KR101427573B1 - Cosmetic composition for improving skin acne containing ginseng fruit extracts - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving acne skin containing an extract of ginseng fruit, and more particularly, to a composition containing ginseng fruit extract having a unique composition and composition in the ginseng root as an active ingredient to control sebaceous gland secretion, The present invention relates to a composition for improving acne and skin troubles.

Description

[0001] The present invention relates to a cosmetic composition for improving acne skin containing ginseng fruit extract as an active ingredient,

The present invention relates to a cosmetic composition for improving acne skin containing an extract of ginseng fruit, and more particularly, to a composition containing ginseng fruit extract having a unique composition and composition in the ginseng root as an active ingredient to control sebaceous gland secretion, The present invention relates to a cosmetic composition for improving acne skin, which is effective for improving acne and skin troubles.

Ginseng (Panax ginseng C.A. Meyer) is a herb that belongs to the genus Ogawa and Ginseng. It has been used in Korea, China and Japan for over 2,000 years and has been used empirically to prevent disease and prolong the life span. The efficacy and effect of ginseng which has been known so far can be determined by the action of the central nervous system, the anticarcinogenic action, the anticancer activity, the immune function regulating action, the anti-diabetic action, the hyperfunction of liver function, the improvement of cardiovascular disorder, (Korean Ginseng & Tobacco Research Institute, 56-112, 1996). In addition, it has been reported that the antioxidant activity and antioxidant activity of osteoporosis are improved.

Ginsenoside, which is a representative physiologically active ingredient of ginseng, is distributed evenly on the ground and underground of ginseng. Especially, ginsenoside (root), ginseng leaf and ginseng berries have not only ginsenoside content but also composition (Attele AS et al, Biochem Pharmacol, 58: 1685-1693, 1999). In particular, the fruit of ginseng has been reported to exhibit superior antidiabetic efficacy to ginseng root based on the contents of ginseng roots and other ginsenosides (Dey L. et al., Phytomedicine, 10, 600 -605, 2003).

Recently, consumers are interested in natural cosmetics. At the same time, many cosmetics using herbal materials have been launched, and ginseng has been studied as a plant material having important skin effect of cosmetics. However, most of them are ginseng root extract or ginseng root ginsenoside Side components and ginseng polysaccharides are used only by the ingredients of ginseng fruit is a fact.

The ginseng fruit is considered to be more valuable than ginseng since ancient times and has been harvested for the purpose of obtaining seeds. Ginseng fruit is grown only once in 4 years of ginseng cultivation period. When seeds are cultivated in 3 year olds, it is difficult to produce good seedling ginseng because seeds are too small. If seeds are grown for 5 years or more, seeds are faithful and good, The quality of red ginseng can be greatly reduced when red ginseng is produced because it is not only inhibited development but also is inadequate in the tissue. When the number of seedlings is increased to more than 2 times during the cultivation period, the yield and the quality of red ginseng are significantly lowered (the latest Korean ginseng "cultivation", Korea Ginseng & Tobacco Research Institute, 130-131, 1996).

In general, sebaceous glands in the skin, including the scalp and face, play a role in preventing moisturization and microbial invasion of the skin. However, excessive secretion of sebum promotes hair loss, worsens acne, promotes enlargement of pores caused by acne, And many other diseases are caused.

The hypersecretion of sebaceous glands is caused by various causes, and the most important one is the activation of sebaceous glands by the amount of dihydrotestosterone (DHT), which is one of the hormones involved in promoting sebaceous secretion in the activity of sebaceous glands, Is overdone. That is, testosterone (T) is converted into dihydrotestosterone, one of the male hormones, by 5-alpha-reductase type 2 in cells in the hair loss, (5-α-reductase type 1) in the skin or the sebaceous glands. The testosterone is converted to dihydrotestosterone by the action of 5-α-reductase type 1, (J. Invest Dermatol 105: 209-214 Diane et al.).

Acne and hair loss, which are skin troubles other than simple sebum hypersecretion, are further exacerbated by the minute inflammatory reaction of the skin. If you look at the development of acne, excess sebum accumulates in the hair follicle, activating acne bacteria and causing inflammation.

It has antioxidant effect with excellent antioxidant effect, inhibiting the production of prostaglandin, especially prostaglandin E2 (PGE2) in cell inflammation reaction, inhibiting the production of nitric oxide (NO) by iNOS, It is possible to alleviate the skin troubles.

However, there are not many raw materials that can reduce the secretion of sebum as a skin-safe natural material through fundamental causes, and at the same time, relieve inflammation.

Accordingly, the inventors of the present invention found that when extracts of ginseng fruit were investigated for cosmetics having skin effect using the ginseng fruit which is above the ginseng fruits, they found skin effects due to ingredients and compositions different from general ginseng and red ginseng, It was completed.

Accordingly, an object of the present invention is to provide a cosmetic composition for anti-inflammation which can improve sebum control effect and anti-inflammatory effect, that is, the production of an inflammatory factor by containing the ginseng fruit extract as an active ingredient, thereby improving skin troubles including acne .

In order to achieve the above object, the present invention provides an anti-inflammatory cosmetic composition having an effect of controlling sebum and an inflammation reaction and containing acne and skin troubles by containing ginseng fruit extract as an active ingredient.

In the present invention, it was possible to provide a cosmetic composition having an effect of improving acne and skin troubles by containing ginseng root and ginseng fruit extract having different composition and composition among ginseng root as an active ingredient.

FIG. 1 is a graph showing ginsenoside components of the ginseng fruit extract of Example 1 and the ginseng extract of Comparative Example 1. FIG.
Fig. 2 is a graph showing the active oxygen scavenging activity of the test substance.
FIG. 3 is a graph showing the amount of collagen produced in the ginseng fruit extract of Example 1. FIG.
4 is a graph showing the inhibitory effect of MMP-1 on the biosynthesis of ginseng fruit extract of Example 1. Fig.
FIG. 5 is a graph comparing the nitrogen monoxide inhibitory effects of the ginseng fruit extract and the red ginseng extract of Example 1. FIG.
6 is a graph showing the amount of NO produced by the ginseng fruit extract of Example 1. FIG.
FIG. 7 is a graph showing the amount of NO produced by red ginseng extract. FIG.

