KR101460569B1 - Cosmetic composition comprising of gingenoside F2 for skin wrinkle improvement, skin whitening or anti-acne - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving wrinkles, skin whitening or acne improvement comprising ginsenoside F2 as an active ingredient. The ginsenoside F2 of the present invention has an effect of promoting fibroblast proliferation, increasing collagen synthesis and improving wrinkles through inhibition of elastase; Not only has a skin whitening effect through melanin synthesis inhibitory activity and tyrosinase inhibitory activity; At the same time, acne-causing bacteria Propionebacterium acne ( Propionebacterium Stability of Philo coke acne) and hwanonggyun Alas Reus (Staphylococcus aureus , and the like. Therefore, the composition of the present invention containing it as an effective ingredient can effectively improve skin wrinkles, skin whitening, and acne, and thus can be usefully used as a functional cosmetic composition .

Description

[0001] The present invention relates to a cosmetic composition for improving wrinkles, skin whitening or acne, comprising ginsenoside F2 as an active ingredient,

The present invention relates to a cosmetic composition for improving wrinkles, skin whitening or acne improvement comprising ginsenoside F2 as an active ingredient.

In recent years, cosmetics industry has been expanding as interest in women 's skin beauty has increased. However, the development of technology using natural cosmetics has been a trend in recent years as the technology has developed and the adverse effects on skin caused by synthetic cosmetics are increasing.

With respect to skin whitening, conventionally, substances having ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione, derivatives thereof, or tyrosinase inhibiting activity have been used in combination with cosmetics or medicines. However, The use thereof is limited due to the whitening effect, the safety problem to the skin, the formulation and the stability of the cosmetic composition.

In order to maintain proper skin moisture, studies have been conducted to supply moisture from the outside or to prevent water loss from the body. In fact, many kinds of moisturizers having water retention ability have been developed, It is widely used in the area. However, due to abnormal epidermal cell division and differentiation, skin causes various skin diseases such as dryness of skin, atopy and psoriasis. Such diseases can be slightly alleviated by a conventional moisturizing agent having only water retention function , But it is difficult to expect fundamental healing.

Among the causes of acne, which are mainly found in adolescence, Propionebacterium acne ), and the microorganisms decompose the sebum in the pores into free fatty acids in the anaerobic environment in the clogged pores, and thus the free fatty acids give a severe irritation to the skin to cause inflammation Resulting in the formation of redness and congestion. Representative substances used in the treatment of acne include antihistamines for inhibiting sebum formation, corticosteroids and nonsteroidal anti-inflammatory agents showing antiinflammatory action, tetracycline and erythromycin, which are antibiotics for inhibiting the proliferation of acne, (U.S. Pat. Nos. 4,469,684; 5,045,533).

However, the above-mentioned conventional substances used for the treatment of acne may have serious side effects besides the proliferation inhibiting / treating effect of acne bacteria. In the case of anti-male hormone, it is effective in decreasing androgen, which is a male hormone that affects the occurrence of acne. However, the target of treatment is confined to women, and in the case of steroid anti-inflammatory analgesic, , The use of antibiotics not only removes the useful microorganisms present on the skin but also causes various side effects such as digestive disorders, phototoxic dermatitis, secondary infection, vaginitis, perianal pruritus, etc., and substances such as vitamin A or its derivatives In general, the effect is good, but the lips and mouth mucosa dry, palms keratin peel off, and may cause side effects such as itching of the whole body. Especially when you take too much when pregnant, there is a risk of malformations. In vitro growth by metabolism This time, there are problems that need attention, such as one month after the end of a long period of time and then taking.

Therefore, in order to solve the above problems, there is a need to develop a product having excellent effects such as wrinkle improvement, whitening, acne improvement, and atopic dermatitis improvement, while being safe for the human body even in long term use using functional materials derived from natural materials In fact.

On the other hand, saponin ginsenoside, which is known as the main active ingredient of ginseng (Panax Ginseng CA Meyer), is a component contained in plants such as ginseng, white ginseng, ginseng, ginseng and ginseng cultivating roots, and contains about 20 triols and diols It is classified as senoside. Among them, various activities have been studied centering on ginsenosides such as Rb1, Rb2, Rh1, Rh2, Rg1, Rg2, and Rg3, and studies on cosmetic compositions using these have also been conducted.

In this regard, cosmetics containing black ginseng extract containing a large amount of ginsenoside Rg3 have the effect of improving skin wrinkles (Journal of the Academia-Industrial cooperation Society, 11 (9): 3325-3329, 2010) . In addition, a cosmetic composition containing a single ginsenoside Rg3 has been patented (Korean Patent Laid-Open No. 10-2003-0065882) by clarifying the skin improving effect of ginsenoside Rg3.

However, there has been a patent application (Korean Patent Laid-Open No. 10-2009-0083780) concerning the prevention and treatment of dementia in the conventional studies related to ginsenoside F2, There is no research on

Accordingly, the present inventors focused on ginsenoside F2 and examined its pharmacological effect while searching for a natural substance having no side effects on the body and excellent wrinkle, skin whitening or acne improvement effect.

As a result, ginsenoside F2 has the effect of promoting fibroblast proliferation, increasing collagen synthesis, and improving wrinkles through inhibition of elastase; Not only has a skin whitening effect through melanin synthesis inhibitory activity and tyrosinase inhibitory activity; At the same time, acne-causing bacteria Propionebacterium acne ( Propionebacterium Stability of Philo coke acne) and hwanonggyun Alas Reus (Staphylococcus aureus , and the like. Thus, the present invention has been completed.

