KR20160061601A - Cosmetic composition and pharmaceutical composition containing the extract of Corchorus olitorius L, ginko and chlorella - Google Patents

Abstract

The present invention relates to a composition comprising a compound consisting of a Corchorus olitorius L. extract, a Ginko biloba L. leaf extract, and a chlorella extract. A pharmaceutical composition of the present invention is a composition comprising extracts of Corchorus olitorius L., Ginko biloba L. leaves, and chlorella; and has excellent anti-inflammatory properties and an effect of generating collagen. When used as a pharmaceutical medicine or a cosmetic composition, the composition has substantial properties such as anti-inflammatory properties, alleviating dermatitis, recovering a skin barrier function, and improving ability to retain moisture in the skin.

Description

The present invention relates to a cosmetic composition and a pharmaceutical composition containing the extract of Corchorus olitorius L, ginko and chlorella,

The present invention relates to a cosmetic composition and a pharmaceutical composition containing an extract of Moroheya, ginkgo leaf and chlorella as an active ingredient.

Moroheya is a one-year-old grass originating in tropical Asia and Africa belonging to the Tiliaceae family. The official name is Corchorus olitorius L. '. There are about 40 genera and 400 species distributed in tropical and temperate forests. Among them, 11 genera of 3 genera are distributed in the country. The height is 50 to 120 cm (4 m in the tropics), and the oval oval leaf is distinctive and has a mount. The leaf quality is soft and is similar to jute (Corchorus capsularis L.). Flowers bloom from August to September with large, dark yellow flowers. Moroheya is characterized by a slippery, thick liquid that comes out when cutting leaves. This mucin is an enzyme that combines polysaccharide and protein, called mucin, and is a kind of mucopolysaccharide that is water-soluble vegetable fiber.

Moroheya contains a large amount of this vegetable fiber to improve the constipation and secreted from the gallbladder bile secreted in the small intestine secreted by the release of cholesterol synthesis by lowering the blood cholesterol and bile acid salts and neutral fat to prevent re-absorption This results in lowering triglycerides in the body.

And harmful to the body is absorbed, excreted, and gives vitality to the cells to increase immunity. It also works in the stomach to preserve the moisture in the tissues and maintain the elasticity of the joints and moisturizing.

When there is less mucopolysaccharide in the gastrointestinal mucosa, the mucosa becomes dry and hardens and becomes inelastic. Then the movement of the stomach is dull and the digestion is not absorbed well, because the stomach will actively stimulate the stomach to digestion and absorption is good.

Also, if the stomach is scarred and it is easy to become inflamed, it protects the surface by covering the surface, helping the cells to recover healthily.

Compared with other vegetable ingredients, it also contains nutrients such as potassium, calcium, iron, beta-carotene, vitamins A, B1, B2, C and E to prevent adult diseases and aging. . Moroheiya is a vegetable, so it should be expressed as 'eat', but to say 'drink' means to eat Moroheiya means to drink vitamins.

Beta carotene acts as a main ingredient to prevent active oxygen, anti-aging and itching. Vitamin B has antioxidant, skin protection and skin-soothing effects.

Polysaccharide is known to be effective for moisturizing and skin elasticity. Moro Haya is rich in carotene and various vitamins, potassium, and calcium compared to other green and yellow vegetables, and dietary fiber contains up to 1.7 times as much as other green and yellow vegetables.

Therefore, it is known that the ingestion of Moroheya can obtain osteoporosis prevention effect, anti-cancer and anti-aging effect by these useful ingredients, and has an excellent effect on constipation and skin beauty. (Seok Ho Moon, Characterization of Domestic Morrow Haya and Development of Processing Products Using it, Korea Food Research Institute, 2001).

Ginkgo biloba extract is extracted from Ginko biloba L., a deciduous tree belonging to Ginkgo biloba. Ginkgo biloba extract contains flavonoids with free radical elimination, antioxidation, cell membrane protection, vascular disease reduction, , Blood circulation improvement, blood clot removal, blood aging prevention, skin aging suppression, dry skin protection, and the face contains a variety of substances that prevent wrinkles. In addition, it has antimicrobial activity and antiallergic effect.

