KR101939112B1 - Composition of skin external application containing ginsenoside F1 - Google Patents

Abstract

The present invention relates to a composition for external application for skin containing ginsenoside F1. More specifically, the present invention relates to a composition for external application for skin comprising ginsenoside F1, which has excellent antioxidative power by containing ginsenoside F1, It is possible to provide an effect of improving the scalp and hair condition such as dandruff prevention effect, hair growth effect and anti-whiplash effect as well as overall skin condition improvement effect such as improvement effect, whitening effect, sebum control effect, pore contraction effect, ≪ / RTI >

Description

Composition of skin external application containing ginsenoside F1 "

The present invention relates to a composition for external application for skin containing ginsenoside F1. More specifically, the present invention relates to a composition for external application for skin comprising ginsenoside F1, which has excellent antioxidative power by containing ginsenoside F1, It is possible to provide an effect of improving the scalp and hair condition such as dandruff prevention effect, hair growth effect and anti-whiplash effect as well as overall skin condition improvement effect such as improvement effect, whitening effect, sebum control effect, pore contraction effect, ≪ / RTI >

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

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

Therefore, the present inventors have found that ginsenoside F1 can provide not only whitening and moisturizing effect but also improvement effect of acne, skin trouble and atopic symptoms, and can improve the skin condition such as skin color improvement effect, sebum control and pore shrinkage effect And it is also possible to provide an effect of improving scalp and hair condition such as prevention of dandruff, hair growth and hair loss, and completed the present invention.

Accordingly, an object of the present invention is to provide a composition for external application for skin, which can contain the ginsenoside F1 to improve the overall condition of the skin.

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

In addition, the present invention provides a whitening skin external composition composition containing ginsenoside F1 as an active ingredient.

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

Further, the present invention provides a composition for external application for skin for acne improvement comprising ginsenoside F1 as an active ingredient.

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

The present invention also provides a composition for external application for skin color and skin tone, which contains ginsenoside F1 as an active ingredient.

The present invention also provides a composition for external application for skin pore reduction comprising ginsenoside F1 as an active ingredient.

In addition, the present invention provides a composition for external application for skin sebum control comprising ginsenoside F1 as an active ingredient.

Further, the present invention provides a composition for external application for skin for preventing dandruff comprising 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.

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

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

The composition of the present invention has excellent antioxidative power by containing ginsenoside F1, and it has an excellent antioxidant ability, and also has a skin moisturizing effect, anti-inflammatory effect, skin troubles such as acne and atopy, whitening effect, sebum control effect, It is possible to provide an effect of improving the scalp and hair condition such as dandruff prevention effect, hair growth effect and anti-whiplash effect as well as overall skin condition improvement effect such as improvement of complexion through skin.

Fig. 1 shows the hair growth effect after applying Formulation Example 5 and Comparative Formulation Example 6. Fig.

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

The ginsenoside F1 used in the present invention has a structure represented by the following formula (1).

The ginsenoside F1 of the present invention may be extracted from a plant, synthesized according to a method known in the art, or commercially available one may be used. Also, ginsenoside F1 can be obtained from ginseng extract. The type of ginseng to be used at this time is not particularly limited, and ginseng, red ginseng, white ginseng, taegeuk ginseng and ginseng can be used. The ginseng extract is not only contained in the leached liquid obtained by leaching and transferring from the ginseng but also contained in the concentrate obtained by partially or completely concentrating the leach solution or in the stagnant, premix, seasoned, fluid extract and mulberry produced by drying the concentrate again It contains all of the plant itself as well as chemicals that exert the main effect. Extracts from all parts of ginseng such as stem, root, leaf, flower, and fruit are available and are not limited to any particular part of the extract. In addition, a known method can be used for extracting ginsenoside F1 from ginseng extract.

Specifically, the ginsenoside F1 can be isolated from ginseng after preparing ginseng extract from water or an organic solvent by a method well known in the art. The organic solvent used in the present invention may be selected from the group consisting of ethanol, methanol, butanol, ether, ethyl acetate, chloroform and a mixed solvent of these organic solvents and water, preferably 80% ethanol. At this time, the extraction temperature is preferably 10 to 80 ° C, and the extraction can be performed for 3 to 24 hours. If the extraction temperature and the extraction time are exceeded, the extraction efficiency may be deteriorated or the component may be changed.

The composition 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 ginsenoside F1 is less than 0.001% by weight, the effect and effect of the component are insufficient. If the content is more than 50% by weight, skin safety or shape problems may occur.

Since ginsenoside F1 is a component having excellent antioxidant ability, 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 whitening composition, which can provide an excellent whitening effect by inhibiting tyrosinase activity and inhibiting the production of melanin.

The composition of the present invention can be used as a composition for external application for moisturizing skin, which can enhance the skin barrier function and induce differentiation of dermal keratinocytes. Therefore, it can be effectively used as a composition for external application for skin, which prevents or improves skin dryness, atopic dermatitis, contact dermatitis, psoriasis or the like caused by incompleteness of epidermal differentiation.

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

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

The composition of the present invention can be used as a composition for external application for skin for reducing pores, controlling sebum, and improving skin troubles. It reduces sebum secreted by the skin when applied to the skin, reduces active oxygen and promotes collagen synthesis, , And suppression of skin troubles due to decreased expression of inflammatory factors. In addition, excellent antioxidant power can prevent the generation of skin irritation.

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

The composition of the present invention can be used as a composition for external application for hair growth, which promotes the transition of the resting hair cycle to the growing hair cycle, thereby promoting hair growth and preventing hair loss.