Korean Patent Publication No. 10-2003-0065882 Korean Patent Publication No. 10-2009-0083780

Accordingly, an object of the present invention is to provide a functional cosmetic composition which can effectively improve wrinkles, skin whitening or acne without adverse effects on the living body.

In order to accomplish the object of the present invention as described above, the present invention provides a cosmetic composition for improving wrinkles, skin whitening or acne, comprising ginsenoside F2 as an active ingredient.

In one embodiment of the present invention, the ginsenoside F2 may be contained in an amount of 0.0001% to 20% based on the total weight of the composition.

In one embodiment of the present invention, the composition may have a wrinkle-reducing effect through promoting fibroblast proliferation, increasing collagen synthesis, and inhibiting elastase activity.

In one embodiment of the present invention, the composition may have a skin whitening effect through a melanin synthesis inhibitory activity and a tyrosinase inhibitory activity.

In one embodiment of the present invention, the composition is selected from the group consisting of Propionebacterium acne-causing bacteria Stability of Philo coke acne) and hwanonggyun Alas Reus (Staphylococcus aureus ) can have an acne improvement effect.

The ginsenoside F2 of the present invention has an effect of promoting fibroblast proliferation, increasing collagen synthesis and improving wrinkles through inhibition of elastase; Not only has a skin whitening effect through melanin synthesis inhibitory activity and tyrosinase inhibitory activity; At the same time, acne-causing bacteria Propionebacterium acne ( Propionebacterium Stability of Philo coke acne) and hwanonggyun Alas Reus (Staphylococcus aureus , and the like. Therefore, the composition of the present invention containing it as an effective ingredient can effectively improve skin wrinkles, skin whitening, and acne, and thus can be usefully used as a functional cosmetic composition .

Figure 1 shows the chemical structure of ginsenoside F2.
Fig. 2 shows the cell proliferation effect according to the treatment of ginsenoside F2 at different concentrations.
Fig. 3 shows the inhibitory activity of the elastase activity according to the treatment of ginsenoside F2 by concentration.
FIG. 4 shows melanin synthesis inhibition activity (%) by treatment with ginsenoside F2 at different concentrations.

The present invention is characterized by providing a cosmetic composition for improving wrinkles, skin whitening or acne improvement comprising ginsenoside F2 as an active ingredient.

The ginsenoside F2 of the present invention is a kind of ginseng saponin contained in ginseng, red ginseng, white ginseng, fresh ginseng, ginseng and wild ginseng culture roots, and is a kind of ginseng saponin (20S) -3β- (β-D-Glucopyranosyloxy ) -20- (β-D-glucopyranosyloxy) dammar-24-ene-12β-ol, the molecular formula is C ₄₂ H ₇₂ O ₁₃ , and the molecular weight is 785.01 (see FIG.

The inventors of the present invention have focused on ginsenoside F2 and examined its pharmacological effects while searching for a natural substance having no side effects on living body and excellent wrinkle, skin whitening or acne improvement effect. As a result, Promoting cell proliferation, increasing collagen synthesis, and improving wrinkles through elastase inhibitory activity; Not only has a skin whitening effect through melanin synthesis inhibitory activity and tyrosinase inhibitory activity; At the same time, it was confirmed that the antimicrobial activity against the acne-causing bacteria Propionebacterium acne and the bacteria Staphylococcus aureus is also effective for improving acne.

In the following Example 2 of the present invention, the wrinkle-reducing effect of the ginsenoside F2 of the present invention was confirmed. As a result, in the experimental group treated with the ginsenoside F2 at a concentration of 5% and 10%, a high fibroblast proliferation effect In addition, in the experimental group treated with Jinsephonoside F2 at a concentration of 0.5% or more, it showed a collagen synthesis rate of 50% or more, which effectively promoted collagen synthesis. In addition, the experimental group treated with Jinsephonoside F2 at a concentration of 0.5% or more showed an inhibitory activity of about 77-78%, indicating that ginsenoside F2 is effective for improving skin wrinkles.

In the following Example 3 of the present invention, the skin whitening activity of ginsenoside F2 of the present invention was measured. As a result, it was confirmed that the inhibition rate of melanin synthesis was increased in a concentration-dependent manner in the experimental group treated with ginsenoside F2, And 31%, respectively. Furthermore, it was shown that the inhibitory efficiency of tyrosinase was increased depending on the treatment concentration of ginsenoside F2, and finally it was confirmed that the inhibition rate was about 32%. From these results, it was confirmed that melanin synthesis can be effectively inhibited by inhibiting tyrosinase enzyme of ginsenoside F2, which is useful for skin whitening.

In the following Example 4 of the present invention, the ginsenoside F2 inhibitory effect of the present invention was confirmed, and as a result, it was found that Propionebacterium acne , an acne-causing germ, The concentration of Staphylococcus aureus was rapidly decreased. These results confirmed that the ginsenoside F2 effectively inhibited the acne and inflammation-causing strains of the skin . In addition, clinical studies have shown that acne improvement / reduction of sebum secretion / irritation has been shown to reduce acne improvement as well as sebaceous secretion.