In addition, chlorella is a single-celled organism belonging to the green algae that grow in fresh water. It is a kind of plankton, rich in nutrients such as proteins, chlorophyll, vitamins, minerals and amino acids. It has been studied and became famous. Algae (algae) is a generic term for plants of the lower family, which grows mostly in water or water, and assimilates into chlorophyll and lives on its own. Chlorella is widely distributed on the earth from the tropics to the earth and can be collected from lakes, ponds and ponds. Chlorella is a single-celled plant with one individual cell formed, round or oval in shape. The size can be seen only by microscope at 2 ~ 10 / 1,000mm and it grows with amazing vitality. In Japan, Chlorella products are popular enough to be number one in health food sales for decades. In Japan, the world's longest longevity country, 73% of the population over the age of 50 is a health food that is popular enough to take chlorella products.

However, there have been no patents and studies to prepare the composition from the extracts of three medicinal herbicidal compounds having the above-mentioned medicinal effects, and thus the inventors of the present invention have made a pharmaceutical composition prepared from the above three medicinal ingredient extracts.

Korean Patent No. 10-1085019

The present invention provides a composition comprising the above extracts of Morey Haya, Gingko leaf and Chlorella, and the provided composition is intended to be provided in pharmaceutical or cosmetic formulations.

In order to solve the above problems, the present invention is characterized by providing a composition comprising a mixture composed of Moroheya, ginkgo leaf and chlorella extract.

In addition, the composition of the present invention may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, creams, gels, patches, sprays, ointments, alerts, lotions, liniments, pastes or cataplasms The composition can be formulated in the form of external preparations and sterile injectable solutions.

The pharmaceutical composition of the present invention provides a composition having an anti-inflammatory and collagen-producing effect by providing a composition comprising extracts of Morrow Haya, Ginkgo biloba and Chlorella, wherein the composition is used as a pharmaceutical preparation or a cosmetic preparation It has excellent anti-inflammatory effect, improvement of dermatitis, restoration of skin barrier function and improvement of skin moisturizing effect.

Fig. 1 shows the anti-inflammatory effect according to Experimental Example 1 of the composition of the present invention.
Figures 2 and 3 show excellent inhibition of TNF- [alpha], IL-1 [beta] cytokines according to Experimental Example 2 of the composition of the present invention.
Figure 4 shows the excellent inhibitory effect of PGE ₂ according to Experimental Example 2 of the composition of the present invention.

Corochus olitorius L. (Jew's Marrow, Egypt name: mulukhi-yya) used as an active ingredient of the present invention is a plant belonging to the genus Pinus densiflora and Jute.

In the present invention, the Ginkgo biloba leaf extract is a synthetic component composed of a flavonoid of a leaf and an aromatic oxycarboxylic acid, and may be used as a synthetic component such as fibrin glicoside, gingconic acid, ginkgoetin, zingoide A, B, C, M, Contains active ingredients.

In the present invention, the cosmetic composition contains an extract of chlorella.

The mixed extracts of Moroheya, Ginkgo biloba and Chlorella extracts were prepared by drying Morrow Haya leaves, Ginkgo biloba and Chlorella, and then extracting them using ethanol after filtering. Then, they were filtered with a filter paper to remove solids and concentrated using a vacuum concentrator But it is possible to use one or more of methanol, propylene glycol, 1,3-butylene glycol, ethyl acetate, diethyl ether and dimethyl sulfoxide as an extraction solvent in place of ethanol. Supercritical extraction And the like.

The mixed extract of Moroheya, Ginkgo leaf and chlorella used in the present invention can be prepared, for example, by a process including the following steps.

1) an aqueous solution containing 1 to 10 times water, 1 to 4 carbonic anhydrides or hydrated lower alcohols, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol in combination with moroheya, ginkgo leaf and chlorella Adding one solvent;

2) extracting the solvent-added Moroheya, ginkgo leaf and chlorella of step 1) at 4-100 ° C for 1 hour to 5 days;

3) filtering the extract of step 2), aging the solution at 3 to 5 ° C for 7 to 10 days, and then filtering;

4) drying the filtrate of step 3) with a rotary evaporator

In this case, when 1,3-butylene glycol is used as the extraction solvent, it is difficult to dry using the rotary evaporator, so that the drying loss is adjusted to 1% (w / v) after the extraction step 2) It is also necessary.