In the present invention, the ginseng fruit extract may be mixed in an amount of 0.001 to 50% by weight based on the total weight of the composition, depending on the form of the cosmetic composition. The ginseng fruit extract is preferable when the effect of mixing 0.001 wt% or more of the ginseng fruit extract is considered, and it is a preferable value when 50 wt% or less of the ginseng fruit extract is considered considering the formulation.

The ginseng fruit extract according to the present invention is prepared by extracting the flesh and the skin of ginseng fruit which have been separated from seeds by sunlight drying or hot air drying and extracting with water or ethanol.

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

The cosmetic composition of the present invention can be used in the form of a cosmetic or dermatological composition comprising a lipid, an organic solvent, a solubilizing agent, a thickening agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, , Cosmetics such as fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredient commonly used in cosmetics May contain adjuvants conventionally used in the field of dermatology. Such adjuvants are introduced in amounts commonly used in the cosmetics or dermatological fields.

The cosmetic composition comprising the ginseng fruit extract of the present invention is not particularly limited in its formulation, and examples thereof include softening lotion, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, Cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil and body essence.

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

[Example 1] Preparation of ginseng fruit extract

1) Ginseng fruit pretreatment: The raw ginseng fruit was harvested and the seeds were separated and removed. Then, the ginseng fruit and the peel were dried in daylight or hot air to prepare ginseng fruit raw material.

2) Preparation of ginseng fruit extract: 1 kg of dried ginseng fruit was refluxed by adding 3 L of water, and then filtered and concentrated under reduced pressure at 40 to 45 ° C. to obtain 300 g of ginseng fruit extract.

[Comparative Example 1] Production of ginseng muscle root extract

Example 1 was prepared in the same manner as in Example 1, except that ginseng root (root) was used instead of ginseng fruit.

[Test Example 1] Comparison of components of ginseng fruit extract

<Analysis of Ginsenoside (Ginseng Saponin) Components of Ginseng Fruit and Ginseng Root>

Ginseng root extract and ginseng root extract were prepared in Example 1 and Comparative Example 1, respectively. Then, the extract was treated with ether to remove fat-soluble components, then crude saponin was extracted with butanol (BuOH) Senoside component analysis was carried out, and the results are shown in FIG. 1 and Table 1.

Example 1 Comparative Example 1 Crude saponin content
(Dry weight) 33.42% 16.70% PD / PT ratio 0.73 3.23

The ginseng fruit extract prepared in Example 1 had about twice the crude ginsenoside content of the ginseng root extract prepared in Comparative Example 1, and the ginsenoside was divided into PD (protopanaxadiol) -ginsenoside Rb1, Rb2 , Rc and Rd "and PT (Protopanaxatriol) system -" Ginsenoside Re, Rg1 and Rg2 ", the ginseng fruit and ginseng root showed distinct differences and characteristics in the composition of 0.73 and 3.23 respectively .

<Mineral composition analysis of ginseng fruit extract>

In order to distinguish the ginseng fruit extract prepared in Example 1 from the ginseng, which is different from "ginseng", mineral components including vitamins were analyzed. The results are shown in Table 2 below.

ingredient content ingredient content Potassium (mg / 100g) 5865.57 Magnesium (mg / 100 g) 354.38 Calcium (mg / 100g) 819.26 Zinc (mg / 100g) 178.49 Iron (mg / 100g) 59.31 Vitamin A (μg / 100 g, RE) 213.11 Phosphorus (mg / 100 g) 187.17 Vitamin B1 (mg / 100g) 12.29 Vitamin B2 (mg / 100g) 8.45 Vitamin B6 (mg / 100g) 10.50 Vitamin C (mg / 100g) 4.91 Vitamin E (mg / 100 g, α-TE) 23.61 Vitamin K (μg / 100g) 232.12 Niacin (mg / 100 g, NE) 5.76 Pantothenic acid (mg / 100g) 5.87 Folic acid (μg / 100g) 349.97

As described above, the constituent characteristics of the ginseng fruit used in the present invention is that the ginseng saponin is contained in a larger amount than that of the ginseng root, and the saponin composition properties are opposite to each other. Also, unlike the ginseng root, the content of 16 vitamins and minerals is abundant , And based on this, the following experiments on skin efficacy were conducted.

[Test Example 2] Effect of suppressing skin aging and improving wrinkles

1) Antioxidative effect of ginseng fruit extract

Antioxidant effect was investigated by comparing the ability to remove reactive oxygen species (ROS) from cells by ultraviolet (UV) irradiation. As a positive control, trolox, which is generally used to compare antioxidative effects, was used and compared with white and red ginseng extracts (FIG. 2). In addition, the test group was not used as a control group.

2, the ginseng fruit extract of Example 1 according to the present invention significantly inhibited the active oxygen generated by UV in the human HaCaT keratinocyte monolayer culture system (human HaCaT keratinocytes monolayer culture system) . This is an excellent activity similar to that of trolox used as an activity index of antioxidants, and is in contrast to white ginseng and red ginseng extracts which have no significant scavenging activity. That is, it was confirmed that the extract of the ginseng fruit according to Example 1 of the present invention has the effect of preventing wrinkle formation, elasticity degradation, pigmentation, and the like by significantly erasing active oxygen which is a cause of skin aging.

2) Inhibitory activity against DNA oxidative damage by single cell gel electrophoresis (SCGE, Comet assay)

After normal human fibroblasts were cultured for 24 hours, the ginseng fruit extract of Example 1 and vitamin E were treated for each concentration. After 12 hours, the positive control group, H ₂ O ₂ (final concentration: 10 ^-3 M) The result of staining with electrophoresis was observed with a fluorescence microscope. At this time, the length (um) of the tail as a result of DNA cleavage in each of 25 cells was analyzed using an image analyzer, KOMET 3.1 (Kinetic Imaging, England) Respectively.

