity of mice to a final concentration of 500 M, thereby irreversibly inhibiting cyclooxygenase (COX) activity remaining in the cells. Then, 100 μl of the suspension was added to each well of the 96 well cell culture tube and incubated for 2 hours in an incubator at 37 ° C. with 5% CO ₂ to attach macrophages to the container surface. Subsequently, the attached macrophages were washed three times with PBS and then used for the inflammation improvement effect test. After cultured for 12 hours by adding RPMI medium containing 1% (w / v) of LPS to 5 × 10 ⁴ cells / ml of the cultured macrophages, prostaglandin production was induced and 100 μl of ginsenoside F1 was liberated. Prostaglandins were quantified using enzyme immunoassay (ELISA).

At this time, the production inhibitory activity of the prostaglandin of ginsenoside F1 is set to 100% of the difference between the prostaglandin produced in the LPS treated group and the untreated group, and the percentage of prostaglandin decreased by treating the LPS and the sample together. Compared with the control group, and determined, and the results (prostaglandin production inhibitory effect) are shown in Table 19 below.

Blank (BlanF1) 100% Control group (Aspirin treated group) 25.0% Gin Senocide F1 19.2%

As shown in Table 19, it can be seen that even when treated with ginsenoside F1 as a control group treated with aspirin, the effect of inhibiting the production of prostaglandins is very high.

Through this ginsenoside F1 of the present invention it can be seen that it can provide an excellent inflammation improving effect.

It can also be seen that ginsenoside F1 can prevent and improve skin trouble by inhibiting the expression of prostaglandin, a skin inflammatory factor.

2. Inhibitory Effect of IL-8 Production

One day prior to the experiment, normal human sF1in F1eratinocyte (NHEF1, obtained from Lonza) was dispensed in a 96 well plate at 5x10 ⁴ cell number / well, followed by 37 ° C, 5% CO ₂ incubator. Incubated for 24 hours at. After 24 hours, the cells were washed twice with PBS and replaced with serum-free F1Beratinocyte basement media (F1BM). Each well was treated with ginsenoside F1 according to the concentrations shown in Table 20 for 30 minutes, followed by PGSA (10 µg / ml), PGSA (50 µg / ml), and PGSA (50 µg / ml) + LPS (1). [Mu] g / ml) was treated respectively. Here, PGSA (peptidoglycan from S. aureus ) is a peptidoglycan (peptidoglycan) extracted from staphylococcus aureus, a major component of the Gram-positive (+) cell wall and bacterial membrane components are known to cause inflammation, In particular, in staphylococci, about 90% of patients with atopic dermatitis have been reported to cause secondary infection. Lypopolysaccaride (LPS) is a major component of the cell membrane of Gram-negative bacteria and is known to be a major cause of inflammation.

After incubation for 24 hours at 37 ℃, 5% CO ₂ incubator, the culture medium was taken and subjected to ELISA for Interleukin-8 (InterleuF1in-8, IL-8), the results are shown in Table 20 below. ELISA used the experimental method of the manufacturer (BD science).

division IL-8 secretion (pg / ml) Untreated Control 935.12 PGSA (10 μg / ml) 4812.60 PGSA (50 μg / ml) 5895.08 PGSA (50 μg / ml) + LPS (1 μg / ml) 6814.91 Ginsenoside F1 (5 ppm) 1034.92 Ginsenoside F1 (25 ppm) 1189.03 Ginsenoside F1 (50 ppm) 1302.84

In Table 20, ginsenoside F1 was found to significantly reduce and inhibit the secretion of IL-8 increased by PGSA and LPS. Therefore, it can be seen that the topical skin composition of the present invention can provide an excellent anti-inflammatory effect by significantly reducing the secretion of IL-8 increased by PGSA and LPS.