In the following Example 5 of the present invention, a clinical trial was conducted to confirm the effect of the ginsenoside F2 of the present invention on improving atopic dermatitis. As a result, about 90% of the effect of using ginsenoside F2 as 1% And it was confirmed that there was a significant effect on atopic dermatitis.

Through these results, the present inventors have experimentally proved that ginsenoside F2 is effective for improving wrinkles, skin whitening, acne and atopic dermatitis.

The ginsenoside F2 may be a commercially available ginsenoside F2, or it may be prepared by a method commonly used in the art.

In the following Example 1 of the present invention, methanol was added to ginseng and the mixture was refluxed to obtain a methanol extract. Methanol was evaporated to remove the residue. The residue was suspended in water and extracted three times with ether. The remaining aqueous layer was washed with butanol / ethyl acetate (10: 1). The remaining water layer was extracted three times with water saturated butanol to obtain a fraction containing saponin. The saponin extract obtained above was dissolved in a sodium acetate buffer solution and reacted by adding β-glucosidase (EC 3.2.1). Then, the reaction solution was heated to stop the enzyme reaction, and the enzyme and the sugar were removed by butanol (BU) extraction method. The obtained sample was analyzed by HPLC and the chromatogram of the ratio of Rb1 / Rg1 was changed from 1.2 to 9.1 . 218 mg of fraction 7 (BU-7) was obtained from a 600 mg butanol fraction thus converted, using a silica gel column (mobile phase, chloroform: methanol: water = 7: 3: 1). BU-7 fractions were separated using ODS column chromatography (acetone: water = 1: 1) to obtain detailed fractions. ODS column chromatography and C18 column chromatography, the ginsenoside-F2 was finally separated and recrystallized.

When the composition of the present invention is prepared with a cosmetic composition, the composition of the present invention may contain not only the above-mentioned ginsenoside F2 but also components commonly used in cosmetic compositions such as antioxidants, stabilizers , Customary adjuvants such as solubilizers, vitamins, pigments and flavoring agents, and carriers.

In addition to the above-mentioned ginsenoside F2, the composition of the present invention may be mixed with a wrinkle remover, a skin lightening agent or an anti-inflammatory agent which has been used in the past so long as it reacts with ginsenoside F2 and does not impair the skin protecting effect.

Examples of products to which the cosmetic composition of the present invention can be added include cosmetics such as astringent lotion, softening longevity lotion, nutrition lotion, various creams, essences, packs, foundation and the like, cleansing, cleanser, soap, .

As a specific formulation of the cosmetic composition of the present invention, there may be mentioned a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, It includes formulations such as soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, latex, lipstick, makeup base, foundation, press powder, loose powder, eye shadow.

According to a preferred embodiment of the present invention, the content of ginsenoside F2 of the present invention is 0.0001-20% by weight, preferably 0.5-20%, more preferably 1.0-20% by weight based on the total weight of the composition . If the content of ginsenoside F2 is less than 0.0001% by weight, the effect of wrinkle improvement, whitening or acne improvement by ginsenoside F2 is greatly reduced, and if it exceeds 20% by weight, skin irritation may be caused, May occur.

Meanwhile, the cosmetic composition according to the present invention may be formulated by containing ginsenoside F2 in the inside of a nanoliposome and stabilizing it. When the ginsenoside F2 is contained in the inside of the nanoliposome, the components of ginsenoside F2 are stabilized, thereby solving problems such as precipitation, discoloration, and discoloration during formulation, solubility and transdermal absorption rate of the component can be increased, It is possible to maximize the efficacy expected from the extract.

In the present invention, nanoliposome refers to a liposome having a typical liposome form and having an average particle diameter of 10 to 500 nm. According to a preferred embodiment of the present invention, the average particle diameter of the nanoliposome is 50 to 300 nm. When the average particle diameter of the nanoliposome is more than 300 nm, improvement of skin penetration and improvement of formulation stability is very weak among technological effects to be achieved in the present invention.

The nanoliposomes used to stabilize the extract according to the present invention may be prepared by a mixture comprising a polyol, an oily component, a surfactant, a phospholipid, a fatty acid and water.

The polyol used in the nanoliposome of the present invention is not particularly limited and is preferably a polyol such as propylene glycol, dipropylene glycol, 1,3-butylene glycol, glycerin, methylpropanediol, isopropylene glycol, pentylene glycol, erythritol, , Sorbitol, and mixtures thereof. The amount thereof is 10 to 80% by weight, preferably 30 to 70% by weight, based on the total weight of the nanoliposome.

The oil component used in the preparation of the nanoliposome of the present invention may be selected from a variety of oils known in the art and is preferably a hydrocarbon oil such as hexadecane and paraffin oil, Vegetable oils such as silicon oil such as dimethicone and cyclomethicone, sunflower oil, corn oil, soybean oil, avocado oil, sesame oil and fish oil, ethoxylated alkyl ether oils, propoxylated alkyl ether oils ₄₀ is a fatty alcohol, and mixtures thereof -, phytosphingosine, sphingosine, and scan non-ping the same scan pinggo cannabinoid lipid, cholesterol side-by celebrity, sitosterol cholesteryl sulfate, cholesteryl sulfate, cytokines, C _10. The amount thereof may be 1.0 to 30.0% by weight, and preferably 3.0 to 20.0% by weight based on the total weight of the nanoliposome.