Inhibitory effect of ginseng fruit extract and vitamin E on DNA oxidative damage Test substance Tail length (탆) Inhibition rate (%) H ₂ O ₂ 115.6 - H ₂ O ₂ + Example 1 (10 ppm) 75.8 34.4 H ₂ O ₂ + Example 1 (50 ppm) 65.7 43.2 H ₂ O ₂ + Example 1 (100 ppm) 52.1 54.9 H ₂ O ₂ + vitamin E 10 ppm 70.6 38.9

In the results of Table 3, the ginseng fruit extract of Example 1 by cell gel electrophoresis showed a significant oxidative DNA damage inhibitory activity, and thus the ginseng fruit extract according to the present invention showed hydroxyl radical (.OH) , Which is a DNA oxidative damage inhibitor that suppresses DNA single strand break caused by DNA damage.

3) Type I procollagen assay (Type I Procollagen assay)

Human fibroblasts were cultured in a 12-well plate incubator at a concentration of 10 ⁵ and then replaced with a medium containing 1 ppm and 10 ppm of the ginseng fruit extract of Example 1. On the third day of culture, the cells were harvested and the amount of type I procollagen produced by ELISA was quantitated. The result was calculated by comparing the value obtained with the control group not including the ginseng fruit extract of Example 1 as 100, and TGF-b was used as a positive control group. Ginseng root extract was also compared with white ginseng extract and red ginseng extract.

In the normal human fibroblast monolayer culture system, the ginseng fruit extract of Example 1 markedly promoted the formation of type I procollagen as compared with the control. That is, the ginseng fruit extract of Example 1 suppressed the reduction of collagen production caused by aging of human skin, and showed an effect of improving wrinkles (FIG. 3).

4) MMP-1 expression inhibition assay

Human fibroblasts were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , irradiated with 40 mJ / cm 2 of ultraviolet B, and then replaced with a medium containing 1 ppm and 10 ppm of the ginseng fruit extract of Example 1. On the second day of culture, the cells were harvested and the amount of MMP-1 (matrix metalloproteinase I) produced by ELISA was quantitated. The result was calculated by comparing the value measured with the ultraviolet control group containing no test substance as 100, and TGF-b was used as the positive control group. Ginseng root extract was also compared with white ginseng extract and red ginseng extract.

In the human normal fibroblast monolayer culture system, the ginseng fruit extract of Example 1 according to the present invention significantly inhibited the expression of MMP-1 induced by UVB 40 mJ / cm 2. Thus, the ginseng fruit extract of Example 1 according to the present invention inhibits the biosynthesis of MMP-1, a skin tissue degrading enzyme caused by internal aging or external environmental factors, thereby inhibiting skin aging and improving wrinkles (Fig. 4).

5) Effect of inhibiting biosynthesis of cyclooxygenase-2 (COX-2) by ultraviolet light

Human fibroblasts were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , and irradiated with UVA A at 15 J / cm 2. Then, the extracts of ginseng fruit of Example 1 and ginseng root extract of Comparative Example 1 were applied at 0.1 ppm, 1 ppm and 10 ppm Containing medium. On the second day of culture, cells were harvested and the amount of cyclooxygenase-2 (COX-2) produced using the Westren blot method was quantitated. The results are shown in Table 4 below. The results are shown in Table 4 below. [Table 4] &lt; EMI ID = 16.1 &gt;

Test substance (concentration: ppm) COX-2 biosynthesis (%)
Example 1 10 11 One 38 0.1 63
Comparative Example 1 10 72 One 91 0.1 99 Control group 100

From the results shown in Table 4, the ginseng fruit extract of Example 1 reduced the biosynthesis of cyclooxygenase-2 by ultraviolet light in a concentration-dependent manner, and the skin tissue degradation by prostaglandin E2 (PGE2), the product of COX- . &Lt; / RTI &gt;

6) Inhibitory effect of UV-induced tumor necrosis factor alpha (TNF-α) on biosynthesis

Human keratinocytes were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , and irradiated with ultraviolet ray B at 30 mJ / cm 2. Then, the extracts of ginseng fruit of Example 1 and ginseng root extract of Comparative Example 1 were applied at 0.1 ppm, 10 ppm. Cells were harvested 6-24 hours after culture and the amount of tumor necrosis factor alpha (TNF-a) produced was quantitated using ELISA (Pharmingen 555212) method, The results are shown in Table 5 below.

Test substance (concentration: ppm) TNF-α biosynthesis (%) Example 1 10 16 One 38 0.1 72 Comparative Example 1 10 74 One 85 0.1 92 Control group 100

From the results shown in the above Table 5, it can be seen that the ginseng fruit extract of Example 1 according to the present invention can be produced by the biosynthesis of tumor necrosis factor alpha by reducing the biosynthesis of tumor necrosis factor alpha (TNF-α) It was confirmed that skin aging can be prevented in advance.

7) Skin wrinkle improvement effect on human skin

For the 40 subjects who had facial wrinkles of 30-50 years of age, the nutritional creams of Example 2 containing the ginseng fruit extract of Example 1 and the composition of Comparative Example 2 not containing the extract of Example 1 having the composition shown in the following Table 6 were prepared The skin wrinkle-improving effect was compared and evaluated (unit: wt%).

ingredient Example 2 Comparative Example 2 Ginseng fruit extract 0.1 - Wax 10 10 Polysorbate 60 1.5 1.5 Piggy 60 hardened castor oil 2 2 Sorbitan sesquioleate 0.5 0.5 Liquid paraffin 10 10 Squalane 5 5 Caprylic / capric triglyceride
(Estasan: Uniqema (manufacturer)) 5 5 glycerin 5 5 Butylene glycol 3 3 Propylene glycol 3 3 Triethanolamine 0.2 0.2 Preservative, coloring, fragrance Suitable amount Suitable amount Purified water Balance Balance

Example 2 was used for the face side of the subject, and Comparative Example 2 was used for the right side for 3 months. The skin condition of both sides of the facial area before the use of the cream was measured, and then the same part was remeasured 3 months after using the cream to measure the change of the skin wrinkles. The skin wrinkles were measured with a Visiometer system (C + K company) by removing the wrinkles of the eye tail region with a replica in a constant temperature and humidity chamber at 24 ° C and a relative humidity of 40%. The amount of change in the skin wrinkles was calculated according to the following formula (1).