The composition of the present invention can be used as a composition for external application for skin preventing anti-whiplash, which can significantly increase the expression of MITF in melanocytes, thereby inhibiting white moth and promoting gray hair induction.

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

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

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

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

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

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

[Referential Example 1]

The ginsenoside F1 for testing the efficacy of the composition of the present invention was purchased from Ambo Laboratories.

[Test Example 1] Inhibitory effect on reactive oxygen species (ROS) production

The keratinocytes isolated from human epidermal tissue (F1 eratinocyte) were added to each well of a 24-well plate at 5 x 10 ⁴ for 24 hours. After 16 hours, the ginsenoside F1 was treated with 1%. For comparison, the control group did not treat ginsenoside F1. After 2 hours, the culture solution was removed, and 100 μl of phosphate buffered saline (PBS) was added to each well. This keratinocyte was irradiated with ultraviolet rays of 30 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UVB15W, SanF1yo DennF1i, Japan), then PBS was removed, ≪ / RTI > The ginsenoside F1 was again treated and the amount of active oxygen species increased by ultraviolet stimulation was determined at certain time intervals. The amount of ROS was determined by referring to Tan's method of measuring the fluorescence of DCF-DA (dichlorofluorescin diacetate) oxidized by ROS (Tan et al., 1998, J. Cell Biol. Vol. 141, pp1423-1432) The ratios of the vehicle-treated control to the ROS were calculated. The results are shown in Table 1 below.

Test substance UVB 30 mJ / cm ² Elapsed time after irradiation 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, the ginsenoside F1 according to the present invention effectively inhibits the production of ROS which is known to cause skin cell damage by ultraviolet rays, and the amount of ROS after ultraviolet stimulation is almost the same level And the antioxidant effect is excellent enough to inhibit the production of ROS. Therefore, it was confirmed that the ginsenoside F1 according to the present invention can prevent the wrinkling of pores by inhibiting oxidation and preventing aging, and can prevent skin irritation, thereby improving skin troubles.

Test Example 2: Effect of tyrosinase inhibition

The tyrosinase enzyme was extracted from Mushroom and was obtained from Sigma (SIGMA). First, the substrate, tyrosine, is dissolved in distilled water to make a 0.3 mg / ml solution. The solution is put into a test tube in an amount of 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 .

0.2 ml of the sample solution prepared by mixing the ginsenoside F1 of the present invention at an appropriate concentration into the ethanol solution was added to the reaction solution, and the mixture was allowed to react in the thermostatic chamber at 37 ° C for 10 minutes. At this time, the control group was prepared by adding 0.2 ml of a solvent instead of each sample solution, and ascorbic acid was used as a positive control group. 0.1 ml of tyrosinase solution of 2500 units / ml was added to the reaction solution, followed by reaction at 37 ° C for 10 minutes. The reaction tube was immersed in ice water to quench the reaction, and absorbance at a wavelength of 475 nm was measured with a photoelectron spectrometer. The results are shown in Table 2 below. Each tyrosinase inhibitory effect was calculated by the following equation (1).

Test substance Inhibition rate of tyrosinase (%) Control group (no addition) 0 Ascorbic acid 52 Gin Senocide F1 71

From the results shown in Table 2, it can be seen that the cosmetic composition of Formulation Example 1 containing ginsenoside F1 according to the present invention has a much higher inhibitory effect on tyrosinase than ascorbic acid, which is a known tyrosinase inhibitor, and thus has excellent whitening effect.

[Formulation Example 1 and Comparative Formulation Example 1]

Nutritive creams were prepared in a conventional manner according to the composition of Table 3 below (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-10pentastearate and behenyl alcohol and sodium stearoyl lactylate 1.00 1.00 Arachidyl behenyl alcohol and arachidyl glucoside 1.00 1.00 Cetylaryl Alcohol and Cetearyl Glucoside 2.00 2.00 PEG-100 Stearate & Glycerololate & Propylene Glycol 1.50 1.50 Caprylic / carboxy triglyceride 11.00 11.00 Cyclomedicone 6.00 6.00 Preservative, incense Suitable amount Suitable amount Triethanolamine 0.1 0.1

[Test Example 3] Melanin production inhibitory effect using B16 / F10 melanoma cells A sample containing 0.001 wt% of each of Formulation Example 1 and Comparative Formulation Example 1 and a sample containing kojiic acid instead of ginsenoside F1 in Formulation Example 1 Was added to the culture medium of B16 / F10 melanoma cells (Korean Cell Line Bank) at a constant concentration for 3 days, and the culture solution was removed, washed with PBS, and the cells were dissolved with 1N NaOH and absorbance was measured at 405 nm. Cells to which the test substance was not added were used as a control group, and the melanin formation inhibition degree of each test substance was measured by comparing with the melanin content in the control group. The inhibition rate of melanin production was calculated according to Equation (2), and the results are shown in Table 4.

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

From the results shown in Table 4, it can be seen that the cosmetic composition of Formulation Example 1 containing ginsenoside F1 according to the present invention has a much higher inhibitory effect on melanin formation than kojic acid, which is a known melanin-producing inhibitor, and thus has excellent whitening effect.

[Test Example 4] Measurement of skin moisturizing effect

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

20 men and women in the 40s and 50s classified as dry skin were divided into two groups of 10 persons for each of Formulation Example 1 and Comparative Formulation Example 1 to apply the nutritional cream twice daily for 4 weeks to the face. (24 ° C, relative humidity 40%) after 1 week, 2 weeks, and 4 weeks after application, and after 2 weeks (total 6 weeks) (Corneometer CM825, C + F1 Electronic Co., Germany). The results are shown in Table 5 below. The results in Table 5 are based on the skin moisture meter measured just before the start of the test, and the increase in the measured value after a certain period of treatment is expressed as a percentage.