On the other hand, the mixed extracts of Moroheya, Ginkgo biloba, and Chlorella of the present invention include not only the extracts obtained by the above-mentioned extraction methods, but also the extracts obtained through ordinary purification processes. For example, by separation using an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatographies (made for separation by size, charge, hydrophobicity or affinity), and the like, Fraction is also considered to be included in the extract of the present invention.

In addition, the mixed extract of Moroheya, Ginkgo biloba, and Chlorella of the present invention may be produced in powder form by an additional process such as vacuum distillation, freeze drying, spray drying and the like.

In the present invention, a mixed extract of Moroheya, ginkgo leaf and chlorella can be added to various cosmetic formulations. By adding an appropriate amount of such extract, artificial cosmetics or skin irritation caused by external stimuli such as ultraviolet rays and dusts can be alleviated In addition, since natural substances are used, there is no other skin irritation due to the addition of an anti-inflammatory agent as in conventional cosmetics, and it is possible to provide cosmetics safe for the skin.

The external preparation for skin according to the present invention may be a cosmetic composition or a pharmaceutical composition.

When the composition for external application for skin is a pharmaceutical composition, it may further include a pharmaceutically acceptable carrier which is already known and used. In other words, it may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions, and the like.

The pharmaceutical composition containing the extract according to the present invention can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, external preparations such as syrups and aerosols, and sterilized injection solutions, The present invention provides a pharmaceutical composition for skin application, which is preferably a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta or a cataract.

Examples of carriers, excipients and diluents that may be contained in the composition containing the extract include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, Calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose ), Lactose, gelatin and the like.

In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.

Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The composition for external application for skin of anti-inflammatory, which comprises the mixed extract of Moroheya, ginkgo leaf and chlorella, of the present invention is an external preparation for skin which can be applied to the skin, and is a cream, gel, patch, spray, ointment, warning agent, lotion, liniment A pharmaceutical composition in the form of a topical, topical or cataplasmatic external preparation for skin, but is not limited thereto.

When the composition for external use according to the present invention is a cosmetic composition, it can be used variously in cosmetics and cleanser for improving the skin containing the mixed extract of Moroheya, ginkgo leaf and chlorella of the present invention as an active ingredient. Examples of products to which the present composition can be added include cosmetics such as various creams, lotions, and skins, and cleansing, cleansing agents, soaps, treatments, essences, packs and the like.

The cosmetic composition of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

The cosmetic of the present invention may be blended with other essential ingredients, if necessary, in combination with the essential ingredients. Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.

The above cosmetic composition may contain an acceptable carrier in a cosmetic preparation. Here, the acceptable carrier in the cosmetic preparation is a compound or composition which is already known and used in the cosmetic preparation, or which is a compound or composition to be developed in the future, and which has no toxicity, instability or irritability It says.

 The carrier may be included in the skin topical composition of the present invention in an amount of from about 1% by weight to about 99.99% by weight, preferably from about 90% by weight to about 99.99% by weight of the composition, based on the total weight thereof. However, since the ratio varies depending on the above-described formulation in which the composition for external application for skin of the present invention is prepared, and its specific application site (face, neck, etc.) or its preferable application amount, And should not be construed as limiting the scope of the invention.

Examples of the carrier include alcohols, oils, surfactants, fatty acids, silicone oils, humectants, moisturizers, viscosifiers, emulsifiers, stabilizers, sunscreens, coloring agents and perfumes.

As an embodiment of the present invention, the composition for external application for skin according to the present invention may contain glycerin, butylene glycol, propylene glycol, polyoxyethylene hardened castor oil, ethanol, triethanolamine and the like in addition to the above extract, , Purified water, and the like may be included as needed.

In particular, the composition for external application for skin may be prepared in the form of a general emulsified formulation and a solubilized formulation by a conventional manufacturing method in addition to the manufacturing method specifically disclosed in the present invention.

