KR100602684B1 - Cosmetic composition comprising extract of erigeron breviscapus for whitening - Google Patents

Abstract

The present invention relates to a cosmetic composition for whitening containing Erigeron breviscapus extract as an active ingredient, the forage extract according to the present invention inhibits melanin production and inhibits tyrosinase activity in melanin producing cells, and also excellent antioxidant activity In addition, since it shows little cytotoxicity, it may be usefully used as a cosmetic composition for skin whitening.

Forget-me-not, whitening, cosmetics

Description

Cosmetic composition comprising extract of Erigeron breviscapus for whitening}

The present invention relates to a cosmetic composition for whitening containing Erigeron breviscapus extract.

The present invention also relates to an antioxidant composition containing the forget-me-not extract.

Skin is a very important tissue that has direct contact with the external environment, protects the human body and has biochemical and physical functions. This tissue is largely divided into three parts: the epidermis, the dermis and the hypodermis. Melanin is a pigment in the epidermal layer of the skin, made from melanocytes in the basal layer of the epidermis and migrates to epidermal cells called keratinocytes. There are no enzymes in the body that break down melanin, but they are removed by falling off the skin as keratinocytes fall off the epidermis.

In the melanin synthesis process, in the melanocyte vesicles in melanocytes, tyrosinase is first converted into dopaquinone via dopa (DOPA) by an enzyme called tyrosinase, and autooxidation and enzyme reaction from dopaquinone is performed. As a result, melanin, a blackish brown copolymer, is produced via DOPAchrome. The melanin thus produced is transferred to keratinocytes through the melanosomes, where it is lost to the surface of the skin along with the keratinization process for about 28 days.

Melanin absorbs the light energy of sunlight UV to protect skin organs under the dermis from UV damage, and it protects the skin from external harmful factors such as trapping harmful oxygen and free radicals generated in skin skin. Play a role. In addition, the skin color of a person is determined by the amount of melanin in the skin cells. People with high melanin pigments have brown or black skin, while those with low melanin pigments have white skin.

However, abnormally low production of melanin causes skin lesions such as vitiligo and, conversely, excessive synthesis or keratinization of melanin due to daylight, hormonal changes, inflammation, or drugs can damage the skin if it is not completely lost. In addition, the pigment of the skin is deposited to form blemishes, freckles, and even from such lesions may cause skin cancer. Therefore, in order to prevent such a pigmentation phenomenon, it is necessary to inhibit a part of melanogenesis process to reduce melanin production.

Today, Asian women prefer white and clean skin like white jade and make it an important standard of beauty. Therefore, the development of whitening agent for treatment of abnormal skin pigmentation and satisfying beauty needs is being actively developed.

At present, in the development of a whitening agent for skin beauty, a method of reducing the generated melanin pigment by decolorization and a method of inhibiting the activity of tyrosinase, an enzyme that forms melanin pigment, are known. However, whitening agents using tocopherols and vitamins used to reduce the melanin pigment are known to have a slight decolorizing effect of the melanin pigment. Therefore, attention is being paid to inhibitors that inhibit the production of melanin by inhibiting the activity of tyrosinase.

In the field of conventional cosmetics, as a whitening component, for example, substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, hydroquinone, vitamin C (L-Ascorbic acid) and these Derivatives of and various plant extracts have been used.

However, the stability in the prescription system is poor, the color is degraded, or the use thereof is limited due to the occurrence of off-flavor, efficacy at the biological level, unclear effect and safety problems. Kojic acid inhibits enzymatic activity by adsorbing copper ions present in the active site of tyrosinase, but has been discontinued as a cosmetic ingredient because it is found to cause instability, skin side effects, and liver cancer in recent animal experiments. Vitamin C and its derivatives are difficult to use as raw materials for cosmetics due to their oxidative instability, and hydroquinone has excellent whitening effect on skin but allergic properties, toxicity to melanocytes, and permanent depigmentation to skin. It is highly polarized and has been recently defined as a carcinogenic substance and its use is banned and only a limited concentration is allowed in each country. In addition, arbutin is a derivative in which glucopyranoside is combined with hydroquinone, and has little side effects when using hydroquinone and inhibits the synthesis of melanin pigments without toxicity to humans, thereby increasing melanin pigmentation. Applicability has been suggested as a therapeutic agent for the skin disease, which has the disadvantage of being partially degraded by skin enzymes.