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

The results of Table 5 indicate that when Comparative Formulation Example 1 was applied, the moisture increase rate was about 30% until 4 weeks of application, but the amount of skin water decreased after stopping application, while the ginsenoside F1 It can be confirmed that the skin moisture increase rate is mostly 30% or more even after the application is stopped. Thus, it can be seen that the composition of the present invention containing ginsenoside F1 has excellent skin moisturizing effect.

[Test Example 5] Measurement of promoting effect of keratinocyte differentiation

In order to examine the differentiation promoting effect of kinsenoside F1 on keratinocytes, the amount of CE (cornified envelope) produced upon differentiation of keratinocytes was measured by absorbance as described below.

First, keratinocytes isolated from the epidermis of newborn infants were adhered to the bottom by placing them in a culture flask, and the cells were treated with 5 ppm of ginsenoside F1 at a concentration of 70 to 80% The cells were cultured for 5 days until they were grown. At this time, low calcium (0.03 mM) and high calcium (1.2 mM) treatment groups were negative control and positive control, respectively. Then, the cultured cells were harvested, washed with PBS (Phosphate buffered saline), and then suspended in 10 mM Tris-HCl buffer (Tris-HCl, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiothreitol) pH 7.4) was added, sonication, boiling, and centrifugation. The precipitate was suspended again in 1 ml of PBS and the absorbance at 340 nm was measured. Separately, a part of the solution after the sonication was taken to measure the protein content and used as a standard in 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 ginsenoside F1 was treated, it was confirmed that the effect of promoting differentiation of keratinocytes was excellent.

[Test Example 6] Measurement of recovery effect on skin barrier function

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 conducted. Ten adult male and female adult rats were injured on the skin barrier using a tape stripping method, and were applied for 7 days while applying the two groups of Formulation Example 2 and Comparative Formulation Example 2 prepared in the composition shown in Table 7 below The recovery rate of TWEL was measured and compared with Vapometer (Delfin, Finland) once a day. Here, Comparative Formulation Example 2 is a vehicle as a negative control. The experimental results are shown in Table 8 below. The results in Table 8 were compared before treatment before barrier damage and after treatment before barrier damage on the basis of 100%.

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 from the above Table 8, when Comparative Formulation Example 2 containing no ginsenoside F1 was treated, the transdermal water loss was gradually increased over time, whereas when the ginsenoside F1 was treated, The rate of transdermal water loss is returned to normal and the barrier damage is recovered.

[Test Example 7]

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

The face was soaked in a constant temperature and humidity chamber, and then adapted for 30 minutes. Initial values were measured using LDPI. First, the initial blood flow in the lower part of the forehead of 30 women with cold hands and feet was measured by LDPI. Table 9 below shows the result of comparing the blood flow measured with the initial measurement value (skin blood flow change) measured after using Formulation Example 1 and Comparative Formulation Example 1 for one week to the subject.

LDPI results before and after using cosmetics - skin blood flow Test substance Percent change of skin blood flow after 1 week use (%) Formulation Example 1 15 Comparative Formulation Example 1 5

From the results shown in Table 9, it was confirmed that the cosmetic composition according to the present invention significantly increased skin blood flow compared to Comparative Formulation Example 1 which did not contain ginsenoside F1, and that blood circulation was improved by promoting blood circulation . This ultimately suggests that the cosmetic composition containing the ginsenoside F1 according to the present invention can contribute to effectively transmit nutrients of the skin and inhibit and retard skin aging.

[Test 8] Effect of improving skin tone

In order to examine the skin tone improvement effect of Formulation Example 1 and Comparative Formulation Example 1, 30 subjects were used (one evening / one day application for a total of one week) and then facial stage DM-3 (Moritex, Japan) To evaluate the degree of skin tone improvement. The improvement rate of the skin tone was determined by the brightness and the color change value of the skin and the brightness and the color change value of the skin. The results are shown in Table 10 below. The larger the change in brightness and color, the better the skin tone.

Test substance Skin tone improvement rate (%) Brightness (mean ± standard deviation) Color (mean ± standard deviation) Formulation Example 1 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 which does not contain ginsenoside F1 according to the present invention shows no significant skin tone-improving effect, whereas Formulation Example 1 containing ginsenoside F1 as an active ingredient is used It was confirmed that the skin tone after the treatment was greatly improved.

[Test Example 9] Pore reduction effect

1. Collagen biosynthesis promotes pore reduction effect

The effect of promoting collagen biosynthesis by ginsenoside F1 according to the present invention was measured in comparison with TGF-β. First, fibroblasts were seeded at 10 ⁵ per well in 24 wells and cultured until they grew to about 90%. The cells were cultured in serum-free DMEM medium for 24 hours, and then treated with 10 ng / ml of each of ginsenoside F1 and TGF-β of the present invention dissolved in serum-free medium for 24 hours in a CO ₂ incubator. The supernatant was removed and the procolagen (I) ELISA kit (procollagen type (I)) was used to determine whether procollagen was increased or decreased. The results are shown in Table 11, and the combined performance of the collagen was set to 100 in the non-treatment group.

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

From the results of Table 11, it was confirmed that the ginsenoside F1 according to the present invention exhibits a superior collagen combining performance higher than that of the positive control group TGF-β. Therefore, it was confirmed that the ginsenoside F1 according to the present invention can increase collagen production around the pores, thereby reducing the enlarged pores.