When the cosmetic composition is manufactured from a cosmetic composition, the cosmetic composition of the emulsified formulation includes nutritional lotion, cream, essence and the like. It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

In addition, suitable cosmetic formulations may include, for example, solutions, gels, solid or paste anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) In the form of a follicular dispersing agent, cream, skin, lotion, powder, ointment, spray or conical stick. It can 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 composition for external application for skin of the present invention may further comprise at least one component selected from the group consisting of fatty substances, organic solvents, solubilizing agents, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, foaming agents, As used herein means any emulsion or emulsion which is commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Adjuvants commonly used in the cosmetics or dermatological fields, such as other ingredients. And, the above ingredients can be introduced in amounts commonly used in the field of dermatology.

In addition, any of the above components may be blended within the range not to impair the objects and effects of the present invention, but is preferably 0.01 to 5% by weight based on the total weight, Preferably 0.01 to 3% by weight.

The cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture or the like. The ingredients contained in the cosmetic composition of the present invention may contain, in addition to the above-mentioned extracts, effective ingredients commonly used in cosmetic compositions, for example, conventional additives such as stabilizers, solubilizers, vitamins, And a carrier.

The composition of the present invention can be prepared in any formulations conventionally produced in the art, and examples thereof include emulsions, creams, lotions, packs, foundations, lotions, essences, and hair cosmetics.

Specifically, the cosmetic composition of the present invention can be used as 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, a nutrition cream, a moisturizing cream, a hand cream, Packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol chibasic esters.

In the present invention, in the case of compositions for preventing hair loss or promoting hair, it is possible to use various formulations for preventing hair loss or promoting hair, such as shampoos, rinses, treatments, essences, hair packs and hair essences . ≪ / RTI >

Hereinafter, the present invention will be described in more detail with reference to the following examples and experimental examples. However, the following examples and experimental examples are only for illustrating the present invention, and thus the scope of the present invention is not limited thereto.

Example 1: Preparation of Moroheya, Gingko leaf, and chlorella mixed extracts

100.0 g of each of Moroheya, Ginkgo leaf and chlorella which had been washed with purified water and dried was pulverized and then added to 1.2 L of water and extracted at 65 ° C for 5 hours. Then, the mixture was filtered through a 300-mesh filter cloth and allowed to stand at 4 ° C for 5 days, And filtered through filter paper No. 5 of Watts. The filtrate was dried at 65 DEG C with a rotary evaporator to obtain 13.00 g of dry weight.

Experimental Example 1: Anti-inflammatory activity test of mixed extract of Moroheya, Gingko leaf and chlorella of the present invention

Various inflammatory mediators are involved in the pathogenesis of inflammation, and the causes of the disease vary. Treatment of macrophages with LPS (lipopolysaccharide), known as endotoxin, increases NO biosynthesis by inducible NOS (iNOS) expression and induces an inflammatory response. NO is a pathological secondary signaling substance secreted by many cells, regulates the activity of COX and acts synergistically to the inflammatory response.

By measuring NO produced by LPS induced by NOS in Raw264.7 cells, the inhibitory effect of the extract on inflammation can be determined.

Raw264.7 cells, which are mouse macrophage cells, were treated with artificial LPS, which is an inflammation inducing substance, to evaluate whether they have an inflammation inhibitory effect.

96 a well plate for dispensing such that the 2 × 10 ⁵ cell per well and then, overnight incubation at 24 hours 37 ℃ while, 5% CO ₂ incubator, and then haejugo replaced with fresh medium, processing the LPS 1 ug / mL, and at the same time the The mixed extract of Moroheya, Ginkgo leaf and chlorella (ethanol extract) obtained according to Example 1 was added at a concentration ranging from 1 ug / mL to 100 ug / mL per concentration and cultured for 24 hours.

After incubation, the supernatant was taken and centrifuged at 13,000 rpm for 3 minutes to collect supernatant, reacted with Griess reagent reagent 1: 1 and absorbance at 570 nm.