Any surfactant known in the art may be used as the surfactant used in the preparation of the nanoliposome of the present invention. For example, anionic surfactants, cationic surfactants, amphoteric surfactants and nonionic surfactants can be used. Preferred are anionic surfactants and nonionic surfactants. Specific examples of anionic surfactants include alkyl acyl glutamates, alkyl phosphates, alkyl lactylates, dialkyl phosphates and trialkyl phosphates. Specific examples of nonionic surfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters, alkyl polyglycosides, polyglyceryl esters and sugar esters. Particularly preferred surfactants are polysorbates belonging to nonionic surfactants. The amount thereof may be 0.1 to 10% by weight, preferably 0.5 to 5.0% by weight, based on the total weight of the nanoliposome.

The phospholipid, another component used in the preparation of the nanoliposomes of the present invention, is a lipophilic lipid that is used with both affinity lipids and is composed of natural phospholipids (e.g., egg yolk lecithin or soy lecithin, sphingomyelin) Dipalmitoyl phosphatidylcholine or hydrogenated lecithin), preferably lecithin. In particular, unsaturated lecithin or saturated lecithin derived from natural origin extracted from soybean or egg yolk is preferable. Normally, the amount of phosphatidylcholine is 23 to 95% and the amount of phosphatidylethanolamine is 20% or less in natural derived lecithin. In the production of the nanoliposome of the present invention, the amount of the phospholipid to be used is 0.5 to 20.0% by weight, preferably 2.0 to 8.0% by weight based on the total weight of the nanoliposome.

Fatty acid to be used in nano-liposome preparation of the present invention is a higher fatty acid, preferably a C ₁₂ - ₂₂ as a saturated or unsaturated fatty acid of the alkyl chain, e.g., lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid and linoleic acid . The amount thereof may be 0.05 to 3.0% by weight, preferably 0.1 to 1.0% by weight, based on the total weight of the nanoliposome.

The water used in the preparation of the nanoliposome of the present invention is generally deionized distilled water, and the amount thereof may be 5.0-40 wt% based on the total weight of the nanoliposome.

The preparation of nanoliposomes can be accomplished through a variety of methods known in the art, but most preferably is made by applying a mixture comprising the ingredients to a high pressure homogenizer. The preparation of a nanoliposome by a high pressure homogenizer can be carried out under various conditions (for example, pressure, number of times, etc.) depending on a desired particle size, and preferably at a high pressure homogenizer So that the nanoliposome can be produced.

The cosmetic composition for skin improvement according to the present invention may contain 0.1 to 20.0% by weight based on the weight of the ginsenoside F2 nanoliposome.

The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease comprising ginsenoside F2 as an active ingredient.

The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.

The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.

The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

In one embodiment of the present invention, the ginsenoside F2 of the present invention may be included in an amount of 0.0001 to 20% by weight based on the total weight of the composition.

In another embodiment of the present invention, the pharmaceutical composition of the present invention may be a composition for external application for skin.

The pharmaceutical composition for dermatological use of the present invention is characterized in that the acne-causing bacteria, Propionebacterium acne) and hwanonggyun of Staphylococcus aureus (Staphylococcus aureus) as an external preparation for skin creams, gels, patches, sprays, ointments, alert agents, lotions, shall have an antimicrobial effect with respect to the cement agent, pasta claim or kata plasma claim Of the present invention can be used as a pharmaceutical composition in the form of external preparation for skin, but the present invention is not limited thereto.

Skin inflammatory diseases for which the pharmaceutical composition of the present invention can exhibit a therapeutic effect include acne, hypersensitive dermatitis, atopic dermatitis, insect allergies, food allergies, drug allergies and urticaria, But is not limited thereto.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1>

Gin Senocide  Isolation and preparation of F2

1 kg of ginseng was refluxed with 1000 ml of methanol to obtain a methanol extract. Methanol was evaporated to remove the residue. The residue was suspended in 100 ml of water and extracted three times with 100 ml of ether. The remaining aqueous layer was washed with butanol / ethyl acetate (10: 1) mixed solution (100 ml) three times. The remaining water layer was extracted with 100 ml of water-saturated butanol three times to obtain a fraction containing saponin.

The resulting saponin extract was dissolved in 10 mM sodium acetate buffer (pH 5.0) at a concentration of 2%, 4.0 unit / ml of β-glucosidase (EC 3.2.1) was added thereto, and the mixture was incubated at 37 ° C. for 48 hours Lt; / RTI &gt; The enzyme reaction was stopped by boiling the reaction solution, and enzymes and saccharides were removed by butanol (BU) extraction method. The obtained sample was analyzed by HPLC to obtain a chromatogram in which the ratio of Rb1 / Rg1 was changed from the initial 1.2 to 9.1. 218 mg of fraction 7 (BU-7) was obtained from a 600 mg butanol fraction thus converted, using a silica gel column (mobile phase, chloroform: methanol: water = 7: 3: 1). BU-7 fractions were separated using ODS column chromatography (acetone: water = 1: 1) to obtain detailed fractions. ODS column chromatography and C18 column chromatography, the ginsenoside-F2 was finally separated and recrystallized.

< Example  2>

Gin Senocide  Measuring wrinkle improvement effect of F2

In order to confirm the wrinkle-reducing effect of the ginsenoside F2 of the present invention prepared in Example 1, the effect of promoting fibroblast proliferation and promoting collagen synthesis was measured.