In addition to these whitening substances, research has been conducted to find whitening active ingredients in natural products, especially plants. Among them, white skin (Japanese Patent Application Laid-Open No. 55-44375, No. 64-26507, No. 64-83009, No. 1-25687) No. 5-139950, Korean Patent Publication Nos. 92-002109, 97-021273, Licorice (Japanese Patent Laid-Open No. 60-214721, No. 60-214728, No. 63-23809, No. 64- 63506, Hei 1-149706, and Korean Patent Nos. 92-002109, 97-025601), Peony (Japanese Patent Laid-Open No. 61-246109 and Korean Laid-Open Patent No. 92-002111), Cinnamon Patent Nos. 63-30403, Hep. 5-139954), Gosam (Japanese Patent Laid-Open No. 64-26507 and Korean Patent Laid-Open No. 92-002110), Brown root (Japanese Laid-Open Patent Nos. 60-214727, Small 64-16709 ), Angelica (Japanese Patent Laid-Open No. 56-92211, Hei 4-26610), Mokdanpi (Japanese Laid-Open Patent No. 60-214721, No. 61-50915, No. 61-246109), Banha (Japanese Laid-Open Patent Publication) Pyeong 2-207028, for A number of plant extracts and herbal extracts such as Korean Patent Publication No. 96-033442) and Aloe (Japanese Patent Laid-Open No. 52-44375, Hei 2-207030, Hei 5-139950) act on tyrosinase to produce melanin. Although it has been found to suppress, they also have a lot of problems in the use of more than the effective concentration in cosmetics or pharmaceuticals in terms of safety, discoloration, etc., and still does not show an excellent effect.

The inventors of the present invention, while researching among the natural medicinal plants for a whitening cosmetic composition that is safe to the skin and low discoloration potential, the forget-me-not extract, which is a plant native to the high mountains of Tibet, China, reduces melanin production and inhibits tyrosinase activity. It has an antioxidant effect, and completed the present invention by revealing that it can be usefully used as a cosmetic composition for skin whitening.

The present invention is to provide a cosmetic composition for whitening containing the forget-me-not extract ( Erigeron breviscapus ).

The present invention also provides an antioxidant composition containing the forget-me-not extract.

In order to achieve the above object, the present invention provides a cosmetic composition for whitening containing the forget-me-not extract ( Erigeron breviscapus ).

More preferably, the extract is characterized by thermal extraction of the forget-me-not with a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, most preferably with methanol.

In addition, the whitening effect of the composition is characterized by the inhibition of melanin production in melanocytes, the inhibition of tyrosinase activity.

The present invention also provides an antioxidant composition containing the forget-me-not extract.

The forget-me-not ( Erigeron breviscapus ) It is a perennial wild plant in Yunnan, Sichuan and Guizhou provinces in southern China. Forget-me-not is active in increasing cardiovascular flow, decreasing pressure in the cerebrovascular vessels, and enhancing immune function of macrophage cells in the body, thereby treating diseases such as apoplexy, coronary artery disease, and angina sequelae. Effective (US Pat. No. 6840080).

The forget-me-not extract according to the present invention may be prepared as follows.

Wash the forget-me-not with water and dry it in the shade. The dried forget-me-not is placed in an extraction vessel and extracted with a suitable amount of extraction solvent at a suitable temperature for a certain time. The extract can be filtered and concentrated.

As the extraction solvent, a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof may be used, and preferably methanol is used.

The amount of the extraction solvent is 2 to 10 times, preferably 4 times, the dry weight of the herbal medicine.