As shown in FIG. 1, when the mixed extracts of Moroheya, Gingko leaf and chlorella were treated at the concentrations of 1, 10, 50, and 100 ug / mL, The amount of nitric oxide was low in a concentration - dependent manner, and it was found that the mixed extract of Moroheya, Ginkgo leaf and chlorella had anti - inflammatory effect.

Experimental Example 2: Measurement of TNF-α and IL-1β Cytokine Levels of the Mixed Extract of Morrow Haya, Gingko Leaf and Chlorella

RAW 264.7 cells were cultured in a 24-well plate at a rate of 5 × 10 ⁵ per well, and cultured in a 24-well plate. The cells were mixed with 1 μg / mL of LPS, a mixture of Moroheya, The extracts (ethanol extracts) were each administered at a concentration ranging from 1 ug / mL to 100 ug / mL. After incubation for 24 hours at 37 ° C and 5% CO ₂ , the cell culture was taken and TNF-α and IL-1β contents were measured using an enzyme immunoassay kit (EIA) kit. Cell culture was added to a 96-well plate with a capture antibody for each cytokine. After 24 hours of reaction, the diluted TNF-α, IL-1β antibody and horseradish peroxidase Secondary antibodies conjugated with horseradish peroxidase were reacted at room temperature for 2 hours each. After the reaction, the plate was washed 5 times with 1 × phosphate buffered saline tween (1 × PBST), and the enzyme substrate was added thereto for 30 minutes. The reaction was then quenched by the addition of 2N-sulfuric acid. After stopping the reaction, the developed absorbance was measured at a wavelength of 450 nm using a spectrophotometer. The content of TNF-α and IL-1β was calculated using the standard curve obtained from the reaction of the reference material.

As shown in FIG. 2 and FIG. 3, in the group treated with the mixed extracts of Moroheya, Ginkgo biloba and Chlorella at a concentration of 1, 10, 50 and 100 ug / mL, α, and IL-1β levels were significantly lowered in a concentration-dependent manner. It was found that the extracts of Moroheya, Ginkgo leaf and chlorella showed excellent inhibitory effect on TNF-α and IL-1β cytokines which cause inflammation.

Experimental Example 3: Preparation of prostaglandin E of a mixed extract of Moroheya, ginkgo leaf and chlorella ₂ (Prostaglandin E _2, PGE ₂ ) Production measurement

For 24 hours in each well (well) 1 × 10 ⁵ into each 37 ℃, 5% CO ₂ culture condition per the suspended Raw 264.7 cells in a DMEM medium containing 10% FBS 96- well plates (96-well plate) Lt; / RTI > Raw 264.7 cells were washed once with phosphate buffered saline (PBS) and replaced with fresh DMEM medium. Then, 1 ug / mL of LPS and the mixed extracts of Moroheya, Ginkgo biloba and Chlorella of Example 1, mL to 100 < RTI ID = 0.0 > pg / mL < / RTI > for 24 hours. Dilute the supernatant was added to the PGE ₂ antibody plates (PGE ₂ antibody plate) and cultured for 18 hours or more at room temperature by treating the PGE ₂ -AchE teuraenseo (trancer). After the culture was incubated for about 7 hours at room temperature by treatment with PGE ₂ antibody plates (PGE ₂ antibody plate) for a 0.05% Tween in PBS to wash 5 times 1 min, elreom reagent (Ellman's reagent). After measuring the absorbance at 405 nm, the value of PGE ₂ production was converted by substituting the value into a calibration curve prepared with PGE ₂ standard solution. The inhibitory activity of PGE ₂ production in the LPS-only treated group was determined by non-linear regression analysis to determine the IC ₅₀ (concentration at which 50% inhibition of PGE ₂ production) of each test substance was determined. Were compared.

As shown in FIG. 4, the group treated with the mixed extracts of Moroheya, Ginkgo leaf and chlorella at the concentrations of 1, 10, 50 and 100 ug / mL showed a higher amount of PGE ₂ than the control group Concentration - dependently. It was found that the extracts of Moroheya, Ginkgo leaf and chlorella showed excellent inhibitory effect on the inflammatory PGE ₂ .