2. Pore reduction effect

In order to examine the pore reduction effect of Formulation Example 1 and Comparative Formulation Example 1, the following evaluation was made. Twenty male and female subjects with large pore sizes were selected and divided into two groups of 10 persons. The nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were applied to the faces for each group for 4 weeks each day. The evaluation of the effectiveness of the pore reduction was made by a visual evaluation of experts by taking pictures before and after the experiment. The results are shown in Table 12 below (rating scale: 0 - no reduction at all; 5 - very shrunk).

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

From the results of Table 12, Comparative Formulation Example 1 shows no effect of reducing pores, but in the case of Formulation Example 1, the effect of reducing pores is visible to the naked eye. Ginsenoside F1 according to the present invention reduces pore size And the effect was excellent.

Test Example 10 Effect of suppressing sebum secretion

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

The ratio of [ ¹⁴ C] testosterone to [ ¹⁴ C] dihydrotestosterone (DHT) in HEF1293-5αR2 cells was measured to confirm 5α-reductase inhibitory activity. HEF1293 cells were transfected with p3 x FLAG-CMV-5αR2 and cultured in 2.5 × 10 ⁵ cells per well in a 24-well plate (ParF1 et al., 2003, JDS Vol. 31, pp. 191-98). The next day, the medium was changed to a new medium supplemented with an enzyme substrate and an inhibitor. 0.05 [mu] Ci [ ^14C ] testosterone (Amersham Pharmacia biotech, UF1) was used as the substrate of the medium.

To confirm the inhibition of 5α-reductase activity, ginsenoside F1 was added and cultured in a 5% CO ₂ incubator at 37 ° C. for 2 hours. At this time, no ginsenoside F1 was used as a negative control and finasteride was used as a positive control. Thereafter, the culture medium was collected, and the steroid was extracted with 800 쨉 l of ethyl acetate. The upper organic solvent layer was separated, dried, and the remaining residue was again dissolved in 50 쨉 l of ethyl acetate to obtain a silica plastic sheet F1ieselgel 60 F154 ) Using ethyl acetate-hexane (1: 1) as the solvent.

After drying the plastic sample in air, the bath system was used to measure the amount of isotope. The dried plastic sheet and x-ray film were put together in a bath cassette, and after 1 week, the testosterone remaining in the film and isotopes of dihydrotestosterone The conversion and inhibition rates were calculated according to the following equations (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 shown in Table 13, testosterone is converted into dihydrotestosterone, which binds to the receptor protein in the cytoplasm to enter the nucleus, activates the sebaceous gland, accelerates differentiation, and induces 5α-reductase Was effectively inhibited by ginsenoside F1, thereby blocking the conversion of testosterone to dihydrotestosterone. It showed an inhibitory effect superior to that of finasteride, which is known to inhibit 5α-reductase activity. Thus, it was confirmed that the ginsenoside F1 effectively inhibited the activity of 5? -Reductase, thereby suppressing sebaceous hyperpermeability.

2. Inhibitory effect of sebum secretion

To evaluate the inhibitory effect of sebum secretion of Formulation Example 1 and Comparative Formulation Example 1, the following evaluation was made. 30 men and women who felt that sebum secretion was high were selected and nutritional creams of Formulation Example 1 and Comparative Formulation Example 1 were made daily for four weeks in designated areas. The results are shown in Table 14 below. The results are shown in Table 14 below. The results are shown in Table 14 below. The results are shown in Table 14 below. Respectively.

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 can be seen that Formulation Example 1 containing ginsenoside F1 according to the present invention as an active ingredient can effectively inhibit excess sebum secreted from Comparative Formulation Example 1 which does not contain ginsenoside F1 as an active ingredient.

[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 contains no effective ingredient for acne skin improvement, and comparative formulation example 4 is a standard for antibacterial activity. , Which contains erythromycin, which is widely used as an acne remedy.

The manufacturing method of Formulation Example 3 and Comparative Formulation Examples 3 to 4 is as follows. The ingredients of phase A in Table 15 were completely dissolved and the components of phase B were completely dissolved in a separate melting tank, and the phase B was added to the phase A to be mixed and solubilized. The ingredients of the C phase were added thereto in accordance with the mixing ratios shown in Table 15, mixed and homogenized and then filtered to prepare the 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 hardened 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] Test for antibacterial activity against acne bacterium Each of the cosmetic compositions prepared by the formulation of Formulation Example 3 and Comparative Formulation Examples 3 to 4 was mixed with propionibacterium acnes (ATCC 6919: medium-BHI broth Broth) were tested for their antibacterial activity.

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

(1) Preparation of test strain

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

(2) Dilution solution preparation

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

(3) Preparation of sample

The undiluted solution of the cosmetic composition prepared from Formulation Example 3 and Comparative Formulation Examples 3 to 4 was used as a sample.

(4) Antimicrobial activity test

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

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

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

The results of the antimicrobial activity test against acne bacteria are shown in Table 16 below. The MIC was 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 Maximum concentration (no antimicrobial activity) Comparative Formulation Example 4 5.7 > 100 ppm

In the case of Formulation Example 3, the concentration of ppm was remarkably smaller than that of Comparative Formulation Example 4 using erythromycin, which is a known acne treatment agent, It can be confirmed that the composition containing the side F1 has much better antibacterial activity against the test bacteria.