The extraction method may be an extraction method such as heat extraction, reflux cooling extraction, ultrasonic extraction, etc., preferably may be extracted 1 to 5 times as the extraction method of the heat extraction.

The deposition temperature is 30 to 70 ℃, more preferably 40 to 60 ℃, most preferably maintained at about 50 ℃ is most effective in the prevention of rot, discoloration and odor.

The deposition time is 5 to 20 hours, preferably 10 hours.

After obtaining the forage extract, solids are removed using a filter paper, the suspension is centrifuged, and the supernatant can be filtered under reduced pressure.

The filtered extract is concentrated by removing methanol in the extract using nitrogen.

The forget-me-not extract according to the present invention has a whitening effect of reducing melanin production by B16 melanoma cells (Table 1) and inhibiting tyrosinase activity (Table 2). In addition, there is an antioxidant effect of scavenging free radicals (Table 3) and less cytotoxicity against B16 melanoma cells (Table 4). Therefore, the forget-me-not extract of the present invention can be effectively used as a cosmetic composition for whitening that suppresses excessive melanin production. In addition, it can be usefully used as an antioxidant composition.

In the composition of the present invention, the forage extract is preferably contained in an amount of 0.005 to 50% by weight based on the total weight of the composition, but may be used in the above or below the range according to the intended use in the range of whitening effect and no toxicity.

The composition of the present invention may further contain at least one active ingredient exhibiting the same or similar function in addition to the forget-me-not extract.

Cosmetics prepared by the cosmetic composition containing the forget-me-not extract of the present invention as an active ingredient can be prepared in the form of a general emulsion formulation and solubilized formulation. Cosmetics of the emulsion formulations include nutrient cosmetics, creams, essences, etc., cosmetics of the solubilized formulations are soft cosmetics. In addition to cosmetics containing the forget-me-not extract of the present invention by containing a dermatologically acceptable medium or base may be prepared in the form of adjuvants that can be topically or systemically applied commonly used in the field of dermatology.

Suitable cosmetic formulations include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionics (liposomes), nonionics obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase. It may be provided in the form of a vesicle dispersant, in the form of a cream, skin, lotion, powder, ointment, spray or conceal stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition of the present invention, in addition to the forage extract, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water Commonly used in ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field, such as any other ingredient. In addition, the above components may be introduced in an amount generally used in the dermatology field.

Examples of products to which the cosmetic composition of the present invention may be added include, for example, cosmetics such as astringent cosmetics, soft cosmetics, nourishing cosmetics, various creams, essences, packs, foundations, and the like, cleansing agents, soaps, treatments, and essences. Etc.

Specific formulations of the cosmetic composition of the present invention include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, nutrition essence, pack, Formulations such as soaps, shampoos, cleansing foams, cleansing lotions, cleansing creams, body lotions, body cleansers, emulsions, press powders, loose powders, eye shadows and the like.

In addition, the method of use is generally oral, parenteral use, can be used to apply to the skin during parenteral use, it is possible to select the preferred method according to the formulation.

Hereinafter, preferred examples and experimental examples are presented to help understand the present invention. However, the following Examples and Experimental Examples are provided only to more easily understand the present invention, and the contents of the present invention are not limited by the Examples.

Example: Preparation of Forage Skin Extract

The forget-me-not of this example was obtained from the high mountain region of China.

The collected plants were washed with water to remove foreign substances and dried. 50 g of the dried plant was immersed in 200 ml of 100% methanol and warmed in a 50 ° C. water bath for 10 hours. The obtained extract was cooled to room temperature and filtered through a filter paper to give a filtrate. Methanol was evaporated from the filtrate using nitrogen, concentrated and then dissolved in 10% methanol to prepare an extract.

Experimental Example 1: Measurement of the whitening effect

1-1: B16 Melanoma Cell Culture

B16 melanoma cells were supplied and used at the Suwon Forest Research Institute. B16 melanoma cells are cell lines producing melanin and are commonly used for whitening screening.