In the equation (1), &quot; Tdi &quot; is the measured site value at D90 and &quot; Tdo &quot; is the measured site value at D0.

As a result of calculation according to the above formula (1), skin wrinkles at the site using Comparative Example 2 showed a decrease of 6.5 ± 4% (mean ± standard deviation), while skin wrinkles at the site using Example 2 were 39 ± 11% Of the total wrinkles.

8) Skin elasticity improvement effect in human body

The skin elasticity improving effect of Example 2 and Comparative Example 2 prepared in the composition of Table 6 was measured. The nutritional creams of Example 2 and Comparative Example 2 were applied to the face for 2 weeks each day for 12 weeks in each group. The skin elasticity meter (Cutometer SEM 575, C + K Electronic Co., Germany). The results are shown in Table 7 below. The results shown in Table 7 indicate the viscoelasticity of the skin elasticity measuring device.

Test substance Skin elasticity effect Example 2 0.46 + 0.12 Comparative Example 2 0.29 ± 0.09

As shown in the results of Table 7, when the nutritional cream of Example 2 containing the ginseng fruit extract of Example 1 was used, skin elasticity was improved by about 150%.

[Test Example 3] Skin whitening effect

1) Inhibitory effect of melanin on melanin production in rat

To investigate the melanin inhibitory effect of the ginseng fruit extract of Example 1, mouse pigment cells were used.

First, a mouse-Ab cell (Dooley, TP et al, Skin Pharmacol, 7, pp. 188-200) derived from C57BL / 6 mice was inoculated into DMEM (Dulbeccos modified Eagles media) The cells were cultured in a medium supplemented with 100 nM 2-O-tetradecanoyphorbol-13-acetate and 1 nM cholera toxin at 37 ° C and 5% CO ₂ . Cells were cultured in a 24-well plate incubator at a concentration of 10 ⁵ cells / well (cells / well). Then, the cells were incubated for 3 consecutive days with 1 ppm and 10 ppm of ginseng Fruit extract was added and cultured. At this time, red ginseng extract and hydroquinone were used as positive control. Then, the culture solution was removed, washed with PBS, and the cells were dissolved with 1 N sodium hydroxide. The absorbance at 400 nm was measured, and the inhibition rate of melanin production was calculated according to the following formula (2) (Dooley's method).

Test substance Melanin production inhibition rate (%) Example 1 (1 ppm) 14.0 Example 1 (10 ppm) 34.4 Red ginseng extract (10ppm) 5.3 Hydroquinone (control group) 41.1

As shown in Table 8, the ginseng fruit extract of Example 1 according to the present invention showed a better melanin production inhibitory rate than the red ginseng extract and showed a similar degree of inhibition of melanin production to hydroquinone. Thus, it was confirmed that the ginseng fruit extract of Example 1 according to the present invention exhibited excellent whitening effect.

2) Whitening effect on human skin

The following experiment was conducted to examine the whitening effect of the ginseng fruit extract of Example 1 on human skin.

First, 12 healthy boys were exposed to a 1.5 cm diameter opaque tape at the upper part of the subject's body, and then they were exposed to ultraviolet rays (UVB) of about 1.5 to 2 times the minimum erythema dose of each subject. To induce blackening of the skin.

The ginseng fruit extract of Example 1, which is the test substance after irradiation with ultraviolet rays, and hydroquinone were each applied, and one state was observed for 10 weeks without applying anything. The color of skin was measured with a colorimeter CR2002 (Minolta Co., Japan) every 1 week.

Then, the difference (ΔL *) between the start time of application of the respective test materials and the completion time of the application of the test materials was calculated according to the following equation (3). On the other hand, the whitening effect is judged by comparison of DELTA L * between the sample application site and the control site. When the DELTA L * value is 2, the whitening of the deposited pigment is noticeable. .

Test substance The degree of skin color brightness (DELTA L *) Example 1 1.76 ± 0.21 Hydroquinone (positive control) 1.90 + 0.13 Vehicle (negative control) 0.50 0.16

As shown in Table 9, it was confirmed that the ginseng fruit extract of Example 1 according to the present invention showed a degree of skin color brightness similar to that of hydroquinone. This is because the ginseng fruit extract improves pigmentation caused by ultraviolet light and brightens skin color.

[Test Example 4] Skin moisturizing effect and atopic symptom improvement effect

1) induction of differentiation of human keratinocytes

The amount of CE (cornified envelope) produced upon differentiation of keratinocyte and human skin cell line (HaCaT) was measured to investigate the promoting effect of ginseng fruit extract of Example 1 according to the present invention on cell differentiation.

The primary cultured human keratinocytes were placed in a culture flask and adhered to the bottom. The test substances described in the following Table 10 were added to the culture medium by concentration and the cells were cultured for 5 days until the cells reached 70-80% Lt; / RTI &gt; The cells were harvested and washed with PBS (phosphate buffered saline). Tris-HCl buffer (Tris-HCl buffer, pH 7.4) containing 2% SDS (sodium dodecyl sulfate) and 20 mM DTT (Dithiotime- -HCl, pH 7.4), sonication, boiling, and centrifugation were 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 was used as a standard for evaluation of the degree of cell differentiation. The test results are shown in Table 10 below, in which the low calcium (0.03 mM) treatment group and the high calcium (1.2 mM) treatment group were used as negative / positive control groups and the test substance was added to low calcium concentration.