[Test Example 15] Itching relief evaluation

One day prior to the experiment, keratinocytes (cell name: HaCaT obtained from ATCC) were dispensed into 96 well plates at ⁴ × 10 ⁴ cells / well, followed by 24 hours in a 37 ° C., 5% CO ₂ incubator. Incubated. After 24 hours, wash 96 well plates twice with HanF1s'Balanced Salt solution (HBSS) buffer, and then add reaction buffer (2μΜ Fluo-4-AM, 20% pluronic acid, 2.5mM probenecid) to the cells. gave. After reaction at 37 ° C., 5% CO ₂ incubator for 30 minutes and room temperature for 30 minutes, the cells were washed twice with HBSS buffer and treated with ginsenoside F1 at the concentrations (%) shown in Table 21 below.

After reaction for 10 minutes to process the 2U / ml trypsin (Trypsin) or 5μM PAR-2 activation peptide (SLIGF1V) and was measured in Ca ^{2 +} concentration in the cell 80 seconds. In Ca ^{2 +} concentration in the measurement cell presentation flex 3: it was used (FlexStation3 Molecular Device, USA). After the ginsenoside F1 and 2U / ml Trypsin or 5μM PAR-2 active peptide (SLIGF1V) treatment, the flex was measured for 80 seconds to determine the difference between the minimum and maximum values. Inhibition rate (%) for the cellular influx of calcium ions compared to the difference between the minimum and maximum values when treated with 2U / ml trypsin or 5μM PAR-2 active peptide (SLIGF1V) is shown in Table 21 below.

Ginsenoside F1 Concentration % Inhibition of intracellular influx of calcium ions Trypsin (2U / ml) PAR-2 active peptide (5 [mu] M) 0.05% 25 29 0.1% 33 38 0.5% 39 43 1.0% 47 52

As can be seen in Table 21, the influx of intracellular calcium ions by trypsin or PAR-2 active peptide (SLIGF1V) is reduced by the treatment of ginsenoside F1, and the concentration of ginsenoside F1 is increased. It was confirmed that the influx of calcium ions was significantly reduced.

Therefore, the external preparation composition for skin of the present invention can provide an excellent antipruritic effect by effectively inhibiting PAR-2 activity causing itching.

Formulation Example 4 and Comparative Formulation Example 5

To prepare a shampoo in the composition of Table 22. 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 fragrance, and a hair conditioner were added and mixed, followed by cooling to room temperature under stirring.

ingredient Formulation Example 4 Comparative Formulation Example 5 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 Gin Senocide F1 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 Example 16 Anti-dandruff force evaluation

The antidandruff force of Formulation Example 4 and Comparative Formulation Example 5 were measured and compared. At this time, the test strain is dandruff Pytirosporum Ovale ( Pityrosporum) Obalae (ATCC 12078)).

Antibacterial activity was measured by the skin disk diffusion method. The sF1in disc diffusion method (SDDM) is a method that is closest to a consumer's habit of use, and is a method of measuring antibacterial activity on guinea pig skin similar to human skin.

Specifically, after peeling off guinea pig skin (guinea pig sF1in) by treating with 70% ethanol to dry uniformly, a cut skin disk was used. The sterilized skin discs were soaked in dilute shampoo solution for 3 minutes and then washed with running water. Then, the skin disk was placed on a solid medium inoculated with the test bacteria and then cultured. After incubation, the relative antimicrobial activity was measured by measuring the size of the clear zone in which the growth of bacteria was suppressed. The experimental results are shown in Table 23 below as average values of the results of three measurements.

Anti-dandruff division Inhibitor size (mm) Formulation Example 4 Comparative Formulation Example 5 Dandruff inhibitor ring size 3.1 0

Looking at the Table 23, in the case of Comparative Formulation Example 5 that does not contain ginsenoside F1, there is no effect of reducing dandruff. In the case of Formulation Example 4 containing ginsenoside F1 effectively reducing dandruff, it can be confirmed that there is an excellent anti-dandruff effect.

Test Example 17 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 4 and Comparative Formulation Example 5 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 Examples 4 and Comparative Formulation Example 5, 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 5 below and the results are shown in Table 24 below.

Formulation Example 4 Comparative Formulation Example 5 Dandruff reduction rate (%) 56.2
63.1
55.9
72.3
69.7
53.2
66.7
51.0
68.5
46.9
72.6
64.8 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 61.7 3.4 SD 8.4 2.1

As can be seen in Table 24, it can be seen that in the case of Formulation Example 4 containing ginsenoside F1 exhibits an excellent anti-dandruff effect.