Experimental Example 4. Effect of enhancing collagen synthesis of Moroheya, Ginkgo leaf and chlorella mixture extract

Human fibroblasts (Human Skin Fibroblast) for 37 ℃ to maintain a constant humidity of 10% in a 5% CO ₂ incubator FBS, Penicillin (50 U / mL ), Streptomycin DMEM ( containing (50 U / mL) Dulbecco's Modified Eagle's Medium, Gibco, USA). Cells were seeded at a density of 1 × 10 ⁵ cells / mL into a 24-well plate (500 μL) and cultured for 24 hours. The medium containing the mixed extracts of Moroheya, Ginkgo leaf and chlorella at various concentrations was added to the cells and cultured in a 5% CO ₂ incubator at 37 ° C for 48 hours. After 48 hours, the amount of collagen newly synthesized was measured by measuring 20 uL of each supernatant of the medium using a Procollagen Type I C-Peptide EIA Kit (PICP, Takara, Cat No. MK101). The amount of PICP was converted to ng / 1 × 10 ⁵ cells, and the results are shown in Table 1 below.

Sample concentration Amount of collagen synthesis
(ng / 1 x 10 < ⁵ > cells) Control group (purified water) 99 Moroheya, ginkgo leaf, chlorella mixed extract


0.1% 99 One% 124 5% 138 10% 169

Experimental Results As shown in Table 1, when the mixed extracts of the present invention were applied at concentrations of 0.1, 1, 5, and 10 ug / mL, 99, 124, 138, and 169 ng / ^It was confirmed that collagen production was increased by ⁵ cells, and no cytotoxicity was observed at all the concentrations.

Example 2

According to the results of the experiment, cosmetic lotion, cream and lotion type cosmetic compositions containing Morohayaya, ginkgo leaf and chlorella mixed extract in a certain weight were prepared in the following composition. The cosmetic composition of each formulation is not necessarily limited to the composition of the examples in the following table.

Example 2.1 - lotion

ingredient Weight% (w / w) 1,3-butylene glycol 5.0 Citric acid 0.1 Polyoxyethylene oleyl ether 0.8 ethanol 20.0 Spices 0.15 antiseptic Suitable amount Moroheya, ginkgo leaf, chlorella mixed extract 15 Purified water Balance

Example 2.2 - Cream

ingredient Weight% (w / w) Macadamia oil 2.0 Squalane 3.0 Stearic acid 7.0 Vaseline 3.0 1,3-butylene glycol 7.0 Polyoxyethylene sorbitan monostearate 1.3 Self emulsifying glycerin monostearate 2.7 glycerin 4.0 Spices 0.25 antiseptic Suitable amount Moroheya, ginkgo leaf, chlorella mixed extract 15 Purified water Balance

Example 2.3 and Comparative Prescription Example 2.3 - Lotion

ingredient Example 2 2.3% (w / w) Comparative Example 2 2.3 wt% (w / w) Stearic acid 0.5 0.5 vaseline 1.0 1.0 Squalane 2.5 2.5 Liquid paraffin 6.0 6.0 1,3-butylene glycol 8.0 8.0 ethanol 10.0 10.0 Spices 0.2 0.2 antiseptic Suitable amount Suitable amount Moroheya, ginkgo leaf, chlorella mixed extract 15.0 - Distilled water Balance Balance

Example 2.4 and Comparative Prescription Example 2.4 - Essence

ingredient Example 2.4 Weight% (w / w) Comparative Prescription Example 2.4 Weight% (w / w) Cetostearyl alcohol 1.5 1.5 Self emulsifying monostearic acid 1.5 1.5 Wax 0.5 0.5 Squalane 5.0 5.0 Isocetyl octanoate 3.0 3.0 Dimethylsiloxane 0.3 0.3 Sorbitan monostearate 0.5 0.5 Polyethylene glycol monostearate 7.0 7.0 glycerin 4.0 4.0 Propylene glycol 0.1 0.1 Carboxy polymer 0.3 0.3 Triethanolamine 0.2 0.2 antiseptic Suitable amount Suitable amount Spices Suitable amount Suitable amount flash Suitable amount Suitable amount Moroheya, ginkgo leaf, chlorella mixed extract 15.0 - Purified water to 100 to 100