Test substance The ability to differentiate in keratinocytes (%) Control group The low calcium treatment group (Ca ^{2 +} 0.03 mM) 100 The calcium treatment group (Ca ^{2 +} 1.2 mM) 210 Red ginseng extract (100 ppm) 104 Example 1 1 ppm 115 10 ppm 120 100 ppm 124

From the results of Table 10, it can be confirmed that the extract of ginseng extract of Example 1 according to the present invention promotes differentiation in keratinocytes.

2) Expression of transglutaminase in human skin cell line

Human skin cell lines were plated in a 96 well plate incubator at 5 x 10 ^{4 per} well for 24 h. After the adhered skin cell line was treated with the test substance, after 2 days, the medium was removed and stored in a -20 ° C refrigerator. Cells treated with the substance were treated with acetone: ethanol (1: 1, v / v) stored at -20 ° C for 30 minutes at 4 ° C, followed by freezing-thawing twice Cells were fixed. (1% bovine serum albumin), transglutaminase antibody (primary antibody) and time P anti-mouse antibody (secondary) were performed. The color development was performed using OPD o-phennyldiamine). Absorbance was measured at 490 nm and calibration was performed by measuring the background at 630 nm. The results are shown in Table 11 below, in which the low calcium (0.03 mM) treatment group and the high calcium (1.2 mM) treatment group were used as negative / positive control groups and the test substance was added to low calcium concentration.

Test substance Expression amount (%) of transglutaminase
Control group
The low calcium treatment group (Ca ^{2 +} 0.03 mM) 100 The calcium treatment group (Ca ^{2 +} 1.2 mM) 140 Red ginseng extract (100 ppm) 131
Example 1
1 ppm 135 10 ppm 175 100 ppm 210

From the results of Table 11, it can be seen that the extract of Ginseng Extract of Example 1 according to the present invention showed an increase of about twice as much as that of the control group, and the amount thereof was higher than that of the control group. Thus, it can be seen that the extract of ginseng fruit of Example 1 according to the present invention improves the expression of transglutaminase.

3) Increased production of epidermal lipid (total ceramide) in human skin

Carbopol ETD 2020 0.5%, TEA 0.45%, test substance 5%, and 100 with deionized water were used to evaluate the effect of the ginseng fruit extract of Example 1 on the increase of lipid synthesis in human skin. Twelve adult men and women were exposed to the diameter of a 50ml polypropylene cornical tube on their forearms and then sprayed with 20μl of the control group and the ginseng fruit extract of Example 1 twice daily, Were divided into two groups, and the lipid was extracted and analyzed by the following method on the 3rd and 11th days. The forearms were washed with tap water and tape stripped once with Scotch 810 Magic tape and cut into 1 ml of cyclic nucleic acid / water using a 50 ml polypropylene cornic tube cut into reservoirs, After the addition of cyclohexane / ethanol (4: 1), the mixture was mixed for about 1 minute and transferred to a new tube. 1 ml of cyclohexane / ethanol (1: 1) was treated in the same manner as above and collected in a tube. Each sample was dissolved in 500 μl of chloroform / methanol (2: 1) by using a nitrogen gas at 50 ° C., and then analyzed by silica gel TLC (Automated TLC sampler-4) (Thin Layer Chromatography) plate and developed with developing solvent of composition ratio shown in Table 12 using AMD (Automated Multiple Development).

chloroform Methanol water Acetic acid Hexane Diethyl ether Petroleum ether Solvent migration distance
(cm) Primary 40 10 One 3.0 Secondary 190 9 One 6.5 Third 2 12 3 7.5 Fourth 100 Top

After developing the TLC plate, the size of each band was measured at a wavelength of 570 nm using a TLC scanner-II (CAMAG: densitometer). The amount of ceramide produced in each treatment group in which the amount of ceramide produced in the non-treated group was converted to 100% is shown in Table 13 below.

Test substance After 3 days treatment (%) After 11 days treatment (%) Control group Untreated group 100 100 Vehicle 103 98 Glycerin (5%) 102 94 Example 1 (5%) 108 116

As shown in Table 13, the ginseng fruit extract of Example 1 showed an increase of about 5 to 8% compared with the negative control group (control group / vehicle) 3 days after the treatment, and about 6% of the control group (glycerin) Respectively. After 11 days, the lipid productivity was about 16% higher than the negative control and 22% higher than the positive control. This demonstrates the increase of ceramide by the ginseng fruit extract of Example 1 according to the present invention.

4) Increase skin moisture in human body

Fifty female adults and 50 female adults representing skin dryness were divided into two groups, and each group was applied to the face of Example 2 and Comparative Example 2 twice daily for 4 weeks. (24 ° C, relative humidity: 40%) after one week, two weeks, and four weeks after application and before two weeks (six weeks after completion of application) Skin moisture was measured and the results are shown in Table 14 below. The test result is a percentage of the increment of the measured value after a certain period of treatment based on the coneometer value measured just before the start of the test.

Test substance Moisture growth rate (%) 1 week Two weeks 4 weeks 6 weeks Example 2 34 41 42 33 Comparative Example 2 30 34 34 15

Connio meter is a skin moisture analyzer that measures the amount of water present in the skin by measuring the electrical conductivity of the skin. As shown in Table 14, in the group to which the substance of Example 2 containing the extract of ginseng extract of Example 1 was applied, the skin moisture amount was further increased as compared with the control group to which Comparative Example 2 was applied. In addition, the values of the skin moisture measured 2 weeks after the application of the nutritional cream of Example 2 were stopped (total of 6 weeks) were similar to those after 1 to 2 weeks, It was found that the skin moisture by the application of the ginseng fruit extract according to the present invention was continuously maintained during the period.