[Test Example 12] Lipogenesis inhibition test

3T3-L1 cells, a fibroblast cell line of mice, were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) (Dulbecco's modified eagles medium, GIBCO BRL, Life Technologies) Lt; ⁵ > cells / well in a well culture plate. After 2 days, the cells were replaced with fresh DMEM (containing 10% FBS) medium and cultured for 2 days. The cultured cells were then induced to differentiate into DMEM (containing 10% FBS) containing 1 μg / ml insulin, 0.5 mM IBMX and 0.25 μM dexamethasone, and the gensenosides F1 and caffeine After 2 days of treatment, the cells were replaced with insulin-containing DMEM and cultured for 5 days. After 5 days, the cells were replaced with normal medium (DMEM, containing 10% FBS) and cultured while observing until the cells morphologically changed into adipocytes.

In order to evaluate the inhibitory effect of ginsenoside F1 on the fat accumulation in adipocytes, S-III staining (S4136, sigma-aldrich) was carried out using 3T3-L1 adipocytes differentiated as above. The adipocytes were fixed with 4% paraformaldehyde (pH 7.2) in phosphate buffer at room temperature, washed with PBS (phsphate buffered saline), stained with SIII and then photographed and compared visually. The control group was fed with the test substance or the test substance without the reference substance, and 50 μM of caffeine was treated with another comparative group. The degree of inhibition of fat accumulation was graded by dividing the degree of staining by +++, +, +, -, which means that the degree of staining is greater toward +++. The results are shown in Table 17.

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

As shown in Table 17, it was found that the ginsenoside F1 used in the present invention had an effect of inhibiting lipid synthesis which is superior to caffeine, which is a known lipid synthesis inhibitor, in addition to a small amount of fat accumulated in adipocytes have. Therefore, lipid synthesis is inhibited, and sebum is reduced, so that occurrence of acne can be suppressed.

[Test Example 13] Test for improvement of acne and reduction of sebum secretion and stimulation

30 persons who had acne were divided into 3 groups of 10 persons, and subjects corresponding to each group were allowed to use the cosmetic composition prepared in Formulation Example 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 shown in Table 18 below with an average score of 10 persons.

The acne extermination period was based on the number of days of disappearance readings, and the results were based on the results after 1 month with or without recurrence of acne. The reduction of sebaceous secretion was from 1 point to 5 points, 1 point was 'No', 3 points were 'Normal' and 5 points were 'Very agree'. The experimental results are shown in Table 18 below with an average score of 10 persons. The presence or absence of skin irritation was evaluated as (number of stimuli) / (total number of test subjects).

Inflammatory acne improvement Acne-bloating period Recurrence of acne Decrease sebaceous glands Presence or absence of stimulation Formulation Example 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, it can be seen that Formulation Example 3 has no recurrence of acne compared to Comparative Formulation Example 3, and has excellent effects on improvement of acne as a whole. On the other hand, Comparative Formulation Example 4 containing the antibacterial activity reference material shows the effect of improving acne, but there is a fear of skin irritation for long-term use due to strong stimulus in use, but the composition according to the present invention is not stimulated appear.

[Test Example 14] Inflammation improving effect

1. Inhibitory effect of prostaglandins

The anti - inflammatory effect was evaluated by the inhibitory effect of prostaglandin production. The effect was measured on macrophages using ginsenoside F1. First, aspirin was added to the macrophage cells collected from the abdominal cavity of mice to a final concentration of 500 M to irreversibly inhibit the cyclooxygenase (COX) activity remaining in the cells. Then, the suspension was added to each well of a 96-well cell culture tube in an amount of 100 μl, followed by culturing in an incubator of 5% CO ₂ and 37 ° C for 2 hours to adhere macrophages to the surface of the container. Then, the adhered macrophages were washed three times with PBS and used for the inflammation improving effect test. RPMI medium containing 1% (w / v) of LPS was added to the cultured macrophage 5x10 ⁴ cells / ml, followed by incubation for 12 hours. Then, production of prostaglandin was induced and treated with 100 μl of ginsenoside F1 Prostaglandins were quantitated by enzyme immunoassay (ELISA).

At this time, the inhibitory activity on the production of prostaglandin in the ginsenoside F1 was determined by setting the difference of the prostaglandins produced in the LPS-treated group and the untreated group to 100%, and treating the LPS and the sample together to obtain a reduced prostaglandin percentage (Inhibitory effect of prostaglandin production) is shown in Table 19 below.

Blank (BlanF1) 100% Control group (aspirin treatment group) 25.0% Gin Senocide F1 19.2%

As shown in the above Table 19, it can be seen that the effect of inhibiting the production of prostaglandin is very high even when the ginsenoside F1 is treated as in the control group treated with aspirin. Thus, it can be seen that the ginsenoside F1 of the present invention can provide an excellent inflammation improving effect.

Also, it can be seen that ginsenoside F1 inhibits the expression of prostaglandin, which is a skin inflammation factor, to prevent and improve skin troubles.

2. IL-8 production inhibitory effect

The skin keratinized epithelial cells (normal human sF1in F1 eratinocyte, NHEF1, available from Lonza) were dispensed on a 96 well plate at 5 × 10 ⁴ cells / well, and incubated at 37 ° C. in a 5% CO ₂ incubator For 24 hours. Twenty-four hours later, the cells were washed twice with PBS and replaced with serum-free F1 eratinocyte basement media (F1BM). Each of the wells was treated with the concentration of ginsenoside F1 according to the concentrations shown in the following Table 20 and reacted for 30 minutes. Then, PGSA (10 μg / ml), PGSA (50 μg / ml), PGSA (50 μg / Mu] g / ml), respectively. Here, peptidoglycan from S. aureus (PGSA ) is a peptidoglycan extracted from Staphylococcus aureus, which is a major constituent of the cell wall of Gram-positive (+) bacteria, and the cell membrane components of bacteria are known to cause inflammation, Especially in staphylococcus, about 90% of atopic patients are reported to cause secondary infection by this bacterium. LPS (lypopolysaccaride) is a major constituent of Gram negative (-) bacterial cell membrane and is known to be the main cause of inflammation.