The medium for culturing the cells was prepared aseptically by preparing and sterilizing DMEM medium containing 10% FBS, 1% penicillin / streptomycin. Cell cultures were incubated in an incubator at 37 ° C., 5% CO 2. Melanoma cells were cultured and passaged when the 100 mm Petri dishes were about 90% full.

1-2: melanin inhibition measurement using absorbance measurement

In order to measure the whitening effect of the forget-me-not extract, the change in the amount of melanin produced by treatment of B16 melanoma cells with the extract or arbutin was measured. Cells were seeded on the 6-well plate and treated with the concentration of 200 μg / ml of the forget-me-not extract or arbutin prepared in Example, followed by cell recovery using trypsin-EDTA. 1N NaOH containing 10% DMSO (Dimethylsulfoxide) was added to the recovered cells and reacted at 90 ° C. for 1 hour to be sufficiently dissolved in melanin. Then, the absorbance was measured at a wavelength of 405 nm using an ELISA reader. Melanin production was calculated using the following equation 1, the experimental results are shown in Table 1.

Melanin Formation (%) = Treatment Absorbance / No Treatment Absorbance × 100

Sample Name No treatment Arbutin Forget-me-not extract Melanin Production (%) 100 71.4 55.0

As shown in Table 1, arbutin, known as a general whitening substance, has a melanin content of 71.4% at a concentration of 200 µg / ml, compared to the untreated group, while the forage extract is 55.0%, about 1.5 times more than arbutin. It was confirmed that it has a high melanin production inhibitory effect.

1-3: Determination of tyrosinase activity inhibition

The following experiment was conducted to determine if the forget-me-not extract affects tyrosinase enzyme activity, which plays an important role in producing melanin.

20 μl of 0.1 M phosphate buffer (pH 6.8) was added to the 96-well plate, 200 μg / ml of forget-me-not extract, 50 μl of kojic acid or distilled water as a comparative group, and 10 μl of mushroom-derived tyrosinase. (Sigma) was added. After adding 20 μl of 5 mM L-DOPA as a substrate and reacting for 2 minutes at 37 ° C., the absorbance was measured at a wavelength of 475 nm using an ELISA reader.

The tyrosinase activity was calculated using Equation 2 below, and Table 2 shows the koji acid widely used as a whitening agent and tyrosinase activity inhibited by the extract.

Tyrosinase Activity (%) = Treated Absorbance / Untreated Absorbance × 100

Sample Name Untreated Kojisan Forget-me-not extract Tyrosinase Activity (%) 100 2.6 54.0

As shown in Table 2, tyrosinase showed only 2.6% of the total activity when treated with kojic acid, while 54.0% of the activity was treated when treated with forage extract. Forget-me-not extract showed a lower effect of inhibiting tyrosinase activity compared to kojic acid, but it was found that it inhibits tyrosinase enzyme activity by about 50% compared to no treatment.

Experimental Example 2: Antioxidant Activity Experiment

In order to check whether the forget-me-not extract has antioxidant activity, it was generally measured by using a method of measuring antioxidative activity using DPPH (1,1-diphenyl-2-picrylhydrazyl) solution, and experimented with vitamin C, which is known as an antioxidant. The relative level of activity of the extract was compared.

100 μl of 200 μg / ml of the forget-me-not extract or vitamin C and the DPPH solution, which is an antioxidant measurement solution, were reacted for 30 minutes at 37 ° C., and then the absorbance was measured at a wavelength of 517 nm using an ELISA reader.

Antioxidant activity was calculated using the following equation (3), the experimental results are shown in Table 3.

Antioxidant Activity (%) = (1-treated Absorbance / Untreated Absorbance) × 100

Sample Name Vitamin c Forget-me-not extract Antioxidant activity (%) 66.2% 36.9%

As shown in Table 3, it was found that the antioxidant activity of the forget-me-not extract was 36.9%, indicating that it had antioxidant power. It is interesting to note that the forget-me-not extract has lower antioxidant activity than vitamin C, but its stability is high and its activity is maintained even after a long time.