5) Improvement of atopic dermatitis

The effect of improving atopic symptoms was measured as follows. Fifty patients between the ages of 10 and 50 who were diagnosed with atopic symptoms (itch, eruption or severe dryness) or who had atopy-like symptoms by visual observation were divided into two groups and provided with Example 2 and Comparative Example 2 And applied to the area showing atopic symptoms twice a day for 12 weeks. The efficacy evaluation was conducted after 12 weeks' treatment, and the degree of improvement was judged subjective by the questionnaire. In the questionnaire, symptoms of atopic dermatitis were divided into "itching", "dryness", "keratinization", "dandruff", "erythema", "swelling", "skin crack" The degree of improvement of each symptom was evaluated and then the average of the scores was evaluated to evaluate the improvement effect of atopic symptoms. The results are shown in Table 15 below.

Test substance Number of respondents (in) Significant improvement Improving No change worse Example 2 2 4 14 0 Comparative Example 2 6 8 6 0

As shown in Table 15, it can be seen that the group treated with the formulation containing the extract of Ginseng extract of Example 1 according to the present invention showed more improvement in atopic symptoms overall than the control group.

[Test Example 5] Acne and skin troubles were alleviated

1) sebum control effect: 5-alpha-reductase inhibitory effect

Mature SD (Sproque-Dawley) male rats (7-8 weeks old) were sacrificed with diethyl ether, the abdomen prostate was removed, the connective tissue was removed and a buffer (0.32M sucrose, 0.1 mM dithiothreitol, And 20 mM sodium acetate), and the mixture was finely pulverized and suspended in a pulverizer. The suspension was centrifuged, and the supernatant was taken to partially refine 5-alpha-reductase, and the activity of the enzyme was inhibited.

A portion of the supernatant obtained above is placed in a buffer containing 0.2 M base acid and 0.2 M dibasic acid, and then reacted with a substrate (testosterone) with an isotope, ³ H, to measure the amount of product dihydrootestosterone Respectively.

The reaction solution contained 1 mM dithiothreitol, 40 mM sodium phosphate (pH 6.5), 50 μM NADPH, [1,2,6,7- ³ H] testosterone / testosterone (2.2 × 10 ⁹ mol) and enzyme suspension .

The ginseng fruit extract of Example 1 was dissolved in 10% ethanol and added to 10 μl of 100 μg / 10% ethanol per reaction. Riboflavin was used as a control group in the same volume and riboflavin was used as a positive control.

The reaction was started at the same time as adding the enzyme suspension and proceeded at 37 DEG C for 30 minutes and then extracted with 1 mL of ethyl acetate. 100 mu l of the ethyl acetate phase was applied onto a silica plastic sheet kieselgel 60 FEP 254 60 F254), the developing solvent system was developed with acetate-cyclohexane (1: 1).

After the plastic sheet was air-dried, the bath system was used to measure the amount of isotope. The dried plastic sheet and x-ray film were put in a cassette together with the isotope amounts of testosterone and dihydrotestosterone remaining on the film after one week And the results for each inhibition rate are shown in Table 16 below.

Test substance T (dpm) DHT (dpm) Conversion Rate (%) Inhibition rate (%) Example 1 6878 2355 25.5 39.9 Control group 7520 5536 42.4 - Positive control group 6300 2530 28.7 32.4 (1) T: 3H radioactivity in the testosterone area
(2) DHT: 3H radioactivity in the dihydrotestosterone region
(3) Conversion rate: Radio activity in the DHT area / Total radio activity
(4) Inhibition rate: 100 x (Conversion rate of control group - Conversion rate of sample) / Conversion rate of control group

From Table 16, it can be seen that the ginseng fruit extract has an inhibitory effect on 5-alpha reductase.

2) Inhibitory effect on lipogenesis

The extent of lipid synthesis in the sebaceous glands was evaluated by assessing the extent of uptake of carbon elements required for lipid synthesis and assessing the degree of lipid synthesis inhibition.

In other words, the amount of inhibition was evaluated by biopsy of hamster ear tissues having a large number of sebaceous glands, and comparing and quantifying the test substance with the control group using a radioactivity measuring device for 6 hours on a C14 labeled medium. Test substances were used by adding 0.01% test substance solution to the medium using the right ear tissue of the hamster, and 0.01% physiological saline was added to the medium using the hamster left ear tissue as a control group. The results are shown in the following Table 17 .

On the other hand, in Table 17, the inhibition rate is calculated according to the following equation (4), and the value is an average value for four hamsters.

Test substance Inhibition rate (%) Example 1 47 The positive control (retinoic acid) 52

From the results shown in Table 17, it can be seen that the ginseng fruit extract of Example 1 according to the present invention has an effect of inhibiting lipid biosynthesis.

3) Effect of Cotton Reduction

The ginseng fruit extract of Example 1 used in the present invention was dissolved in 1,3-butylene glycol so as to be a 1% solution and used as a test substance for rabbit clinical experiment in order to confirm the cottonseed reduction effect.

In the ear of a white rabbit, 0.2 ml of 50% oleic acid / paraffin oil, which is a skin inducing substance, was applied to the ear and rubbed with a cotton swab. Twice daily (50% oleic / paraffinic oil) and in the afternoon (substance) and applied for one month, the total duration of the experiment. The cottonseed occurs from about 10 days afterwards. As a result of applying 0.2 ml of the ginseng fruit extract of Example 1 in the afternoon, the number and size of the cottonseed gradually decreased after 2 weeks.

Each tissue was biopsied and the keratin was peeled off and the size of the cotton was measured under a microscope. The results are shown in Table 18 below.

Test substance Average skin size measurement (0.1 mm) N = 3 Control group (untreated group) 1.78 Oleic acid (OA) 3.5 Oleic acid (OA) + Example 1 2.3

From the results of Table 18, it can be seen that the rabbit skin treated with the ginseng fruit extract of Example 1 according to the present invention was significantly reduced. That is, it was estimated that the sebum extract was reduced by the ginseng fruit extract of Example 1, and as a result, the skin and keratinization were reduced, and the skin texture was reduced.

4) Effect of suppressing sebum secretion

In order to measure the oily skin mitigation effect, 20 women aged 20-45 years, 20 men with oily skin, 10 men and 2 women in the forehead region, After 2, 3, and 4 weeks of application, the skin sebum secretion amount was measured using a skin moisture analyzer (Sebumeter SM810). Experiments were carried out at 20 ° C and 20% relative humidity. The measurement results are shown in Table 19. The results were expressed as the average value of 10 subjects. In the case of oily skin, the measurement value is 220 or more, and in the case of normal skin, it is between 100 and 220.