After culturing in a 5% CO ₂ incubator at 37 ° C for 24 hours, the culture was taken and ELISA for Interleukin-8 (IL-8) was performed. The results are shown in Table 20 below. The ELISA used the experimental method of BD science.

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

In Table 20, it was confirmed that ginsenoside F 1 significantly reduced and suppressed the secretion of IL-8 increased by PGSA and LPS. Therefore, it can be seen that the composition for external application for skin of the present invention can provide a superior anti-inflammatory effect by significantly reducing the secretion of IL-8 increased by PGSA and LPS.

[Test Example 15] Evaluation of itching relief

The day before the experiment, keratinocytes (cell line name: HaCaT available from ATCC) were dispensed in a 96 well plate at ⁴ × 10 ⁴ cells / well and cultured in a 5% CO ₂ incubator at 37 ° C. for 24 hours Lt; / RTI > Twenty-four hours later, the 96-well plate was washed twice with HBSS (HanF1's Balanced Salt Solution) buffer, and the reaction buffer (2 μM Fluo-4-AM, 20% pluronic acid, 2.5 mM probenecid) gave. After incubation at 37 ° C in a 5% CO ₂ incubator for 30 minutes and at room temperature for 30 minutes, the cells were treated with ginsenoside F1 at a concentration (%) as shown in Table 21, followed by washing twice with HBSS buffer.

After 10 minutes of reaction, the cells were treated with 2 U / ml trypsin or 5 μM PAR-2 active peptide (SLIGF1V) and the change in intracellular Ca ²⁺ concentration was measured for 80 seconds. Measurement of intracellular Ca ²⁺ concentration was performed using FlexStation 3 (Molecular Device, USA). The difference between the minimum value and the maximum value obtained by measuring the flex for 80 seconds after the treatment with ginsenoside F1 and 2U / ml trypsin or 5 μM PAR-2 active peptide (SLIGF1V) was obtained, The percent inhibition (%) of intracellular influx of calcium ions compared to the difference between the minimum and maximum values in the treatment of 2 U / ml trypsin or 5 μM PAR-2 active peptide (SLIGF1V) is shown in Table 21.

Ginsenoside F1 concentration Inhibition rate of intracellular influx of calcium ions (%) Trypsin (2 U / 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 activating peptide (SLIGF1V) decreases with the treatment of ginsenoside F1, and as the concentration of ginsenoside F1 increases, It was confirmed that the inflow of calcium ions was remarkably reduced. Therefore, the composition for external application for skin of the present invention can effectively provide an excellent antinociceptive effect by effectively inhibiting the itching-inducing PAR-2 activity.

[Formulation Example 4 and Comparative Formulation Example 5]

Shampoo was prepared with the composition shown in Table 22 below. Specifically, the surfactant and ethylene glycol distearate were added to purified water, heated to 80 DEG C to dissolve uniformly, and then gradually cooled to 40 DEG C with stirring. To the mixture were added the active ingredient according to the present invention and a preservative, A viscosity adjusting agent, a perfume, and a hair conditioning agent were mixed and stirred, followed by cooling to room temperature.

ingredient Formulation Example 4 Comparative Formulation Example 5 Ammonium lauryl sulfate 10 10 Polyoxyethylene lauryl ammonium sulfate 5 5 Cocoamidopropyl betaine 2 2 Ethylene glycol distearate 1.5 1.5 Cocoyl monoethanolamide 0.8 0.8 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 evaluation The dandruff strengths of Formulation Example 4 and Comparative Formulation Example 5 were measured and compared. At this time, as a test strain, dicembryum Pityrosporum Ovalae (ATCC 12078) was used.

The skin disc diffusion method was used to measure the antibacterial activity. The sF1in disc diffusion method (SDDM) is the closest method to the consumer's habits and is a method of measuring the antimicrobial activity of guinea pig skin similar to human skin.

Specifically, guinea pig sF1in was peeled off, treated with 70% ethanol, uniformly spread, and then skin disks cut to a certain size were used. The sterilized skin discs were immersed in the shampoo dilution solution for 3 minutes and then rinsed with flowing water. Then, the skin discs were placed on a solid medium inoculated with the test bacteria, and then the test bacteria were cultured. After the culture, the size of the clear zone in which the growth of the bacteria was inhibited was measured, and the relative antibacterial activity was measured. The results of the experiment are shown in Table 23 as the average value of the results of three measurements.

Anti-dandruff evaluation division Restraining Size (mm) Formulation Example 4 Comparative Formulation Example 5 Strain Reduction Size 3.1 0

In Table 23, Comparative Example 5, which does not contain ginsenoside F1, has no effect of reducing dandruff. In the case of Formulation Example 4 containing ginsenoside F1, it is confirmed that effective anti-dandruff effect is obtained because it effectively reduces dandruff.

[Test Example 17] Dandruff reduction test

Twenty-four men aged 19 to 35 years with a relatively large number of dandruff were selected and the dandruff reduction rate was measured after using the shampoo of each of Formulation Example 4 and Comparative Example 5 for 2 months in the following manner for 12 months in the following manner .