Experimental Example 6: Recovery effect of skin barrier function

In order to examine the skin barrier function recovery effect of the cosmetic composition containing the mixed extract of Moroheya, ginkgo leaf and chlorella of the present invention, the following experiment was conducted. Experiments were carried out using a taping stripping method to damage the skin barrier, and skin moisture loss (TEWL, transdermal water loss) was measured with an evaporimeter EPI from servomed (Sweden) to reach 4.0 g / m ² / h , The creams of Formulation Example 1 and Comparative Formulation Examples 1 to 4 prepared in Example 1 were applied to an area of 5 cm ² , respectively, and the amount of skin moisture loss was measured after 1 hour, 2 hours, 4 hours, and 8 hours , And the extent to which the barrier function was restored was evaluated by evaluating the degree of decrease. At this time, a known lipid mixture (ceramide: 2: 1: 1 mixture of cholesterol: fatty acid) was used as a positive control.

sample 0 hours 1 hours 2 hours 4 hours 8 hours Example 2.3 80 50 21 16 13 Comparative Prescription Example 2.3 80 76 58 34 21 Positive control group 80 67 57 30 24

As a result of the experiment, as shown in the results of Table 6, in Example 2.3, which is a cosmetic containing the mixed extract of Moroheya, Ginkgo leaf and Chlorella of the present invention, the recovery effect of the skin barrier function And it was confirmed that it is excellent.

Experimental Example 7: Improvement of skin moisturizing effect

In order to examine the skin moisturizing effect of the cosmetic composition containing the mixed extract of Moroheya, Ginkgo leaf and chlorella of the present invention, the following experiment was conducted. Twenty patients in the 20s to 40s without skin disease were divided into two groups each consisting of 20 patients per group, and the nutritional cream of the cream of Prescription Example 1 and Comparative Prescription Examples 1 to 3 prepared in Example 1 above Twice a day for one month. The skin conductivity was measured by using a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under the constant temperature and humidity conditions (24 ° C, 40% humidity) before the start of application, and after 1 week, 2 weeks and 4 weeks The rate of increase in skin conductivity (%) was measured and the rate of increase was evaluated.

sample One week after 2 weeks Four weeks Example 2.4 60 74 77 Comparative Prescription Example 2.4 23 35 44

As a result of the experiment, as shown in the results of Table 7, in Example 2.4, which is a cosmetic containing the mixed extract of Moroheya, Ginkgo leaf and Chlorella of the present invention, the rate of skin conductivity increase was much superior to Comparative Example 2.4. In general, skin conductivity is proportional to the amount of skin moisture. Therefore, the above results show that the cosmetic composition containing Moroheya, Gingko leaf, and chlorella mixed extracts has a higher skin pure content than the cosmetic ingredients not containing such extract.

Experimental Example 8: Effectiveness test of a mixed extract of Moroheya, Ginkgo leaf and chlorella with atopic mice

In order to confirm the atopy effectiveness of the mixed extracts of Moroheya, Gingko leaf and Chlorella, Nc / Nga mouse, a well known atopic animal model, was used. After anesthetizing female 6-week-old female Nc / Nga mice, the inner and outer ears of the ear were gently peeled off with a gauze tape such as a bandage to remove fine hairs and keratin, and 100 μL of 1% DNCB was applied to the inside and outside of the ear. To the ear. Each mouse was thoroughly coated with a mixture of Moroheya, ginkgo leaf and chlorella in the atopy-induced Nc / Nga mouse, and blood was drawn from the eyes for 2 weeks. Serum for ELISA was prepared to confirm the IgE concentration, The efficacy of the extracts was also confirmed by comparing the levels of IgE to those of the control group. In addition, the number of scratches was calculated and the most effective ratio was selected for Moroheya, Gingko leaf, Chlorella mixed extract and control group. In order to observe the effectiveness of the mixed extracts of Moroheya, Ginkgo biloba and Chlorella in atopy-induced mice, DNCB and mite extract were treated with ear to induce atopy, and blood was collected and the concentration of IgE was measured by ELISA. The method for calculating the atopic improvement effect is based on the average IgE concentration and the average number of scratches in the mice of the group not treated with any of the mixed extracts, IgE concentration and average number of scratches in%.