<2-1> Fibroblast Proliferative ability  Measure

Dermatological atrophy is a typical aging phenomenon after 70 years of age. Changes in the dermis are caused by changes in the number of fibroblasts and of substances with large molecular weights in the extracellular matrix due to a decrease in their ability to synthesize them. In this experiment, fibroblast proliferative capacity was measured using CCD986sk, a human skin fibroblast, in order to measure the wrinkle improving effect.

Fibroblast CCD986sk was cultured in DMEM medium containing fetal bovine serum (FBS) for 24 hours at 37 ° C by dispensing fibroblasts at a concentration of 5 × 10 ³ cells in each well of a 96-well microplate. 100 μl of the ginsenoside F2 obtained by the method of Example 1 was added to the fibroblast culture medium at the concentration (0.1%, 0.5%, 1%, 2%, 5%, 10% ₂ incubator at 37 캜 for 72 hours, and then 50% of 0.2% MTT solution was added to each well and left for 4 hours. After removing the supernatant, DMSO (dimethyl sulfoxide) of 150 was added and formazan was dissolved by shaking for 10 minutes. The absorbance of the formazan dissolved in DMSO was measured at 570 nm using a microplate reader. This was compared with the absorbance of the control group to promote fibroblast proliferation.

Cell proliferation effect (%) = (absorbance of experimental group-absorbance of control group) / absorbance of control group x 100

As a result, as shown in Fig. 2, in the experimental group treated with ginsenoside F2, skin cell proliferation was promoted, and in the experimental group treated with 2% of ginsenoside F2, the effect of fibroblast proliferation was 114.5% I can see that it appears. Particularly, the effect was increased in the concentration-treated group of 2% or more, and it was confirmed that the treated group of 5% and 10% showed a high fibroblast proliferation effect of about 120%.

<2-2> Measurement of collagen synthesis amount

Collagen is the main constituent of the skin and is necessary for constituting the skin, bone, ligament, cartilage and tooth. In addition, collagen has various functions such as mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, induction of cell division and differentiation. It is also reduced by aging and photoaging by ultraviolet irradiation, which is closely related to the wrinkling of the skin. Therefore, in this experiment, the degree of collagen synthesis according to the treatment of ginsenoside F2 was measured as an index of the wrinkle improving effect.

Each of the ginsenosides F2 obtained by the method of Example 1 was prepared by concentration (0.1%, 0.5%, 1%, 2%, 5%, 10%) and dissolved in PBS buffer solution. The culture medium of the CCD986sk cell line cultured in the same manner as described in < 2-1 > was replaced with DMEM medium supplemented with no serum, and then added to CCD986sk cell culture plate to a final concentration of 0-50 μg / , And cultured for 48 hours. After culturing, each cell culture solution was collected and the amount of procollagen type I C-peptide present in the culture solution collected using a collagen protein measurement kit (Takara Shuzo Co., Ltd., Japan) was measured, Respectively. The primary collagen antibody was applied to a 96-well microplate, and the collected CCD986sk cell culture solution was added and reacted at 37 ° C for 3 hours. After removing the cell culture medium, the cells were washed three times with PBS buffer solution. After adding the coloring reagent to each well, the reaction was allowed to proceed at room temperature for 15 minutes. Then, 1 N sulfuric acid solution was added to stop the reaction and the absorbance was measured at 450 nm using an ELISA reader. The absorbance value obtained by the above method was substituted into the standard curve obtained from the standard solution, and the amount of collagen production in the CCD986sk cell culture fluid to which each sample was added was calculated. The synthesis rate of collagen production was obtained from the following equation, and the results are shown in Table 1 below.

Collagen synthesis rate (%) = [(Ax100) / B] -100

(A: collagen production amount of the culture solution to which the sample is added, and B: collagen production amount of the culture solution to which no sample is added)

Measurement of collagen synthesis amount according to the concentration of ginsenoside F2 Concentration (%) of ginsenoside F2 Collagen synthesis rate
(% of control) 0.1 45.04 0.5 50.42 One 51.18 2 52.03 5 52.12 10 53.09

As shown in Table 1, the collagen synthesis rate of 45.04% of the experimental group treated with 0.1% of ginsenoside F2 was high, and the collagen synthesis ratio of 50.42% of the experimental group treated with 0.5% It was confirmed that Jinsephonoside F2 effectively promotes collagen synthesis.

&Lt; 2-3 > Elastase activity Low performance

Elastase is an enzyme that decomposes elastin. If excess elastase is produced, excessive decomposition of elastin causes skin to sag as it sags. Therefore, it inhibits the activity of elastase to prevent skin aging. can do. Therefore, in this experiment, the degree of inhibition (inhibition) of elastase activity by treatment with ginsenoside F2 was measured as another index of the wrinkle-reducing effect.

Elastase activity inhibition was measured using porcine pancreas elastase (pig pancreas elastase, Sigma Co., Ltd USA) and the substrate N-succinyl- (L-Ala) 3-p-nitroanilide (Sigma Co., Ltd USA) Enzyme reaction was investigated. In detail, 0.5 ml of ginsenoside F2 solution was put into a test tube, and 0.5 ml of porcine pancreas elastase (1 U / ml) solution dissolved in 100 mM Tris-HCl buffer (pH 8.0) was added thereto. The solution was then dissolved in DMSO to prepare 3.2 μM N-succinyl- (L-Ala) 3-ρ-nitroanilide (0.1 ml) was added to the reaction mixture, followed by reaction at room temperature for 20 minutes. Elastase activity was measured by measuring the amount of ρ-nitroanilide produced from the substrate using a spectrophotometer at 410 nm. The inhibitory activity of the elastase activity was expressed by the absorbance reduction rate of the addition of the sample solution and the non-addition solution.