Before the start of the test, the dandruff was smeared with ordinary shampoo, and the dandruff accumulated for 2 days after shedding was collected. The dandruff weight was sampled once every two days with the shampoo of each of Formulation Example 4 and Comparative Example 5, The weight of accumulated dandruff was compared and evaluated. At this time, the accumulated dandruff was collected directly from the scalp using a vacuum suction device, and the dandruff reduction rate was calculated according to the following equation (5). 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 from the above Table 24, Formulation Example 4 containing ginsenoside F1 shows excellent dandruff prevention effect.

[Test Example 18] Test for preventing scalp itchiness

Twenty-four male and female twenty-four to twenty-five to fifty-year-olds who felt severe scalp itching were selected and divided into two groups of twelve persons. Each shampoo of Formulation Example 4 and Comparative Example 5 was used once every three days for two weeks. Then, scalp itching The preventive 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

Usually - 3 points

Poor - 2 points

Very bad - 1 point

division Formulation Example 4 Comparative Formulation Example 5 The itching effect of scalp 4.1 2.3

As can be seen from the above Table 25, it can be seen that Formulation Example 4 containing ginsenoside F1 has a better effect of preventing scalp itching.

[Test Example 19] Effect of proliferation of dermal papilla cells

The keratin protein that constitutes the hair is produced in the hair follicle keratinocyte (F1 eratinocyte), and the keratinocyte is differentiated from the dermal papilla cell. In order to evaluate the activity of the dermal papilla cells of the present composition, DP6 (rat immortalized dermal papilla cell) cell line was used in the present invention (Wendy Filsell, Journal of Cell Science 107, 1761-1772 (1994)). This dermal papilla cell line was obtained by microdissection in the hair roots of male PVG rat beard and was cultured in DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS (Fetal bovine serum) USA) for 24 hours in an incubator maintained at 37 ° C in 5% CO ₂ . DP6 was added to a 96-well plate and cultured in a 37 ° C incubator for 24 hours. Then, the main ginsenosides F1 were treated at concentrations of 5 ppm, 10 ppm and 20 ppm, respectively. After 24 hours of drug treatment, cell proliferation was measured using WST-1 kit (Roche). The results are shown in Table 26 below.

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

As can be seen from Table 26, the proliferation of the dermal papilla cells was increased when ginsenoside F1 was treated, which is significantly increased in a concentration-dependent manner.

Test Example 20 Evaluation of Potassium Ion Channel Activity Increase Effect

Minoxidil, a treatment for alopecia, is known as a potential mitochondrial potassium ion channel opener (F1 _ATP channel opener), and is a representative drug used for the treatment of androgenetic alopecia. In order to evaluate the mechanism of minoxidil, cytotoxicity was inhibited by treatment with tolbutamide (SIGMA AlDRICH, T0891) blocking the F1 _ATP channel in the fibroblasts constituting the dermis dermis, The restored test method was used.

To evaluate the function of the present composition as an F1 _ATP channel opener, the fibroblast cell line NIH3T3 (mouse embryonic fibroblast cell line) was used in the present invention. This cell line is a cell line obtained by natural immortalization of fibroblasts isolated from NIH Swiss mouse embryo by the 3T3 protocol. The cells were cultured in DMEM (Gibco BRL, Gaithersburg, MD, USA) containing 10% FBS for 24 hours in an incubator maintained at 37 ° C in 5% CO ₂ . NIH3T3 was added to a 96-well plate, cultured in a 37 ° C incubator for 24 hours, treated with 2.5 mM tolbutamide, and 10 minutes after the positive control, minoxidil 10 μM and ginsenoside F1 were added at concentrations of 2.5 ppm, 5 ppm and 10 ppm And 48 hours after drug treatment, cell proliferation was measured using WST-1 kit (Roche). The results are shown in Table 27 below.

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

As can be seen from Table 27, when the ginsenoside F1 was treated, proliferation of the fibroblast was restored and the cell proliferative capacity was increased depending on the concentration of the treated ginsenoside F1. Is treated with 10 ppm, it can be confirmed that the proliferation of the cells is restored to the level when minoxidil is treated.

[Formulation Example 5 and Comparative Formulation Example 6]

A cream-based composition was prepared according to the composition of Table 28 below in a conventional manner (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] Hairiness effect of ginsenoside F1 Hair of the back of an ICR mouse was epilated with a depilator, and hair was completely removed using a depilatory cream (bit cream). The other two groups except the control group (normal group) were applied to the skin once a day by 200ul of 1% DNCB to induce skin inflammation for 3 days. After 3 days, the cream base of Formulation Example 5 and Comparative Formulation Example 6 was applied once a day to the groups excluding the control group, and the degree of hair growth in each group was observed, and the results are shown in FIG.

1, the control group showed no hair growth in the epilated area during the 15-day observation period, but showed only a slight hair growth effect in the group treated only with the cream base without the ginsenoside F1, In the group treated with cream, it was confirmed that hair removal effect was remarkable in the entire hair removal part.

[Test Example 22] Wool effect of ginsenoside F1

The mice were treated with dinitrochlorobenzene (DNCB) to induce skin irritation by setting the stress condition in the skin to maximally reduce the hair growth, To evaluate the hair regrowth efficacy of the composition, the creams of Formulation Example 5 and Comparative Formulation Example 6 containing ginsenoside F1 were treated and the length of the hair growth was measured according to the days of treatment and compared with the control group. The results are shown in Table 29 below.