Experimental Example 3: Toxicity Test

In order to determine whether the forget-me-not extract can be used as a whitening cosmetic composition, a cytotoxicity experiment was conducted. Cells were seeded in 96-well plates and treated with 200 μg / ml of forget-me-not extract and incubated in an incubator at 37 ° C., 5% CO 2 for 2 days. After incubation, the medium was removed, and 100 μl of MTT (3- (4,5-dimethythiazol-2-yl) -2,5-diphenyltetrazoliu, bromide) solution was added and reacted at 30 ° C. for 4 hours. After the reaction was completed, the solution was removed, and 100 μl of DMSO was added to dissolve the stained cells, and the absorbance was measured at a wavelength of 540 nm using an ELISA reader. Cell viability was calculated using Equation 4 below, and the results are shown in Table 4.

Cell viability (%) = (1-treated absorbance / untreated absorbance) × 100

Sample Name No treatment Arbutin Forget-me-not extract Cell survival rate (%) 100 98.3 90.8

As shown in Table 4, the cell viability was found to be 90.8% almost no cytotoxicity in spite of the high concentration of 200 ㎍ / ㎖ compared to the untreated treatment without the forage extract.

Examples of preparations for the compositions of the present invention are illustrated below.

Formulation Example 1 Flexible Lotion

Formulation example of the flexible lotion containing the forget-me-not extract was prepared as shown in Table 5.

Raw material Content (% by weight) Forget-me-not extract 10.00 1,3-butylene glycol 1.00 Disodium ID 0.05 Allantoin 0.10 Dipotassium glycylizate 0.05 Citrix Acid 0.01 Sodium citrate 0.02 Glyceres-26 1.00 Arbutin 2.00 Hydrogenated Castor Oil 1.00 ethanol 30.00 Preservative a very small amount coloring agent a very small amount Flavor a very small amount Purified water Remaining amount

Formulation Example 2: Nutrition Cream

Formulation of nutrition cream containing the forget-me-not extract was prepared as shown in Table 6.

Raw material Content (% by weight) Forget-me-not extract 10.0 1,3-butylene glycol 7.0 glycerin 1.0 D-panthenol 0.1 Plant extracts 3.2 Magnesium Aluminum Silicate 0.3 PEG-40 Stearate 1.2 Stearic acid 2.0 Polysorbate 60 1.5 Lipophilic glyceryl stearate 2.0 Sorbitan sesquioleate 1.5 Cetearyl Alcohol 3.0 Mineral oil 4.0 Squalane 3.8 Carlylic / Capric Triglycerides 2.8 vegetable oil 1.8 Dimethicone 0.4 Dipotassium glycylizate a very small amount Allantoin a very small amount Sodium hyaluronate a very small amount Tocopheryl Acetate Quantity Triethanolamine Quantity Preservative Quantity Flavor Quantity Purified water Remaining amount

As described above, the forget-me-not extract according to the present invention refers to the forget-me-not extract extracted using a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the production of melanin by B16 melanoma cells Inhibits tyrosinase activity. In addition, the forget-me-not extract is also an antioxidant activity, because it shows little cytotoxicity, it can be usefully used as a cosmetic composition for skin whitening.

Claims (6)

A cosmetic composition for skin whitening containing the extract of Erigeron breviscapus . The cosmetic composition for skin whitening according to claim 1, wherein the extract is extracted with a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The cosmetic composition for skin whitening according to claim 1, wherein the extract is heat-extracted with methanol. The cosmetic composition for skin whitening according to claim 1, wherein the skin whitening effect of the composition is due to inhibition of melanin production. The cosmetic composition for skin whitening according to claim 1, wherein the skin whitening effect of the composition is due to inhibition of tyrosinase activity. Antioxidant composition containing the forget-me-not extract.