As a result, as shown in Fig. 3, the experiment group treated with the above-mentioned ginsenoside F2 at the concentration of 0.1% showed an inhibition of elastase of about 66%. Especially, in the experimental group treated with the concentration of ginsenoside F2 of 0.5% or more, it was confirmed that the effect of the inhibition of elastase was 77 ~ 78%, which is highly effective.

< Example  3>

Gin Senocide  Of F2 Skin whitening  Active measurement

In order to confirm the skin whitening activity of the ginsenoside F2 of the present invention prepared in Example 1, melarin synthesis inhibitory activity and tyrosinase inhibitory activity were measured.

The main function of melanin is to protect the skin from damage by removing harmful radicals. Melanin is produced by ultraviolet rays in melanogenesis cells through the complex oxidation and condensation reaction of tyrosine after being converted into a dopa and dopachrome by an enzyme called tyrosinase. The produced melanin is transferred to the skin cells, and the melanin is lost with epidermal detachment, and the circulating action appears to disappear. This process of melanin production is a naturally occurring phenomenon and does not result in overproduction of melanin in normal human skin. However, when the skin responds to external stimuli such as ultraviolet rays, environmental pollution, stress, or the like, melanin is excessively produced. If excessive melanin is not released out of the skin and remains in the skin, pigmentation will occur. One of the most important features of the melanin generation mechanism is that tyrosinase is involved in only one enzyme. If tyrosinase activity is inhibited and melanin production is prevented, a whitening effect can be expected.

Therefore, in this experiment, it was firstly determined whether ginsenoside F2 inhibits melanin synthesis (production), and it was confirmed whether the inhibitory effect of melanin synthesis is actually inhibited by the activity of tyrosinase enzyme activity.

<3-1> Melanin synthesis inhibitory activity

Melanin synthesis inhibition activity was tested in order to confirm that the ginsenoside F2 of the present invention had activity in skin whitening.

Clone-M3 cells, a mouse-derived melanocyte, produce melanin. Melanin is biosynthesized in the melanosome, an organelle in the melanocytes in the basal layer of the epidermis, which can be quantified by measuring the absorbance of the visible light region. After the inoculation with CloneM-3 cells, each of the ginsenoside F2 samples was treated for 3 days, and the medium was removed. The cells were washed with PBS, 1 ml of 1N NaOH was added per well, and the mixture was melted to melt the melanin component And the absorbance was measured at 400 nm, which is the blue visible ray region having the shortest wavelength among the visible light region band. Clon-M3 cells were treated with the concentration of ginsenosides by concentration, and after 24 hours, the inhibition rate of melanin formation was measured.

As a result, as shown in FIG. 4, it was confirmed that the inhibition rate of melanin synthesis was increased in a concentration-dependent manner in the experimental group treated with ginsenoside F2, and finally, the inhibition rate reached about 31%.

<3-2> Tyrosinase ( Tyrosinase ) Inhibitory activity

In Example 3-1, the ginsenoside F2 of the present invention was found to have melanin synthesis inhibitory activity. In this experiment, it was confirmed that the melanin synthesis inhibitory activity was well known as an enzyme promoting the production of melanin pigment And inhibition of tyrosinase activity.

The tyrosinase inhibitory effect was measured using the dopachrome method. Mushroom Tyrosinase, 225 μl of 2.5 mM L-tyrosine, 225 μl of 0.4 M hepes buffer (pH 6.8), and 300 μl of ethanol solution or sample (1 mg / ml) Before and after incubation for 15 min, absorbance was measured at 475 nm and the degree of inhibition was examined. The degree of inhibition of tyrosinase was calculated as follows, and the results are shown in Table 2 below.

Tyrosinase inhibition (%) = (D-C) - (B-A) /

(A and B are the absorbance before and after the culture of the sample solution, respectively, and C and D are the absorbance values before and after the culture of the unfilled solution (reference solution) .These tyrosinase inhibitory effects are 100 It means complete inhibition when it appears, and zero inhibition at all.)

Tyrosinase inhibition according to the concentration of ginsenoside F2 Concentration (%) of ginsenoside F2 Tyrosinase inhibition (%) 0.1 16.15 0.5 21.21 One 29.93 2 34.55 5 34.36 10 35.03

As shown in Table 2, tyrosinase inhibition efficiency was increased depending on the treatment concentration of ginsenoside F2 of the present invention, and finally, it was confirmed that the inhibition rate was about 32%.

In conclusion, it was confirmed that ginsenoside F2 effectively inhibited melanin synthesis by inhibiting tyrosinase enzyme, which is useful for skin whitening.

< Example  4>

Gin Senocide  Measuring the inhibitory effect of F2 on acne formation

In order to confirm the effect of the ginsenoside F2 of the present invention prepared in Example 1 on the inhibition of the formation of acne, the ginseng root of propionebacterium acne acne and Staphylococcus aureus , which is a pathogen, were evaluated for anti-acne / anti-sebaceous / anti-irritation effects.