Days to be treated (days) Fur Length (cm) Formulation Example 5 Treatment Comparative Formulation Example 6 Treatment 0 0.1 0.1 6 0.2 0.1 9 0.5 0.1 12 1.1 0.1

As shown in Table 29, the hair length of the group treated with Formulation Example 5 showed a statistically significant difference (p < 0.01) as compared with the group treated with Comparative Formulation Example 6, The flesh of the group treated with the cream base of Formulation Example 5 was longer than that of the group treated with the cream base of Comparative Formulation Example 6 which did not contain ginsenoside F1 , Indicating that Ginsenoside F1 promoted hair regeneration even under hair regrowth inhibition conditions caused by skin stress, thereby confirming that Ginsenoside F1 has a regenerating function against hair loss under stress .

[Test Example 23] Test for promoting melanin production of ginsenoside F1

Melanocytes (melan-a) were added to a 24-well microtiter plate at 50,000 cells / well in RPMI medium supplemented with 5% fetal bovine serum, 100 IU penicillin G and 0.2 μM TPA Respectively. The next day, the cells were treated with 0.1% DMSO as a negative control, 100 μM IBMX as a positive control, and then cultured at 37 ° C. for 3 days as a test substance at a final concentration of 10, 50 ppm. After incubation, the wells were washed with PBS, and 100 μl of 1N NaOH was added thereto, and melanin in the cells was dissolved. The absorbance of dissolved melanin was measured at 405 nm using a microplate reader. The results of comparing melanin production promoting effect of ginsenoside F1 with the control group are shown in Table 30 below.

sample Amount of melanin synthesis (%) DMSO (0.1%) 100 IBMX (100 [mu] M) 120 Ginsenoside F1 (10 ppm) 112 Ginsenoside F1 (50 ppm) 134

Table 30 shows that ginsenoside F1 promotes melanin synthesis of melanocyte, resulting in increased melanin production, and shows excellent melanin production promoting effect.

[Test Example 24] Promotion of MITF and tyrosinase expression in melanocytes of ginsenoside F1

The cells were divided into 6-well microtiter plates at a density of 500,000 cells / well using a 501mel cell line. DMSO 0.1% as a negative control, IBMX 100 μM as a positive control group, and ginsenoside F1 was treated at 10 ppm, followed by culturing at 37 DEG C for 24 hours, 48 hours, and 72 hours to obtain a protein. The proteins thus obtained were subjected to western blotting using MITF and tyrosinase antibodies. Protein extraction and Western blotting were routinely performed by standard methods used by those skilled in the art. After Western blotting, the result was compared with the negative control group of 100, and the results are shown in Table 31 below.

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

As shown in Table 31, it can be confirmed that ginsenoside F1 increases MITF and tyrosinase protein expression in melanocytes.

[Test Example 25] Evaluation of prevention of white hair and in vitro induction of in vivo ginsenoside F1 using white algae-stimulated albumin mice

Vitiligo mice (C57bl / 6- Mitf ^mi-vit ) were purchased from The JacF1son Lab in the USA. The anti-whiplash effect test method using the mouse in which the white moth is promoted is as follows. Dorsal hairs of 12-week-old mice were removed by depilation. However, the width of the hair removal area was adjusted to be the same for each individual. From the day after the hair was pulled out, twice a day, the anti-whiplash material was applied on the hairless area. A mixture of EtOH: 1,3-BG: DW = 3: 2: 5 (volume ratio) was used as the vehicle of the anti-whiplash material. The carrier was used as a negative control, and a solution containing 50 mM of IBMX As a control group, a solution containing 2.5% of ginsenoside F1 was used as a test group. After about 3 weeks, when the difference in antibacterial effect between the substances was visually recognized, newly-emerged hairs were collected and the amount of melanin in the hair was measured. The amount of melanin in hair was measured using the protein hydrolyzing enzyme Esperase (Novozyme). Reaction buffer was prepared by dissolving Esperase in a buffer (50 mM Tris-HCl, 5 mM DTT, pH 9.3) to a concentration of 1 NPU / ml. Five milligrams of mouse hair was added to 1 ml of reaction buffer and allowed to react for 13 hours at 37 ° C with shaking at 1,000 rpm. Then, the hair and the reaction solution were separated by an instant centrifugation. The reaction solution thus obtained is placed in a 96-well plate, and the amount of melanin in the reaction solution can be measured by measuring the absorbance at a wavelength of 405 nm. The results of visual inspection and the determination of melanin in the hair when the negative control, the positive control, and the test group material were treated with the white algae-stimulated albino mouse model are shown in Table 32, Is 100, and is compared with this value.

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

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

[Test Example 26] Evaluation of antibacterial activity of ginsenoside F1

An antibacterial experiment was conducted to evaluate the antibacterial activity of ginsenoside F1. Specific experimental methods are as follows.

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

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

In a 96-well 96-well plate, add 16 μl of each sample to the first line, and add 184 μl of the diluted solution. In the remaining wells, 100 μl of the dilution solution was added. 100 μl of the mixed solution of No. 1 line was mixed well and 100 μl of the mixture was added to the No. 2 line. After mixing well, 100 μl of the mixture was added to the No. 3 line and diluted 2-fold.

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

After 48 hours, the microbial proliferation 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), unit% Sedona Monas
Aruzinosa Staphylococcus aureus Escherichia coli Candida Albikans Aspergillus niger Gin Senocide F1 0.1 0.06 0.122 > 4 > 1

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

Claims (3)

A cosmetic composition for improving atopy, comprising as an active ingredient ginsenoside F1 represented by the following formula (1).
[Chemical Formula 1]

delete The method according to claim 1,
Wherein the ginsenoside F1 is contained in an amount of 0.001 to 50% by weight based on the total weight of the composition.

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