Test Example 18 Scalp Itching Prevention Test

Twenty-four men and women aged 25 to 45 who feel severely itching scalp were divided into two groups of 12 people, each of whom used the shampoos of Formulation Example 4 and Comparative Formulation Example 5 once every three days for two weeks. The prevention effect was evaluated through the following evaluation criteria and the results are shown in Table 25 below.

[Evaluation standard]

Very good-5 points

Excellent-4 points

Normal-3 points

Poor-2 points

Very bad-1 point

division Formulation Example 4 Comparative Formulation Example 5 Itching effect of scalp 4.1 2.3

As can be seen in Table 25, it can be seen that the case of Formulation Example 4 containing ginsenoside F1 exhibits an effect of preventing better scalp itching.

[Test Example 19] hair follicle dermal papilla cell proliferation effect

The keratin proteins that make up hair are produced in hair root keratinocytes (F1eratinocytes), which are differentiated from dermal papilla cells. In order to evaluate the activity of the dermal papilla cells of the composition, a rat immortalized dermal papilla cell (DP6) cell line was used in the present invention (Wendy Filsell, Journal of Cell Science 107, 1761-1772 (1994)). This dermal papilla cell line was cultured by microdissection from the hair roots of male PVG rat whiskers and cultured with DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, MD, USA) was incubated for 24 hours in an incubator maintained at 5% CO ₂ , 37 ℃. DP6 was placed in a 96 well plate and incubated in a 37 ° C. incubator for 24 hours and then treated with Ginsenoside F1 at concentrations of 5 ppm, 10 ppm and 20 ppm, respectively. After 24 hours of drug treatment, cell proliferation was measured using the WST-1 kit (Roche). The results are shown in Table 26 below.

division Cell proliferation (%) Untreated Control 100 Ginsenoside F1 (5 ppm) 119 Ginsenoside F1 (10 ppm) 125 Ginsenoside F1 (20 ppm) 148

As can be seen in Table 26, the treatment of ginsenoside F1 increases the growth of dermal papilla cells, which can be confirmed to increase significantly in a concentration-dependent manner.

Test Example 20 Evaluation of Potassium Ion Channel Activity Increase Effect

Hair loss treatment of minoxidil is known as mitochondrial potential potassium ion channel openers (F1 _ATP channel opener), a representative drug used in the treatment of androgenetic alopecia. In order to evaluate the mechanism of minoxidil, the treatment of tollamide (SIGMA AlDRICH, T0891), which blocks the F1 _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 an F1 _ATP channel opener, a mouse embryonic fibroblast cell line (NIH3T3) cell line was used in the present invention. 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 F1 at concentrations of 2.5 ppm, 5 ppm and 10 ppm, respectively. After 48 hours of drug treatment, cell proliferation was measured using the WST-1 kit (Roche). The results are shown in Table 27 below.

division Cell proliferation (%) Untreated Control 100 Minoxidil 132 Ginsenoside F1 (2.5 ppm) 115 Ginsenoside F1 (5 ppm) 121 Ginsenoside F1 (10 ppm) 127

As can be seen in Table 27, when the ginsenoside F1 is treated, the proliferation of fibroblasts is recovered, and the cell proliferation ability is increased depending on the concentration of the treated ginsenoside F1. When treated with 10ppm it can be confirmed that the proliferation of cells is restored to the level when the minoxidil treatment.

Formulation Example 5 and Comparative Formulation Example 6

To the cream-based composition was prepared in a conventional manner according to the composition of Table 28 (unit: wt%).

Compounding ingredient Formulation Example 5 Comparative Formulation Example 6 Labrafac 23 23 Tween 80 10.5 10.5 Glycerol monostearate 4.0 4.0 Stearic acid 7.0 7.0 Cetyl alcohol 2.0 2.0 Propylene glycol 9.0 9.0 Glycerol Monooleate 3.5 3.5 Gin Senocide F1 One - water 40 41

Test Example 21 Hair Growth Effect of Ginsenoside F1

After the hair on the back of the ICR mouse was depilated with a hair remover, hair was completely removed using a hair removal cream (beet cream). The skin inflammation was induced for 3 days by applying 200ul of the skin to 1% DNCB once a day for the other 2 groups except the control group (normal group). After 3 days, the cream base of Formulation Example 5 and Comparative Formulation Example 6 was applied to the group except the control group once a day, and the degree of hair growth of each group was observed, and the results are shown in FIG. 1.