Effect of suppressing sebum secretion Test substance Before application 1 week 2 weeks 3 weeks 4 weeks Comparative Example 2 245 240 239 235 230 Example 2 245 236 231 218 209

From the results shown in Table 19, it was confirmed that the nutritional cream of Example 2 containing the ginseng fruit extract of Example 1 according to the present invention had an effect of suppressing excessive sebum secretion and had sebum control effect.

5) Anti-inflammatory effect: Inhibitory effect of prostaglandin E2 (PGE2) production

In order to examine the inhibitory effect of the ginseng fruit extract according to the present invention on the production of prostaglandin E2, the following experiment was carried out (Reference: Jeffrey, KH, Anglea, SW, Zoe, S., and Rhia CM Intracellular Measurement of Prostaglandin E2: Effect of Anti-inflammatory Drugs on Cyclooxygenase Activity and Prostanoid Expression, Analytical Biochemistry, 1999, 271, 18-28).

First, the fibroblasts isolated from the human epidermal tissue were cultured in a CO ₂ incubator for 24 hours, then replaced with a medium containing no FBS (fetal bovine serum), and cultured for 2 hours. Then, The extracts were mixed at a concentration of 1 μg / mL, 10 μg / mL and 100 μg / mL, respectively, and then cultured in a CO ₂ incubator at 37 ° C. and 5% (v / v) for 2 hours .

Then, calcium iodate A23187 (Calcium ionophore A23187) and arachidonic acid were treated at a concentration of 50 쨉 m for 5 minutes as a prostaglandin E2 stimulus source, and the cells were hemolyzed and the absorbance was measured at 450 nm in an ELISA reader to quantitate prostaglandin E2 , And the results are shown in Table 20 below. The inhibition rates shown in Table 20 below were calculated based on the difference in the amounts of prostaglandin E2 produced in the fibroblasts treated with the ginseng fruit extract of Example 1 and the ginseng fruit extract treated with the ginseng extract of Example 1 The mean value was obtained by repeating 5 times.

Test substance density
(쨉 g / mL) Prostaglandin E2 (pg / 10 ⁵ cells) % Inhibition Negative control group - 25.2 ± 4.4 - Arachidonic acid - 385.1 + - 9.1 -
Arachidonic acid + Example 1 One 289.6 ± 6.5 24.8 10 246.4 ± 8.6 36.0 100 195.6 ± 14.3 49.2

From the results of Table 20, it can be confirmed that the extract of ginseng fruit of Example 1 according to the present invention can inhibit the production of prostaglandin E2 by a stimulus source.

6) Anti-inflammatory effect: inhibition of biosynthesis of cyclooxygenase-2 (COX-2) induced by Lipopolysaccharide (LPS)

Human fibroblasts were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , and after the LPS treatment, the ginseng fruit extract of Example 1 was replaced with a medium containing 1 ppm, 10 ppm and 100 ppm. On the second day of culture, cells were harvested and the amount of cyclooxygenase-2 (COX-2) produced using the Westren blot method was quantitated. The comparison with the value measured with a densitometer with the control group as 100 is shown in Table 21 below. The control group was an untreated group in which the ginseng fruit extract of Example 1 was cultured without treatment.

Test substance (concentration: ppm) COX-2 biosynthesis (%) Example 1 100 15 10 35 One 71 Control group 100

As can be seen from the results of Table 21, the ginseng fruit extract of Example 1 according to the present invention inhibited the inflammatory reaction by reducing the biosynthesis of cyclooxygenase-2 induced by LPS, which is an inflammatory reaction inducer, And showed efficacy to alleviate skin troubles.

7) Anti-inflammatory effect: Inhibition effect of LPS (Lipopolysaccharide) -induced tumor necrosis factor alpha (TNF-α)

Human keratinocytes were cultured in a 12-well plate incubator at a concentration of 10 ⁵ , and after the treatment with LPS, the medium containing 1 ppm, 10 ppm and 100 ppm of the ginseng fruit extract of Example 1 was replaced. After incubation for 6-24 hours, the cells were harvested and the amount of tumor necrosis factor alpha (TNF-a) produced was quantitated using the ELISA (Pharmingen 555212) method. The comparison with the value measured with the control group as 100 is shown in Table 22. &lt; tb &gt;&lt; TABLE &gt; The control group was an untreated group in which the ginseng fruit extract of Example 1 was cultured without treatment.

Test substance (concentration: ppm) TNF-α biosynthesis (%) Example 1 100 14 10 41 One 69 Control group 100

As can be seen from the results of Table 22, the ginseng fruit extract of Example 1 according to the present invention decreases the biosynthesis of tumor necrosis factor alpha (TNF-α) by inflammation inducing factor LPS in a concentration-dependent manner, It was possible to suppress the inflammatory reaction and to contribute to the improvement of skin troubles.

8) NO (nitric oxide) inhibition effect induced by LPS on macrophages

In order to examine the anti-inflammatory effect of the ginseng fruit extract according to the present invention, LPS-induced NO (nitric oxide) inhibition experiments were performed on macrophages.

Macrophages (Raw 264.7 cells) were cultured in 5% CO ₂ medium supplemented with 10% serum. 96 after the ball cultivated in the plate culture medium to 2 × 10 ⁵ different concentrations, and each process and then stimulated with LPS (1 ug / ml) Example 1 of the ginseng fruit extract and ginseng extract retained for 1 hour at 37 ℃ NO The degree of production was measured and the effect of inhibiting NO induced by LPS was compared. The NO production amount was measured by the Griess reaction method (Minghetti, L. et al., 1991, Glia 19. 152-160), and the results are shown in FIG.