<4-1> Acne-causing bacteria Propione bacterium Acne Propionebacterium acne ) And Staphylococcus Aureus ( Staphylococcus aureus )of  Antimicrobial activity measurement

The antimicrobial activity against acne and fungi was measured in order to measure the inhibitory effect of ginsenoside F2 of the present invention prepared in Example 1 on acne formation. Acne bacteria and fungi were cultivated on a brain heart infusion (BHI) medium for 18 hours. Then, in a medium supplemented with a stepwise diluted sample in a 96-well plate, the acne-causing bacteria such as Propionibacterium Propionebacterium acne and Staphylococcus aureus , respectively, were separately inoculated at a concentration of 5% (10 μl). After inoculation, the cells were cultured at 37 ° C for 24 hours. The bacterial counts were measured by microscopic observation of the strains and the antibacterial activity was measured.

Measuring the antimicrobial activity of Propionibacterium acnes and Staphylococcus aureus according to the concentration of ginsenoside F2 Concentration of ginsenoside F2
(%) Propionebacterium acne
(ppm) Propionebacterium acne
(ppm) 0.1 <21 <10 0.5 <15 <10 One <10 <8 2 <7 <2 5 <2 <2 10 <1 <2

As a result, as shown in Table 3 above, it was confirmed that the concentration of ginsenoside F2 was significantly higher than that of Propionebacterium acne-causing bacteria acne and Staphylococcus aureus , which is a pathogenic fungus, were found to be abruptly reduced, thereby allowing ginsenoside F2 to effectively inhibit acne and inflammation-causing strains of the skin I could confirm.

<4-2> Improve acne / decrease sebum secretion / measure irritation

Fifteen people with acne were allowed to use the cosmetic composition prepared 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 results of the experiment are shown in Table 4 below with an average score of 12 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 4 below with an average score of 15 persons. The presence or absence of skin irritation was evaluated as (number of stimuli) / (total number of test subjects). The results are shown in Table 2 below.

Improvement of acne according to the concentration of ginsenoside F2, sebaceous gland secretion and stimulation result Concentration of ginsenoside F2 (%) Acne improvement Decrease sebaceous glands Presence or absence of stimulation 0.1 1.3 3.2 0/15 0.5 2.3 3.5 0/15 One 2.4 4.4 1/15 2 3.7 4.7 2/15 5 3.8 4.5 2/15 10 4.2 4.8 3/15

As a result, as shown in Table 4, it was confirmed that when the cosmetic for improving acne was developed using ginsenoside F2, it was found to be effective in improving acne. In addition, it was confirmed that there was a partial decrease in sebum secretion, and it was confirmed that stimulation was observed in some portions only at very high concentrations for skin irritation. Acquired 4 points or more for acne improvement and 4 points or more for sebaceous gland secretion were obtained, showing excellent effect on acne improvement. Also, it was confirmed that when the concentration of ginsenoside F2 was about 1% or more at the most suitable concentration, the effect for improving acne was excellent.

< Example  5>

Gin Senocide  Measuring the improvement effect of F2 on atopic dermatitis

In order to confirm the effect of the ginsenoside F2 of the present invention prepared in Example 1 on atopic dermatitis, a clinical test was conducted.

Ten out of 20 adults with atopic dermatitis were applied with 1% concentration of ginsenoside F2, and the remaining 10 were applied with water as a control. The test was performed for 10 days by applying 1 ml or 1 g twice daily to cosmetic cotton and applying to the affected area. After 10 days of application, the improvement effect was quantified in four steps: very effective, slightly effective, no effect, and worse. The results are shown in Table 5 below.

The effect of ginsenoside F2 on atopic dermatitis sample Atopic dermatitis improvement Average(%) Water (control) Very good 0 0 good 0 0 no effect 9 90 worse One 10 F2 1% Very good 2 20 good 7 70 no effect One 10 worse 0 0

As a result, as shown in Table 5, about 90% of ginsenoside F2 was used as 1%, and it was confirmed that there was a significant effect on atopic dermatitis. Most of them were not effective in comparative prescriptions using water. Therefore, it was confirmed that the use of ginsenoside F2 of the present invention at 1% or more is effective for atopic dermatitis.

< Example  6>

Gin Senocide  For the treatment of skin inflammation diseases containing F2 Cosmetics  Composition manufacturing

<6-1> Flexible longevity skin )

The softening skin containing the ginsenoside F2 of the present invention was prepared in a conventional manner at the following proportions.

ingredient Production Example 1
(Unit: wt%) Production Example 2
(Unit: wt%) Gin Senocide F2 0.3 12.0 glycerin 5.0 5.0 1,3-butylene glycol 3.0 3.0 REG 1500 1.0 1.0 Allantoin 0.1 0.1 DL-Panthenol 0.3 0.3 EDTA-2Na 0.02 0.02 Benzophenone-9 0.04 0.04 Sodium hyaruronate 5.0 5.0 ethanol 10.0 10.0 Octyldodec-16 0.2 0.2 Polysorbate 20 0.2 0.2 Unicide - Yu 13 0.01 0.01 Distilled water Balance Balance Sum 100 100

<6-2> Nourishing lotion ( lotion )

A nutritional lotion containing the ginsenoside F2 of the present invention was prepared in a conventional manner at the following proportions.