(%) = Average IgE concentration in the untreated group / mean IgE concentration in the treated group of Morrow Haya, Ginkgo biloba, and chlorella extracts in the group treated with the mixed extract of Morrow Haya, Concentration × 100

(%) = Number of scratches in the untreated group / number of scratches in the group treated with the mixed extract of Moroheya, gingko leaf and chlorella × 100

As a result, the IgE concentration and the number of scratching were 54.8 ± 11 (%) and 49.6 ± 9 (%) in the untreated group, 157.5 ± 5 (%) and 139.4 ± 5 9 (%) showed the atopy improvement effect. Therefore, compared to the untreated group, the mixed extract of Moroheya, Gingko leaf and Chlorella showed much better atopic dermatitis improvement effect.

Experimental Example 9: Improvement of atopic dermatitis

In order to confirm the improvement effect of atopic dermatitis, the cosmetic composition was prepared as shown in Table 9 below for 20 weeks for all male and female adults with atopic dermatitis, and the cosmetic composition of Moroheya, Gingko leaf, 0.5 g each twice a day on a cotton swab and applied to the affected area. After two weeks of application, the improvement effect was quantified in four stages: very effective, slightly effective, no effect, and worsening. Fifteen out of 20 patients were given cosmetic preparations containing Moroheya, Gingko leaf, and Chlorella mixed extract as control, and 5 patients were given comparative prescriptions for cosmetics containing the same amount of purified water.

As shown in Table 8, among the total 15 patients, the effectiveness was improved to 3, and the effect was slightly improved to 10, which was about 87% of the total. In general, subjects with atopic dermatitis with severe itching remained calm at 3 days and most of the lesions disappeared at 7 days.

sample Improvement effect Number of people (persons) ratio (%) Moroheya, gingko leaf, chlorella mixed extract cosmetic Very effective 3 20 Slight effect 10 67 no effect 2 13 worse 0 0 Control group
(Purified water administration group) Very effective 0 0 Slight effect 0 0 no effect 4 80 worse One 20

Composition of cosmetics to test atopic dermatitis Raw material name Example Control Example Moroheya, ginkgo leaf, chlorella mixed extract 12.00 - Vegetable hydrogenated oil 8.00 8.00 Vaseline 10.00 10.00 glycerin 10.00 10.00 Polyoxyethylene sorbitan morostearate 0.10 0.10 Antioxidant 3.00 3.00 Butylene glycol 1.00 1.00 Citric acid 0.10 0.10 antiseptic Suitable amount Suitable amount Spices Suitable amount Suitable amount Purified water Balance Balance

Claims (6)

≪ / RTI > moroheya, gingko leaf and chlorella extract.
The method according to claim 1,
The composition may be in the form of a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, cream, gel, patch, spray, ointment, Wherein the composition is formulated in the form of an injection solution.
The method according to claim 1,
Wherein the composition has a skin wrinkle-improving effect, a skin moisturizing effect, a skin soothing effect, an anti-inflammatory effect or an atopic dermatitis-improving effect.
The cosmetic composition according to claim 1, wherein the composition has a skin wrinkle-improving effect, a skin moisturizing effect or a skin soothing effect.
1) adding a solvent selected from the group consisting of water, alcohols, acetone, ethyl acetate, butyl acetate and 1,3-butylene glycol to Morrow Haya, Gingko leaf and chlorella;
2) extracting the solvent-added Moroheya, ginkgo leaf and chlorella through the above step 1) at 4 to 100 캜;
3) filtering, aging and aging the extract through step 2) to prepare a filtrate; And
4) drying the filtrate of step 3) with a rotary evaporator; ≪ / RTI >
6. The method of claim 5,
Wherein the step 2) is conducted for 1 to 5 days, and the step 3) is left at 3 to 5 占 폚 for 7 to 10 days.

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