As shown in the results of FIG. 5, the ginseng fruit extract of Example 1 according to the present invention effectively inhibited NO production as an inflammatory factor (cytokine), and its effect was much better than that of red ginseng extract. Therefore, the ginseng fruit extract of Example 1 according to the present invention is expected to contribute to the effect of healing wounds due to skin troubles through the control of inflammation and inhibiting wrinkle formation.

9) Acne and skin trouble improvement effect

The subjects were 10 women with severe skin troubles including acne. The subjects were soaked in soap for 30 minutes in the temperature and humidity chamber, and the skin trouble was measured by comparing the number of acne and inflammation .

The subjects were allowed to use the nutritional creams of Example 2 and Comparative Example 2 for 4 weeks, respectively, and the degree of acne and skin troubles were compared. The results are shown in Table 23 below.

Example 1
(5 people) Comparative Example 1
(5 people) Skin Trouble Improvement Ratio 80% 20%

As shown in the results of Table 23, when Example 2 containing the ginseng fruit extract of Example 1 according to the present invention was applied for 4 weeks, reduction in the number of existing acnes and improvement of inflammatory accompanying trouble were shown.

[Test Example 6] Skin color fastness improving effect

1) NO production in human umbilical vein endothelial cell (HUVEC) of ginseng fruit extract

Endothelial nitric oxide synthase (eNOS) is present in the vascular endothelial cells of humans and its activity is increased to produce NO (nitric oxide) to expand blood vessels and promote blood circulation. After culturing human vascular endothelial cells, the amount of NO produced by treating ginseng fruit extract (FIG. 6) and common red ginseng extract (FIG. 7) on the medium was compared. In other words, vascular endothelial cells were attached at a density of 2.5 × 10 ⁴ cells / well in a gelatin-coated 24-well plate incubator, and then cultured in a growth medium for 12 hours. The endothelial cells of the vascular endothelium were pretreated for 12 hours with the group treated with the ginseng fruit extract and red ginseng extract of Example 1 according to the present invention and the group without treatment. Endothelial cells were treated with 10 μmol / L DAF-FM diacetate (Molecular Probe, OR) in M199 medium without FBS for 30 min at 37 ° C. The vascular endothelial cells were washed three times with M199 medium without FBS, and then placed in a parallel plate flow chamber and stimulated with light separated from a mercury lamp. The excitation wavelength is 488 nm, and NO-coupled DAF exhibits fluorescence at 515 nm. The fluorescence intensity was measured with a confocal laser microscope (Atto Bioscience, USA), and the brightness of the fluorescence was analyzed with Image-Pro Plus v4.5 software (Media Cybernetics, San Diego, CA, USA) Respectively.

6 and 7, the ginseng fruit extract of Example 1 according to the present invention showed a NO production rate of 500 to 1300% compared to that of the non-treated group, and the extract of general red ginseng showed 100 to 200% NO (50 to 100 ug / ml of the same concentration). Thus, the ginseng fruit extract of Example 1 according to the present invention showed superior NO production ability in vascular endothelial cells as compared with the red ginseng extract. The excellent NO (nitric oxide) production ability of the ginseng fruit extract of Example 1 can expand the capillary blood vessels and promote blood circulation, thereby smoothly supplying nutrients to the skin, suppressing skin aging, and improving skin color Contributing to the efficacy.

2) Improvement of color of human skin (face)

In order to evaluate the effect of accelerating the blood circulation in the skin of the composition containing the fruit extract of 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 the skin. It is a highly sensitive device that can measure not only the velocity and quantity of blood in the capillaries of the skin but also the flow in the small arteries and the parenchyma .

The face was soaked with soap and allowed to adjust for 30 minutes. The initial value was measured using an IR (Infrad) camera with LDPI and a skin temperature measuring instrument. The initial blood flow in the lower part of the forehead was measured by LDPI and the initial temperature of the forehead, under the eyes and cheeks was measured using an IR camera.

The blood flow and skin temperature were compared with initial measurements after using Example 2 for a week, and the results are shown in Tables 24 and 25 below.

LDPI results before and after use Face Before use After 1 week use medium 0.952 1.169

IR results before and after use forehead Under the eyes cheek Before use 32.26 32.02 30.44 After 1 week use 33.87 33.69 33.57

As described above, as a result of using the Example 2 containing the extract of the ginseng fruit according to the present invention, it was confirmed that the blood color was improved due to the peripheral blood circulation improving effect on the faces of the subjects, This suggests that when the fruit extract contains the nutrients of the skin, it can contribute to inhibiting and delaying the aging of the skin.

[Formulation Example 1] Softening lotion (skin lotion)

Compounding ingredient Content (% by weight) Purified water Balance Ginseng fruit extract 0.1 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Phage-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, fragrance Suitable amount

[Formulation Example 2] Nutritional lotion (Milk lotion)

Compounding ingredient Content (% by weight) Purified water Balance Ginseng fruit extract 0.1 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Montana 202 (Manufacturer: Seppic) 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount

[Formulation Example 3] Massage cream

Compounding ingredient Content (% by weight) Purified water Balance Ginseng fruit extract 0.1 Wax 10.0 Polysorbate 60 1.5 Piggy 60 hardened castor oil 2.0 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Montana 202 (Manufacturer: Seppic) 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservative, coloring, fragrance Suitable amount

[Formulation Example 4] Pack

Compounding ingredient Content (% by weight) Purified water Balance Ginseng fruit extract 0.1 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 Pingyi-12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservative, coloring, fragrance Suitable amount

Claims (3)

A cosmetic composition for improving acne skin containing ginseng fruit extract as an active ingredient. [Claim 7] The cosmetic composition for improving acne skin according to claim 1, wherein the ginseng fruit extract is prepared by extracting ginseng fruit and ginseng fruit by separating and removing seeds from sun or dry air, followed by extraction with water or ethanol. The cosmetic composition according to claim 1, wherein the cosmetic composition is selected from the group consisting of softening agents, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, , Body oil, and body essence. &Lt; RTI ID = 0.0 &gt; 11. &lt; / RTI &gt;

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