ingredient Production Example 3
(Unit: wt%) Production Example 4
(Unit: wt%) Gin Senocide F2 0.3 12.0 Glyceryl stearate SE 1.5 1.5 Cetearyl alcohol 1.5 1.5 lanolin 1.5 1.5 Polysorbate 60 1.3 1.3 Sorbitastarate 0.5 0.5 Hardened vegetable oil 4.0 4.0 Mineral oil 5.0 5.0 Trioctanoin 2.0 2.0 Dimethicone 0.8 0.8 Tocopheryl acetate 0.5 0.5 Carboxyvinyl polymer 0.12 0.12 glycerin 5.0 5.0 1,3-butylene glycol 3.0 3.0 Sodium hyaruronate 5.0 5.0 Triethanolamine 0.12 0.12 Unicide - Yu 13 0.02 0.02 Distilled water Balance Balance Sum 100 100

<6-3> Nourishing cream

A nutritional cream containing the ginsenoside F2 of the present invention was prepared in a conventional manner at the following proportions.

ingredient Production Example 5
(Unit: wt%) Production Example 6
(Unit: wt%) Gin Senocide F2 0.3 12 Pro-type glycerin monostearate 1.5 1.5 Cetearyl alcohol 1.5 1.5 Stearic acid 1.0 1.0 Polysorbate 60 1.5 1.5 Sorbitastarate 0.6 0.6 Isostearyl isoserate 5.0 5.0 Squalane 5.0 5.0 Mineral oil 35.0 35.0 Dimethicone 0.5 0.5 Hydroxyethylcellulose 0.12 0.12 glycerin 6.0 6.0 Triethanolamine 0.7 0.7 Unicide - Yu 13 0.02 0.02 Distilled water Balance Balance Sum 100 100

<6-4> Pack

A pack containing the ginsenoside F2 of the present invention was prepared in a conventional manner at the following proportions.

ingredient Production Example 7
(Unit: wt%) Production Example 8
(Unit: wt%) Gin Senocide F2 0.3 12 glycerin 10.0 10.0 Betaine 5.0 5.0 REG 1500 2.0 2.0 Allantoin 0.1 0.1 DL-Panthenol 0.3 0.3 EDTA-2Na 0.02 0.02 Benzophenone-9 0.04 0.04 Hydroxyethylcellulose 0.1 0.1 Sodium hyaruronate 80. 80. Carboxyvinyl polymer 0.2 0.2 Triethanolamine 0.18 0.18 Octyldodecanol 0.3 0.3 Octyldodec-16 0.4 0.4 ethanol 6.0 6.0 Unicide - Yu 13 0.01 0.01 Distilled water Balance Balance Sum 100 100

< Example  7>

Stability test of formulation

For the formulation stability test of the cosmetic composition of the present invention, the cosmetic composition formulated in Example 6 was placed in an opaque glazed container and stored for 12 weeks in a constant temperature bath kept at 45 DEG C, The degree of discoloration was measured after keeping for 12 weeks in a constantly kept, fully shaded refrigerator. The degree of discoloration of the product is classified into the following six grades (0: no change, 1: very little separation (discoloration), 2: little separation (discoloration), 3: (Discoloration)).

 Skin stability test Temperature Degree of discoloration of the formulation Production Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 Production Example 7 Production Example 8 45 0 0 0 0 0 0 0 0 4 0 0 0 0 0 0 0 0

As a result, as shown in Table 10, the cosmetic composition of Production Example does not cause discoloration and separation phenomenon, and it can be seen that there is no abnormality in the stability of the formulation of cosmetic agent containing ginsenoside F2 according to the present invention.

< Example  8>

Skin safety experiment

In order to test skin safety for the composition of the present invention, the following safety test was conducted on the cosmetic composition formulated in Example 6 above.

<8-1> Primary skin irritation test

The cosmetic preparation was applied to the skin of male white rabbits with hair on the dorsal skin removed once a day, five times a week to a thickness of 2 mg / cm 2 or more. The application was carried out for 2 weeks, and hair removal was carried out every week at the final application day. The test was performed at 24, 48, and 72 hours after application with the primary irritation test using erythema and edema as an index. As a result of the test, it was judged to be negative because no erythema and edema were observed in all experimental groups.

<8-2> Skin Cumulative Irritation Test

The cosmetic preparation was applied to the skin of male white rabbits with hair on the dorsal skin removed once a day, five times a week to a thickness of 2 mg / cm 2 or more. The application was carried out for 2 weeks, and hair removal was carried out every week at the final application day. The determination was carried out using the erythema and edema as the indicators by the primary irritation method on the day of each application and the day after the last application. As a result of the test, skin irritation caused by repeated application of the preparation was not observed in all the test groups and it was judged as negative.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (5)

A cosmetic composition for improving wrinkles, whitening skin and improving acne,
The composition contains ginsenoside F2 as an active ingredient,
Wherein the composition has an anti-acne improving effect through antimicrobial activity against Propionebacterium acne which is an acne-causing microbe and Staphylococcus aureus which is a microbe . The method according to claim 1,
Wherein the ginsenoside F2 is contained in an amount of 0.0001 to 20% by weight based on the total weight of the composition. The method according to claim 1,
Wherein the composition has a wrinkle-reducing effect through promoting fibroblast proliferation, increasing collagen synthesis, and inhibiting elastase. The method according to claim 1,
Wherein the composition has a skin whitening effect through melanin synthesis inhibitory activity and tyrosinase inhibitory activity. delete

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