Referring to FIG. 1, the control group did not show the hair growth phenomenon in the depilated portion during the observation for 15 days, but showed a slight hair growth effect in the group treated only with the cream base containing no ginsenoside F1, and included the ginsenoside F1. In the cream-treated group, it can be seen that there is a significant hair growth effect in the entire depilated area.

Test Example 22 Wool Effect of Ginsenoside F1

By treating the hair of the back hair by the method of Test Example 21 by treating dinitrochlorobenzene (DNCB) to induce skin irritation by setting the stress conditions in the skin to maximize the growth of the hair, the present invention In order to evaluate the hair regeneration efficacy of the composition, the cream of Formulation Example 5 and Comparative Formulation Example 6 containing ginsenoside F1 was treated, and the length of the hair with the number of treatment days was measured and compared with the control group. The results are shown in Table 29 below.

Days of Treatment (Days) Hair length (cm) Formulation Example 5 Treatment Comparative Formulation 6 Treatment 0 0.1 0.1 6 0.2 0.1 9 0.5 0.1 12 1.1 0.1

Referring to Table 29, the hair length of the group treated with Formulation Example 5 showed a statistically significant difference (p <0.01) compared to the group treated with Comparative Formulation Example 6, and the length increased according to the number of treatment days. On the 12th day, the hair length of the group treated with the cream base of Formulation Example 5 was longer than the group treated with the cream base of Comparative Example 6 containing no ginsenoside F1. It can be confirmed.

It is determined that ginsenoside F1 promotes hair regeneration even under the condition of hair regeneration that is formed under skin stress, and it can be seen that ginsenoside F1 has a regeneration function for hair that has been lost under stress.

Test Example 23 Melanin Promoting Effect Test of Ginsenoside F1

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 F1 at a final concentration of 10, 50 ppm as a test substance, 0.1% DMSO as a negative control, 100μM IBMX as a positive control, and then incubated 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 F1 with the control is shown in Table 30 below.

sample Melanin Synthesis (%) DMSO (0.1%) 100 IBMX (100 μM) 120 Ginsenoside F1 (10 ppm) 112 Ginsenoside F1 (50 ppm) 134

In Table 30, ginsenoside F1 promotes melanin synthesis of melanocytes, which increases melanin production, and shows an excellent melanin production promoting effect.

[Test Example 24] Effect of Ginsenoside F1 on the Expression of MITF and Tyrosinase in Melanosite

Dispense 500,000 cells / well in a 6-well microtiter plate using a 501mel cell line, DMSO 0.1% for the negative control, IBMX 100μM for the positive control, and ginsenosides for the test group. The F1 was treated with 10 ppm and 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 31 below by comparing the negative control to 100.

MITF Tyrosinase IBMX Gin Senocide F1 IBMX Gin Senocide F1 24hr 121 98 160 146 48hr 98 110 149 150 72hr 224 118 298 188

Looking at Table 31, it can be seen that ginsenoside F1 increases the expression of MITF and tyrosinase protein in melanocytes.

[Test Example 25] Evaluate the effect of ginsenoside F1 on white hair and induce black hair in vivo using leukemia-promoting vitiligo mice

Vitiligo mice (C57bl / 6- Mitf ^{mi - vit} ) were purchased from The JacF1son Lab in the United States. Experimental method for the effect of white hair prevention material using the mouse to promote the development of white hair is as follows. Dorsal hairs of 12 week old mice were removed by depilation. However, the area of the hair removal site was adjusted in the same manner for each individual. From the day after the hair was removed, the anti-hair white matter was applied to the area where the hair was removed twice a day. As a vehicle for the anti-harvest material, a mixture of EtOH: 1,3-BG: DW = 3: 2: 5 (volume ratio) was used, and the carrier was used as a negative control, and the solution added with IBMX 50 mM was positive. As a control group, a solution containing 2.5% of ginsenoside F1 was used as a test group. After about three weeks, when the difference in anti-white hair efficacy between each substance was visually identified, newly grown hairs were collected and the amount of melanin in the hair was measured. The amount of melanin in the hair was measured using a proteinase enzyme, Esperase (Novozyme). A reaction buffer was prepared by dissolving esperase at a concentration of 1 NPU / ml in a buffer (50 mM Tris-HCl, 5 mM DTT, pH 9.3). 5 mg of mouse hair was added to 1 ml of the reaction buffer, followed by reaction for 13 hours while shaking at a speed of 1,000 rpm at 37 ° C., and the hair and the reaction solution were separated by instant centrifugation. The reaction solution thus obtained was placed in a 96 well plate, and the absorbance at 405 nm was measured to determine the amount of melanin in the reaction solution. In the case of treatment with a negative control group, a positive control group, and a test group material in a leukemia-promoted vitiligo mouse model, the results of visual observation and measurement of melanin quantification in hair are shown in Table 32. Is 100 and compared with this value.

Negative control group IMBX Gin Senocide F1 % Against control 100 105.9 109.6

According to Table 32, it can be seen that ginsenoside F1 can inhibit the white hair and increase the amount of melanin in the hair to promote the induction of black hair in the mouse in vivo which promoted the development of white hair.

Test Example 26 Evaluation of Antibacterial Activity of Ginsenoside F1

Antimicrobial experiments were conducted to evaluate the antimicrobial activity of ginsenoside F1. Specific experimental methods are 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 the sample 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. 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, 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 growth was checked by suspension and microscope to determine the minimum inhibitory concentration (MIC) value. The results are shown in Table 33 below.

Sample Minimum Inhibitory Concentration (MIC),% Sedonas
Aruginosa Staphylococcus aureus Escherichia Coli Candida Albicans Aspergillus Niger Gin Senocide F1 0.1 0.06 0.122 > 4 > 1

As shown in Table 33, it can be seen that ginsenoside F1 exhibits antimicrobial activity against various strains, through which it can be predicted that ginsenoside F1 can act as a natural preservative or antimicrobial agent in the composition.

Claims (20)

The external preparation composition for skin containing ginsenoside F1 represented by the following formula (1) as an active ingredient.
[Formula 1]

According to claim 1, wherein the ginsenoside F1 is an external composition for skin, characterized in that contained in an amount of 0.001 to 50% by weight relative to the total weight of the composition. The composition for applying the external of the skin according to claim 1, wherein the composition is for whitening. The composition for applying the external of the skin according to claim 1, wherein the composition is for moisturizing. According to claim 1, wherein the composition is an external composition for skin, characterized in that for enhancing the skin barrier function. According to claim 1, wherein the composition is an external composition for skin, characterized in that for inducing differentiation of keratinocytes of the skin. According to claim 1, wherein the composition is an external composition for skin, characterized in that for improving acne. The composition for applying the external of the skin according to claim 1, wherein the composition is for antibacterial use. The composition for external application of skin according to claim 1, wherein the composition is for anti-inflammatory. The composition for applying the external of the skin according to claim 1, wherein the composition inhibits lipid formation. The external preparation of the skin of claim 1, wherein the composition is for improving atopic dermatitis. According to claim 1, wherein the composition is an external composition for skin, characterized in that for improving the color tone and skin tone. The external preparation composition for skin according to claim 1, wherein the composition is for shrinking pores. The external preparation composition for skin according to claim 1, wherein the composition is for controlling sebum. The external preparation composition for skin according to claim 1, wherein the composition is for improving skin troubles. The external preparation composition for skin according to claim 1, wherein the composition is for defense against skin irritation. According to claim 1, wherein the composition is an external topical composition, characterized in that for preventing dandruff. According to claim 1, wherein the composition for external application to the skin, characterized in that for hair growth. According to claim 1, wherein the composition for external application to the skin, characterized in that for preventing white hair. The external preparation composition for skin according to claim 1, wherein the ginsenoside F1 is used as a natural preservative.

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